Difference between revisions of "Os02g0743400"
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The expression of OsPIN1 in different tissues was analyzed by reverse transcriptase PCR (RT-PCR) and a β-glucuro- nidase (GUS) assay of transgenic plants expressing an OsPIN1 promoter/GUS fusion construct. RT-PCR analysis showed that OsPIN1 mRNA was transcribed in all tissues tested. The expression of the gene was higher in leaf, flower and seed than in root, stem and root collar. To confirm the expression patterns, a binary vector containing the GUS gene driven by the OsPIN1 promoter was constructed and used for rice transformation. The transgenic plants showed GUS expression in the vascular tissues of root, stem, anther, leaf and embryo of the seed. Cross-sections showed that GUS is also expressed in the primordia of adventitious and lateral roots, suggesting that OsPIN1 may be involved in root development. | The expression of OsPIN1 in different tissues was analyzed by reverse transcriptase PCR (RT-PCR) and a β-glucuro- nidase (GUS) assay of transgenic plants expressing an OsPIN1 promoter/GUS fusion construct. RT-PCR analysis showed that OsPIN1 mRNA was transcribed in all tissues tested. The expression of the gene was higher in leaf, flower and seed than in root, stem and root collar. To confirm the expression patterns, a binary vector containing the GUS gene driven by the OsPIN1 promoter was constructed and used for rice transformation. The transgenic plants showed GUS expression in the vascular tissues of root, stem, anther, leaf and embryo of the seed. Cross-sections showed that GUS is also expressed in the primordia of adventitious and lateral roots, suggesting that OsPIN1 may be involved in root development. | ||
| − | To investigate the function of OsPIN1 in rice, transgenic plants overexpressing or underexpressing OsPIN1 were produced using 35S-driven OsPIN1 and OsPIN1 RNA interfering (RNAi) constructs. Two transgenic lines were selected to represent the overexpressing or underexpressing groups. They were designated as 35S1 and 35S2, and RNAi1 and RNAi2, respectively. At the seedling stage, 14 days after germination, plant height, primary root length, adventitious root number and lateral root number on the primary root were investigated in wild-type and transgenic plants. The tiller number was counted at the maximum tillering stage (80 days after germination) under field conditions. The results showed that overexpression of the OsPIN1 gene significantly increased the primary root length and lateral root number, while underex- pression of OsPIN1 resulted in a reduced number of adventitious roots and a significantly increased number of tillers. Moreover, the tiller angles in the RNAi plants exceeded 30 degrees, which was much higher than the wildtype plants | + | To investigate the function of OsPIN1 in rice, transgenic plants overexpressing or underexpressing OsPIN1 were produced using 35S-driven OsPIN1 and OsPIN1 RNA interfering (RNAi) constructs. Two transgenic lines were selected to represent the overexpressing or underexpressing groups. They were designated as 35S1 and 35S2, and RNAi1 and RNAi2, respectively. At the seedling stage, 14 days after germination, plant height, primary root length, adventitious root number and lateral root number on the primary root were investigated in wild-type and transgenic plants. The tiller number was counted at the maximum tillering stage (80 days after germination) under field conditions. The results showed that overexpression of the OsPIN1 gene significantly increased the primary root length and lateral root number, while underex- pression of OsPIN1 resulted in a reduced number of adventitious roots and a significantly increased number of tillers. Moreover, the tiller angles in the RNAi plants exceeded 30 degrees, which was much higher than the wildtype plants. |
| + | |||
| + | To verify whether OsPIN1 is involved in polar auxin | ||
| + | transport, seedlings of the wild-type and transgenic plants were | ||
| + | exposed to 0.1 μM α-naphthylacetic acid (α-NAA) and 0.5 μM | ||
| + | NPA. The numbers of adventitious roots at the 7, 14 and 21- | ||
| + | day-old seedling stages were recorded (Table 2). Results | ||
| + | showed that the suppression of the OsPIN1 gene inhibited the | ||
| + | formation of adventitious roots. Thus the number of adventitious | ||
| + | roots in the OsPIN1 RNA interference (RNAi) transgenic | ||
| + | plants was significantly less than that in wild-type plants (Table | ||
