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通过cDNA表达文库筛选对盐生植物结缕草中的耐盐基因进行系统挖掘。

Systematic mining of salt-tolerant genes in halophyte-Zoysia matrella through cDNA expression library screening.

作者信息

Chen Yu, Zong Junqin, Tan Zhiqun, Li Lanlan, Hu Baoyun, Chen Chuanming, Chen Jingbo, Liu Jianxiu

机构信息

Jiangsu Key Laboratory for Bioresources of Saline Soils, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, China; College of Ago-grassland Science, Nanjing Agricultural University, Nanjing 210095, China.

Jiangsu Key Laboratory for Bioresources of Saline Soils, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, China.

出版信息

Plant Physiol Biochem. 2015 Apr;89:44-52. doi: 10.1016/j.plaphy.2015.02.007. Epub 2015 Feb 12.

Abstract

Though a large number of salt-tolerant genes were identified from Glycophyte in previous study, genes involved in salt-tolerance of halophyte were scarcely studied. In this report, an important halophyte turfgrass, Zoysia matrella, was used for systematic excavation of salt-tolerant genes using full-length cDNA expression library in yeast. Adopting the Gateway-compatible vector system, a high quality entry library was constructed, containing 3 × 10(6) clones with an average inserted fragments length of 1.64 kb representing a 100% full-length rate. The yeast expression library was screened in a salt-sensitive yeast mutant. The screening yielded dozens of salt-tolerant clones harboring 16 candidate salt-tolerant genes. Under salt-stress condition, these 16 genes exhibited different transcription levels. According to the results, we concluded that the salt-tolerance of Z. matrella might result from known genes involved in ion regulation, osmotic adjustment, as well as unknown pathway associated with protein folding and modification, RNA metabolism, and mitochondrial membrane translocase, etc. In addition, these results shall provide new insight for the future researches with respect to salt-tolerance.

摘要

尽管在先前的研究中已从甜土植物中鉴定出大量耐盐基因,但对盐生植物耐盐相关基因的研究却很少。在本报告中,一种重要的盐生植物草坪草——结缕草,被用于利用酵母中的全长cDNA表达文库对耐盐基因进行系统挖掘。采用与Gateway兼容的载体系统,构建了一个高质量的入门文库,包含3×10⁶个克隆,平均插入片段长度为1.64 kb,全长率达100%。在一个盐敏感酵母突变体中筛选酵母表达文库。筛选得到了数十个耐盐克隆,其中包含16个候选耐盐基因。在盐胁迫条件下,这16个基因表现出不同的转录水平。根据结果,我们得出结论,结缕草的耐盐性可能源于参与离子调节、渗透调节的已知基因,以及与蛋白质折叠和修饰、RNA代谢和线粒体膜转位酶等相关的未知途径。此外,这些结果将为未来关于耐盐性的研究提供新的见解。

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