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Os08g0398400

2014-06-02T18:06:37Z

Panpan Liu: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
[[File:1.jpg|left|thumb|150px|"Figure1.Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference <ref name="ref1"/>"]]
[[File:2.jpg|right|thumb|150px|"Figure2.Phylogenetic analysis of OsHIR1 and its published plant homologues.(from reference <ref name="ref1"/>"]]
[[File:3.jpg|left|thumb|300px|"Figure3.The mRNA and protein levels of OsHIR1 0, 2, 4 and 6 days after inoculation of Xanthomonas oryzae pv. oryzae (Xoo) race LN44 or mock treatment by a leaf-clipping method. Ten μg total RNA and 10 μg total protein were loaded onto each lane.(from reference <ref name="ref1"/>"]]
[[File:4.jpg|left|thumb|300px|"Figure4.Expression of OsLRR1 and OsHIR1 in an OsLRR1 overexpressing rice line. Real-time RT-PCR analysis was performed to compare the relative gene expression (expression in untransformed control was set to 1). Error bars show the standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:5.jpg|right|thumb|300px|"Figure5.Semi-quantitative analysis of OsHIR1 and OsLRR1 electron microscopy signals in the untransformed control and the OsLRR1 overexpressing rice line. The immunogold-labeled signal counting was described in Methods. Error bars show the standard errors (N = 10). * in (b) and (c) indicates that the difference is significant (p < 0.05, Student’s t-test) between the transformants and the untransformed wild type.(from reference <ref name="ref1"/>"]]
[[File:6.jpg|left|thumb|600px|"Figure6.Double labeling of OsHIR1 and OsLRR1. Two independent photos were shown to illustrate the co-localization of OsHIR1 (15 nm gold particles) and OsLRR1 (6 nm gold particles) to the plasma membrane. PM: Plasma membrane; CW: Cell wall.(from reference <ref name="ref1"/>"]]
[[File:7.jpg|right|under|250px|"Figure7.Hypersensitive response lesions in some OsHIR1 transgenic plants. Three weeks after germination, white necrotic lesions located randomly at the margins and tips of leaves (red arrows) were observed in about 20% of the OsHIR1 transgenic plants. Such a phenomenon was not found in untransformed wild type (Col-0), empty vector transgenic control (Col-0/V7), or OsLRR1 transgenic plants (Col-0/OsLRR1).(from reference <ref name="ref1"/>"]]
[[File:8.jpg|left|under|250px|"Figure8.Lactophenol-trypan blue staining showing spontaneous cell death. Leaves of 3-week-old plants were stained with lactophenol-trypan blue to detect dead cells. Spontaneous cell death found on the leaves of OsHIR1 and OsLRR1 transgenic plants were indicated by black arrows. Bars = 100 μm(from reference <ref name="ref1"/>"]]
[[File:9.jpg|left|under|400px|"Figure9.Disease symptoms after pathogen inoculation. Sixweek-old seedlings of the untransformed wild type (Col-0), the empty vector-transformed control (Col-0/V7), and the OsLRR1 (Col-0/OsLRR1) and OsHIR1 transgenic lines (Col-0/OsHIR1) were challenged with Pst DC3000. The symptoms were recorded 5 days after inoculation.(from reference <ref name="ref1"/>"]]
[[File:10.jpg|right|thumb|300px|"Figure10.Pathogen titers 5 days after pathogen inoculation. Rosette leaves were collected from inoculated plants for pathogen titer determination. Statistical analysis using ANOVA followed by Fisher’s LSD Test (p < 0.05) reveals 3 groups: 1, the untransformed wild type and the vector-only control; 2, OsLRR1 transgenic plants; and 3, OsHIR1 transgenic plants. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:11.jpg|left|under|300px|"Figure11.Expression of defense marker genes without (mock) inoculation. Real-time RT-PCR was performed using reverse-transcribed RNA samples. Relative expression levels of PR1 and PR2 in all plants were compared to the mock-inoculated untransformed wild type parent (Col-0; expression level set to 1). Both the expressions of PR1 and PR2 can be categorized into different groups using ANOVA followed by Fisher’s LSD Test (p < 0.05). In (d), the gene expression in mock-treated Col-0 was used just to set the reference for gene expression and was not included in the statistical analysis. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:12.jpg|right|under|300px|"Figure12.Expression of defense marker genes with Pst DC3000 inoculation.(from reference <ref name="ref1"/>"]]
[[File:13.jpg|left|thumb|600px|"Figure13.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) or 50 μM Cd in a controlled environment with a 16 h light/8 h dark photoperiod at 22/18 °C. Root length was analyzed using the Image J software at 12 days after planting (DAP). The data are presented as the mean ± SD of three independent experiments (total n = 30 seedlings per treatment).Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:14.jpg|right|under|200px|"Figure14.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) for 2 weeks. The data are presented as the mean ± standard deviation (SD; n = 250). Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:15.jpg|right|under|200px|"Figure15.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 50 μM Cd for 2 weeks.(from reference <ref name="ref2"/>"]]
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 11), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>
</references>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T18:03:40Z

Panpan Liu: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
[[File:1.jpg|left|thumb|150px|"Figure1.Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference <ref name="ref1"/>"]]
[[File:2.jpg|right|thumb|150px|"Figure2.Phylogenetic analysis of OsHIR1 and its published plant homologues.(from reference <ref name="ref1"/>"]]
[[File:3.jpg|left|thumb|300px|"Figure3.The mRNA and protein levels of OsHIR1 0, 2, 4 and 6 days after inoculation of Xanthomonas oryzae pv. oryzae (Xoo) race LN44 or mock treatment by a leaf-clipping method. Ten μg total RNA and 10 μg total protein were loaded onto each lane.(from reference <ref name="ref1"/>"]]
[[File:4.jpg|left|thumb|300px|"Figure4.Expression of OsLRR1 and OsHIR1 in an OsLRR1 overexpressing rice line. Real-time RT-PCR analysis was performed to compare the relative gene expression (expression in untransformed control was set to 1). Error bars show the standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:5.jpg|right|thumb|300px|"Figure5.Semi-quantitative analysis of OsHIR1 and OsLRR1 electron microscopy signals in the untransformed control and the OsLRR1 overexpressing rice line. The immunogold-labeled signal counting was described in Methods. Error bars show the standard errors (N = 10). * in (b) and (c) indicates that the difference is significant (p < 0.05, Student’s t-test) between the transformants and the untransformed wild type.(from reference <ref name="ref1"/>"]]
[[File:6.jpg|left|thumb|600px|"Figure6.Double labeling of OsHIR1 and OsLRR1. Two independent photos were shown to illustrate the co-localization of OsHIR1 (15 nm gold particles) and OsLRR1 (6 nm gold particles) to the plasma membrane. PM: Plasma membrane; CW: Cell wall.(from reference <ref name="ref1"/>"]]
[[File:7.jpg|right|under|250px|"Figure7.Hypersensitive response lesions in some OsHIR1 transgenic plants. Three weeks after germination, white necrotic lesions located randomly at the margins and tips of leaves (red arrows) were observed in about 20% of the OsHIR1 transgenic plants. Such a phenomenon was not found in untransformed wild type (Col-0), empty vector transgenic control (Col-0/V7), or OsLRR1 transgenic plants (Col-0/OsLRR1).(from reference <ref name="ref1"/>"]]
[[File:8.jpg|left|under|250px|"Figure8.Lactophenol-trypan blue staining showing spontaneous cell death. Leaves of 3-week-old plants were stained with lactophenol-trypan blue to detect dead cells. Spontaneous cell death found on the leaves of OsHIR1 and OsLRR1 transgenic plants were indicated by black arrows. Bars = 100 μm(from reference <ref name="ref1"/>"]]
[[File:9.jpg|left|under|300px|"Figure9.Disease symptoms after pathogen inoculation. Sixweek-old seedlings of the untransformed wild type (Col-0), the empty vector-transformed control (Col-0/V7), and the OsLRR1 (Col-0/OsLRR1) and OsHIR1 transgenic lines (Col-0/OsHIR1) were challenged with Pst DC3000. The symptoms were recorded 5 days after inoculation.(from reference <ref name="ref1"/>"]]
[[File:10.jpg|right|thumb|300px|"Figure10.Pathogen titers 5 days after pathogen inoculation. Rosette leaves were collected from inoculated plants for pathogen titer determination. Statistical analysis using ANOVA followed by Fisher’s LSD Test (p < 0.05) reveals 3 groups: 1, the untransformed wild type and the vector-only control; 2, OsLRR1 transgenic plants; and 3, OsHIR1 transgenic plants. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:11.jpg|left|under|300px|"Figure11.Expression of defense marker genes without (mock) inoculation. Real-time RT-PCR was performed using reverse-transcribed RNA samples. Relative expression levels of PR1 and PR2 in all plants were compared to the mock-inoculated untransformed wild type parent (Col-0; expression level set to 1). Both the expressions of PR1 and PR2 can be categorized into different groups using ANOVA followed by Fisher’s LSD Test (p < 0.05). In (d), the gene expression in mock-treated Col-0 was used just to set the reference for gene expression and was not included in the statistical analysis. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:12.jpg|right|under|300px|"Figure12.Expression of defense marker genes with Pst DC3000 inoculation.(from reference <ref name="ref1"/>"]]
[[File:13.jpg|left|thumb|600px|"Figure13.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) or 50 μM Cd in a controlled environment with a 16 h light/8 h dark photoperiod at 22/18 °C. Root length was analyzed using the Image J software at 12 days after planting (DAP). The data are presented as the mean ± SD of three independent experiments (total n = 30 seedlings per treatment).Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:14.jpg|right|under|200px|"Figure14.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) for 2 weeks. The data are presented as the mean ± standard deviation (SD; n = 250). Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:15.jpg|right|under|200px|"Figure15.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 50 μM Cd for 2 weeks.(from reference <ref name="ref2"/>"]]
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 11), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>
</references>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T18:01:02Z