| + | 2). NPA treatment had a similar effect on the wild-type plants | ||
| + | (Table 2), suggesting that underexpression of the OsPIN1 gene | ||
| + | may have affected the auxin transport process. Exogenously | ||
| + | applied NAA on the RNAi transgenic plants partially complemented | ||
| + | the inhibition effect of the underexpression of OsPIN1, | ||
| + | confirming that OsPIN1 is involved in polar auxin transport. | ||
| + | |||
[[File: OsPIN1 expression in different tissues.jpg]] | [[File: OsPIN1 expression in different tissues.jpg]] | ||
Revision as of 14:29, 3 June 2014
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Contents
Annotated Information
Function
OsPIN1 was expressed in the vascular tissues and root primordial in a manner similar to AtPIN1. Adventitious root emergence and development were significantly inhibited in the OsPIN1 RNA interference (RNAi) transgenic plants, which was similar to the phenotype of NPA (N-1-naphthylphalamic acid, an auxintransport inhibitor)-treated wild-type plants. α-naphthylacetic acid (α-NAA) treatment was able to rescue the mutated phenotypes occurring in the RNAi plants. Overexpression or suppression of the OsPIN1 expression through a transgenic approach resulted in changes of tiller numbers and shoot/root ratio. Taken together, these data suggest that OsPIN1 plays an important role in auxindependent adventitious root emergence and tillering[1][2][3].
Five adventitious roots were formed at the coleoptile node of 7-day-old seedlings in both the RNAi transgenic and the wild-type plants. In the second week, numbers of adventitious roots developed from the first node of the wildtype plants, while no new adventitious roots were initiated until the third week in the RNAi transgenic plants. Moreover, the number of adventitious roots in the RNAi transgenic plants was significantly less than in the wild-type plants.
Results showed that adventitious root primordia were initiated in 7-day-old seedlings of both wild-type and OsPIN1 RNAi transgenic plants, indicating that the formation of adventitious root primordia at the first node was not affected by the underexpression of OsPIN1. However, in the 14-day-old seedlings, new adventitious roots had developed in the wildtype plants, while they were not visible in the OsPIN1 RNAi transgenic plants. The adventitious root primordial in these RNAi plants stayed at the same size as those in the 7-day-old seedlings, indicating that the emergence of adventitious root primordia was arrested in these RNAi seedlings. Data showed that the development of adventitious roots in RNAi seedlings ceased at the sixth stage.
Expression
The expression of OsPIN1 in different tissues was analyzed by reverse transcriptase PCR (RT-PCR) and a β-glucuro- nidase (GUS) assay of transgenic plants expressing an OsPIN1 promoter/GUS fusion construct. RT-PCR analysis showed that OsPIN1 mRNA was transcribed in all tissues tested. The expression of the gene was higher in leaf, flower and seed than in root, stem and root collar. To confirm the expression patterns, a binary vector containing the GUS gene driven by the OsPIN1 promoter was constructed and used for rice transformation. The transgenic plants showed GUS expression in the vascular tissues of root, stem, anther, leaf and embryo of the seed. Cross-sections showed that GUS is also expressed in the primordia of adventitious and lateral roots, suggesting that OsPIN1 may be involved in root development. To investigate the function of OsPIN1 in rice, transgenic plants overexpressing or underexpressing OsPIN1 were produced using 35S-driven OsPIN1 and OsPIN1 RNA interfering (RNAi) constructs. Two transgenic lines were selected to represent the overexpressing or underexpressing groups. They were designated as 35S1 and 35S2, and RNAi1 and RNAi2, respectively. At the seedling stage, 14 days after germination, plant height, primary root length, adventitious root number and lateral root number on the primary root were investigated in wild-type and transgenic plants. The tiller number was counted at the maximum tillering stage (80 days after germination) under field conditions. The results showed that overexpression of the OsPIN1 gene significantly increased the primary root length and lateral root number, while underex- pression of OsPIN1 resulted in a reduced number of adventitious roots and a significantly increased number of tillers. Moreover, the tiller angles in the RNAi plants exceeded 30 degrees, which was much higher than the wildtype plants.
