Functional Research Of Candidate Gene Of The Novel Blast Resistance Gene Pi39 In Rice

Rice is an important food crop.Rice blast is one of the three major diseases,which seriously affects rice yield.Breeding for disease resistance is the most economical and effective method for controlling rice blast.Due to the variation and diversity of rice blast fungus isolates,resistant varieties tend to lose resistance to rice blast after several years of planting,it is important to explore new resistance genes.The new rice blast resistance gene Pi39 was derived from the local cultivar Q15 in Yunnan with broad-spectrum resistance.In the early stage,the physical map of Pi39has been constructed,and the functional complementary vector of 10 candidate genes has been constructed,and genetic transformation has been carried out.The research is to remain two candidate genes.This study is mainly to screen the candidate genes and their functions of Pi39.The main contents are as follows:?1?Pi39C and Pi39E functional complementary vectors were constructed successfully by using long-segment PCR amplification and blunt-end ligation,and carried out rice genetic transformation.Positive transgenic plants were identified the results showed that 18 transgenic positive plants of Pi39C were obtained,and 44transgenic positive vaccines were obtained from Pi39E.?2?The transgenic T₁ plants with functional complementation of 12 candidate genes were positively identified,which were inculated artificially by Magnaporthe oryzae in vivo and vitro,respectively the candidate gene Pi39E₁ was selected as the resistance gene Pi39 by observing their phenotype.?3?RNAi vector of Pi39E₁ was constructed and then genetic transformation of rice Q15 was carried out,which are currently in the differentiation stage of genetic transformation,which will provide materials for subsequent identification of resistance to rice blast.The resistance function of Pi39 can also be further verified.?4?Nipponbare and Q15 was inoculated with race 007 in vitro,and samples were taken at 0,12,24,and 48 h after inoculation to extract RNA,and the expression of Pi39E₁ was analyzed.The results showed that the expression level of Pi39E₁ in Q15 was significantly higher than that in Nipponbare,indicating that Pi39E₁ plays a role in the defense mechanism of rice blast in Q15.?5?The pM999-Pi39E₁-GFP transient expression vector was constructed to confirm the location of Pi39E₁ protein expression and lay the foundation for subsequent screening of interaction proteins.