Rice (Oryza sativa) plays a significant role in achieving global food security. However, it suffers from several biotic and abiotic stresses that seriously affect its production. Rice blast caused by hemibiotropic fungal pathogen Magnaporthe oryzae is one of the most widespread and devastating diseases of rice. The crop rice is vulnerable to this pathogen from seedlings to adult plant stages affecting leaves, nodes, collar, panicles and roots. This disease can be effectively managed through host resistance. Of the 100 blast resistance genes, identified and mapped in different genotypes of rice, 19 genes have been cloned and characterized at the molecular level. Most of these genes belong to nucleotide binding sites and leucine rich repeats. Besides more than 350 quantitative trait loci (QTLs) have also been identified in the rice genome. These blast resistance genes and QTLs have been successfully mobilized in the commercial cultivars by using standard plant breeding techniques and also by marker assisted backcross breeding. With the advent of latest molecular biology techniques and our understanding of the basic mechanisms of Magnaporthe-rice pathosystem, the strategies for broadspectrum resistance to M. oryzae can be designed in future.

The dominant rice blast resistance gene Pi54 cloned by map-based cloning approach from indica rice cultivar Tetep confers broad spectrum resistance to Magnaporthe oryzae. In this investigation, an orthologue of Pi54 designated as Pi54of was cloned from Oryza officinalis conferring high degree of resistance to M. oryzae and is functionally validated. We have also characterized the Pi54of protein and demonstrate its interaction with AVR-Pi54 protein. The Pi54of encoded ∼43 kDa small and unique cytoplasmic LRR family of disease resistance protein having unique Zinc finger domain overlapped with the leucine rich repeat regions. Pi54of showed Magnaporthe-induced expression. The phylogenetic and western blot analysis confirmed orthologous nature of Pi54 and Pi54of genes, whereas the identity of protein was confirmed through MALDI-TOF analysis. The in silico analysis showed that Pi54of is structurally more stable than other cloned Pi54 proteins. The molecular docking revealed that Pi54of protein interacts with AVR-Pi54 through novel non-LRR domains such as STI1 and RhoGEF. The STI1 and GEF domains which interact with AVR-Pi54 are also components of rice defensome complex. The Pi54of protein showed differential domain specificity while interacting with the AVR protein. Functional complementation revealed that Pi54of transferred in two rice lines belonging to indica and japonica background imparts enhanced resistance against three highly virulent strains of M. oryzae. In this study, for the first time, we demonstrated that a rice blast resistance gene Pi54of cloned from wild species of rice provides high degree of resistance to M. oryzae and might display different molecular mechanism involved in AVRPi54-Pi54of interaction.

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