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作物杂志,2023, 第2期: 46–50 doi: 10.16035/j.issn.1001-7283.2023.02.007

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

基于PDS基因的番茄VIGS高效沉默体系的优化

郭燕1(), 刘志达1(), 康立茹2, 包婷婷1, 杨旭1, 赵君1, 张之为1()   

  1. 1内蒙古农业大学园艺与植物保护学院，010018，内蒙古呼和浩特
    2内蒙古自治区农牧业科学院，010031，内蒙古呼和浩特
  • 收稿日期:2021-11-16 修回日期:2022-01-03 出版日期:2023-04-15 发布日期:2023-04-11
  • 通讯作者: 张之为，主要从事植物逆境生理研究，E-mail：zzw_217@163.com
  • 作者简介:郭燕，主要从事植物病理学研究，E-mail：1916542008@qq.com；|刘志达，主要从事植物病理学研究，E-mail：954713531@qq.com
  • 基金资助:
    国家自然科学基金(31860495);内蒙古自然科学基金(2019MS03057)

Optimization of Efficient Silencing System of Tomato VIGS Based on PDS Gene

Guo Yan1(), Liu Zhida1(), Kang Liru2, Bao Tingting1, Yang Xu1, Zhao Jun1, Zhang Zhiwei1()   

  1. 1College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot 010018, Inner Mongolia, China
    2Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Hohhot 010031, Inner Mongolia, China
  • Received:2021-11-16 Revised:2022-01-03 Online:2023-04-15 Published:2023-04-11

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摘要：

病毒诱导的基因沉默（VIGS）具有独特优势，被广泛应用于基因功能验证。高效基因沉默体系的建立和优化是对目标基因功能进行研究的前提和基础。以番茄品种money maker和携带八氢番茄红素脱氢酶（PDS）基因序列的TRV病毒载体为材料，研究了农杆菌不同OD值、不同侵染方法以及侵染后不同培养温度对PDS基因沉默株率的影响。结果表明，农杆菌OD值分别为0.50、1.00和1.50时，PDS基因的沉默株率分别为5%、13%和6%；当农杆菌OD值为1.0时，采用注射法、真空浸润法和注射处理24h后再进行真空浸润后的PDS基因沉默株率分别为13%、30%和15%；侵染后的培养温度对于基因沉默株率也有显著影响，在25℃时的沉默株率为13%，22℃时沉默株率达到46%。以上研究表明，农杆菌OD值为1.00时，注射24h后再进行真空浸润，接种后植株在22℃下生长能够提高番茄目标基因的沉默株率。

关键词: 番茄, 病毒诱导的基因沉默, 八氢番茄红素脱氢酶基因, 沉默株率, 优化

Abstract:

Virus-induced gene silencing (VIGS) is widely used for gene function verification due to its unique advantages. The establishment and optimization of an efficient gene silencing system is the premise and basis for the study of target gene function. In this study, the effects of different OD values, different infection methods, and different culture temperatures after being infected of Agrobacterium on the silencing rate of the PDS gene were studied by using the money maker cultivar and the TRV virus vector carrying phytoene desaturase (PDS) gene sequence. The results showed that, when the OD values of Agrobacterium were 0.50, 1.00, and 1.50, the silencing rates of the PDS gene were 5%, 13%, and 6%, respectively. When the OD value was 1.00, the silencing effects was compared by injection method, vacuum infiltration method and vacuum infiltration after 24 hours of injection treatment, the results showed that, the rates of PDS gene silencing were 13%, 30%, and 15%, respectively. The culture temperature after infection also had a significant effects on the gene silencing rate, with 13% of the plants at 25°C and 46% of the silencing plants at 22°C. The above studies showed that, when the OD value of Agrobacterium is 1.00, the vacuum infiltration was carried out after injection for 24 hours, and the growth of plants at 22°C after inoculation could improve the silencing rate of tomato target genes.

Key words: Tomato, Virus-induced gene silencing, Phytoene desaturase gene, Silence strain rate, Optimization

图1

PDS基因沉默后植株的表型

图2

光漂白植株中PDS基因表达量分析 不同小写字母表示差异显著（P < 0.05），下同

图3

农杆菌不同接种浓度对PDS基因沉默株率的影响

图4

农杆菌不同接种方法对沉默株率的影响

图5

不同温度处理对沉默株率的影响

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