Styrax japonicus functional genomics: an efficient virus induced gene silencing (VIGS) system

IF 5.7 1区农林科学 Q1 HORTICULTURE

Horticultural Plant Journal Pub Date : 2024-01-01 DOI:10.1016/j.hpj.2023.05.003

Gangyu Sun , Yiqian Ju , Cuiping Zhang , Lulu Li , Xinqiang Jiang , Xiaoman Xie , Yizeng Lu , Kuiling Wang , Wei Li

{"title":"Styrax japonicus functional genomics: an efficient virus induced gene silencing (VIGS) system","authors":"Gangyu Sun , Yiqian Ju , Cuiping Zhang , Lulu Li , Xinqiang Jiang , Xiaoman Xie , Yizeng Lu , Kuiling Wang , Wei Li","doi":"10.1016/j.hpj.2023.05.003","DOIUrl":null,"url":null,"abstract":"<div>VIGS (Virus-induced gene silencing), a method for posttranscriptional gene silencing, is an effective technique for investigating the activities of genes in plants. Since there is no report for available VIGS system in Styrax japonicus, the application of a VIGS approach that results in a gene knockdown to study gene function is limited. In this study, we compared the characteristics that could affect the viability of VIGS in S. japonicus, including the acetosyringone (AS) concentration, the Agrobacterium's optical density and the inoculation method. The stable reference genes of S. japonicus were selected to validate the gene's knockdown by quantitative PCR. As a result, we successfully constructed 2 VIGS systems based on TRV virus: vacuum with AS concentration of 200 μmol·L−1 and OD600 of 0.5, and friction-osmosis with AS concentration of 200 μmol·L−1 and OD600 of 1.0, which silencing efficiency was 83.33% and 74.19%, respectively. The successfully applied VIGS method provides a rapid and effective reverse gene functional analysis approach in S. japonicus to identify unknown gene functions.</div>","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"10 1","pages":"Pages 252-258"},"PeriodicalIF":5.7000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2468014123000833/pdfft?md5=41dd6228e4e13915ac24028c29a90d11&pid=1-s2.0-S2468014123000833-main.pdf","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Horticultural Plant Journal","FirstCategoryId":"1091","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2468014123000833","RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"HORTICULTURE","Score":null,"Total":0}

引用次数: 1

Abstract

VIGS (Virus-induced gene silencing), a method for posttranscriptional gene silencing, is an effective technique for investigating the activities of genes in plants. Since there is no report for available VIGS system in Styrax japonicus, the application of a VIGS approach that results in a gene knockdown to study gene function is limited. In this study, we compared the characteristics that could affect the viability of VIGS in S. japonicus, including the acetosyringone (AS) concentration, the Agrobacterium's optical density and the inoculation method. The stable reference genes of S. japonicus were selected to validate the gene's knockdown by quantitative PCR. As a result, we successfully constructed 2 VIGS systems based on TRV virus: vacuum with AS concentration of 200 μmol·L⁻¹ and OD₆₀₀ of 0.5, and friction-osmosis with AS concentration of 200 μmol·L⁻¹ and OD₆₀₀ of 1.0, which silencing efficiency was 83.33% and 74.19%, respectively. The successfully applied VIGS method provides a rapid and effective reverse gene functional analysis approach in S. japonicus to identify unknown gene functions.

查看原文本刊更多论文

日本栉水母功能基因组学：高效的病毒诱导基因沉默（VIGS）系统

病毒诱导基因沉默（VIGS）是一种转录后基因沉默方法，是研究植物基因活性的有效技术。由于目前还没有关于日本栉水母（Styrax japonicus）可用 VIGS 系统的报道，因此应用 VIGS 方法导致基因敲除以研究基因功能的应用非常有限。在本研究中，我们比较了可能影响日本草履虫 VIGS 生命力的特征，包括乙酰丁香酮（AS）浓度、农杆菌的光密度和接种方法。我们选择了稳定的日本农杆菌参考基因，通过定量 PCR 验证基因敲除效果。结果，我们成功构建了两个基于TRV病毒的VIGS系统：AS浓度为200 μmol-L-1、OD600为0.5的真空系统和AS浓度为200 μmol-L-1、OD600为1.0的摩擦渗透系统，其沉默效率分别为83.33%和74.19%。VIGS方法的成功应用为日本鹅提供了一种快速有效的反向基因功能分析方法，以确定未知基因的功能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Horticultural Plant Journal Environmental Science-Ecology

CiteScore

9.60

自引率

14.00%

发文量

293

审稿时长

33 weeks

期刊介绍： Horticultural Plant Journal (HPJ) is an OPEN ACCESS international journal. HPJ publishes research related to all horticultural plants, including fruits, vegetables, ornamental plants, tea plants, and medicinal plants, etc. The journal covers all aspects of horticultural crop sciences, including germplasm resources, genetics and breeding, tillage and cultivation, physiology and biochemistry, ecology, genomics, biotechnology, plant protection, postharvest processing, etc. Article types include Original research papers, Reviews, and Short communications.