首页> 美国卫生研究院文献>Annals of Botany >Glutathione Reductase in Leaves of Cowpea: Cloning of Two cDNAs Expression and Enzymatic Activity under Progressive Drought Stress Desiccation and Abscisic Acid Treatment
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Glutathione Reductase in Leaves of Cowpea: Cloning of Two cDNAs Expression and Enzymatic Activity under Progressive Drought Stress Desiccation and Abscisic Acid Treatment

机译:of豆叶片谷胱甘肽还原酶:两个cDNA的克隆干旱胁迫脱水和脱落酸处理下的表达和酶活性

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摘要

• Background and Aims Reactive oxygen species are frequently produced when plants are exposed to abiotic stresses. Among the detoxication systems, two enzymes, ascorbate peroxidase and glutathione reductase (GR) play key roles. GR has also a central role in keeping the reduced glutathione pool during stress thus allowing the adjustments on the cellular redox reactions. The aim of this work was to study the variations in cytosolic and dual-targeted GR gene expression in the leaves of cowpea plants submitted to progressive drought, rapid desiccation and application of exogenous abscisic acid (ABA).• Methods Two cowpea (Vigna unguiculata) cultivars, one drought-resistant (‘EPACE-1’), the other drought-sensitive (‘1183’) were submitted to progressive drought stress by withholding irrigation. Cut-off leaves were air-dried or treated with exogenous ABA. Two GR cDNAs, one cytosolic, the other dual-targeted to chloroplasts and mitochondria were isolated by PCR and cloned in plasmid vectors. Reverse-transcription PCR was used to study the variations in GR gene expression.• Key Results Two new cDNAs encoding a putative dual-targeted and a cytosolic GR were cloned and sequenced from leaves of V. unguiculata. Drought stress induced an up-regulation of the expression of the cytosolic GR gene directly related to the intensity of the stress in both cultivars. The expression of dual-targeted GR was up-regulated by the drought treatment in the susceptible cultivar only. Under a fast desiccation, the ‘1183’ cultivar responded later than the ‘EPACE-1’, although in ‘EPACE-1’ it was the cytosolic isoform which responded and in ‘1183’ the dual-targeted one. Exogenous ABA enhanced significantly the activity and expression levels of GR in both cultivars after treatment for 24 h.• Conclusions These results demonstrate a noticeable activation in both cultivars of the antioxidant metabolism under a progressive water stress, which involves both GR genes in the case of the susceptible cultivar. Under a fast desiccation, the susceptible cultivar responded later than the resistant one, suggesting a weaker capacity of response versus the resistant one. Exogenous ABA probably acts on GR gene expression via a mediated signal transduction pathway.
机译:•背景和目的当植物受到非生物胁迫时,经常会产生活性氧。在脱毒系统中,抗坏血酸过氧化物酶和谷胱甘肽还原酶(GR)这两种酶起着关键作用。 GR在应激期间保持减少的谷胱甘肽池中也起着中心作用,因此可以调节细胞的氧化还原反应。这项工作的目的是研究进行性干旱,快速干燥和外源脱落酸(ABA)施用后的cow豆植物叶片中胞质和双靶GR基因表达的变化。方法二种cow豆(Vigna unguiculata)一个抗旱品种('EPACE-1'),另一个对干旱敏感的品种('1183'),通过不灌溉来承受进行性干旱胁迫。将切下的叶子风干或用外源ABA处理。通过PCR分离出两个GR cDNA,一个胞质,另一个双重靶向叶绿体和线粒体,并克隆到质粒载体中。逆转录PCR被用于研究GR基因表达的变化。•关键结果从V. unguiculata的叶子中克隆了两个新的编码假定的双靶和胞质GR的cDNA。干旱胁迫导致与两个品种胁迫强度直接相关的胞质GR基因表达上调。仅在易感品种中,干旱处理上调了双重靶向GR的表达。在快速干燥下,“ 1183”品种的反应晚于“ EPACE-1”,尽管在“ EPACE-1”中是胞浆同工型,而在“ 1183”中是双靶品种。外源ABA处理24h后,两个品种的GR活性和表达水平显着提高。•结论这些结果表明,在渐进的水分胁迫下,两个品种的抗氧化代谢都有明显的激活,在干旱胁迫下涉及两个GR基因。易感品种在快速干燥下,易感品种对抗病品种的反应晚于抗病品种,这表明其抗病能力较抗病品种弱。外源ABA可能通过介导的信号转导途径作用于GR基因表达。

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