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GSL2 过表达赋予马铃薯对果胶杆菌的抗性。

GSL2 over-expression confers resistance to Pectobacterium atrosepticum in potato.

机构信息

The New Zealand Institute for Plant & Food Research Ltd, Private Bag 4704, Christchurch, 8140, New Zealand.

出版信息

Theor Appl Genet. 2014 Mar;127(3):677-89. doi: 10.1007/s00122-013-2250-2. Epub 2013 Dec 27.

Abstract

Over-expression of the potato Gibberellin Stimulated-Like 2 ( GSL2 ) gene in transgenic potato confers resistance to blackleg disease incited by Pectobacterium atrosepticum and confirms a role for GSL2 in plant defence. The Gibberellin Stimulated-Like 2 (GSL2) gene (also known as Snakin 2) encodes a cysteine-rich, low-molecular weight antimicrobial peptide produced in potato plants. This protein is thought to play important roles in the innate defence against invading microbes. Over-expression of the GSL2 gene in potato (cultivar Iwa) was achieved using Agrobacterium-mediated gene transfer of a plant expression vector with the potato GSL2 gene under the regulatory control elements of the potato light-inducible Lhca3 gene. The resulting plants were confirmed as being transgenic by PCR, and subsequently analysed for transcriptional expression of the Lhca3-GSL2-Lhca3 chimeric potato gene. Quantitative RT-PCR analysis demonstrated that the majority of the transgenic potato lines over-expressed the GSL2 gene at the mRNA level. Based on qRT-PCR results and evaluation of phenotypic appearance, eight lines were selected for further characterisation and evaluated in bioassays for resistance to Pectobacterium atrosepticum (formerly Erwinia carotovora subsp. atroseptica), the causal agent of blackleg in potato. Three independent pathogenicity bioassays showed that transgenic lines with significantly increased transcriptional expression of the GSL2 gene exhibit resistance to blackleg disease. This establishes a functional role for GSL2 in plant defence against pathogens in potato.

摘要

马铃薯赤霉素刺激样 2(GSL2)基因在转基因马铃薯中的过表达赋予其对由果胶杆菌引起的黑胫病的抗性,并证实了 GSL2 在植物防御中的作用。赤霉素刺激样 2(GSL2)基因(也称为 Snakin 2)编码一种富含半胱氨酸的低分子量抗菌肽,在马铃薯植物中产生。这种蛋白质被认为在抵御入侵微生物的先天防御中发挥重要作用。使用农杆菌介导的基因转移,将马铃薯 GSL2 基因置于马铃薯光诱导 Lhca3 基因的调控元件下,在马铃薯(品种 Iwa)中过表达 GSL2 基因。通过 PCR 确认所得植物为转基因植物,并随后分析 Lhca3-GSL2-Lhca3 嵌合马铃薯基因的转录表达。定量 RT-PCR 分析表明,大多数转基因马铃薯系在 mRNA 水平上过度表达 GSL2 基因。基于 qRT-PCR 结果和表型外观评估,选择了 8 条系进行进一步表征,并在生物测定中评估其对果胶杆菌(以前称为胡萝卜软腐欧文氏菌亚种)的抗性,果胶杆菌是马铃薯黑胫病的病原体。三个独立的致病性生物测定表明,GSL2 基因转录表达显著增加的转基因系表现出对黑胫病的抗性。这确立了 GSL2 在马铃薯植物防御病原体中的功能作用。

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