甜瓜自毒相关基因CmGST的克隆及其对自毒胁迫的响应
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引用本文:王景荣,张政达,樊佳茹,张贝贝,王革伏,张志忠.甜瓜自毒相关基因CmGST的克隆及其对自毒胁迫的响应[J].西北植物学报,2019,39(7):1172~1180
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王景荣1,张政达1,樊佳茹1,张贝贝1,王革伏1,2,张志忠1* (1 福建农林大学 园艺学院福建农林大学 戴尔豪西大学联合实验室, 福州 3500022 加拿大戴尔豪西大学 农学院加拿大特鲁罗 N2B 5E3) 
基金项目:福建农林大学创新专项基金(KFA17108A,KFA17347A);
中文摘要:植物谷胱甘肽硫转移酶基因(GST)在清除生物和非生物胁迫产生的氧化损伤中扮演着重要的角色,为探究GST在根系分泌物介导的甜瓜自毒胁迫中的响应机制,该实验在转录组测序基础上,以甜瓜叶片cDNA为模板,采用RT PCR技术获得了一个根系分泌物介导的甜瓜自毒作用密切相关的GST基因,命名为CmGST(GenBank 登录号:AYU66762.1);对CmGST基因进行了相关生物信息学分析,同时对自毒胁迫过程中,CmGST的差异表达情况、谷胱甘肽硫转移酶(GST)活性和谷胱甘肽(GSH)含量进行了相关测定。结果显示:(1)CmGST基因含有完整开放阅读框(ORF)654 bp,编码217个氨基酸;CmGST蛋白为酸性亲水蛋白,且较为稳定,具有GST家族Tau亚家族的2个典型的保守结构域;CmGST蛋白与黄瓜的亲缘关系较近。(2)亚细胞定位结果表明CmGST定位于细胞质中。(3)qRT PCR分析结果表明,CmGST在甜瓜根系和幼苗中均有表达,在正常生长的甜瓜幼苗中表达量相对稳定;自毒胁迫后在叶中的表达先降后升,后期迅速增加;在根系中变化趋势类似,但程度较为缓和。(4)GST活性和GSH含量分析表明,叶片中GST活性变化趋势与CmGST基因表达变化基本吻合;在根系中,GST活性呈现先升再降最后又升高的趋势;随胁迫时间增加,GSH含量在根系和叶片中均都有不同程度的增加,但根部含量低于叶片。研究认为,在自毒胁迫过程中,CmGST基因在甜瓜植株根和叶片均有相应表达水平的变化,参与了对根系分泌物介导的自毒胁迫响应。
中文关键词:甜瓜  谷胱甘肽S 转移酶基因  自毒作用  根系分泌物
 
Cloning and Expression of CmGST Gene Related to Autotoxicity Stress in Cucumis melo L.
Abstract:The plant glutathione S transferase gene (GST) plays an important role in scavenging oxidative damage caused by biotic and abiotic stresses. In order to explore the response mechanism of GST in allelopathic autotoxicity mediated by root exudates, we obtained a GST gene closely related to root exudates mediated autotoxicity in melon by RT PCR based on transcriptome sequencing and using melon leaf DNA as template, named CmGST (GenBank login number: AYU66762.1). In this study, the CmGST gene was analyzed by bioinformatics, and the differential expression of CmGST, the activity of glutathione S transferase (GST) and the content of glutathione (GSH) in the process of autotoxic stress were determined. The results showed that: (1) the CmGST gene contains a complete open reading frame (ORF) of 654 bp, encoding 217 amino acids. CmGST was an acidic hydrophilic protein and was relatively stable. It has two typical conserved domains of GST family Tau subfamily. CmGST protein had close relationship with Cucummis sativus. (2) Subcellular localization indicated that CmGST was located in the cytoplasm. (3) The results of q PCR analysis showed that CmGST was expressed in root and seedling of melon, and the expression level was relatively stable in normal growth melon seedlings. The expression of CmGST in leaves decreased firstly and then increased rapidly after autotoxicity stress, and the change trend was similar in roots, but the degree was relatively mild. The analysis of GST activity and GSH content showed that the change trend of GST activity in leaves was basically consistent with the change of gene expression of CmGST. In roots, GST activity increased firstly, then decreased, and finally increased. (4) With the increase of stress time, GSH content in roots and leaves increased in varying degrees, but the content in root was lower than that in leaves. The study indicated that the expression level of CmGST in roots and leaves of melon plants changed during the process of autotoxic stress, and CmGST participated in the response to autotoxic stress mediated by root exudates.
keywords:Cucumis melo L.  CmGST gene  autotoxicity stress  root exudates
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