Os01g0149500
Please input one-sentence summary here.
Contents
Annotated Information
Function
The plant genome contains a large number of disease resistance (R) genes that have evolved through diverse mechanisms. Pit confers race-specific resistance against the fungal pathogen Magnaporthe grisea, and is a member of the nucleotide-binding site leucine-rich repeat (NBS-LRR) family of R genes.
Expression
The structure of the predicted Pit protein is typical of NBSLRR proteins (Figures 3a, S1 and S3). Pit contains conserved motifs indicative of an NBS domain, and the putative LRR domain (with 18 imperfect repeats) matches the cytoplasmic LRR consensus sequence. The Pit protein encoded by the Nipponbare allele (PitNpb) differs from that encoded by the K59 allele (PitK59) at four amino acid positions: 143 (R fi G), 176 (M fi I), 720 (A fi T) and 780 (M fi V) (Figures 3a, S1 and S3). Two of the substitutions (R143G and M176I) are located in the Nterminal half of the protein, whereas the others are in the LRR domain. A series of chimeric transgenes are used to examine which substitution(s) is important for determining Pit resistance(Figure 3b). First, the LRR region of PitK59 is replaced with that of PitNpb (A720T/M780V), because a single amino acid difference in the LRR domain has been shown to affect resistance. However, the PitK59 (A720T/M780V) lines conferred resistance (‘R + RM’) to 92% of the transgenic Nipponbare plants, suggesting that the two substitutions in the LRR region are not important for Pit function. Next, mutating the corresponding amino acids in PitNpb (R143G and M176I) to analyze the two substitutions in the N-terminal half of PitK59. A 24% reduction in resistance was observed in the transgenic plants as a result. Whereas the M176I mutation, located between the second CC motif (CC2) and the P loop, conferred resistance to 87% of the plants, R143G, located in CC2, conferred resistance to only 55% of the plants. These results suggest that the R143G mutation affected the resistance. Indeed, transgenic populations harboring the R143G mutation [i.e. PitK59 (R143G/M176I), PitK59 (R143G), Kpro-PitNpb and Ubi-PitNpb] obviously showed higher proportions of RS phenotypes (more than 18%) compared with the other populations. To examine whether these RS phenotypes are caused by the lower expression of chimeric transgenes, RT-PCR analysis was performed (Figure 3c). Both PitK59 (R143G/M176I) and PitK59 (R143G) transgenic plants showed higher levels of transgene expression than K59, irrespective of their phenotypes: i.e. transgenic plants with the RS phenotype also expressed the transgenes at high levels. These results further suggest that the R143G mutation may have destabilized the resistance function. However, considering the result that 80% of gPitNpb plants showed the complete susceptible (S) phenotype (Figure 3b), Pit requires another factor for its activity. As noted above, the structures of PitK59 and PitNpb differ in their promoter and 5¢-UTR regions; moreover, the efficiency of translation is controlled by various features (e.g. 5¢-UTR). Therefore, experiments were taken to examine the effects of the promoter and 5¢-UTR from each gene on Pit function using the constructs Npro5¢-PitK59, Npro-PitK59, Kpro-PitNpb and Ubi-PitNpb (Figure 3b). Npro5¢-PitK59, which contained 1.9 kb of the promoter and the 1.3-kb 5¢-UTR of PitNpb fused to the remainder of PitK59, conferred resistance to only 31% of the T0 plants. Similarly, only 30% of T0 plants expressing Npro-PitK59, in which the 1.9-kb promoter of PitNpb was fused directly with the coding sequence and terminator of PitK59, to eliminate possible effects of the 5¢-UTR of PitNpb, exhibited a resistant phenotype. These results negate the possibility that the extended 5¢-UTR of PitNpb affects Pit function. However, surprisingly, Kpro-PitNpb, which carried the PitNpb coding sequence driven by the PitK59 promoter consisting of the 3¢ fragment of Renovator (1040 bp) and the 5¢-UTR (254 bp) of PitK59, conferred resistance to 78% of the transgenic plants (Figure 3b). Finally, 71% of plants overexpressing the PitNpb open-reading frame (ORF) (Ubi-PitNpb) exhibited resistance (Figure 3b). The expression levels of PitK59 or PitNpb in gPitNpb, Npro5¢-PitK59, Npro-PitK59 and Ubi-PitNpb were correlated with their resistance (Figure 3d,e). These results clearly indicate that the promoter activity conferred by the Renovator retrotransposon fragment, rather than the amino acid substitutions, plays an important role in Pit resistance by enhancing its transcription. Notably, the increased resistance displayed by the plants expressing Kpro-PitNpb, compared with those expressing Npro-PitK59, indicates that promoter activity plays a dominant role in PitK59 function.