To verify whether OsPIN1 is involved in polar auxin transport, seedlings of the wild-type and transgenic plants were exposed to 0.1 μM α-naphthylacetic acid (α-NAA) and 0.5 μM NPA. The numbers of adventitious roots at the 7, 14 and 21- day-old seedling stages were recorded (Table 2). Results showed that the suppression of the OsPIN1 gene inhibited the formation of adventitious roots. Thus the number of adventitious roots in the OsPIN1 RNA interference (RNAi) transgenic plants was significantly less than that in wild-type plants (Table 2). NPA treatment had a similar effect on the wild-type plants (Table 2), suggesting that underexpression of the OsPIN1 gene may have affected the auxin transport process. Exogenously applied NAA on the RNAi transgenic plants partially complemented the inhibition effect of the underexpression of OsPIN1, confirming that OsPIN1 is involved in polar auxin transport.
Evolution
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Labs working on this gene
State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Kaixuan Road, Hangzhou, Zhejiang 310029, PR China
References
- ↑ Xu M, Zhu L, Shou H, Wu P. A PIN1 family gene, OsPIN1, involved in auxin-dependent adventitious root emergence and tillering in rice. Plant Cell Physiol. 2005 Oct;46(10):1674-81.
- ↑ Benková E, Michniewicz M, Sauer M, Teichmann T, Seifertová D, Jürgens G, Friml J. Local, efflux-dependent auxin gradients as a common module for plant organ formation. Cell. 2003 Nov 26;115(5):591-602.
- ↑ Blilou I1, Xu J, Wildwater M, Willemsen V, Paponov I, Friml J, Heidstra R, Aida M, Palme K, Scheres B.The PIN auxin efflux facilitator network controls growth and patterning in Arabidopsis roots. Nature. 2005 Jan 6;433(7021):39-44.
Structured Information
| Gene Name |
Os02g0743400 |
|---|---|
| Description |
Auxin transport protein REH1 |
| Version |
NM_001054630.1 GI:115448630 GeneID:4330700 |
| Length |
3075 bp |
| Definition |
Oryza sativa Japonica Group Os02g0743400, 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 | |
| Location |
Chromosome 2:32045781..32048855 |
| Sequence Coding Region |
32046245..32046311,32046396..32046472,32046764..32046921,32047015..32047100,32047215..32047488 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008395:32045781..32048855 source=RiceChromosome02 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008395:32045781..32048855 source=RiceChromosome02 