Panpan Liu: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
[[File:1.jpg|left|thumb|150px|"Figure1.Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference <ref name="ref1"/>"]]
[[File:2.jpg|right|thumb|150px|"Figure2.Phylogenetic analysis of OsHIR1 and its published plant homologues.(from reference <ref name="ref1"/>"]]
[[File:3.jpg|left|thumb|300px|"Figure3.The mRNA and protein levels of OsHIR1 0, 2, 4 and 6 days after inoculation of Xanthomonas oryzae pv. oryzae (Xoo) race LN44 or mock treatment by a leaf-clipping method. Ten μg total RNA and 10 μg total protein were loaded onto each lane.(from reference <ref name="ref1"/>"]]
[[File:4.jpg|left|thumb|300px|"Figure4.Expression of OsLRR1 and OsHIR1 in an OsLRR1 overexpressing rice line. Real-time RT-PCR analysis was performed to compare the relative gene expression (expression in untransformed control was set to 1). Error bars show the standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:5.jpg|right|thumb|300px|"Figure5.Semi-quantitative analysis of OsHIR1 and OsLRR1 electron microscopy signals in the untransformed control and the OsLRR1 overexpressing rice line. The immunogold-labeled signal counting was described in Methods. Error bars show the standard errors (N = 10). * in (b) and (c) indicates that the difference is significant (p < 0.05, Student’s t-test) between the transformants and the untransformed wild type.(from reference <ref name="ref1"/>"]]
[[File:6.jpg|left|thumb|600px|"Figure6.Double labeling of OsHIR1 and OsLRR1. Two independent photos were shown to illustrate the co-localization of OsHIR1 (15 nm gold particles) and OsLRR1 (6 nm gold particles) to the plasma membrane. PM: Plasma membrane; CW: Cell wall.(from reference <ref name="ref1"/>"]]
[[File:7.jpg|right|under|250px|"Figure7.Hypersensitive response lesions in some OsHIR1 transgenic plants. Three weeks after germination, white necrotic lesions located randomly at the margins and tips of leaves (red arrows) were observed in about 20% of the OsHIR1 transgenic plants. Such a phenomenon was not found in untransformed wild type (Col-0), empty vector transgenic control (Col-0/V7), or OsLRR1 transgenic plants (Col-0/OsLRR1).(from reference <ref name="ref1"/>"]]
[[File:8.jpg|left|under|250px|"Figure8.Lactophenol-trypan blue staining showing spontaneous cell death. Leaves of 3-week-old plants were stained with lactophenol-trypan blue to detect dead cells. Spontaneous cell death found on the leaves of OsHIR1 and OsLRR1 transgenic plants were indicated by black arrows. Bars = 100 μm(from reference <ref name="ref1"/>"]]
[[File:9.jpg|left|under|300px|"Figure9.Disease symptoms after pathogen inoculation. Sixweek-old seedlings of the untransformed wild type (Col-0), the empty vector-transformed control (Col-0/V7), and the OsLRR1 (Col-0/OsLRR1) and OsHIR1 transgenic lines (Col-0/OsHIR1) were challenged with Pst DC3000. The symptoms were recorded 5 days after inoculation.(from reference <ref name="ref1"/>"]]
[[File:10.jpg|right|thumb|300px|"Figure10.Pathogen titers 5 days after pathogen inoculation. Rosette leaves were collected from inoculated plants for pathogen titer determination. Statistical analysis using ANOVA followed by Fisher’s LSD Test (p < 0.05) reveals 3 groups: 1, the untransformed wild type and the vector-only control; 2, OsLRR1 transgenic plants; and 3, OsHIR1 transgenic plants. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:11.jpg|left|thumb|300px|"Figure11.Expression of defense marker genes without (mock) inoculation. Real-time RT-PCR was performed using reverse-transcribed RNA samples. Relative expression levels of PR1 and PR2 in all plants were compared to the mock-inoculated untransformed wild type parent (Col-0; expression level set to 1). Both the expressions of PR1 and PR2 can be categorized into different groups using ANOVA followed by Fisher’s LSD Test (p < 0.05). In (d), the gene expression in mock-treated Col-0 was used just to set the reference for gene expression and was not included in the statistical analysis. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:12.jpg|right|thumb|300px|"Figure12.Expression of defense marker genes with Pst DC3000 inoculation.(from reference <ref name="ref1"/>"]]
[[File:13.jpg|left|thumb|600px|"Figure13.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) or 50 μM Cd in a controlled environment with a 16 h light/8 h dark photoperiod at 22/18 °C. Root length was analyzed using the Image J software at 12 days after planting (DAP). The data are presented as the mean ± SD of three independent experiments (total n = 30 seedlings per treatment).Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:14.jpg|right|thumb|250px|"Figure14.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) for 2 weeks. The data are presented as the mean ± standard deviation (SD; n = 250). Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:15.jpg|right|thumb|250px|"Figure15.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 50 μM Cd for 2 weeks.(from reference <ref name="ref2"/>"]]
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 11), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>
</references>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T17:58:48Z

Panpan Liu: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
[[File:1.jpg|left|thumb|150px|"Figure1.Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference <ref name="ref1"/>"]]
[[File:2.jpg|right|thumb|150px|"Figure2.Phylogenetic analysis of OsHIR1 and its published plant homologues.(from reference <ref name="ref1"/>"]]
[[File:3.jpg|left|thumb|300px|"Figure3.The mRNA and protein levels of OsHIR1 0, 2, 4 and 6 days after inoculation of Xanthomonas oryzae pv. oryzae (Xoo) race LN44 or mock treatment by a leaf-clipping method. Ten μg total RNA and 10 μg total protein were loaded onto each lane.(from reference <ref name="ref1"/>"]]
[[File:4.jpg|left|thumb|300px|"Figure4.Expression of OsLRR1 and OsHIR1 in an OsLRR1 overexpressing rice line. Real-time RT-PCR analysis was performed to compare the relative gene expression (expression in untransformed control was set to 1). Error bars show the standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:5.jpg|right|thumb|300px|"Figure5.Semi-quantitative analysis of OsHIR1 and OsLRR1 electron microscopy signals in the untransformed control and the OsLRR1 overexpressing rice line. The immunogold-labeled signal counting was described in Methods. Error bars show the standard errors (N = 10). * in (b) and (c) indicates that the difference is significant (p < 0.05, Student’s t-test) between the transformants and the untransformed wild type.(from reference <ref name="ref1"/>"]]
[[File:6.jpg|left|thumb|600px|"Figure6.Double labeling of OsHIR1 and OsLRR1. Two independent photos were shown to illustrate the co-localization of OsHIR1 (15 nm gold particles) and OsLRR1 (6 nm gold particles) to the plasma membrane. PM: Plasma membrane; CW: Cell wall.(from reference <ref name="ref1"/>"]]
[[File:7.jpg|right|thumb|250px|"Figure7.Hypersensitive response lesions in some OsHIR1 transgenic plants. Three weeks after germination, white necrotic lesions located randomly at the margins and tips of leaves (red arrows) were observed in about 20% of the OsHIR1 transgenic plants. Such a phenomenon was not found in untransformed wild type (Col-0), empty vector transgenic control (Col-0/V7), or OsLRR1 transgenic plants (Col-0/OsLRR1).(from reference <ref name="ref1"/>"]]
[[File:8.jpg|left|thumb|250px|"Figure8.Lactophenol-trypan blue staining showing spontaneous cell death. Leaves of 3-week-old plants were stained with lactophenol-trypan blue to detect dead cells. Spontaneous cell death found on the leaves of OsHIR1 and OsLRR1 transgenic plants were indicated by black arrows. Bars = 100 μm(from reference <ref name="ref1"/>"]]
[[File:9.jpg|left|thumb|300px|"Figure9.Disease symptoms after pathogen inoculation. Sixweek-old seedlings of the untransformed wild type (Col-0), the empty vector-transformed control (Col-0/V7), and the OsLRR1 (Col-0/OsLRR1) and OsHIR1 transgenic lines (Col-0/OsHIR1) were challenged with Pst DC3000. The symptoms were recorded 5 days after inoculation.(from reference <ref name="ref1"/>"]]
[[File:10.jpg|right|thumb|300px|"Figure10.Pathogen titers 5 days after pathogen inoculation. Rosette leaves were collected from inoculated plants for pathogen titer determination. Statistical analysis using ANOVA followed by Fisher’s LSD Test (p < 0.05) reveals 3 groups: 1, the untransformed wild type and the vector-only control; 2, OsLRR1 transgenic plants; and 3, OsHIR1 transgenic plants. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:11.jpg|left|thumb|300px|"Figure11.Expression of defense marker genes without (mock) inoculation. Real-time RT-PCR was performed using reverse-transcribed RNA samples. Relative expression levels of PR1 and PR2 in all plants were compared to the mock-inoculated untransformed wild type parent (Col-0; expression level set to 1). Both the expressions of PR1 and PR2 can be categorized into different groups using ANOVA followed by Fisher’s LSD Test (p < 0.05). In (d), the gene expression in mock-treated Col-0 was used just to set the reference for gene expression and was not included in the statistical analysis. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:12.jpg|right|thumb|300px|"Figure12.Expression of defense marker genes with Pst DC3000 inoculation.(from reference <ref name="ref1"/>"]]
[[File:13.jpg|left|thumb|600px|"Figure13.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) or 50 μM Cd in a controlled environment with a 16 h light/8 h dark photoperiod at 22/18 °C. Root length was analyzed using the Image J software at 12 days after planting (DAP). The data are presented as the mean ± SD of three independent experiments (total n = 30 seedlings per treatment).Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:14.jpg|right|thumb|250px|"Figure14.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) for 2 weeks. The data are presented as the mean ± standard deviation (SD; n = 250). Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:15.jpg|right|thumb|250px|"Figure15.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 50 μM Cd for 2 weeks.(from reference <ref name="ref2"/>"]]
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 11), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>
</references>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T17:54:59Z

