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绢毛苣γ-谷氨酰半胱氨酸合成酶基因的分子克隆与特性分析

Molecular cloning and characterization of a gamma-glutamylcysteine synthetase gene from Chorispora bungeana.

作者信息

Wu Jianmin, Qu Tao, Chen Shuyan, Zhao Zhiguang, An Lizhe

机构信息

MOE Key Laboratory of Arid and Grassland Ecology, School of Life Sciences, Lanzhou University, Lanzhou, 730000, China.

出版信息

Protoplasma. 2009 Mar;235(1-4):27-36. doi: 10.1007/s00709-008-0026-3. Epub 2008 Dec 12.

Abstract

Chorispora bungeana Fisch. and C.A. Mey (C. bungeana) is a rare alpine subnival plant species that is highly tolerant of freezing temperatures. Glutathione (GSH) is a major player in various metabolic processes involved in plant growth and development and stress responses. A recent study has found that the inhibition of GSH synthesis decreases the chilling tolerance of C. bungeana callus (J. Wu et al.: Cryobiology 57:9-17, 2008). We have isolated and characterized a full-length cDNA encoding gamma-glutamylcysteine synthetase (Cb gamma ECS), the key enzyme of GSH synthesis, from the leaves of C. bungeana, with the aim of furthering our understanding of the role of GSH at the molecular level. Cb gamma ECS was found to encode a 524-amino acid protein with moderate to high nucleotide sequence similar to previously reported plant gamma-ECS genes. Cb gamma ECS transcripts were detected in the calluses, roots and leaves of C. bungeana, but there was no tissue-specific expression. The transcripts of Cb gamma ECS accumulated both rapidly and at high levels when C. bungeana was treated with chilling (4 degrees C), freezing (-4 degrees C), several other environmental stresses (such as heat, salinity, osmotic and heavy metals), abscisic acid and hydrogen peroxide. These results suggest that Cb gamma ECS may participate in the cellular responses against multiple environmental stimuli in C. bungeana.

摘要

Bunge千里光(Chorispora bungeana Fisch. and C.A. Mey,简称C. bungeana)是一种稀有的高山亚冰雪带植物物种,对低温具有高度耐受性。谷胱甘肽(GSH)在植物生长发育和应激反应所涉及的各种代谢过程中起着重要作用。最近的一项研究发现,抑制GSH合成会降低C. bungeana愈伤组织的耐寒性(J. Wu等人:《低温生物学》57:9 - 17,2008)。我们从C. bungeana的叶片中分离并鉴定了一个编码γ-谷氨酰半胱氨酸合成酶(CbγECS)的全长cDNA,CbγECS是GSH合成的关键酶,目的是在分子水平上进一步了解GSH的作用。发现CbγECS编码一个524个氨基酸的蛋白质,其核苷酸序列与先前报道的植物γ-ECS基因具有中度到高度的相似性。在C. bungeana的愈伤组织、根和叶中检测到CbγECS转录本,但没有组织特异性表达。当C. bungeana受到低温(4℃)、冷冻(-4℃)、其他几种环境胁迫(如热、盐度、渗透和重金属)、脱落酸和过氧化氢处理时,CbγECS的转录本迅速且大量积累。这些结果表明,CbγECS可能参与了C. bungeana对多种环境刺激的细胞反应。

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