Expression Analysis And Cloning Of Genes Related To Disease-resistance For Japonica Variety Yunyin

Analyzing disease-resistance genes and developing high-yieldhigh-quality, long-lasting disease-resistant rice varieties have become one of the priorities of breeding by genetic engineering and molecular biology techniques. Based on the proteomic analysis of near-isogenic lines derived from blast resistant variety Yunyin (japonica variety) combined with NCBI, three genes that may be associated with the disease resistance have been found:The first gene OsPRX2 (Genbank sequence number Os02g0537700) was related to heat and bacteria-stress, the second gene OsHAD4 (Genbank sequence number Os04g0490800) was associated with photorespiration that coding halo acid dehalogenase and oxidative-stress, the third gene OsGST5 (Genbank sequence number Os05g0116100) was coding Valley glutathione transferase. Those three genes would be cloned and carried out expression analysis in this research.The main research contents and results were as follows:(1) Those three genes were all response to Magnaporthe oryzae and there expression significantly different between blast resistant and susceptible near-isogenic lines (NILs), verified by real-time quantitative PCR (qRT-PCR).(2) The coding region and the promoter region sequences of those three genes were cloned, the homologous sequences were blasted between blast resistant and susceptible NILs, indicated that both the coding region and the promoter region of OsPRX2, OsGST5 were the same. The similarity of coding region sequence of OsHAD4 between blast resistant and susceptible NILs was 99%, but the difference promoter region sequence was 61.01%.(3) In order to initially validate the function of those three genes, the full-length cDNA sequence were cloned using RACE technology respectively. The corresponding protein sequence were analyzed and constructed phylogenetic tree compared with with maize, Arabidopsis, etc. from NCBI-by biological information analysis.(4) The vectors including over-expression and RNA interference expression were constructed and transferred into blast resistance variety Yunyin, so as to carry out their functional identification and resistant mechanisms research.