应用于茄子的病毒诱导的基因沉默体系的建立与优化  

Establishment and optimization of virus-induced gene silencing in eggplants

作　　者：刘军[1] 杨艳 周晓慧[1] 姜悬云 鲍生有[1] 庄勇[1] LIU Jun;YANG Yan;ZHOU Xiao-hui;JIANG Xuan-yun;BAO Sheng-you;ZHUANG Yong(Institute of Vegetable Crops,Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory of Horticultural Crop Genetic hnprovement,Nanfing 210014,China)

机构地区：[1]江苏省农业科学院蔬菜研究所/江苏省高效园艺作物遗传改良重点实验室,江苏南京210014

出　　处：《江苏农业学报》2018年第4期880-886,共7页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31672145);国家自然科学青年基金项目(31701932);江苏省重点研发计划项目(BE2016329)

摘　　要：本研究以八氢番茄红素脱氢酶(PDS)基因作为指示基因,探讨不同接种方式、培养环境、菌液数量浓度和接种时间对烟草脆裂病毒(TRV)诱导茄子植物内源PDS mRNA沉默效率的影响,进而测试该沉默体系在不同基因型茄子上的沉默效率。结果表明,在昼夜温度25℃/20℃,16 h光周期条件下,-25 k Pa压强下真空抽气2min渗透侵染露白后2~3 d的种子能够获得最佳沉默效率;与传统注射压迫法相比真空渗透萌芽种子能够更快、更明显出现沉默表型,缩短试验周期,且该方法在不同基因型茄子中都获得较高沉默效率,说明基于真空侵染萌芽种子法的VIGS体系可应用于茄子基因功能研究。In this study,phytoene desaturase（ PDS） was used as an indicator to explore effects of different inoculation methods,temperatures,microbial concentrations,inoculation time on silencing efficiency of eggplant endogenous PDS mRNA induced by tobacco rattle virus（ TRV）. Moreover,silencing efficiency of PDS in eggplants with different phenotypes was tested. The results showed that higher silencing efficiency was induced under the conditions of-25 k Pa vacuum infiltration for two minutes,the seeds germinated for two to three days,and plant growth temperature at 25 ℃ day and 20 ℃night. Compared to other techniques such as leaf infiltration,the new inoculation method could obtain phenotype earlier and more obviously and shorten assay period. In addition,the phytoene desaturase gene in different genotypes of eggplants were successfully silenced using the optimized virus-induced gene silencing（ VIGS） system,indicating that the VIGS system based on vacuum infiltration of germinated seeds could be used for investigation of gene function in eggplants.

关 键 词：茄子 VIGS 真空侵染 萌芽期种子 

分 类 号：S641.1[农业科学—蔬菜学]