Evolution
Compared with the non-functional allele PitNpb, the functional allele PitK59 contains four amino acid substitutions, and has the LTR retrotransposon Renovator inserted upstream. Pathogenesis assays using chimeric constructs carrying the various regions of PitK59 and PitNpb suggest that amino acid substitutions might have a potential effect in Pit resistance; more importantly, the upregulated promoter activity conferred by the Renovator sequence is essential for Pit function. It is said that transposon-mediated transcriptional activation may play an important role in the refunctionalization of additional 'sleeping'R genes in the plant genome.
Labs working on this gene
National Agricultural Research Center, National Agriculture and Food Research Organization (NARO), 1-2-1 Inada, Jo-etsu, Niigata 943 0193, Japan
References
Please input cited references here.
Structured Information
| Gene Name |
Os01g0149500 |
|---|---|
| Description |
Similar to Resistance gene analog PIC23 (Fragment) |
| Version |
NM_001048560.1 GI:115434533 GeneID:4324511 |
| Length |
3143 bp |
| Definition |
Oryza sativa Japonica Group Os01g0149500, 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 1:2682222..2685364 |
| Sequence Coding Region |
2682222..2683988 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008394:2682222..2685364 source=RiceChromosome01 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008394:2682222..2685364 source=RiceChromosome01 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atgggcacagttttggatgctctcgcttggaaattcttggaaaagctgggtcaacttattgaggatgaggttatcatgacattaagtgtgaaaaggggtattgagagcctgaagaaaaatctggaattcttcaacgctgttcatgaggatgctgaggctctggccatggaggatccagggatagattcatggtggaagaacatgagagatgttatgtttgatgtcgatgacatcgtagatcttttcatggttcattcacagaagctcttgctgccaccaaggccggtatgttgcaatcagcccttgttctctagctttgcaaagttttcatttgatcacatgattgccaaaagaattgataatataaatgaaaagtttgaagaaattaaaatgaacaaagagatgtttggcttggagagaaccaacgggcaacaaatccagataacaattgttgacagaagtcagacatctcctgtagatgaacttgaggtggttggagaagatatcagaagggctattgatgacatcgtcaaaatgattgttagtagtaactacaatgaaagcaggtcaactgtttttgggatccaggggatgggaggcatcggtaagacaacattagctcaaaagatatacaatgaacagaggataagggagaaattccaagttcatatatggttgtgcatttctcagaattacacagagaccagtttgttaaaacaggcaatcagaatggctggaggaatttgtgatcagctggagactaagactgagcttctaccgcttctagtggacactattagagggaagagtgtttttcttgtgctggatgatgtttggaaatctgatgtatggattgatcttcttcgtttgcctttcttgagaggtttgaattctcatatccttgtgacctcaagaaacctggatgttttggttgaaatgcatgcaacttatacccacaaggtaaacaaaatgaatgattgcgatggcctagaactgctgatgaagatgtcattagggccatacgagcaaagcagagaattcagtggtgttggatatcaaattgtcaaaaagtgtgatggccttcctcttgccatcaaagttgttgcaggtgttctatctactaaaagaacaagggcagaatgggagagtatccgagatagcaagtggtccattcatggacttcccagagaactaggaggccctctgtacttaagctacagcaacttaccccctgaacttaaacaatgctttctctggtgtgctttgttgccgtcaaattttgtgatacgtcgtgatgctgtcgcttactggtgggttgctgaaggtttcgtgacggaagtgcatgggtactcaatacatgaggttgctgaagagtactaccatgagctaatcagaaggaatcttctgcagccacgaccagagtttgtagacaaaggcgagtcaacaatgcatgatctattgaggtcacttggtcaatttttgacaaaggatcactccatattcatgaacatggaatatagcaaagccttaccaaatctacgccacttatgtatcagcaatgatgtagaagaaatacctgctatagaaaagcaaaagtgcttgaggagcctcctagtttttgataacaaaaacttcatgaaaattaataaggacattttcagagaactcaagcatatccgtgtcttagttctcagtggaacaagcatccaaatcataccagagtcagtgggaaacttcttgttactgaggcttctagatctaagctat</cdnaseq> |
| Protein Sequence |
<aaseq>MGTVLDALAWKFLEKLGQLIEDEVIMTLSVKRGIESLKKNLEFF NAVHEDAEALAMEDPGIDSWWKNMRDVMFDVDDIVDLFMVHSQKLLLPPRPVCCNQPL FSSFAKFSFDHMIAKRIDNINEKFEEIKMNKEMFGLERTNGQQIQITIVDRSQTSPVD ELEVVGEDIRRAIDDIVKMIVSSNYNESRSTVFGIQGMGGIGKTTLAQKIYNEQRIRE KFQVHIWLCISQNYTETSLLKQAIRMAGGICDQLETKTELLPLLVDTIRGKSVFLVLD DVWKSDVWIDLLRLPFLRGLNSHILVTSRNLDVLVEMHATYTHKVNKMNDCDGLELLM KMSLGPYEQSREFSGVGYQIVKKCDGLPLAIKVVAGVLSTKRTRAEWESIRDSKWSIH GLPRELGGPLYLSYSNLPPELKQCFLWCALLPSNFVIRRDAVAYWWVAEGFVTEVHGY SIHEVAEEYYHELIRRNLLQPRPEFVDKGESTMHDLLRSLGQFLTKDHSIFMNMEYSK ALPNLRHLCISNDVEEIPAIEKQKCLRSLLVFDNKNFMKINKDIFRELKHIRVLVLSG TSIQIIPESVGNFLLLRLLDLSY</aaseq> |
| Gene Sequence |
<dnaseqindica>1377..3143#gtgcggtggggggcatttcatcgaacaccattcgcgcatcgctctccccgagcgaaccgcattcgttacgttacgttacgtgcgagatgaatcaacttgtcaggtacaaacacaaccggagaatttgcttttgtttatactggagttatacgatgtgatcctgatccaggctcagttcgacctagcccctgcttatctttgcttgtgggtcgttgtcgccgtctttggttacgctaggagaagcatctctctttcttgcgtaggtaatcaactatcttaatttttcttttgtgtcttgattatgtgcaacaccgagtgatgcgaatgggatgtgttcctttcgatgttttttctatataagctctgaaacttggacttttgcaacaaatggttcgggtgcagatgagctgtttgacgaaatgcccaggagattcctgcgtgcgttgcatgcccagcggcggtttgactctatttttcctttagtatggtgtctgctctctggtgatctaatgtaccattttctcttaaccttgttttacccaatcttaatgcaaagatgcataaagagaactctgcatattcctgttttctcttaaccttgtttcagagaaccttgttttctctgaactctgcaaaagattttctgcatattcctgaaaaaaggagaactctgctccggtgcttttaactagacttataaaggctttggcataggattcctgtattggttgatgcaaaatctgctggtaacagtcagctaaattcgcttaaaaagtacacttctttgggtcttcttgctagtgcaggtagagaagcaccatggccttcctggtttcttaggtggtaaacaggcaaagaaagtaaagcaccaagtggcgccattgctgcaatgaaatcaaattattattagcaagtggatgaaaggaaaagaatacacagcctggaagagctttcaacttgaatccccttgcaattgtgtaacactacttgtccttttcacattgcatcatcgttgaaacatttggattttaatacaaaaatcattgtgggtagattttgcctcaaaaagtgctttcatactgttttaattttgtcaggtttttcagacgttttgcaatagagaatgtcaatgtccattgtgacaaaggcagttggtggcaagaaagggaggagaatctataacagctctcttctgcaatctgcattgccctctctgttctcatcaggttttagccacactgcttcaatatctctttccatcttccgtatttaagtgatcggcactcagcagctctgtgagatatctcttctgggatctgcagtttcaagcatctcccataggagagatccagtgggaatccgtaggttctagcactatgggcacagttttggatgctctcgcttggaaattcttggaaaagctgggtcaacttattgaggatgaggttatcatgacattaagtgtgaaaaggggtattgagagcctgaagaaaaatctggaattcttcaacgctgttcatgaggatgctgaggctctggccatggaggatccagggatagattcatggtggaagaacatgagagatgttatgtttgatgtcgatgacatcgtagatcttttcatggttcattcacagaagctcttgctgccaccaaggccggtatgttgcaatcagcccttgttctctagctttgcaaagttttcatttgatcacatgattgccaaaagaattgataatataaatgaaaagtttgaagaaattaaaatgaacaaagagatgtttggcttggagagaaccaacgggcaacaaatccagataacaattgttgacagaagtcagacatctcctgtagatgaacttgaggtggttggagaagatatcagaagggctattgatgacatcgtcaaaatgattgttagtagtaactacaatgaaagcaggtcaactgtttttgggatccaggggatgggaggcatcggtaagacaacattagctcaaaagatatacaatgaacagaggataagggagaaattccaagttcatatatggttgtgcatttctcagaattacacagagaccagtttgttaaaacaggcaatcagaatggctggaggaatttgtgatcagctggagactaagactgagcttctaccgcttctagtggacactattagagggaagagtgtttttcttgtgctggatgatgtttggaaatctgatgtatggattgatcttcttcgtttgcctttcttgagaggtttgaattctcatatccttgtgacctcaagaaacctggatgttttggttgaaatgcatgcaacttatacccacaaggtaaacaaaatgaatgattgcgatggcctagaactgctgatgaagatgtcattagggccatacgagcaaagcagagaattcagtggtgttggatatcaaattgtcaaaaagtgtgatggccttcctcttgccatcaaagttgttgcaggtgttctatctactaaaagaacaagggcagaatgggagagtatccgagatagcaagtggtccattcatggacttcccagagaactaggaggccctctgtacttaagctacagcaacttaccccctgaacttaaacaatgctttctctggtgtgctttgttgccgtcaaattttgtgatacgtcgtgatgctgtcgcttactggtgggttgctgaaggtttcgtgacggaagtgcatgggtactcaatacatgaggttgctgaagagtactaccatgagctaatcagaaggaatcttctgcagccacgaccagagtttgtagacaaaggcgagtcaacaatgcatgatctattgaggtcacttggtcaatttttgacaaaggatcactccatattcatgaacatggaatatagcaaagccttaccaaatctacgccacttatgtatcagcaatgatgtagaagaaatacctgctatagaaaagcaaaagtgcttgaggagcctcctagtttttgataacaaaaacttcatgaaaattaataaggacattttcagagaactcaagcatatccgtgtcttagttctcagtggaacaagcatccaaatcataccagagtcagtgggaaacttcttgttactgaggcttctagatctaagctat</dnaseqindica> |
| External Link(s) |
