浙江大学学报（农业与生命科学版）2025，Vol.51Issue(1)：89-101,13.DOI:10.3785/j.issn.1008-9209.2024.12.181
石蒜同源基因沉默体系构建及鳞茎发生基因功能验证
Establishment of a homologous gene silencing system and gene function verification of bulb formation in Lycoris Herb.
摘要
Abstract
In this study,we explored the construction of a homologous gene silencing system for Lycoris insularis,which is a rich wild resource in Zhejiang Province with bulbs as explants,and through this system,the biological functions of Lycoris insularis bulb genes were verified.The 1 706 bp fragment of LiCWIN gene was cloned from the bulb by using specifically designed primers.After double enzyme digestion,the virus-induced gene silencing(VIGS)recombinant vector was constructed by connection with tobacco rattle virus(TRV)vector TRV2-GFP.After the transformation of Agrobacterium tumefaciens,the damaged and stimulated bulbs were infected with the bacterial solution.After 2 d of infection,the green fluorescence was observed in the vascular tissues of bulbs,and the expression level of LiCWIN gene was significantly decreased in the silenced group,as determined by real time fluorogenic quantitative polymerase chain reaction(qRT-PCR).After 40 d of infection,compared with the empty vector control group,the number of bulblets in LiCWIN gene silencing group decreased,and their formation was delayed.The above results showed that VIGS system successfully silenced functional genes in the bulbous tissues of Lycoris insularis and had a lasting effect on the later development of bulblets.After silencing of LiCWIN gene,the formation of bulblets was blocked.This study can lay a foundation for the study of key gene functions of Lycoris bulb formation and development.关键词
石蒜属/基因功能/瞬时验证/病毒诱导的基因沉默/体系构建/CWIN基因/小鳞茎发生Key words
Lycoris Herb./gene function/instantaneous verification/virus-induced gene silencing(VIGS)/system construction/CWIN gene/bulblet formation分类
农业科技引用本文复制引用
肖潇,夏宜平,崔柳,张佳平,洪震,任梓铭..石蒜同源基因沉默体系构建及鳞茎发生基因功能验证[J].浙江大学学报（农业与生命科学版）,2025,51(1):89-101,13.基金项目
浙江省基础公益研究计划项目(ZCLTGN24C1601) （ZCLTGN24C1601）
浙江省农业新品种重大专项花卉育种专项(2021C02071-6) （2021C02071-6）
国家自然科学基金项目(32101571). （32101571）
