Establishment and Identification of TRV-mediated Tomato spotted wilt orthotospovirus N Gene Silencing System in Pepper

PurposeTo explore the impact of Tomato spotted wilt orthotospovirus (TSWV) nucleocapsid protein gene (N) on pepper disease resistance.

MethodXiangyan 11, a pepper susceptible to TSWV, was used as the research object. Pepper phytoene desaturase (CaPDS) gene and TSWV N gene were cloned by homologous sequence. The CaPDS gene silencing vector (pTRV2-CaPDS) and TSWV N gene silencing vector (pTRV2-N) of pepper were constructed by using Tobacco rattle virus (TRV) and virus-induced gene silencing (VIGS) technology. Agrobacterium GV3101 was inoculated with pepper. The silencing efficiency of the recombinant vector was detected by qRT-PCR.

ResultThree weeks after inoculation with pTRV2-CaPDS, the new leaves appeared albino, which indicated that the construction of pTRV2-CaPDS silencing vector was successful. The qRT-PCR results showed that the pTRV2-N vector can efficiently silence the N gene, and its expression is only 6.79%±2.56%, indicating that after silencing the N gene, Xiangyan 11 is not susceptible to TSWV.

ConclusionpTRV2-N silencing vector was successfully constructed. After silencing the TSWV N gene, pepper had certain resistance to invading TSWV.