Panpan Liu: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
[[File:1.jpg|left|thumb|150px|"Figure1.Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference <ref name="ref1"/>"]]
[[File:2.jpg|right|thumb|150px|"Figure2.Phylogenetic analysis of OsHIR1 and its published plant homologues.(from reference <ref name="ref1"/>"]]
[[File:3.jpg|left|thumb|300px|"Figure3.The mRNA and protein levels of OsHIR1 0, 2, 4 and 6 days after inoculation of Xanthomonas oryzae pv. oryzae (Xoo) race LN44 or mock treatment by a leaf-clipping method. Ten μg total RNA and 10 μg total protein were loaded onto each lane.(from reference <ref name="ref1"/>"]]
[[File:4.jpg|left|thumb|300px|"Figure4.Expression of OsLRR1 and OsHIR1 in an OsLRR1 overexpressing rice line. Real-time RT-PCR analysis was performed to compare the relative gene expression (expression in untransformed control was set to 1). Error bars show the standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:5.jpg|right|thumb|300px|"Figure5.Semi-quantitative analysis of OsHIR1 and OsLRR1 electron microscopy signals in the untransformed control and the OsLRR1 overexpressing rice line. The immunogold-labeled signal counting was described in Methods. Error bars show the standard errors (N = 10). * in (b) and (c) indicates that the difference is significant (p < 0.05, Student’s t-test) between the transformants and the untransformed wild type.(from reference <ref name="ref1"/>"]]
[[File:6.jpg|left|thumb|600px|"Figure6.Double labeling of OsHIR1 and OsLRR1. Two independent photos were shown to illustrate the co-localization of OsHIR1 (15 nm gold particles) and OsLRR1 (6 nm gold particles) to the plasma membrane. PM: Plasma membrane; CW: Cell wall.(from reference <ref name="ref1"/>"]]
[[File:7.jpg|right|thumb|250px|"Figure7.Hypersensitive response lesions in some OsHIR1 transgenic plants. Three weeks after germination, white necrotic lesions located randomly at the margins and tips of leaves (red arrows) were observed in about 20% of the OsHIR1 transgenic plants. Such a phenomenon was not found in untransformed wild type (Col-0), empty vector transgenic control (Col-0/V7), or OsLRR1 transgenic plants (Col-0/OsLRR1).(from reference <ref name="ref1"/>"]]
[[File:8.jpg|left|thumb|250px|"Figure8.Lactophenol-trypan blue staining showing spontaneous cell death. Leaves of 3-week-old plants were stained with lactophenol-trypan blue to detect dead cells. Spontaneous cell death found on the leaves of OsHIR1 and OsLRR1 transgenic plants were indicated by black arrows. Bars = 100 μm(from reference <ref name="ref1"/>"]]
[[File:9.jpg|left|thumb|300px|"Figure9.Disease symptoms after pathogen inoculation. Sixweek-old seedlings of the untransformed wild type (Col-0), the empty vector-transformed control (Col-0/V7), and the OsLRR1 (Col-0/OsLRR1) and OsHIR1 transgenic lines (Col-0/OsHIR1) were challenged with Pst DC3000. The symptoms were recorded 5 days after inoculation.(from reference <ref name="ref1"/>"]]
[[File:10.jpg|right|thumb|300px|"Figure10.Pathogen titers 5 days after pathogen inoculation. Rosette leaves were collected from inoculated plants for pathogen titer determination. Statistical analysis using ANOVA followed by Fisher’s LSD Test (p < 0.05) reveals 3 groups: 1, the untransformed wild type and the vector-only control; 2, OsLRR1 transgenic plants; and 3, OsHIR1 transgenic plants. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:11.jpg|right|thumb|300px|"Figure11.Expression of defense marker genes without (mock) inoculation. Real-time RT-PCR was performed using reverse-transcribed RNA samples. Relative expression levels of PR1 and PR2 in all plants were compared to the mock-inoculated untransformed wild type parent (Col-0; expression level set to 1). Both the expressions of PR1 and PR2 can be categorized into different groups using ANOVA followed by Fisher’s LSD Test (p < 0.05). In (d), the gene expression in mock-treated Col-0 was used just to set the reference for gene expression and was not included in the statistical analysis. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:12.jpg|right|thumb|300px|"Figure12.Expression of defense marker genes with Pst DC3000 inoculation.(from reference <ref name="ref1"/>"]]
[[File:13.jpg|right|thumb|600px|"Figure13.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) or 50 μM Cd in a controlled environment with a 16 h light/8 h dark photoperiod at 22/18 °C. Root length was analyzed using the Image J software at 12 days after planting (DAP). The data are presented as the mean ± SD of three independent experiments (total n = 30 seedlings per treatment).Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:14.jpg|right|thumb|250px|"Figure14.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) for 2 weeks. The data are presented as the mean ± standard deviation (SD; n = 250). Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:15.jpg|right|thumb|250px|"Figure15.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 50 μM Cd for 2 weeks.(from reference <ref name="ref2"/>"]]
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 11), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>
</references>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T17:48:26Z

Panpan Liu: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
[[File:1.jpg|left|thumb|150px|"Figure1.Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference <ref name="ref1"/>"]]
[[File:2.jpg|left|thumb|150px|"Figure2.Phylogenetic analysis of OsHIR1 and its published plant homologues.(from reference <ref name="ref1"/>"]]
[[File:3.jpg|left|thumb|300px|"Figure3.The mRNA and protein levels of OsHIR1 0, 2, 4 and 6 days after inoculation of Xanthomonas oryzae pv. oryzae (Xoo) race LN44 or mock treatment by a leaf-clipping method. Ten μg total RNA and 10 μg total protein were loaded onto each lane.(from reference <ref name="ref1"/>"]]
[[File:4.jpg|left|thumb|300px|"Figure4.Expression of OsLRR1 and OsHIR1 in an OsLRR1 overexpressing rice line. Real-time RT-PCR analysis was performed to compare the relative gene expression (expression in untransformed control was set to 1). Error bars show the standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:5.jpg|left|thumb|300px|"Figure5.Semi-quantitative analysis of OsHIR1 and OsLRR1 electron microscopy signals in the untransformed control and the OsLRR1 overexpressing rice line. The immunogold-labeled signal counting was described in Methods. Error bars show the standard errors (N = 10). * in (b) and (c) indicates that the difference is significant (p < 0.05, Student’s t-test) between the transformants and the untransformed wild type.(from reference <ref name="ref1"/>"]]
[[File:6.jpg|center|thumb|600px|"Figure6.Double labeling of OsHIR1 and OsLRR1. Two independent photos were shown to illustrate the co-localization of OsHIR1 (15 nm gold particles) and OsLRR1 (6 nm gold particles) to the plasma membrane. PM: Plasma membrane; CW: Cell wall.(from reference <ref name="ref1"/>"]]
[[File:7.jpg|right|thumb|250px|"Figure7.Hypersensitive response lesions in some OsHIR1 transgenic plants. Three weeks after germination, white necrotic lesions located randomly at the margins and tips of leaves (red arrows) were observed in about 20% of the OsHIR1 transgenic plants. Such a phenomenon was not found in untransformed wild type (Col-0), empty vector transgenic control (Col-0/V7), or OsLRR1 transgenic plants (Col-0/OsLRR1).(from reference <ref name="ref1"/>"]]
[[File:8.jpg|right|thumb|250px|"Figure8.Lactophenol-trypan blue staining showing spontaneous cell death. Leaves of 3-week-old plants were stained with lactophenol-trypan blue to detect dead cells. Spontaneous cell death found on the leaves of OsHIR1 and OsLRR1 transgenic plants were indicated by black arrows. Bars = 100 μm(from reference <ref name="ref1"/>"]]
[[File:9.jpg|left|thumb|300px|"Figure9.Disease symptoms after pathogen inoculation. Sixweek-old seedlings of the untransformed wild type (Col-0), the empty vector-transformed control (Col-0/V7), and the OsLRR1 (Col-0/OsLRR1) and OsHIR1 transgenic lines (Col-0/OsHIR1) were challenged with Pst DC3000. The symptoms were recorded 5 days after inoculation.(from reference <ref name="ref1"/>"]]
[[File:10.jpg|right|thumb|300px|"Figure10.Pathogen titers 5 days after pathogen inoculation. Rosette leaves were collected from inoculated plants for pathogen titer determination. Statistical analysis using ANOVA followed by Fisher’s LSD Test (p < 0.05) reveals 3 groups: 1, the untransformed wild type and the vector-only control; 2, OsLRR1 transgenic plants; and 3, OsHIR1 transgenic plants. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:11.jpg|left|thumb|300px|"Figure11.Expression of defense marker genes without (mock) inoculation. Real-time RT-PCR was performed using reverse-transcribed RNA samples. Relative expression levels of PR1 and PR2 in all plants were compared to the mock-inoculated untransformed wild type parent (Col-0; expression level set to 1). Both the expressions of PR1 and PR2 can be categorized into different groups using ANOVA followed by Fisher’s LSD Test (p < 0.05). In (d), the gene expression in mock-treated Col-0 was used just to set the reference for gene expression and was not included in the statistical analysis. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:12.jpg|left|thumb|300px|"Figure12.Expression of defense marker genes with Pst DC3000 inoculation.(from reference <ref name="ref1"/>"]]
[[File:13.jpg|right|thumb|600px|"Figure13.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) or 50 μM Cd in a controlled environment with a 16 h light/8 h dark photoperiod at 22/18 °C. Root length was analyzed using the Image J software at 12 days after planting (DAP). The data are presented as the mean ± SD of three independent experiments (total n = 30 seedlings per treatment).Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:14.jpg|left|thumb|250px|"Figure14.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) for 2 weeks. The data are presented as the mean ± standard deviation (SD; n = 250). Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:15.jpg|right|thumb|250px|"Figure15.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 50 μM Cd for 2 weeks.(from reference <ref name="ref2"/>"]]
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 11), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>
</references>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T17:09:38Z

Panpan Liu: /* Function */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
[[File:1.jpg|right|thumb|150px|"Figure1.Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference <ref name="ref1"/>"]]
[[File:2.jpg|right|thumb|150px|"Figure2.Phylogenetic analysis of OsHIR1 and its published plant homologues.(from reference <ref name="ref1"/>"]]
[[File:3.jpg|right|thumb|150px|"Figure3.The mRNA and protein levels of OsHIR1 0, 2, 4 and 6 days after inoculation of Xanthomonas oryzae pv. oryzae (Xoo) race LN44 or mock treatment by a leaf-clipping method. Ten μg total RNA and 10 μg total protein were loaded onto each lane.(from reference <ref name="ref1"/>"]]
[[File:4.jpg|right|thumb|150px|"Figure4.Expression of OsLRR1 and OsHIR1 in an OsLRR1 overexpressing rice line. Real-time RT-PCR analysis was performed to compare the relative gene expression (expression in untransformed control was set to 1). Error bars show the standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:5.jpg|right|thumb|150px|"Figure5.Semi-quantitative analysis of OsHIR1 and OsLRR1 electron microscopy signals in the untransformed control and the OsLRR1 overexpressing rice line. The immunogold-labeled signal counting was described in Methods. Error bars show the standard errors (N = 10). * in (b) and (c) indicates that the difference is significant (p < 0.05, Student’s t-test) between the transformants and the untransformed wild type.(from reference <ref name="ref1"/>"]]
[[File:6.jpg|right|thumb|150px|"Figure6.Double labeling of OsHIR1 and OsLRR1. Two independent photos were shown to illustrate the co-localization of OsHIR1 (15 nm gold particles) and OsLRR1 (6 nm gold particles) to the plasma membrane. PM: Plasma membrane; CW: Cell wall.(from reference <ref name="ref1"/>"]]
[[File:7.jpg|right|thumb|150px|"Figure7.Hypersensitive response lesions in some OsHIR1 transgenic plants. Three weeks after germination, white necrotic lesions located randomly at the margins and tips of leaves (red arrows) were observed in about 20% of the OsHIR1 transgenic plants. Such a phenomenon was not found in untransformed wild type (Col-0), empty vector transgenic control (Col-0/V7), or OsLRR1 transgenic plants (Col-0/OsLRR1).(from reference <ref name="ref1"/>"]]
[[File:8.jpg|right|thumb|150px|"Figure8.Lactophenol-trypan blue staining showing spontaneous cell death. Leaves of 3-week-old plants were stained with lactophenol-trypan blue to detect dead cells. Spontaneous cell death found on the leaves of OsHIR1 and OsLRR1 transgenic plants were indicated by black arrows. Bars = 100 μm(from reference <ref name="ref1"/>"]]
[[File:9.jpg|right|thumb|150px|"Figure9.Disease symptoms after pathogen inoculation. Sixweek-old seedlings of the untransformed wild type (Col-0), the empty vector-transformed control (Col-0/V7), and the OsLRR1 (Col-0/OsLRR1) and OsHIR1 transgenic lines (Col-0/OsHIR1) were challenged with Pst DC3000. The symptoms were recorded 5 days after inoculation.(from reference <ref name="ref1"/>"]]
[[File:10.jpg|right|thumb|150px|"Figure10.Pathogen titers 5 days after pathogen inoculation. Rosette leaves were collected from inoculated plants for pathogen titer determination. Statistical analysis using ANOVA followed by Fisher’s LSD Test (p < 0.05) reveals 3 groups: 1, the untransformed wild type and the vector-only control; 2, OsLRR1 transgenic plants; and 3, OsHIR1 transgenic plants. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:11.jpg|right|thumb|150px|"Figure11.Expression of defense marker genes without (mock) inoculation. Real-time RT-PCR was performed using reverse-transcribed RNA samples. Relative expression levels of PR1 and PR2 in all plants were compared to the mock-inoculated untransformed wild type parent (Col-0; expression level set to 1). Both the expressions of PR1 and PR2 can be categorized into different groups using ANOVA followed by Fisher’s LSD Test (p < 0.05). In (d), the gene expression in mock-treated Col-0 was used just to set the reference for gene expression and was not included in the statistical analysis. The error bars indicate standard errors (N = 3).(from reference <ref name="ref1"/>"]]
[[File:12.jpg|right|thumb|150px|"Figure12.Expression of defense marker genes with Pst DC3000 inoculation.(from reference <ref name="ref1"/>"]]
[[File:13.jpg|right|thumb|150px|"Figure13.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) or 50 μM Cd in a controlled environment with a 16 h light/8 h dark photoperiod at 22/18 °C. Root length was analyzed using the Image J software at 12 days after planting (DAP). The data are presented as the mean ± SD of three independent experiments (total n = 30 seedlings per treatment).Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:14.jpg|right|thumb|150px|"Figure14.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 150 μM As(V) for 2 weeks. The data are presented as the mean ± standard deviation (SD; n = 250). Asterisks represent significant differences for each mean value of OsHIR1-overexpressing plants compared to the control (*P < 0.05 and **P < 0.01, t test).(from reference <ref name="ref2"/>"]]
[[File:15.jpg|right|thumb|150px|"Figure15.Arabidopsis seeds of OsHIR1-overexpressing lines and control (empty vector) were germinated and grown on ½ MS agar plates with 1.5 % sucrose in the absence or presence of 50 μM Cd for 2 weeks.(from reference <ref name="ref2"/>"]]
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 11), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>
</references>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

