School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China.
Bengbu Medical College, Bengbu, 233000, People's Republic of China.
Plant Cell Rep. 2017 Sep;36(9):1457-1476. doi: 10.1007/s00299-017-2168-2. Epub 2017 Jun 27.
The G1-like gene from the Lycium chinense was cloned and transferred into N. tabacum. Evidence showed that endogenous JA accumulation was crucial to LcGR gene expression in cadmium-stressed L. chinense. Glutathione reductase (GR) plays a vital role in glutathione-ascorbate metabolism and is a key enzyme in maintaining the redox state in plants. Jasmonic acids (JA) are important hormones regulating protective responses against bacteria and mechanic damage in plants. At present, the relationship between the endogenous JA accumulation, the glutathione (GSH) content and GR gene expression in plants under cadmium (Cd) stress has not been elucidated. This study primarily aims to explore their interconnected relations. First, we isolated the GR1-like gene from Lycium chinense (LcGR). Real-time PCR showed that gene LcGR and allene oxide cyclase (LcAOC) (a JA synthesis gene) expression in L. chinense plants was significantly enhanced by CdCl and reduced by CdCl cotreatment with 12,13-epoxy-octadecenoic acid (EOA), a JA synthesis inhibitor. Meanwhile, the JA content in plants strongly increased under Cd stress and decreased under Cd + EOA treatment, which was in accordance with expression pattern of LcAOC. The function of gene LcGR was confirmed in vitro with E. coli expression system. The subcellular localization in chloroplasts of LcGR gene was proved in Nicotiana tabacum leaves with transient transfection system of Agrobacterium tumefaciens. Furthermore, the overexpression of gene LcGR in the transgenic tabacum led to great Cd-tolerance and higher GSH accumulation. Overall, the results showed that the endogenous JA accumulation in Cd-stressed plants affects the GR expression which is crucial to the GSH accumulation and GSH-dependent tolerance to cadmium in LcGR transformants.
从宁夏枸杞中克隆并转入烟草的 G1 样基因。证据表明,内源性 JA 积累对镉胁迫下宁夏枸杞 LcGR 基因表达至关重要。谷胱甘肽还原酶 (GR) 在谷胱甘肽-抗坏血酸代谢中发挥重要作用,是维持植物氧化还原状态的关键酶。茉莉酸 (JA) 是调节植物对细菌和机械损伤的保护反应的重要激素。目前,尚未阐明植物在镉 (Cd) 胁迫下内源 JA 积累、谷胱甘肽 (GSH) 含量和 GR 基因表达之间的关系。本研究主要旨在探讨它们之间的相互关系。首先,我们从宁夏枸杞中分离出 GR1 样基因。实时 PCR 显示,CdCl 处理可显著增强宁夏枸杞植物中基因 LcGR 和丙二烯氧化物环化酶 (LcAOC)(一种 JA 合成基因)的表达,而 JA 合成抑制剂 12,13-环氧十八碳烯酸 (EOA) 则降低了其表达。同时,植物在 Cd 胁迫下 JA 含量强烈增加,而在 Cd + EOA 处理下降低,这与 LcAOC 的表达模式一致。通过大肠杆菌表达系统证实了基因 LcGR 的功能。通过农杆菌瞬时转染系统在烟草叶片中证明了 LcGR 基因在叶绿体中的亚细胞定位。此外,在转基因烟草中过表达基因 LcGR 导致对 Cd 的耐受性增强和 GSH 积累增加。总之,结果表明,Cd 胁迫植物中的内源性 JA 积累影响 GR 表达,这对 LcGR 转化体中 GSH 积累和 GSH 依赖的 Cd 耐受性至关重要。