Host-induced gene silencing of the Verticillium dahliae thiamine transporter protein gene(VdThit)confers resistance to Verticillium wilt in cotton  

作　　者：Qi Wang Guoqiang Pan Xingfen Wang Zhengwen Sun Huiming Guo Xiaofeng Su Hongmei Cheng 

机构地区：[1]National Key Laboratory of Agricultural Microbiology/Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing100081,China [2]Xinjiang Jinfengyuan Seeds Co.,Ltd.,Aksu 843100,China [3]College of Agronomy,Hebei Agricultural University,Baoding 071001,China [4]Institute of Microbiology Applications,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China

出　　处：《Journal of Integrative Agriculture》2024年第10期3358-3369,共12页农业科学学报（英文版）

基　　金：supported by the National Key Research and Development Program of China(2022YFD1200300);the National Natural Science Foundation of China(32072376 and 32372515);Winall Hi-tech Seed Co.,Ltd.,China(GMLM2023);the Nanfan Special Project of Chinese Academy of Agricultural Sciences(CAAS)(ZDXM2303 and YBXM2415);the Natural Science Foundation of Hebei Province,China(C2022204205);the Agricultural Science and Technology Innovation Program of CAAS。

摘　　要：Verticillium wilt(VW),induced by the soil-borne fungus Verticillium dahliae(Vd),poses a substantial threat to a diverse array of plant species.Employing molecular breeding technology for the development of cotton varieties with heightened resistance to VW stands out as one of the most efficacious protective measures.In this study,we successfully generated two stable transgenic lines of cotton(Gossypium hirsutum L.),VdThitRNAi-1 and VdThit-RNAi-2,using host-induced gene silencing(HIGS)technology to introduce double-stranded RNA(dsRNA)targeting the thiamine transporter protein gene(VdThit).Southern blot analysis confirmed the presence of a single-copy insertion in each line.Microscopic examination showed marked reductions in the colonization and spread of Vd-mCherry in the roots of VdThit-RNAi cotton compared to wild type(WT).The corresponding disease index and fungal biomass of VdThit-RNAi-1/2 also exhibited significant reductions.Real-time quantitative PCR(qRT-PCR)analysis demonstrated a substantial inhibition of VdThit expression following prolonged inoculation of VdThit-RNAi cotton.Small RNA sequencing(sRNA-Seq)analysis revealed the generation of a substantial number of VdThit-specific siRNAs in the VdThit-RNAi transgenic lines.Additionally,the silencing of VdThit by the siVdThit produced by VdThit-RNAi-1/2 resulted in the elevated expression of multiple genes involved in the thiamine biosynthesis pathway in Vd.Under field conditions,VdThit-RNAi transgenic cotton exhibited significantly enhanced disease resistance and yield compared with WT.In summary,our findings underscore the efficacy of HIGS targeting VdThit in restraining the infection and spread of Vd in cotton,thereby potentially enabling the development of cotton breeding as a promising strategy for managing VW.

关 键 词：Verticillium dahliae thiamine transporter host-induced gene silencing(HIGS) RNAi 

分 类 号：S435.621.2[农业科学—农业昆虫与害虫防治]