烟粉虱CRISPR/Cas9系统的建立及其在BtTRP基因温度耐受性功能验证中的应用

doi:10.16409/j.cnki.2095-039x.2018.02.012

中国生物防治学报 ›› 2018, Vol. 34 ›› Issue (2): 254-258.DOI: 10.16409/j.cnki.2095-039x.2018.02.012

烟粉虱CRISPR/Cas9系统的建立及其在BtTRP基因温度耐受性功能验证中的应用

王原¹, 吕志创², 万方浩^1,2,3

1. 青岛农业大学农学与植物保护学院, 青岛 266109;
2. 中国农业科学院植物保护研究所/植物病虫害生物学国家重点实验室, 北京 100193;
3. 农业部外来入侵生物预防与控制研究中心, 北京 100193

收稿日期:2017-06-07 出版日期:2018-04-08 发布日期:2018-04-13
通讯作者: 万方浩,研究员,E-mail:wanfanghao@caas.cn。
作者简介:王原,硕士,E-mail:1486331498@qq.com
基金资助:
国家自然科学基金（31100269）；国家重点研发计划（2016YFC1200603）；山东省“泰山学者”建设工程专项经费；公益性行业（环保）科研专项（201409061）

The Establishment of the CRISPR/Cas9 System in Bemisia tabaci and the Application in the Identification of Temperature Tolerance Function of Transient Receptor Potential Gene

WANG Yuan¹, LÜ Zhichuang², WAN Fanghao^1,2,3

1. College of Agriculture and Plant Protection, Qingdao Agricultural University, Qingdao 266109, China;
2. State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
3. Center for Management of Invasive Alien Species Ministry of Agricultulture, Beijing 100193, China

Received:2017-06-07 Online:2018-04-08 Published:2018-04-13

摘要/Abstract

摘要： 烟粉虱之所以能在全世界范围内迅速扩散并成为重要害虫，其关键因素之一就是对温度变化的抵抗力很强。随着环境的变化，动物体中的很多细胞表面受体和离子通道感受器都会做出相应的反应，影响动物的行为。其中，昆虫对外界环境温度的感知就与瞬时感受器阳离子通道（transient receptor potential，TRP）有关。本研究探索了一种简单高效的试验方法，即通过CRISPR/cas9系统来诱导烟粉虱的基因突变，探究烟粉虱BtTRP基因在温度感受方面的功能。文中阐述了向烟粉虱MED隐种红眼期伪蛹注射BtTRP靶基因sgRNA和Cas9 mRNA，高效诱导靶标基因发生突变并进行高温和低温胁迫试验，进而检测其击倒和恢复时间。结果表明，注射后的烟粉虱对外界温度变化敏感度显著下降，从而不能进行温度适应性调节，导致对高低温环境的耐受性均有明显下降。

关键词: 烟粉虱MED隐种, 基因突变, 温度感受, TRP基因

Abstract: Whiteflies could quickly spread around the world and become the important pests, and one of the main reasons is its strong resistance to the temperature change. As the change of environment, a lot of cell surface receptors and ion channels in the body could be stimulated, then affected the animal's behavior. Among them, the transient receptor potential (TRP) is a key regulation factor in the insect perception of the environment temperature. This research mainly explored a simple and highly effective method, generating and detecting mutations of TRP gene in Bemisia. tabaci through CRISPR/cas9 system, to identify the function of TRP gene in temperature feeling of B. tabaci Here, we expounded the experimental process of injecting sgRNA in red eye phase pupa of B. tabaci MED, being induced highly efficient mutagenesis of desired target genes, and detected statistical time of heat knockdown or chill coma recovery. The result showed that the sensitivity of the temperature change of injected B. tabaci MED declined significantly, and couldn't adjust temperature adaptability, leading to the decrease of temperature tolerance under hot or cold stress environments.

Key words: Bemicia tabaci MED, gene mutations, thermoreceptor, TRP gene

中图分类号:

S436.3

王原, 吕志创, 万方浩. 烟粉虱CRISPR/Cas9系统的建立及其在BtTRP基因温度耐受性功能验证中的应用[J]. 中国生物防治学报, 2018, 34(2): 254-258.

WANG Yuan, LÜ Zhichuang, WAN Fanghao. The Establishment of the CRISPR/Cas9 System in Bemisia tabaci and the Application in the Identification of Temperature Tolerance Function of Transient Receptor Potential Gene[J]. journal1, 2018, 34(2): 254-258.

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烟粉虱CRISPR/Cas9系统的建立及其在BtTRP基因温度耐受性功能验证中的应用

The Establishment of the CRISPR/Cas9 System in Bemisia tabaci and the Application in the Identification of Temperature Tolerance Function of Transient Receptor Potential Gene

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