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Figure 1. Identification of OsMYB3R-2 transgenic rice and its expression pattern. A, Northern-blot assay of rice transgenic plants. Total RNA isolated from wild-type (WT) or transformed plants underwent hybridization with a [a-32P]dCTP-labeled probe of OsMYB3R-2 cDNA as described in “Materials and Methods.” B, Real-time RTPCR of the expression of OsMYB3R-2 in antisense lines. C and D, Southernblot assay of transformed rice plants. Genomic DNA isolated from wild-type or transformed plants was digested with EcoRI (E) or HindIII (H). The blot was hybridized with the open reading frame of the GUS gene labeled with [a-32P]dCTP. OL3, OL5, OL7, and OL8 and AL1, AL2, AL4, and AL5 represent overexpression (O) and antisense (A) lines of OsMYB3R-2 transgenic rice. E, Expression pattern of OsMYB3R-2 in vivo. GUS staining shows expression pattern of OsMYB3R-2 in vivo in various tissues from the T1 generation of OsMYB3R-2 promoter::GUS transgenic rice. a, Root; b, young internode; c, mature internode; d, node; e, mature leaf; f, lamina joint; g, leaf sheath; h, flower; i, immature seed.

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Working model of long-day flowering signal pathway
in rice. OsMADS50 promotes transition to reproductive stage by
suppressing expression of OsLFL1 that is a repressor of Ehd1.
OsMADS56 inhibits flowering by binding to OsMADS50. Ehd1
is also controlled by other genes, e.g. OsID1 and Ghd7.

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Gross morphology of wildtype (left) and gid1-1 (right) plants. Scale bar, 10 cm. Inset: higher magnification of gid1-1. Scale bar, 1 cm

Os08g0398400

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Panpan Liu: /* Function */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

[[File:1.jpg|right|thumb|150px|"Figure1.Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference <ref name="ref1"/>"]]===Expression===
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 1a), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T16:11:16Z

Panpan Liu: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

[[File:1.jpg|right|thumb|150px|"Schematic representation of the conserved structural domains in OsHIR1 and its homologues.(from reference (from reference <ref name="ref1"/>"]]===Expression===
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 1a), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

File:1.jpg

2014-06-02T16:02:56Z

Panpan Liu: uploaded a new version of "File:1.jpg"

Gross morphology of GID1-overexpressor and control plants. Scale
bar, 50 cm.

File:1.jpg

2014-06-02T16:02:55Z

Panpan Liu: uploaded a new version of "File:1.jpg"

Gross morphology of GID1-overexpressor and control plants. Scale
bar, 50 cm.

File:1.jpg

2014-06-02T16:02:55Z

Panpan Liu: uploaded a new version of "File:1.jpg"

Gross morphology of GID1-overexpressor and control plants. Scale
bar, 50 cm.

Os08g0398400

2014-06-02T15:34:34Z

Panpan Liu: /* Evolution */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 1a), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.<ref name="ref1"/>

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T15:34:08Z

Panpan Liu: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 1a), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).<ref name="ref1"/>

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.<ref name="ref2"/>

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T15:33:16Z

Panpan Liu: /* Function */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).<ref name="ref1"/>
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.<ref name="ref2"/>

===Expression===
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 1a), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T15:28:50Z

Panpan Liu: /* Function */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).[1]
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.[2]

===Expression===
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 1a), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T15:26:52Z

Panpan Liu: /* References */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.

===Expression===
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 1a), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T15:25:16Z

Panpan Liu: /* References */

Please input one-sentence summary here.

==Annotated Information==
===Function===
OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1.HIR1 (OsHIR1) as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1).
OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.

===Expression===
OsHIR1 was identified as a putative interacting partner ofOsLRR1. The OsHIR1 protein exhibits high similarity (from 84% to 96% identity) to homologues from dicots and monocots (Figure 1a), including maize, barley, wheat, pepper, and A. thaliana. For all the close homologues of OsHIR1, computational analysis reveals a putative N-myristoylation site at the N-terminus, followed by a transmembrane domain that is embedded within a Band 7-domain, which covers most of the OsHIR1 protein (Figure 1). In an unrooted phylogenetic tree (Figure 2), HIR proteins can be further divided into two branches: dicots and monocots. Among HIR homologues
from monocots, the OsHIR1 shares the highest similarity with the maize ZmHIR1 (96% identity).

To show that OsHIR1 is related to the plant defense response, we investigated whether its gene expression is responsive to pathogen challenge. Northern and western blot analyses showed that both the mRNA and protein levels of OsHIR1 increased after the rice plant was inoculated with the pathogen Xoo LN44 (Figure 3). On the other hand, no such change was observed after mock treatment (Figure 3).

A transgenic line that exhibited a high level of OsLRR1 gene expression was chosen for subsequent electron microscopy analysis (Figure 4). Interestingly, in addition to the elevated level of OsLRR1 mRNA, the expression of the OsHIR1 gene in the OsLRR1
transgenic line was also enhanced (Figure 4).

Immuno-gold electron microscopy studies showed that not only the signal density of the OsLRR1 proteins, but also that of the OsHIR1 proteins, in the plasma membrane, was increased in the OsLRR1 overexpressing line by at least two folds, when compared to the untransformed control (Figure 5). On the other hand, there was no significant difference (Student’s t-test, p < 0.05) between the number of OsHIR1 signals in the tonoplast of the OsLRR1 overexpressing line and that in the untransformed control. These results indicated that OsLRR1 enhanced the plasma membrane localization of OsHIR1.

Proximal occurrences of large and small gold particles were detected in the plasma membrane (Figure 6), supporting the notion that OsLRR1 and OsHIR1 co-localized and interacted in the plasma membrane.
Ectopic expression of the OsHIR1 can cause spontaneous hypersensitive response lesions in the leaves of transgenic A. thaliana. To perform a rapid gain-of-function test of QsHIR1,transgenic A. thaliana plants ectopically expressing OsHIR1 were generated. Three weeks after germination, the leaves of about 20% of the OsHIR1 transgenic plants (Col-0/OsHIR1) exhibited white spontaneous HR lesions located randomly at the margins and tips (Figure 3a, red arrows). As negative controls, the untransformed wild type (Col-0) and transgenic plants with the empty vector (Col-0/V7) exhibited normal growth. Transgenic plants expressing OsLRR1 (Col-0/OsLRR1) did not exhibit visible differences in the size, shape, or color of the leaves, when compared to the negative controls (Figure 7).To further observe the effect of OsHIR1 on cell death, lactophenol-trypan blue staining was performed using the leaves of the transgenic A. thaliana. The expression of OsHIR1 caused extensive spontaneous cell death (Figure 8,black arrows). On the other hand, the expression of OsLRR1 only resulted in very mild spontaneous cell death (Figure 8). This explains the lack of visible lesions found in OsLRR1 transgenic plants (Figure 7). No spontaneous cell death was observed in the untransformed control and transgenic plants containing the empty vector (Figure 8).

Ectopic expression of OsHIR1 in transgenic A. thaliana enhances resistance to P. syringae pv. tomato DC3000 (Pst DC3000)
When the untransformed wild type (Col-0) or A. thaliana transformed with the empty vector cassette (Col-0/V7) was inoculated with the pathogen Pst DC3000, disease symptoms (yellowing and necrosis) gradually appeared and the infected areas spread out from the original inoculation sites (Figure 9). Such symptoms were alleviated in the transgenic line expressing OsLRR1, consistent with the
results of our previous study . The spread of pathogen infection was also suppressed by the ectopic expression of OsHIR1 (Figure 9). Consistent with these visible symptoms, transgenic plants expressing either OsLRR1 or OsHIR1 exhibited a lower titer of pathogens when compared to Col-0 and the empty vector control (Figure 10).However, the OsHIR1 transgenic lines showed a stronger
effect on lowering the pathogen titer when compared to the OsLRR1 transgenic line (Figure 10).

The expression levels of PR1 and PR2, two defense marker genes in the salicyclic acid pathway related to the defense against biotrophic pathogens such as Pst DC3000, were monitored in both mock- (Figure 11)and pathogen-inoculated (Figure 12) plants. In both mock-treated and pathogen-inoculated plants, the expression levels of PR1 and PR2 were elevated in both OsHIR1 and OsLRR1 transgenic plants when compared to Col-0 and transgenic plants containing the empty vector cassette. However, the OsHIR1 transgenic plants exhibited significantly higher levels of PR1 and PR2 gene induction than the OsLRR1 transgenic plants (p < 0.05).

