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一种来自生菜的新型Em样蛋白LsEm1可增强大肠杆菌和水稻对干旱和盐胁迫的耐受性。

A new Em-like protein from Lactuca sativa, LsEm1, enhances drought and salt stress tolerance in Escherichia coli and rice.

作者信息

Xiang Dian-Jun, Man Li-Li, Zhang Chun-Lan, Li Zhi-Gang, Zheng Gen-Chang

机构信息

College of Agriculture, Inner Mongolia University for Nationalities, Tongliao, 028042, China.

College of Life Science, Inner Mongolia University for Nationalities, Tongliao, 028042, China.

出版信息

Protoplasma. 2018 Jul;255(4):1089-1106. doi: 10.1007/s00709-018-1207-3. Epub 2018 Feb 7.

Abstract

Late embryogenesis abundant (LEA) proteins are closely related to abiotic stress tolerance of plants. In the present study, we identified a novel Em-like gene from lettuce, termed LsEm1, which could be classified into group 1 LEA proteins, and shared high homology with Cynara cardunculus Em protein. The LsEm1 protein contained three different 20-mer conserved elements (C-element, N-element, and M-element) in the C-termini, N-termini, and middle-region, respectively. The LsEm1 mRNAs were accumulated in all examined tissues during the flowering and mature stages, with a little accumulation in the roots and leaves during the seedling stage. Furthermore, the LsEm1 gene was also expressed in response to salt, dehydration, abscisic acid (ABA), and cold stresses in young seedlings. The LsEm1 protein could effectively reduce damage to the lactate dehydrogenase (LDH) and protect LDH activity under desiccation and salt treatments. The Escherichia coli cells overexpressing the LsEm1 gene showed a growth advantage over the control under drought and salt stresses. Moreover, LsEm1-overexpressing rice seeds were relatively sensitive to exogenously applied ABA, suggesting that the LsEm1 gene might depend on an ABA signaling pathway in response to environmental stresses. The transgenic rice plants overexpressing the LsEm1 gene showed higher tolerance to drought and salt stresses than did wild-type (WT) plants on the basis of the germination performances, higher survival rates, higher chlorophyll content, more accumulation of soluble sugar, lower relative electrolyte leakage, and higher superoxide dismutase activity under stress conditions. The LsEm1-overexpressing rice lines also showed less yield loss compared with WT rice under stress conditions. Furthermore, the LsEm1 gene had a positive effect on the expression of the OsCDPK9, OsCDPK13, OsCDPK15, OsCDPK25, and rab21 (rab16a) genes in transgenic rice under drought and salt stress conditions, implying that overexpression of these genes may be involved in the enhanced drought and salt tolerance of transgenic rice. Thus, this work paves the way for improvement in tolerance of crops by genetic engineering breeding.

摘要

胚胎后期丰富(LEA)蛋白与植物的非生物胁迫耐受性密切相关。在本研究中,我们从生菜中鉴定出一个新的类Em基因,命名为LsEm1,它可归类为第1组LEA蛋白,与刺菜蓟Em蛋白具有高度同源性。LsEm1蛋白在C末端、N末端和中间区域分别包含三个不同的20聚体保守元件(C元件、N元件和M元件)。LsEm1 mRNA在开花期和成熟期的所有检测组织中均有积累,在幼苗期根和叶中有少量积累。此外,LsEm1基因在幼苗中也响应盐、脱水、脱落酸(ABA)和冷胁迫而表达。LsEm1蛋白在干燥和盐处理下能有效减少对乳酸脱氢酶(LDH)的损伤并保护LDH活性。过表达LsEm1基因的大肠杆菌细胞在干旱和盐胁迫下比对照表现出生长优势。此外,过表达LsEm1的水稻种子对外源施加的ABA相对敏感,这表明LsEm1基因可能依赖ABA信号通路来响应环境胁迫。基于发芽表现、更高的存活率、更高的叶绿素含量、更多的可溶性糖积累、更低的相对电解质渗漏率以及胁迫条件下更高的超氧化物歧化酶活性,过表达LsEm1基因的转基因水稻植株比野生型(WT)植株表现出更高的耐旱性和耐盐性。在胁迫条件下,过表达LsEm1的水稻株系与WT水稻相比产量损失也更小。此外,在干旱和盐胁迫条件下,LsEm1基因对转基因水稻中OsCDPK9、OsCDPK13、OsCDPK15、OsCDPK25和rab21(rab16a)基因的表达有积极影响,这意味着这些基因的过表达可能参与了转基因水稻耐旱性和耐盐性的增强。因此,这项工作为通过基因工程育种提高作物耐受性铺平了道路。

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