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从耐盐耐镉盐生植物海滨雀稗中克隆的基因的功能鉴定与特性分析

Functional Identification and Characterization of Genes Cloned from Halophyte Seashore Paspalum Conferring Salinity and Cadmium Tolerance.

作者信息

Chen Yu, Chen Chuanming, Tan Zhiqun, Liu Jun, Zhuang Lili, Yang Zhimin, Huang Bingru

机构信息

Department of Turfgrass Science, College of Agro-Grassland Science, Nanjing Agricultural University Nanjing, China.

Department of Plant Biology and Pathology, Rutgers, The State University of New Jersey New Brunswick, NJ, USA.

出版信息

Front Plant Sci. 2016 Feb 9;7:102. doi: 10.3389/fpls.2016.00102. eCollection 2016.

Abstract

Salinity-affected and heavy metal-contaminated soils limit the growth of glycophytic plants. Identifying genes responsible for superior tolerance to salinity and heavy metals in halophytes has great potential for use in developing salinity- and Cd-tolerant glycophytes. The objective of this study was to identify salinity- and Cd-tolerance related genes in seashore paspalum (Paspalum vaginatum), a halophytic perennial grass species, using yeast cDNA expression library screening method. Based on the Gateway-compatible vector system, a high-quality entry library was constructed, which contained 9.9 × 10(6) clones with an average inserted fragment length of 1.48 kb representing a 100% full-length rate. The yeast expression libraries were screened in a salinity-sensitive and a Cd-sensitive yeast mutant. The screening yielded 32 salinity-tolerant clones harboring 18 salinity-tolerance genes and 20 Cd-tolerant clones, including five Cd-tolerance genes. qPCR analysis confirmed that most of the 18 salinity-tolerance and five Cd-tolerance genes were up-regulated at the transcript level in response to salinity or Cd stress in seashore paspalum. Functional analysis indicated that salinity-tolerance genes from seashore paspalum could be involved mainly in photosynthetic metabolism, antioxidant systems, protein modification, iron transport, vesicle traffic, and phospholipid biosynthesis. Cd-tolerance genes could be associated with regulating pathways that are involved in phytochelatin synthesis, HSFA4-related stress protection, CYP450 complex, and sugar metabolism. The 18 salinity-tolerance genes and five Cd-tolerance genes could be potentially used as candidate genes for genetic modification of glycophytic grass species to improve salinity and Cd tolerance and for further analysis of molecular mechanisms regulating salinity and Cd tolerance.

摘要

受盐分影响和重金属污染的土壤限制了甜土植物的生长。鉴定盐生植物中对盐分和重金属具有超强耐受性的基因,在培育耐盐和耐镉甜土植物方面具有巨大的应用潜力。本研究的目的是利用酵母cDNA表达文库筛选方法,鉴定海滨雀稗(Paspalum vaginatum)中与耐盐和耐镉相关的基因,海滨雀稗是一种多年生盐生草本植物。基于Gateway兼容载体系统,构建了一个高质量的入门文库,该文库包含9.9×10⁶个克隆,平均插入片段长度为1.48 kb,全长率达100%。在对盐分敏感和对镉敏感的酵母突变体中筛选酵母表达文库。筛选得到32个耐盐克隆,包含18个耐盐基因和20个耐镉克隆,其中包括5个耐镉基因。qPCR分析证实,18个耐盐基因和5个耐镉基因中的大多数在海滨雀稗中响应盐分或镉胁迫时,转录水平上调。功能分析表明,海滨雀稗的耐盐基因可能主要参与光合代谢、抗氧化系统、蛋白质修饰、铁运输、囊泡运输和磷脂生物合成。耐镉基因可能与参与植物螯合肽合成、HSFA4相关胁迫保护、CYP450复合体和糖代谢的调控途径有关。这18个耐盐基因和5个耐镉基因有可能用作候选基因,用于甜土草本植物的基因改造,以提高其耐盐性和耐镉性,并进一步分析调控耐盐性和耐镉性的分子机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/315b/4746305/a38dba0fe0a1/fpls-07-00102-g0001.jpg

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