OsHIR1‑overexpressing Arabidopsis enhances As and Cd tolerance
The finding that the OsHIR1 gene was upregulated in response to As and Cd treatments raises the question of the function of OsHIR1 associated with both elemental stress responses. Three independent transgenic lines were selected depending on the expression level of the OsHIR1 by RT-PCR and then compared to the control plants under 150 μM As and 50 μM Cd (Figure 13). The OsHIR1-overexpressing lines exhibited no apparent phenotypic differences from the vector control lines under normal conditions. However,
OsHIR1-overexpressing plants showed significantly longer root lengths than those of the control plants under As and Cd conditions (Fig. 4a). To investigate the possible mechanism by which OsHIR1 ubiquitin E3 ligase affects As or Cd uptake in the transgenic line, we compared As and Cd accumulation between the OsHIR1-overexpressing lines and the control plants. The concentrations of As in the shoots and roots of the OsHIR1-overexpressing plants were 14.1 and 46.4 %, respectively, which were lower than those
of the control plants (Figure 14). Cd uptake by the OsHIR1-overexpressing plants was also 3.8 and 12.3 % lower than the control plants in the shoots and roots, respectively(Figure 15). These results suggest that the heterogeneous overexpression of the OsHIR1 gene in Arabidopsis may reduce both As and Cd uptakes by regulating its substrate protein.

===Evolution===
The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant so that it is more prone to HR and hence can react more promptly to limit the invading pathogens’ spread from the infection sites.

OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptakes.

==Labs working on this gene==
*State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, PR China

*Plant Genomics Lab, Department of Applied Plant Sciences, Kangwon National University

==References==
<references>
<ref name="ref1"> Liang Zhou1, Ming-Yan Cheung1, Man-Wah Li1, Yaping Fu2, Zongxiu Sun2, Sai-Ming Sun1, Hon-Ming Lam1* （2010） Plant Biology 10:290</ref>

<ref name="ref2"> Sung Don Lim · Jin Gyu Hwang · A. Reum Han ·Yong Chan Park · Chanhui Lee · Yong Sik Ok ·Cheol Seong Jang （2014）Plant Mol Biol
DOI 10.1007/s11103-014-0190-0</ref>

</references>

==Structured Information==
{{JaponicaGene|
GeneName = Os08g0398400|
Description = Similar to Hypersensitive-induced response protein|
Version = NM_001068279.1 GI:115476295 GeneID:4345499|
Length = 4031 bp|
Definition = Oryza sativa Japonica Group Os08g0398400, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 8|Chromosome 8]]|
AP = Chromosome 8:19099911..19103941|
CDS = 19101742..19101930,19102201..19102321,19102445..19102536,19102609..19102705,19103258..19103613 |
GCID = <gbrowseImage1>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008401:19099911..19103941
source=RiceChromosome08
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatgtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaa</cdnaseq>|
AA = <aaseq>MGQALGLVQVDQSTVAIKESFGKFDEVLEPGCHFLPWCIGKQIA GYLSLRVQQLDVRCETKTKDNVFVNVVASVQYRALAEKASDAFYRLSNTREQIQSYVF DVIRASVPKMNLDDAFEQKNEIAKAVEDELEKAMSMYGYEIVQTLIVDIEPDEHVKRA MNEINAAARLRVAANEKAEAEKILQIKRAEGDAESKYLAGLGIARQRQAIVDGLRDSV LAFSENVPGTSAKDVMDMVLVTQYFDTMKEIGASSKSSSVFIPHGPGAVKDIAAQIRD GQLQAKLI</aaseq>|
DNA = <dnaseqindica>1832..2020#2291..2411#2535..2626#2699..2795#3348..3703#agaatttgccctcctcacttctatacttcaatccccttcctttatttttttttttgggaggggccgaggcaaccgtcgccactccgacgatcccgtcccatccgccgccgccgccggagatttcctcgaggtaagccctaccccgctcctctccagcctctctccccttcggcgggcgcccagcccgagccccgacctcgatttcttggaatttcttggcgagcgtttgtttcttggggtttcctctagattttttcttttttttattattttggggacggatttagctctcgtggaggagattgggcggataatttcgtcggaggggggagggggagggggatgttggaatcgggtggtgggtaatttcgtcgattctccggtccttttccttgagtttagcctgggaattgcggttggttcttggacgcggtttgagttcttggttttttccactttctgtgcggacagatttagggcttgcgctttgcgagtgggggtaaaattcgatggtgtgctgttccagctttgtccacatactgcttggaaatttcaggaggttttttcagtcaagtcaagcacatcttttttgctagactaggccatgagatggaggttaacgtcgaactgtacccacctcagctaaaaataataataattctcaacaacagtatccaagtggctgtattgtttattcttgtttagttatgacgcagatgaatactactcgctaaagattgtgactttcttagttgtcacctcgtcgtgtggagccctgtgggttcttgctcttgacttttatcaaacaatgattgcttggcaagtgtcttgtttgattcgtctgccctttggcttgcatcgccagagtttgctggcttgcgctggcatttaatcaaaagtataatcattcccccaagtataatcaggattagcataatctttagcttcaactatttatatgatatatttggcttgtctgtagtctgtagtggccaagaacatcctgaagcagatttatcctatcaggctatcagcactaagttgatcggcacgtataggttttcttttgcgtcattctttttcattactgcatttctatggtgaattgttttcttttcttttgtatggctacatctattaccctgttgacttctgtgagcctgtgatgtactccctccgggctgataatacttgtcattttggacaagcgcacggtcttcaaaaaacaactttgaccattatttcctattataatatgtaaaaaagtgttaacaaatatatgatcatattaaagtactttttaagaccaatctatatatgtagtcaccatatttgaaagacaaatattttaaaaatgatttatataatcaaatattctaaagtttgacctcacccttgtccaaaacgacaagtattatcagcccggagggagtagtaatcttaggcatttttcagggaatgattacacagtataagtttgagtaatgtgttatcatgatatgcttttcaacaagatatacatattttttgcaacactgtttggtctaagttatctgaaattgcaattgcttgacttttcatttgggatgcgttgggcaatctattgtcctgacatacattcatcttcatttgaagccatggctttatcttttttaattcatatcttttgtttctgtttgtatgtactaaaaacttgaaatatgaatttccattggagaagttaacaccaacggagtataagcaagattatggcaaaacattcagttgttagttgtttcgttagcatatggcaccatttgtaattctgcatttcttcctttctagataagccatgggtcaagcactcggtttggtacaagtagatcaatcgacagtagccatcaaggagtcctttgggaagtttgatgaggttcttgagcctggatgccactttttaccctggtgcatagggaagcagatcgctggatatctttccttgcgtgtgcagcagctggatgtccgttgtgaaacaaagactaaggttccccagtttattttcttaactctggtttcttttatatttcatggcaagtgtctggagcaattgtcaggcatgataaaaaatggttctcaaaattcttgcattttgttccattcgtttactcctttatgataacatactgatgatggttatgtgcacatatctctgttttccttattcttcacttacactaatgcatgatgcatcaaagaataaacccaccttttctgagctctctgacgatgtataccattttgtggatatctgcaggacaatgtttttgtcaatgttgtggcatctgtgcaataccgtgctcttgctgagaaggcatcagacgccttttacaggcttagcaacaccagggagcaaatccagtcgtatgtttttgatggtaaggttcctgtatgctgtgaacattcatgcatccgatgcatttgcagttcatactcttctgattaccttctctatgttttgcctctattgttcaatgctaacagaactttcttcctcgcagtgatcagggctagtgttccaaagatgaacttggatgatgcatttgagcagaagaatgagatcgcaaaagctgtggaagatgagcttgaaaaggtttgtgttcaataattggtggtcatctagagcatagtggagttttatactgacctagtatatactctgtaggcaatgtcaatgtatggttatgagattgtgcaaactcttattgttgatattgaaccagatgagcatgttaagagagcaatgaatgaaatcaacgcaggtaagtattaattaaaacatcaattcattccatttcttcagagacattatttggcttggtgtttatgcttgacatttgagcttgcattggaaacaattttctcctgtttagacatgaaaatgttaaattctctttccgagaaacaaaaccatgtttatattgtctagttttattcaagttttatactgaaacactgaaaattgttgccacaaatatttttaggctttcaacacagtaacactgtaagttatatatagatggaacactgaaaagttatatatagatgtacataacatattatgaatattaacttcgtgaatacttgcaccataacagtcataagtaacacaggcacatgtaactgcaacctattgattacattgtgtgcaacaaaaaatgtgttggtcaactagtcaagtaaatatttcgtgctttttaagggcctgcaacatatatgtttggttggtctggtgtttttctaaagatgagctgtatggctgtaaatgtgctggacggcaacatagttttaactgctgttatttgatgatacagctgctaggctgagggtggcagccaatgagaaagctgaagcagagaagattcttcagatcaagagagctgaaggagatgcagaatccaagtacttggctggtctgggtatcgcaaggcagcgtcaggctatagtggatgggctgagagacagtgttcttgccttctccgagaacgtgcctgggacctctgccaaggatgtcatggatatggttctggttacgcaatacttcgacaccatgaaggagataggagcctcatccaagtcctcttcagtgttcatccctcacgggccaggtgccgtcaaagatatcgctgcgcagataagagatggtcagctccaggccaaattgatctaagaggatggcagggaatggatgttctggatttcttgggcattacgaaagtctgcagtaataccttatctacgtattcgatattttgtgagataaaaacttaggcaaaagaagttctattgtaagtatcacatgcacgcgtgcttcagtaccgtttgttcatagtaattcctgaggttatttccctagacgagtgtcaatatacttcttgaattgcatgtttctgtaaataattcctaagtgttaccgggctgtttggaattatttgcatgttgatgttagtgaccttaagtaatgtcccatcttattatgtgatgtcatgatgatttcc</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001068279.1 RefSeq:Os08g0398400]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 8]]
[[Category:Chromosome 8]]

Os08g0398400

2014-06-02T15:04:47Z

Panpan Liu: /* Labs working on this gene */

Os08g0398400

2014-06-02T15:01:56Z

Panpan Liu: /* Evolution */

Os08g0398400

2014-06-02T14:58:18Z

Panpan Liu: /* Expression */

Os08g0398400

2014-06-02T14:09:56Z

Panpan Liu: /* Function */

Os05g0145000

2014-05-27T14:42:34Z

Panpan Liu: /* Labs working on this gene */

Please input one-sentence summary here.

==Annotated Information==
===Function===
[[File:Functional_mutant.jpg|right|thumb|250px|'''Figure 1.''' ''Functional complementation of ptb1. (a) Plant phenotype of ptb1. (b) Plant phenotype of the PTB1-transformed ptb1 plants (ptb1–PTB1).(c) Panicle phenotype of ptb1. (d) Panicle phenotype of the ptb1–PTB1 transgenic plants. (from reference<ref name="ref1" />).'']]
The domestication-related POLLEN TUBE BLOCKD 1(''PTB1''), a RING-type E3 ubiquitin ligase, positively regulates the rice panicle seed setting rate by promoting pollen tube growth. The ''PTB1'' positively regulates rice PSSR through controlling pollen tube growth. Shuangcheng et al(2013) support the hypothesis that ''PTB1'' is an important maternal sporophytic factor of pollen tube growth and a key modulator of the rice panicle seed setting rate. 
There are many genes that related to the grain yield such as ''GS3/5'', ''GW2/5/8'', ''GL3.1'' and ''TGW6'', but as we know now, no genes regulates PSSR(which is one of the most important yield determinants) have been functionally characterized. Rice PSSR is impressionable to outside environmental conditions, which often cause large rice yield losss and threatens the safety of rice production. Nowadays, compared with the parents, many hybrid rice varieties exhibit sharp decreases in PSSR, the finding of the PBT1 gene has implications for the improvement of rice yield. 
PTB1 has pleiotropic effects on PT growth control. First, it has a decisive role in entering and early growth of the STT.Second, ''PTB1'' functions during the later navigation of PTs in the STT because a lower level of ''PTB1'' expression leads to different numbers of PTs growing in this process.Third, ''PTB1'' promotes growth from the chalazal end of the sporophytic STT to the micropyle because a moderate expression of ''PTB1'' leads to the growth of few PTs in this female location.Finally, PTB1 functions as a promoting agent based on the observation that a higher expression of this gene encourages more PTs to complete their full journey<ref name="ref1" />.

===Expression===
[[File:Expression1.jpg|right|thumb|250px|'''Figure 2.''' ''Expression patterns of PTB1. (a–h) GUS activity in various organs of PPTB1-GUS-transformed rice plants. (a) Growing root, (b) stem node, (c) stem internode, (d) leaf, (e) floret, (f) pistil, (g) shell of unpollinated spikelet and (h) pollen grains. (i) Relative PTB1 transcript levels of several tissues detected by q-PCR analysis. DAP, days after pollination. Values represent the mean±s.e.m. (n?3). Scale bars, 400 mm in (a), 3mm in (b) and (c), 1 cm in (d), 1mm in (e) and (f), 4mm in (g) and 50 mm in (h). (from reference<ref name="ref1" />).'']]
The natural variation in expression of ''PTB1'' which is affected by the promoter haplotype and the environmental temperature, correlates with the rice panicle seed setting rate. The GUS gene test in rice showed that ''PTB1'' was differentially expressed in various rice tissues: the expression of PTB1 was relatively weak in the root, shoot and leaf, and the expression was strong in the stigma of the pistil and the spikelet hulls of the pre-emergence panicle. But , in rice pollen grains,the expression was undetectable. The results of Q-PCR was further confirmed this temporal and spatial expression patterns by showing that PBT1 was expressed prior in the reproductive organs such as the pistil, young panicle,pre-emergence panicle and post-emergence panicle. This pattern corresponded well to the role ''PBT1'' played in the pollen tube growth<ref name="ref1" />. 
The transient expression of PBT1-YFP fusion protein in rice protoplast cells showed that the PBT1 localized preferentially to the cytoplasm and although the PBT1 protein was predicted to contain 4 transmembrane segments, it was weakly expressed on the membrane.

===Evolution===
Please input evolution information here.

You can also add sub-section(s) at will.

==Labs working on this gene==
Rice Research Institute, Sichuan Agricultural University;
State Key Laboratory of Hybrid Rice, Sichuan Agricultural University;
Key Laboratory of Crop Genetic Resources and Improvement, Sichuan Agricultural University, Ministry of Education

==References==
<references>
* <ref name="ref1">
Shuangcheng Li1, Wenbo Li1, Bin Huang1, et al.(2013)Natural variation in PTB1 regulates rice seed setting rate by controlling pollen tube growth.NATURE COMMUNICATIONS, DOI: 10.1038/ncomms3793.
</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os05g0145000|
Description = Similar to Ring finger protein (Fragment)|
Version = NM_001061164.1 GI:115462058 GeneID:4337790|
Length = 5616 bp|
Definition = Oryza sativa Japonica Group Os05g0145000, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 5|Chromosome 5]]|
AP = Chromosome 5:2576595..2582210|
CDS = 2576763..2577185,2577792..2577905,2578448..2578564,2578770..2578835,2579376..2579468 ,2579552..2579690,2581077..2581150,2581225..2581291,2581746..2581784 ,2581880..2581908|
GCID = <gbrowseImage1>
name=NC_008398:2576595..2582210
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008398:2576595..2582210
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atggcgatgcggggggtcgatttcaagtggtacgatggattcttcctctccatgctcgccaccagcctaatcattgtctccatcaactggaagaggtatcgtctctgcgcccacccgttgcacatatggatcgtggttgactacaccaccgtcttcatcttccgccttctcatgttcgtcgataatggccttgccgccggcatgggattggatcttggatggcaacagagatatgctcgtttttgtgggagaattgttgtcttgtcggttcttgtgcttcttctctatccctttctttgggtttggactgtgataggaacattgtggtttagcactgcaagaggctgtttacccgaggaaggacaaaaatggggcttccttatatggctgcttttcagctactgtggtctcgcctgtattgcgtgtgtggctgttggaaagtggctaagccgaaggcatgctctccagcagagggcacaacagggaataccagtctctgaatatggggttttggttgacatgatccgtgtgcctgattgggcatttgaggccgttggcttggaaatgagaggaatgggccaagatactgcatatcatcctggtctttatttaacagctgcccaaagagaggcagttgaggcactgattcaagaactcccgaagttcagactgaaagctgttcctacagactgcagtgagtgtccaatctgcctggaggaattccatgttggcaacgaggtccgtgggctcccgtgcgcgcacaatttccatgtggagtgcatcgaccagtggctccggctgaatgtcaagtgcccccgttgccggtgctccgtcttccccaacctcgacctgagcgcgctcaacaacctccggccatcctccgagccggaccggccgtcggccagcgaggtgacagcggcgacgatggcgcggtacgtgaggtcgtcgcagccggctgggcagagctacctgctgcggctgcaggggctgctgctgcggcaggtggtggttcggcacggcggcggcgacgacatggcgagcgccgagaacggcgcttctcatgtggccgccgcggtgaccgcgccggcgaccaccggcggcgtggagagcgagctgcctagcatagtggttgacggtgggcatcagctgccggatcgctga</cdnaseq>|
AA = <aaseq>MAMRGVDFKWYDGFFLSMLATSLIIVSINWKRYRLCAHPLHIWI VVDYTTVFIFRLLMFVDNGLAAGMGLDLGWQQRYARFCGRIVVLSVLVLLLYPFLWVW TVIGTLWFSTARGCLPEEGQKWGFLIWLLFSYCGLACIACVAVGKWLSRRHALQQRAQ QGIPVSEYGVLVDMIRVPDWAFEAVGLEMRGMGQDTAYHPGLYLTAAQREAVEALIQE LPKFRLKAVPTDCSECPICLEEFHVGNEVRGLPCAHNFHVECIDQWLRLNVKCPRCRC SVFPNLDLSALNNLRPSSEPDRPSASEVTAATMARYVRSSQPAGQSYLLRLQGLLLRQ VVVRHGGGDDMASAENGASHVAAAVTAPATTGGVESELPSIVVDGGHQLPDR</aaseq>|
DNA = <dnaseqindica>5026..5448#4306..4419#3647..3763#3376..3441#2743..2835#2521..2659#1061..1134#920..986#427..465#303..331#gaaaagcttcttcgcttcgccgcctcctccccctctctccggcgcgttcgccgctcgcgatcctcccggcgccgccgccgccgccgcgcgggggccctactcgcgtcgacgccgggagggggacgccgtactccctcctcctcgcgtcgccccgccccgccgccgagtgcgacgccgctctctctctctctctcctcgcgagaaaccctagctggagttgaagtgaggggctcactgaagtgagcggagtgcgggggcgtggatgctgccgcgctcctccggccagatctagctagcaggccatggcgatgcggggggtcgatttcaagtggtactaagtttattctctactccccccttccccgctatggctgtaatatgctttgaagaatctattcatctgttgctttgcttgattggtttcaggtacgatggattcttcctctccatgctcgccaccagcctgtatcctttcatcatttcctcaccatgctttcagttttaattattagaatttcgctcgccgcgcgttcagtttttgttagagtcgattacgttttgcaccagttagattaacctacatttcaatttgcaccggactaggtgcaatgggttgggcgggtgttatgtgaatgcaaagtgaaagtttagactaaacctggtggattttttcttcttttttgacataagaaggtagagccagctaccttattaacttatagccatttcattgacaaaaatagatggttacaaagttacaggaagaaggtacatgaaaaagaggaaggtaaaaaggggaggaaaaaagggggagaatgtacaaaacatggtggaaaaatgaaacaatttacccagtgcaatgtgcattttactgatacattttcatcgattgctattgttcctcctcaactttgtccagaatcattgtctccatcaactggaagaggtatcgtctctgcgcccacccgttgcacatatggatcgtggtcagttcctcaatccaacatctatctcaaatcaaaccgctattcctctatctcactctctcatcttttcccaggttgactacaccaccgtcttcatcttccgccttctcatgttcgtcgataatggccttgccgccggcatgggattgtaagtttataactactatctccttctatttcaacctcatctctcctgtcacaatatacctatggtagacctacaaatggattgtaggggtaaagacagtggttgtcattttcctctgctttgggtgggctgttctgttctgctagacagaacaaatctgtaccattttgaccccaaacttaccactgggtttagttctttttaattatggactagtatgaaatttgcatacatgaaattttgatgtcacccctaaatcgagaaaagcagcctagcaaaaaaggaagaacaacaattgacaatgacatttgtaccctttcgggagaaaaggaaattcctgtaataggcaacctgaccaatttttgtcttgactatcttgattgcaacctcagcctccgctctttttaaacgagcaaattataattaaaatacgtttatgaagggacataaggctaaacagcctctttgcatggttattacctgagcagatgagatataaccttttaaccgtctgctaattttggatgagttcatggcgcaaaacaatatagtttgtagtgcaaaacaatatagtttgtagtttcaacatttcagaagatagaagtctcaatttatttttatttagccattactgtattatttttcttctgtgcaacttactgtcttattttgaagatttgccgtaatatgccattgaaatatttcatatttagatttttgtagtattaccattttaacgtgtaacttcaaagttcaaacagaatatgctcttagaatattttctaagactagctgtatgcccacatgtttgatacagataaaaatgaatagaataagtctgtttgcattttcggaacatatatatgtgattgttgcattgaacctgtaaaccttttccttttctgttacaagagcacttgacaatgtctacacatgtctatttgtaaaatttggtcatcttcgacatttccatgtaaaattgcgaatttagaaagccaaatacaaaaaaaagggaaaaaaataaacaacatttggaaggtagaagtgtcattttcaggaatttaagggagcaattagtaagaaatttagtggaggtgttggtagcagattccctgccaccacactaccttctttctaaaggagtacatataacaaatttgtcattctgtgataaagaaacagcatgcaagaggtgtttctatcattaaagcaacactgtgcaataggacagcacataagtatttcctcttacagttgtaagaggcttgagtatcgcctacaaaatagagttgctaacttgccaatgtccacctttgtatgttggtccttaccttctaaacttttccaaagtatgacgtttgtcctttttgtggctggtaattgtagggatcttggatggcaacagagatatgctcgtttttgtgggagaattgttgtcttgtcggttcttgtgcttcttctctatccctttctttgggtttggactgtgataggaacattgtggtttagcactgcaagaggctgtgtaagttttcttattgttatggtattatgctcatcaccaaatatttagaagttttacttaacatacactcactctatatgtagttacccgaggaaggacaaaaatggggcttccttatatggctgcttttcagctactgtggtctcgcctgtattgcgtgtgtggctgttggaaaggtactgctataagatatgtcggtacaaactgcaatcttgcattggatccgcatattatcgttcatcgatttatccatgattactgtgcaaagattcagtcttcttgagtgtcataccatcagtcatacatactatatctagatgtcatttagattgaagatgttaaggttttagttatagagaattgttatttggcattgatgtattcataatttcatatcttctttgcatcatctagaaatcgtatccatttatttttttcttcttttaaaagcatgaagggtagtgtacgtcttccttcatattgactattagtgcagaagccatactgatagcagaaccctaatcctcatggttgtactggctttagatctttttttagtcagaacttttggtagatttcttgtaattttattttgccttcttattttaataaagcaactgtgggggctttccctacagcgcttttttgttaaaaaaaatgtatatttaaattcacagagggcatatcgccatgttttacttttaattttgttgcagtggctaagccgaaggcatgctctccagcagagggcacaacagggaataccagtctctgaatatggggtaagattttttgttcgttttcgctgctgattttgtccagtgaactgtcaatggtagacgtttgaaaaatatgaaactacagcattgcacgccctttttccatgcttgcctgttcaattcagtttgtgcagcttacttgtctcaactactagctttatatttaagtagggaggatttcaattttttttattgtcatcatctataggttttggttgacatgatccgtgtgcctgattgggcatttgaggccgttggcttggaaatgagaggaatgggccaagatactgcatatcatcctggtctttatttaacagctgcccaagtaagttgctttgcgcaatttgatgaatcgcctttcgacatatcgctagtgtgtgtgttgattgattctgttatttgcttatatttaggcatttagctgtacattcgattcaaggggtcgtactgtctgctagctgttggtataatctagctagacaaaatttccatgtatttatttgctagcgatctcagagatgcaaagaagtctctgaagacttctggatttgtattaaggagcaaaggtcagtacaaaacagagagtatgaggaatagaactcaatagaggcagtagaaaaggacaggaaagaaactaatactaattagcaaccacaatatttttgccccagcttgttgccaagcactggcagatctttttgtgataatctaactgagggtgatgtttcctgatcaaaggcatttagatacctttctttccaatgtttgcccaaatgcatgcatttgggaccattgttgaacactgcagtaacactgaaatccagtgtcattgagcagaatgattttaaattggtcctccttctgcagagagaggcagttgaggcactgattcaagaactcccgaagttcagactgaaagctgttcctacagactgcagtgagtgtccaatctgcctggaggaattccatgttggcaacgaggtaaattacagacaccaagtagagaacttgattgaactgcaattctactagtatttgcacatggtgcttcttcatgtgtttgtatgtggcatgaatccatcaggaaaaaagtccatttaacttattttgccagcaatcaactccctgaactatttcttttacccatttgatcccctaaactacatataaaaacagctcatcttgtttctttgtaacacatctcttttccgtctttatacaaggcatacttaagctaaaatttgtacaaccattgatgacaaaataaactactttacaaaatacttctatcatgaaattctgtatgcagtgttgaatctctatatttaaatttttatcttaggtgtgtctaacctatgcaaatagtgaacaaaaattcaaaaaaagaaaaaagagtaataatgtgtacatgccacaatttcaacgaagaatataacttgtataaagaataaaaaagcatggttagaagtaaagtggagcatttttgttagttcgagggtaaaacaagactagagatggtttatgaagttacgttctccagcatttttaaggactgacaaagacttcttttcccaatccattatgcaggtccgtgggctcccgtgcgcgcacaatttccatgtggagtgcatcgaccagtggctccggctgaatgtcaagtgcccccgttgccggtgctccgtcttccccaacctcgacctgagcgcgctcaacaacctccggccatcctccgagccggaccggccgtcggccagcgaggtgacagcggcgacgatggcgcggtacgtgaggtcgtcgcagccggctgggcagagctacctgctgcggctgcaggggctgctgctgcggcaggtggtggttcggcacggcggcggcgacgacatggcgagcgccgagaacggcgcttctcatgtggccgccgcggtgaccgcgccggcgaccaccggcggcgtggagagcgagctgcctagcatagtggttgacggtgggcatcagctgccggatcgctgaagcgcccggtggacggcgtgatgtagactgatagttatatgtgtggggaaggggaggatcgaagctatgcagcagcatgttttttttttctttgatcttatctcctgtgtgcagagtgtataaatattatgtaaaggagatcattattgcaagcttttgtttcccttt</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001061164.1 RefSeq:Os05g0145000]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 5]]
[[Category:Chromosome 5]]

