Cloning and characterization of a candidate bacterial leaf blight resistance gene, a serine/threonine protein kinase, in rice
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Abstract
Background and Objective: Bacterial leaf blight (BLB), caused by Xanthomonas oryzae pv. oryzae (Xoo), is a major constraint on rice (Oryza sativa L.) production, particularly in Asia, where rice is a dietary staple. Enhancing host resistance through the deployment of resistance genes remains one of the most effective strategies in rice breeding programs. Advances in molecular tools, including marker-assisted selection (MAS), quantitative trait loci (QTL) mapping, and genome-wide association studies (GWAS), have facilitated the identification of at least 46 BLB resistance genes, 11 of which have been functionally characterized. Building on the previous work, 127 putative BLB resistance genes were identified via GWAS. This study aimed to examine the expression and variation of candidate genes to identify genes potentially contributing to BLB resistance.
Methodology: Quantitative PCR (qPCR) was used to evaluate the expression of selected candidate genes in two rice cultivars: IR57514 (a resistant cultivar) and Jao Hom Nin (a susceptible cultivar), following Xoo inoculation. One gene, LOC_Os01g66860, encoding a serine/threonine protein kinase, was selected for further analysis. A phylogenetic analysis was performed to assess evolutionary conservation across the plant kingdom, and the gene was cloned and sequenced from both cultivars.
Main Results: LOC_Os01g66860 was significantly upregulated in the resistant cultivar after pathogen challenge. Phylogenetic analysis confirmed its conservation across plant species. Cloning and sequencing revealed one amino acid substitution between the resistant and susceptible cultivars.
Conclusions: These findings suggest that LOC_Os01g66860 may play a crucial role in BLB resistance. The results provide a foundation for future functional validation and highlight its potential application in rice breeding programs aimed at improving disease resistance. However, a limitation of this study is the need for further functional tests, such as gene knockout or overexpression experiments, to confirm the gene’s specific role in resistance.
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