基于三角梅的病毒诱导基因沉默体系的建立与优化  

Establishment and Optimization of Virus-induced Gene Silencing System in Bougainvillea peruviana‘Thimma’

作　　者：刘珍银 段郅臻 彭婷 王童欣 王健[1] LIU Zhen-yin;DUAN Zhi-zhen;PENG Ting;WANG Tong-xin;WANG Jian(College of Forestry,Key Laboratory of Genetics and Germplasm Innovation of Tropical Forest Trees and Ornamental Plants(Ministry of Education),Key Laboratory of Germplasm Resources Biology of Tropical Special Ornamental Plants of Hainan Province,Hainan University,Haikou 570228;College of Agriculture,Guizhou University,Guiyang 550025)

机构地区：[1]海南大学林学院热带特色林木花卉遗传与种质创新教育部重点实验室、海南省热带特色花木资源生物学重点实验室,海口570228 [2]贵州大学农学院,贵阳550025

出　　处：《生物技术通报》2023年第7期123-130,共8页Biotechnology Bulletin

基　　金：国家自然科学基金项目(32160719,32060365);海南省重点研发项目(ZDYF2020152);海南省研究生创新科研课题(Qhyb2021-33)。

摘　　要：三角梅是紫茉莉科灌木,具有较高的观赏价值,是研究花色叶色调控的理想材料。建立快速高效的基因验证体系,是三角梅基因功能研究的关键。以三角梅‘金心双色’品种为材料,选用八氢番茄红素脱氢酶基因(PDS)为指示基因,探索不同接种部位和基因片段长度对烟草脆裂病毒(TRV)诱导三角梅叶片内源PDS mRNA沉默效果的影响,建立适用于三角梅的病毒诱导基因沉默体系(VIGS)。结果表明,摩擦注射嫩叶法相比顶端嫩茎沉默效果更明显;基因沉默片段长度在336 bp和457 bp均可诱导PDS基因沉默,后者效果更加明显。初步构建了三角梅VIGS体系,为后续基因功能研究提供了技术参考。Bougainvillea peruviana‘Thimma’,belonging to the family Nyctaginaceae,has high ornamental values in landscape.It is an ideal material to study the regulation of flower and leaf coloration.It is vital to establish a rapid and efficient gene verification system in B.peruviana.In this study,phytoene desaturase gene(PDS)was used as an indicator gene,we explored the effects of different inoculation sites and fragment length on the silencing effects of endogenous PDS mRNA in B.peruviana leaves induced by tobacco rattle virus(TRV)and established an optimized virus-induced gene silencing system(VIGS)for B.peruviana.The results showed that the silencing effect of leaf friction injection was more significant than that of tender stems friction injection.Additionally,PDS gene silencing was induced by both the inserted fragment of 336 bp and 457 bp,and the latter one was more significant.The VIGS system of B.peruviana is preliminarily constructed,which can be applied to the following gene function research.

关 键 词：三角梅 八氢番茄红素脱氢酶 烟草脆裂病毒 基因沉默 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]