Os05g0145000

2014-05-27T14:41:19Z

Panpan Liu: /* Labs working on this gene */

Please input one-sentence summary here.

==Annotated Information==
===Function===
[[File:Functional_mutant.jpg|right|thumb|250px|'''Figure 1.''' ''Functional complementation of ptb1. (a) Plant phenotype of ptb1. (b) Plant phenotype of the PTB1-transformed ptb1 plants (ptb1–PTB1).(c) Panicle phenotype of ptb1. (d) Panicle phenotype of the ptb1–PTB1 transgenic plants. (from reference<ref name="ref1" />).'']]
The domestication-related POLLEN TUBE BLOCKD 1(''PTB1''), a RING-type E3 ubiquitin ligase, positively regulates the rice panicle seed setting rate by promoting pollen tube growth. The ''PTB1'' positively regulates rice PSSR through controlling pollen tube growth. Shuangcheng et al(2013) support the hypothesis that ''PTB1'' is an important maternal sporophytic factor of pollen tube growth and a key modulator of the rice panicle seed setting rate. 
There are many genes that related to the grain yield such as ''GS3/5'', ''GW2/5/8'', ''GL3.1'' and ''TGW6'', but as we know now, no genes regulates PSSR(which is one of the most important yield determinants) have been functionally characterized. Rice PSSR is impressionable to outside environmental conditions, which often cause large rice yield losss and threatens the safety of rice production. Nowadays, compared with the parents, many hybrid rice varieties exhibit sharp decreases in PSSR, the finding of the PBT1 gene has implications for the improvement of rice yield. 
PTB1 has pleiotropic effects on PT growth control. First, it has a decisive role in entering and early growth of the STT.Second, ''PTB1'' functions during the later navigation of PTs in the STT because a lower level of ''PTB1'' expression leads to different numbers of PTs growing in this process.Third, ''PTB1'' promotes growth from the chalazal end of the sporophytic STT to the micropyle because a moderate expression of ''PTB1'' leads to the growth of few PTs in this female location.Finally, PTB1 functions as a promoting agent based on the observation that a higher expression of this gene encourages more PTs to complete their full journey<ref name="ref1" />.

===Expression===
[[File:Expression1.jpg|right|thumb|250px|'''Figure 2.''' ''Expression patterns of PTB1. (a–h) GUS activity in various organs of PPTB1-GUS-transformed rice plants. (a) Growing root, (b) stem node, (c) stem internode, (d) leaf, (e) floret, (f) pistil, (g) shell of unpollinated spikelet and (h) pollen grains. (i) Relative PTB1 transcript levels of several tissues detected by q-PCR analysis. DAP, days after pollination. Values represent the mean±s.e.m. (n?3). Scale bars, 400 mm in (a), 3mm in (b) and (c), 1 cm in (d), 1mm in (e) and (f), 4mm in (g) and 50 mm in (h). (from reference<ref name="ref1" />).'']]
The natural variation in expression of ''PTB1'' which is affected by the promoter haplotype and the environmental temperature, correlates with the rice panicle seed setting rate. The GUS gene test in rice showed that ''PTB1'' was differentially expressed in various rice tissues: the expression of PTB1 was relatively weak in the root, shoot and leaf, and the expression was strong in the stigma of the pistil and the spikelet hulls of the pre-emergence panicle. But , in rice pollen grains,the expression was undetectable. The results of Q-PCR was further confirmed this temporal and spatial expression patterns by showing that PBT1 was expressed prior in the reproductive organs such as the pistil, young panicle,pre-emergence panicle and post-emergence panicle. This pattern corresponded well to the role ''PBT1'' played in the pollen tube growth<ref name="ref1" />. 
The transient expression of PBT1-YFP fusion protein in rice protoplast cells showed that the PBT1 localized preferentially to the cytoplasm and although the PBT1 protein was predicted to contain 4 transmembrane segments, it was weakly expressed on the membrane.

===Evolution===
Please input evolution information here.

You can also add sub-section(s) at will.

==Labs working on this gene==
Rice Research Institute, Sichuan Agricultural University
State Key Laboratory of Hybrid Rice, Sichuan Agricultural University
Key Laboratory of Crop Genetic Resources and Improvement, Sichuan Agricultural University, Ministry of Education

==References==
<references>
* <ref name="ref1">
Shuangcheng Li1, Wenbo Li1, Bin Huang1, et al.(2013)Natural variation in PTB1 regulates rice seed setting rate by controlling pollen tube growth.NATURE COMMUNICATIONS, DOI: 10.1038/ncomms3793.
</ref>

