Construction of VIGS Vector of Root-knot Resistant WRKY64 Gene in Rapeseed Addition Line EE

Abstract

The root knot nematode seriously harms the quality and yield of crops. The research of crop varieties resistant to root knot nematode has become one of the important ways to control this disease. Previous studies have found that WRKY64 gene in rapeseed addition line EE plays an important role in resisting Meloidogyne incognita. The construction of WRKY64 gene silencing vector provides material for further study on whether WRKY64 gene expression protein in rapeseed addition line EE plays an important role in resisting infection of M.incognita, and provides certain reference for breeding crop varieties resistant to root-knot nematode and green prevention and control of root-knot nematode. In this study, 156bp of WRKY64 gene was constructed into the pTRV-pTV00 gene silencing vector, and agrobacterium GV3101 was used to infiltrate the rapeseed addition line EE that had been inoculated with Meloidogyne incognita for 7 days. The recombinant vector pTRV-PTV00-WRKY64 of WRKY64 gene was successfully constructed, and the subsequent silencing efficiency test could be carried out. By RT-qPCR quantitative analysis, the silencing efficiency of WRKY64 gene expression was 80% after 9 days of inoculation with the silencing vector. WRKY64 gene silencing vector was successfully constructed and tested on the rapeseed addition line EE, providing material for the subsequent research on WRKY64 gene function and theoretical basis for the prevention and control of root-knot nematode disease.