Simultaneous silencing of Two Target Genes Using Virus-Induced Gene Silencing Technology

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• 英文论文题名：Simultaneous silencing of Two Target Genes Using Virus-Induced Gene Silencing Technology
• 论文作者：ZHU Feng ; ZHOU Yang-kai
• 英文论文作者：ZHU Feng ; ZHOU Yang-kai ; College of Horticulture and Plant Protection ; Yangzhou University
• 年：2016
• 作者机构：College of Horticulture and Plant Protection,Yangzhou University;
• 会议召开时间：2016-08-05
• 会议录名称：中国植物病理学会2016年学术年会论文集
• 英文会议录名称：Proceedings of the Annual Meeting of Chinese Society for Plant Pathology(2016)
• 语种：英文
• 分类号：S432.41
• 学会代码：ZGVS
• 会议名称：中国植物病理学会2016年学术年会
• 会议地点：中国江苏南京
• 主办单位：中国植物病理学会
• 学会名称：中国植物病理学会
• 页数：1
• 文件大小：40k
• 原文格式：O
• 会议级别：全国

摘要

Virus-induced gene silencing(VIGS) is an effective strategy for rapid gene function analysis during plant growth and development.Over the last decade,VIGS has been successfully used to knock down the expression of a single gene in various plant species.However,simultaneous silencing of two target genes using VIGS in plants has been rarely reported.It is well established that the NAC(NAM,ATAF,and CUC) transcription factor and salicylic acid(SA) signal transduction pathway play essential roles in response to biotic stresses.Therefore,in this report,we performed VIGS to silence simultaneously the salicylic acid-binding protein 2(NbSABP2) and NbNAC1 in Nicotiana benthamiana to investigate the gene silencing efficiency of simultaneous silencing of two genes.Overlap extension PCR analysis showed that the combination of NbSABP2 and NbNAC1 was successfully amplified.Bacteria liquid PCR confirmed that the combination of NbSABP2 and NbNAC1 was successfully inserted into the TRV vector.Quantitative real-time PCR results showed that the expression of NbSABP2 and NbNAC1 were significantly reduced in 12 days post silenced plants after TRV infiltration compared with the control(TRV:00) plants.Overall,our results suggest that VIGS can be used to silence simultaneously two target genes only one time infiltration.
Virus-induced gene silencing(VIGS) is an effective strategy for rapid gene function analysis during plant growth and development.Over the last decade,VIGS has been successfully used to knock down the expression of a single gene in various plant species.However,simultaneous silencing of two target genes using VIGS in plants has been rarely reported.It is well established that the NAC(NAM,ATAF,and CUC) transcription factor and salicylic acid(SA) signal transduction pathway play essential roles in response to biotic stresses.Therefore,in this report,we performed VIGS to silence simultaneously the salicylic acid-binding protein 2(NbSABP2) and NbNAC1 in Nicotiana benthamiana to investigate the gene silencing efficiency of simultaneous silencing of two genes.Overlap extension PCR analysis showed that the combination of NbSABP2 and NbNAC1 was successfully amplified.Bacteria liquid PCR confirmed that the combination of NbSABP2 and NbNAC1 was successfully inserted into the TRV vector.Quantitative real-time PCR results showed that the expression of NbSABP2 and NbNAC1 were significantly reduced in 12 days post silenced plants after TRV infiltration compared with the control(TRV:00) plants.Overall,our results suggest that VIGS can be used to silence simultaneously two target genes only one time infiltration.

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