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盐生植物中的基因挖掘:粗茎独行菜耐逆基因的功能鉴定

Gene mining in halophytes: functional identification of stress tolerance genes in Lepidium crassifolium.

作者信息

Rigó Gábor, Valkai Ildikó, Faragó Dóra, Kiss Edina, Van Houdt Sara, Van de Steene Nancy, Hannah Matthew A, Szabados László

机构信息

Biological Research Centre, Institute of Plant Biology, 6726, Szeged, Hungary.

Bayer CropScience, 9052, Ghent, Belgium.

出版信息

Plant Cell Environ. 2016 Sep;39(9):2074-84. doi: 10.1111/pce.12768. Epub 2016 Jun 25.

DOI:10.1111/pce.12768
PMID:27343166
Abstract

Extremophile plants are valuable sources of genes conferring tolerance traits, which can be explored to improve stress tolerance of crops. Lepidium crassifolium is a halophytic relative of the model plant Arabidopsis thaliana, and displays tolerance to salt, osmotic and oxidative stresses. We have employed the modified Conditional cDNA Overexpression System to transfer a cDNA library from L. crassifolium to the glycophyte A. thaliana. By screening for salt, osmotic and oxidative stress tolerance through in vitro growth assays and non-destructive chlorophyll fluorescence imaging, 20 Arabidopsis lines were identified with superior performance under restrictive conditions. Several cDNA inserts were cloned and confirmed to be responsible for the enhanced tolerance by analysing independent transgenic lines. Examples include full-length cDNAs encoding proteins with high homologies to GDSL-lipase/esterase or acyl CoA-binding protein or proteins without known function, which could confer tolerance to one or several stress conditions. Our results confirm that random gene transfer from stress tolerant to sensitive plant species is a valuable tool to discover novel genes with potential for biotechnological applications.

摘要

极端嗜盐植物是赋予耐受性状基因的宝贵来源,可利用这些基因来提高作物的胁迫耐受性。粗叶独行菜是模式植物拟南芥的盐生近缘种,对盐、渗透和氧化胁迫具有耐受性。我们采用改良的条件性cDNA过表达系统,将粗叶独行菜的一个cDNA文库转移到甜土植物拟南芥中。通过体外生长试验和非损伤性叶绿素荧光成像筛选耐盐、渗透和氧化胁迫的植株,鉴定出20个在胁迫条件下表现优异的拟南芥株系。通过分析独立的转基因株系,克隆了几个cDNA插入片段,并证实它们与增强的耐受性有关。例如,全长cDNA编码的蛋白质与GDSL脂肪酶/酯酶或酰基辅酶A结合蛋白具有高度同源性,或者编码功能未知的蛋白质,这些蛋白质可以赋予对一种或几种胁迫条件的耐受性。我们的结果证实,从耐胁迫植物物种向敏感植物物种随机转移基因是发现具有生物技术应用潜力新基因的一种有价值的工具。

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