==Structured Information==
{{JaponicaGene|
GeneName = Os05g0145000|
Description = Similar to Ring finger protein (Fragment)|
Version = NM_001061164.1 GI:115462058 GeneID:4337790|
Length = 5616 bp|
Definition = Oryza sativa Japonica Group Os05g0145000, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 5|Chromosome 5]]|
AP = Chromosome 5:2576595..2582210|
CDS = 2576763..2577185,2577792..2577905,2578448..2578564,2578770..2578835,2579376..2579468 ,2579552..2579690,2581077..2581150,2581225..2581291,2581746..2581784 ,2581880..2581908|
GCID = <gbrowseImage1>
name=NC_008398:2576595..2582210
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008398:2576595..2582210
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atggcgatgcggggggtcgatttcaagtggtacgatggattcttcctctccatgctcgccaccagcctaatcattgtctccatcaactggaagaggtatcgtctctgcgcccacccgttgcacatatggatcgtggttgactacaccaccgtcttcatcttccgccttctcatgttcgtcgataatggccttgccgccggcatgggattggatcttggatggcaacagagatatgctcgtttttgtgggagaattgttgtcttgtcggttcttgtgcttcttctctatccctttctttgggtttggactgtgataggaacattgtggtttagcactgcaagaggctgtttacccgaggaaggacaaaaatggggcttccttatatggctgcttttcagctactgtggtctcgcctgtattgcgtgtgtggctgttggaaagtggctaagccgaaggcatgctctccagcagagggcacaacagggaataccagtctctgaatatggggttttggttgacatgatccgtgtgcctgattgggcatttgaggccgttggcttggaaatgagaggaatgggccaagatactgcatatcatcctggtctttatttaacagctgcccaaagagaggcagttgaggcactgattcaagaactcccgaagttcagactgaaagctgttcctacagactgcagtgagtgtccaatctgcctggaggaattccatgttggcaacgaggtccgtgggctcccgtgcgcgcacaatttccatgtggagtgcatcgaccagtggctccggctgaatgtcaagtgcccccgttgccggtgctccgtcttccccaacctcgacctgagcgcgctcaacaacctccggccatcctccgagccggaccggccgtcggccagcgaggtgacagcggcgacgatggcgcggtacgtgaggtcgtcgcagccggctgggcagagctacctgctgcggctgcaggggctgctgctgcggcaggtggtggttcggcacggcggcggcgacgacatggcgagcgccgagaacggcgcttctcatgtggccgccgcggtgaccgcgccggcgaccaccggcggcgtggagagcgagctgcctagcatagtggttgacggtgggcatcagctgccggatcgctga</cdnaseq>|
AA = <aaseq>MAMRGVDFKWYDGFFLSMLATSLIIVSINWKRYRLCAHPLHIWI VVDYTTVFIFRLLMFVDNGLAAGMGLDLGWQQRYARFCGRIVVLSVLVLLLYPFLWVW TVIGTLWFSTARGCLPEEGQKWGFLIWLLFSYCGLACIACVAVGKWLSRRHALQQRAQ QGIPVSEYGVLVDMIRVPDWAFEAVGLEMRGMGQDTAYHPGLYLTAAQREAVEALIQE LPKFRLKAVPTDCSECPICLEEFHVGNEVRGLPCAHNFHVECIDQWLRLNVKCPRCRC SVFPNLDLSALNNLRPSSEPDRPSASEVTAATMARYVRSSQPAGQSYLLRLQGLLLRQ VVVRHGGGDDMASAENGASHVAAAVTAPATTGGVESELPSIVVDGGHQLPDR</aaseq>|
DNA = <dnaseqindica>5026..5448#4306..4419#3647..3763#3376..3441#2743..2835#2521..2659#1061..1134#920..986#427..465#303..331#gaaaagcttcttcgcttcgccgcctcctccccctctctccggcgcgttcgccgctcgcgatcctcccggcgccgccgccgccgccgcgcgggggccctactcgcgtcgacgccgggagggggacgccgtactccctcctcctcgcgtcgccccgccccgccgccgagtgcgacgccgctctctctctctctctcctcgcgagaaaccctagctggagttgaagtgaggggctcactgaagtgagcggagtgcgggggcgtggatgctgccgcgctcctccggccagatctagctagcaggccatggcgatgcggggggtcgatttcaagtggtactaagtttattctctactccccccttccccgctatggctgtaatatgctttgaagaatctattcatctgttgctttgcttgattggtttcaggtacgatggattcttcctctccatgctcgccaccagcctgtatcctttcatcatttcctcaccatgctttcagttttaattattagaatttcgctcgccgcgcgttcagtttttgttagagtcgattacgttttgcaccagttagattaacctacatttcaatttgcaccggactaggtgcaatgggttgggcgggtgttatgtgaatgcaaagtgaaagtttagactaaacctggtggattttttcttcttttttgacataagaaggtagagccagctaccttattaacttatagccatttcattgacaaaaatagatggttacaaagttacaggaagaaggtacatgaaaaagaggaaggtaaaaaggggaggaaaaaagggggagaatgtacaaaacatggtggaaaaatgaaacaatttacccagtgcaatgtgcattttactgatacattttcatcgattgctattgttcctcctcaactttgtccagaatcattgtctccatcaactggaagaggtatcgtctctgcgcccacccgttgcacatatggatcgtggtcagttcctcaatccaacatctatctcaaatcaaaccgctattcctctatctcactctctcatcttttcccaggttgactacaccaccgtcttcatcttccgccttctcatgttcgtcgataatggccttgccgccggcatgggattgtaagtttataactactatctccttctatttcaacctcatctctcctgtcacaatatacctatggtagacctacaaatggattgtaggggtaaagacagtggttgtcattttcctctgctttgggtgggctgttctgttctgctagacagaacaaatctgtaccattttgaccccaaacttaccactgggtttagttctttttaattatggactagtatgaaatttgcatacatgaaattttgatgtcacccctaaatcgagaaaagcagcctagcaaaaaaggaagaacaacaattgacaatgacatttgtaccctttcgggagaaaaggaaattcctgtaataggcaacctgaccaatttttgtcttgactatcttgattgcaacctcagcctccgctctttttaaacgagcaaattataattaaaatacgtttatgaagggacataaggctaaacagcctctttgcatggttattacctgagcagatgagatataaccttttaaccgtctgctaattttggatgagttcatggcgcaaaacaatatagtttgtagtgcaaaacaatatagtttgtagtttcaacatttcagaagatagaagtctcaatttatttttatttagccattactgtattatttttcttctgtgcaacttactgtcttattttgaagatttgccgtaatatgccattgaaatatttcatatttagatttttgtagtattaccattttaacgtgtaacttcaaagttcaaacagaatatgctcttagaatattttctaagactagctgtatgcccacatgtttgatacagataaaaatgaatagaataagtctgtttgcattttcggaacatatatatgtgattgttgcattgaacctgtaaaccttttccttttctgttacaagagcacttgacaatgtctacacatgtctatttgtaaaatttggtcatcttcgacatttccatgtaaaattgcgaatttagaaagccaaatacaaaaaaaagggaaaaaaataaacaacatttggaaggtagaagtgtcattttcaggaatttaagggagcaattagtaagaaatttagtggaggtgttggtagcagattccctgccaccacactaccttctttctaaaggagtacatataacaaatttgtcattctgtgataaagaaacagcatgcaagaggtgtttctatcattaaagcaacactgtgcaataggacagcacataagtatttcctcttacagttgtaagaggcttgagtatcgcctacaaaatagagttgctaacttgccaatgtccacctttgtatgttggtccttaccttctaaacttttccaaagtatgacgtttgtcctttttgtggctggtaattgtagggatcttggatggcaacagagatatgctcgtttttgtgggagaattgttgtcttgtcggttcttgtgcttcttctctatccctttctttgggtttggactgtgataggaacattgtggtttagcactgcaagaggctgtgtaagttttcttattgttatggtattatgctcatcaccaaatatttagaagttttacttaacatacactcactctatatgtagttacccgaggaaggacaaaaatggggcttccttatatggctgcttttcagctactgtggtctcgcctgtattgcgtgtgtggctgttggaaaggtactgctataagatatgtcggtacaaactgcaatcttgcattggatccgcatattatcgttcatcgatttatccatgattactgtgcaaagattcagtcttcttgagtgtcataccatcagtcatacatactatatctagatgtcatttagattgaagatgttaaggttttagttatagagaattgttatttggcattgatgtattcataatttcatatcttctttgcatcatctagaaatcgtatccatttatttttttcttcttttaaaagcatgaagggtagtgtacgtcttccttcatattgactattagtgcagaagccatactgatagcagaaccctaatcctcatggttgtactggctttagatctttttttagtcagaacttttggtagatttcttgtaattttattttgccttcttattttaataaagcaactgtgggggctttccctacagcgcttttttgttaaaaaaaatgtatatttaaattcacagagggcatatcgccatgttttacttttaattttgttgcagtggctaagccgaaggcatgctctccagcagagggcacaacagggaataccagtctctgaatatggggtaagattttttgttcgttttcgctgctgattttgtccagtgaactgtcaatggtagacgtttgaaaaatatgaaactacagcattgcacgccctttttccatgcttgcctgttcaattcagtttgtgcagcttacttgtctcaactactagctttatatttaagtagggaggatttcaattttttttattgtcatcatctataggttttggttgacatgatccgtgtgcctgattgggcatttgaggccgttggcttggaaatgagaggaatgggccaagatactgcatatcatcctggtctttatttaacagctgcccaagtaagttgctttgcgcaatttgatgaatcgcctttcgacatatcgctagtgtgtgtgttgattgattctgttatttgcttatatttaggcatttagctgtacattcgattcaaggggtcgtactgtctgctagctgttggtataatctagctagacaaaatttccatgtatttatttgctagcgatctcagagatgcaaagaagtctctgaagacttctggatttgtattaaggagcaaaggtcagtacaaaacagagagtatgaggaatagaactcaatagaggcagtagaaaaggacaggaaagaaactaatactaattagcaaccacaatatttttgccccagcttgttgccaagcactggcagatctttttgtgataatctaactgagggtgatgtttcctgatcaaaggcatttagatacctttctttccaatgtttgcccaaatgcatgcatttgggaccattgttgaacactgcagtaacactgaaatccagtgtcattgagcagaatgattttaaattggtcctccttctgcagagagaggcagttgaggcactgattcaagaactcccgaagttcagactgaaagctgttcctacagactgcagtgagtgtccaatctgcctggaggaattccatgttggcaacgaggtaaattacagacaccaagtagagaacttgattgaactgcaattctactagtatttgcacatggtgcttcttcatgtgtttgtatgtggcatgaatccatcaggaaaaaagtccatttaacttattttgccagcaatcaactccctgaactatttcttttacccatttgatcccctaaactacatataaaaacagctcatcttgtttctttgtaacacatctcttttccgtctttatacaaggcatacttaagctaaaatttgtacaaccattgatgacaaaataaactactttacaaaatacttctatcatgaaattctgtatgcagtgttgaatctctatatttaaatttttatcttaggtgtgtctaacctatgcaaatagtgaacaaaaattcaaaaaaagaaaaaagagtaataatgtgtacatgccacaatttcaacgaagaatataacttgtataaagaataaaaaagcatggttagaagtaaagtggagcatttttgttagttcgagggtaaaacaagactagagatggtttatgaagttacgttctccagcatttttaaggactgacaaagacttcttttcccaatccattatgcaggtccgtgggctcccgtgcgcgcacaatttccatgtggagtgcatcgaccagtggctccggctgaatgtcaagtgcccccgttgccggtgctccgtcttccccaacctcgacctgagcgcgctcaacaacctccggccatcctccgagccggaccggccgtcggccagcgaggtgacagcggcgacgatggcgcggtacgtgaggtcgtcgcagccggctgggcagagctacctgctgcggctgcaggggctgctgctgcggcaggtggtggttcggcacggcggcggcgacgacatggcgagcgccgagaacggcgcttctcatgtggccgccgcggtgaccgcgccggcgaccaccggcggcgtggagagcgagctgcctagcatagtggttgacggtgggcatcagctgccggatcgctgaagcgcccggtggacggcgtgatgtagactgatagttatatgtgtggggaaggggaggatcgaagctatgcagcagcatgttttttttttctttgatcttatctcctgtgtgcagagtgtataaatattatgtaaaggagatcattattgcaagcttttgtttcccttt</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001061164.1 RefSeq:Os05g0145000]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 5]]
[[Category:Chromosome 5]]