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atgattacggcggcggacttctaccacgtgatgacggcgatggtgccgttgtacgtggcgatgatactggcgtacgggtcggtgaagtggtggcgcatcttcacgccggaccagtgctccgggatcaaccgcttcgtggcgctcttcgccgtgccgctgctgtcgttccacttcatctccaccaacaacccgtacacgatgaacctccggttcatcgccgccgacacgctgcagaagctgatggtgctggccatgctcacggcgtggagccacctcagccgccgggggagcctcgagtggaccatcacgctcttctccctctccacgctgcccaacacgctcgtcatggggatccctttgctcaagggcatgtacggggagttctccggcagcctcatggtgcagatcgtcgtgctgcagtgcatcatctggtacacgctcatgctcttcatgttcgagtaccgcggcgcccggatgctcatcaccgagcagttcccggacaccgccgccaacatcgcctccatcgtcgtcgacccggacgtcgtgtcgctggacggcaggagggacgccatcgagacggagacggaggtgaaggaggacggcaggatacacgtcaccgtgcgccgctccaacgcgtctcgctcggacatctactcccgccgctccatgggcttctccagcaccacgccgcggccgagcaacctcaccaacgccgagatctactcgctgcagtcgtcgcggaacccgacgccgaggggttcaagcttcaaccacaccgacttctactccatggtgggccgcagctccaacttcggcgcggccgacgcgttcggcgtccgcaccggcgccacgccgcgcccgtccaactacgaggacgacgcgtccaagcccaagtacccgctcccggcgtcgaatgcggcgcccatggcgggccactacccggcgccgaacccggccgtgtcgtcggcgcccaagggcgccaagaaggcggccacgaacgggcaggccaagggcgaggacctccacatgttcgtctggagctccagcgcgtcgcccgtgtccgacgtcttcggcggcggcgcgccagactacaacgacgccgcggcagtcaagtccccccgcaaaatggatggagcgaaggacagggaggactacgtggagcgggacgatttcagcttcgggaacaggggcgtcatggacagggacgcggaggcaggggacgagaaggcggcggcggcggcgggcgccgaccccagcaaggccatggcggcgccgacggcgatgccgccgacgagcgtgatgacccgcctcatcctgatcatggtgtggcgcaagctcatccgcaacccgaacacctactccagcctcatcggcctcatctggtccctcgtctgcttcaggtggaacttcgagatgccggccatcgtcctgaaatccatctcgatcctgtcggacgcggggctcggcatggccatgttcagtctcggtctgttcatggcgctgcagccgcacatcatcgcgtgcgggaacaaggtggcgacgtacgccatggcggtgcggttcctggccgggccggccgtgatggcggcggcgtccttcgccgtcggactccgtggcacgctcctgcacgtcgccattgtccaggcagctctgccccagggcattgtccccttcgtcttcgccaaggagtacagcgtgcaccctagcattctcagcacagctgtcatctttggcatgctcatcgccttgcctatcaccctcgtctactacatcttgcttgggctgtaa</cdnaseq> |
| Protein Sequence |
<aaseq>MITAADFYHVMTAMVPLYVAMILAYGSVKWWRIFTPDQCSGINR FVALFAVPLLSFHFISTNNPYTMNLRFIAADTLQKLMVLAMLTAWSHLSRRGSLEWTI TLFSLSTLPNTLVMGIPLLKGMYGEFSGSLMVQIVVLQCIIWYTLMLFMFEYRGARML ITEQFPDTAANIASIVVDPDVVSLDGRRDAIETETEVKEDGRIHVTVRRSNASRSDIY SRRSMGFSSTTPRPSNLTNAEIYSLQSSRNPTPRGSSFNHTDFYSMVGRSSNFGAADA FGVRTGATPRPSNYEDDASKPKYPLPASNAAPMAGHYPAPNPAVSSAPKGAKKAATNG QAKGEDLHMFVWSSSASPVSDVFGGGAPDYNDAAAVKSPRKMDGAKDREDYVERDDFS FGNRGVMDRDAEAGDEKAAAAAGADPSKAMAAPTAMPPTSVMTRLILIMVWRKLIRNP NTYSSLIGLIWSLVCFRWNFEMPAIVLKSISILSDAGLGMAMFSLGLFMALQPHIIAC GNKVATYAMAVRFLAGPAVMAAASFAVGLRGTLLHVAIVQAALPQGIVPFVFAKEYSV HPSILSTAVIFGMLIALPITLVYYILLGL</aaseq> |
| Gene Sequence |
<dnaseqindica>2545..2611#2384..2460#1935..2092#1756..1841#1368..1641#146..1271#gcgcacacacccaatcaaatgctcctcctccgccgcctcctctcgctgagctgagctgagctgtgaaatagtgccaccgagtgagcgctgagctgagctcgaagcaagagaggaagaggaggaggagggaagagggggggcgaagatgattacggcggcggacttctaccacgtgatgacggcgatggtgccgttgtacgtggcgatgatactggcgtacgggtcggtgaagtggtggcgcatcttcacgccggaccagtgctccgggatcaaccgcttcgtggcgctcttcgccgtgccgctgctgtcgttccacttcatctccaccaacaacccgtacacgatgaacctccggttcatcgccgccgacacgctgcagaagctgatggtgctggccatgctcacggcgtggagccacctcagccgccgggggagcctcgagtggaccatcacgctcttctccctctccacgctgcccaacacgctcgtcatggggatccctttgctcaagggcatgtacggggagttctccggcagcctcatggtgcagatcgtcgtgctgcagtgcatcatctggtacacgctcatgctcttcatgttcgagtaccgcggcgcccggatgctcatcaccgagcagttcccggacaccgccgccaacatcgcctccatcgtcgtcgacccggacgtcgtgtcgctggacggcaggagggacgccatcgagacggagacggaggtgaaggaggacggcaggatacacgtcaccgtgcgccgctccaacgcgtctcgctcggacatctactcccgccgctccatgggcttctccagcaccacgccgcggccgagcaacctcaccaacgccgagatctactcgctgcagtcgtcgcggaacccgacgccgaggggttcaagcttcaaccacaccgacttctactccatggtgggccgcagctccaacttcggcgcggccgacgcgttcggcgtccgcaccggcgccacgccgcgcccgtccaactacgaggacgacgcgtccaagcccaagtacccgctcccggcgtcgaatgcggcgcccatggcgggccactacccggcgccgaacccggccgtgtcgtcggcgcccaagggcgccaagaaggcggccacgaacgggcaggccaagggcgaggacctccacatgttcgtctggagctccagcgcgtcgcccgtgtccgacgtcttcggcggcggcgcgccagactacaacgacgccgcggcagtcaagtccccccgcaaaagtagcaatcttttatcttcaccgtgtactatttgcctgatttggggagcatttcttgggctaatttgtactgactcgtatatttcttttacgtcagtggatggagcgaaggacagggaggactacgtggagcgggacgatttcagcttcgggaacaggggcgtcatggacagggacgcggaggcaggggacgagaaggcggcggcggcggcgggcgccgaccccagcaaggccatggcggcgccgacggcgatgccgccgacgagcgtgatgacccgcctcatcctgatcatggtgtggcgcaagctcatccgcaacccgaacacctactccagcctcatcggcctcatctggtccctcgtctgcttcaggtgcgtacaccccgggcttctcgcaaacccaaacccatctgctcgtacgtgctgtggtggtgcggtgtggtggtctggttctgacaaggcgttcgtctcgttttggcgttgcaggtggaacttcgagatgccggccatcgtcctgaaatccatctcgatcctgtcggacgcggggctcggcatggccatgttcagtctcggtcggtattctcttgtttccatgctaacctcacctcacccccggcagtgtcacgagctgttcccctgagtttgcgatctcgccatgcatgcaggtctgttcatggcgctgcagccgcacatcatcgcgtgcgggaacaaggtggcgacgtacgccatggcggtgcggttcctggccgggccggccgtgatggcggcggcgtccttcgccgtcggactccgtggcacgctcctgcacgtcgccattgtccaggtaaaccggaacggaaaagaacagccatacgcacacactgtcacacgcacagtcacacaccgaaaataaaggtctccgtgtgggtgggcatgcaacctaccgcgcgcagccagccagcgtcgttcttttctcccagcccctcgtgctcgttcgcgtctgctttctctcctgccttttgctctccacggccgcggcactttctacgtttggccagcacgaaagccttttgctttggctaacctttttctctctgttgttcttttgtttggtccaaaccgaacgaacgaacaggcagctctgccccagggcattgtccccttcgtcttcgccaaggagtacagcgtgcaccctagcattctcagcacagcgtaagtactagattctctctcagtttgcactctgtcctgaatcgacgggccactgacattttttgcctttgctcctgctcgcagtgtcatctttggcatgctcatcgccttgcctatcaccctcgtctactacatcttgcttgggctgtaatcgagttgcatgcatgtaaattcctgctcctgacaaccagccatgttaagaagaggggagaagaagacagagctggtacactgtttgcaaagtcaggactctttgatttttcttttcttttctgtatttcttgaagtagaatttgggaggagggggattggaagggagtcaaacgagtcaaggggaggacaggatggtagctagcttagctaggacaatggtgagtcacaaaagagcaccaaaagcaagtacaagtacaaagcttggggggacacaggatccagttcaggtcacagaaacggttcggttttgggaggggattgtgggagttttggttggctgcgctgcgctgacccttgtaaaacggacgccgattctgacaagagatcgaccttgttttgttcgtgtcgtggtattgctgactactactttggaatgattaatctcctactactaattactcc</dnaseqindica> |
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