植物促分裂原活化蛋白激酶研究中的病毒诱导基因沉默

Virus-induced gene silencing in plant MAPK research.

作者信息

Hettenhausen Christian, Baldwin Ian T, Wu Jianqiang

机构信息

Kunming Institute of Botany, Chinese Academy of Sciences, 132 Lanhei Road, Heilongtan, Kunming, 650201, China.

出版信息

Methods Mol Biol. 2014;1171:79-89. doi: 10.1007/978-1-4939-0922-3_7.

DOI:10.1007/978-1-4939-0922-3_7

PMID:24908121

Abstract

Virus-induced gene silencing (VIGS) technology has become more and more widely used in various plant species for rapid screening of gene functions. VIGS does not require time-consuming tissue culture steps that are needed for stable transformation in most plant species and it can be used for studying gene function even in plants that are very difficult to stably transform. Furthermore, VIGS technology provides high gene silencing efficiency (up to 95 %) and specificity. Here, we describe a VIGS protocol that can be used for studying the functions of MAPKs and other genes in a wild tobacco species, Nicotiana attenuata. This method is also suitable for other Nicotiana species and tomato with minor modifications.

摘要

病毒诱导的基因沉默（VIGS）技术已越来越广泛地应用于各种植物物种，用于快速筛选基因功能。VIGS不需要大多数植物物种进行稳定转化时所需的耗时组织培养步骤，甚至可以用于研究极难进行稳定转化的植物中的基因功能。此外，VIGS技术具有高基因沉默效率（高达95%）和特异性。在此，我们描述了一种VIGS方案，可用于研究野生烟草物种——渐狭叶烟草中促分裂原活化蛋白激酶（MAPKs）和其他基因的功能。该方法经少量修改后也适用于其他烟草物种和番茄。