Analysis Of Resistance Functional Of COA OsECH1 To Hirschmanniella Mucronate

Rice is one of the most important food crops in the world.It is the main food for more than half of the world’s population.It is predicted that rice production will need to increase by 25 percent by 2030 to meet population growth.Hirschmanniella spp.are a kind of migratory endoparasitic nematode which is parasitic in the root of rice.It is widely distributed in paddy fields and has become a serious parasitic nematode to rice.The plant-parasitic nematode Hirschmanniella spp.are one of the most serious rice nematodes in Jiangxi Province.In order to control its harmfulness,breeding and planting other disease-resistant varieties is the most economical and effective measure.At present,there are few studies on Hirschmanniella spp.It is of great significance to explore the interaction mechanism between rice and Hirschmanniella spp.and to analyze its inherent molecular mechanism of resistance to Hirschmanniella spp.,which can improve the resistance of Hirschmanniella spp.and reduce the loss of disease.In the early stage of the laboratory,the up-regulated OsECH1 gene was screened by transcriptome analysis of resistant/susceptible rice root tissues infected by H.mucronata.The full length of the gene was cloned,and the overexpression vector pCAMBIA1302-OsECH1,RNAi vector pBWA(V)HS-OsECH1 and prokaryotic expression vector pET28a-OsECH1 were constructed by homologous recombination technology.The results showed that the ORF of OsECH1 gene was 1170 bp in length,encoding 389 amino acids,and it belongs to the PLN02874 superfamily.This family is mainly Enoyl-CoA hydratase and 3-hydroxyisobutyryl-COA hydrolase,which participate in fatty acid β-oxidation reaction.The transgenic rice was infected with Agrobacterium tumefaciens,and the relative expression level of the gene in different rice treatments was verified by qRT-PCR.The experimental results showed that the gene was mainly expressed in the root,but the expression level in the root decreased,and the expression level in the leaf increased after the nematode infection.When IPTG concentration was 0.6 mM and induced at 16℃ for 18 h,the protein expression was the highest.In the purification process,the maximum elution amount was obtained at 200 mM imidazole elution.Pull-down experiments with root total proteins showed that there were 6 proteins interacting with target protein.In conclusion,the interaction mechanism network of this gene is complex when rice is infected by nematodes,and it mainly plays a role in rice leaves.The results will provide the molecular mechanism of rice root nematode resistance.