Abstract
Rice blast seriously threatens rice production worldwide. Utilizing the rice blast resistance gene to breed the rice blast resistant varieties is one of the best ways to control rice blast disease. Using a map-based cloning strategy, here, we cloned a novel rice blast resistance gene, Pi65 from the resistant variety GangYu129 (abbreviated GY129, O. sativa japonica ). Overexpression of Pi65 in the susceptible variety LiaoXing1 (abbreviated LX1, O. sativa japonica ) enhanced rice blast resistance, while knockout of Pi65 in GY129 resulted in susceptible to rice blast disease. Pi65 encodes two transmembrane domains, with 15 LRR domains and one serine/threonine protein kinase catalytic domain, conferring resistance to isolates of M. oryzae collected from northeast China. There are sixteen amino acids differences between the Pi65 resistance and susceptible alleles. Compared with the Pi65 resistant allele, the susceptible allele deleted one LRR domain. Pi65 was constitutively expressed in whole plants, and it could be induce expressed in the early stage of M. oryzae infection . Transcriptome analysis revealed that numerous genes associated with disease resistance were specifically upregulated in GY129 24-hour post inoculation (HPI), on the contrary, the photosynthesis-and carbohydrate metabolism-related genes were particularly downregulated 24 HPI, demonstrating that the disease resistance associated genes has been activated in GY129 (carrying Pi65 ) after rice blast fungal infection, and the cellular basal and energy metabolism was inhibited simultaneously. Our study provides genetic resources for improving rice blast resistance as well as enriches the study of rice blast resistance mechanisms.
Identification of Microsatellite (SSR) and RAPD Markers Linked to Rice Blast Disease Resistance gene in Rice (Oryza sativa L.)