Preliminary Characterization On Candidate Gene Function Of Rice Blast Resistance Gene Pita~2

Rice blast is one of the most devastating diseases of rice, which is a serious threat to rice production. New rice blast resistance genes explored for breeding of disease resistance is an important strategy. Carry out new rice blast resistance genes research on the identification and breeding for disease resistance has important practical significance. On the base of fine mapping and physical mapping of the new rice blast resistance gene Pita~2, in order to identify candidate genes of the Pita~2, the main results are as follows:1. Gene prediction and annotation was conducted in the 75 kb interval by the gene prediction systems. The results showed that five genes served as the candidate genes of the Pita~2-1, Pita~2-2, Pita~2-3, Pita~2-4, and Pita~2-5. Expression pattern of these candidate genes was investigated by RT-PCR, and the results showed that two of the five candidate genes did not express, which may be pseudogenes. The other three genes expressed strongly, which may be functional genes. This study is aid to further narrow the scope of candidate genes Pita~2 and its functional study.2. The four candidate genes were amplified from the genomic DNA by long-range PCR, and the PCR product were inserted into vectors. Each candidate gene were transformed into the blast-susceptible variety Nipponbare. The blast resistance of T0 transgenic plants was confirmed by artificial inoculation of blast fungus. The result shows that the resistant of the transgenic plants did not change significantly. May be Pita~2-2 and Pita~2-5 were not Pita~2.3. Three interference vector and also transform it into the blast-resistant variety PiNo.4. The blast resistance of T0 transgenic plants was confirmed by artificial inoculation of blast fungus. The results showed that the the phenomenon of rice blast disease did not occur significantly. This study will further detail on the identification of the resistance testing of T1 generation.