油棕病毒诱导的基因沉默体系的建立及优化  

Establishment and optimization of oil palm virus-induced gene silencing system

作　　者：李萌晗 王威[1] 邹积鑫 李东栋[1,2] LI Menghan;WANG Wei;ZOU Jixin;LI Dongdong(College of Tropical Crops,Hainan University,Haikou 570228,China;Sanya Nanfan Research Institute,Sanya 572024,China)

机构地区：[1]海南大学热带作物学院,海口570228 [2]海南大学三亚南繁研究所,三亚572024

出　　处：《华中农业大学学报》2023年第2期259-264,共6页Journal of Huazhong Agricultural University

基　　金：海南省重点研发项目(ZDYF2022XDNY148)。

摘　　要：为开展油棕油脂代谢调控相关基因鉴定研究,以油棕八氢番茄红素脱氢酶基因(phytoene desaturase gene,PDS)作为报告基因,探索以病毒载体TRV为载体在油棕胚状体上应用病毒诱导基因沉默(virus-induced gene silencing,VIGS)的可能性,并对油棕胚状体VIGS体系的相关参数进行优化。结果显示:以EHA105为菌种、侵染菌液OD_(600)=0.5、侵染时间5 min、乙酰丁香酮(AS)质量浓度20 mg/L、共培养48 h、侵染后培养时间为12 d能取得最佳的基因沉默效果。在此基础上,利用优化后的VIGS体系对油棕二酰甘油酰基转移酶基因(diacylglycerol acyltransferase gene,DGAT)进行沉默,取得了预期的基因沉默效果。The phytoene desaturase gene(PDS)as a reporter gene was used to study the feasibility of applying virus-induced gene silencing(VIGS)on oil palm embryoid using the TRV as a vector,and to optimize the relevant parameters.The results showed that the effect of gene silencing was the best when EHA105 was used as the strain under the condition that the OD_(600) of infected bacterial solution was 0.5,with infection time of 5 min,acetosyringone(AS)mass concentration of 20 mg/L,co-cultivation of 48 h,and post-infection incubation time of 12 d.On this basis,the VIGS system optimized was used to silence the diacylglycerol acyltransferase gene(DGAT),and the expected gene silencing effect was achieved.

关 键 词：油棕 胚状体 病毒诱导的基因沉默体系 遗传转化 八氢番茄红素脱氢酶 

分 类 号：S565.9[农业科学—作物学]