[1]杨霞,王笑,管荣展,等.多抗性稗草中 ２ 个谷胱甘肽转移酶基因的克隆与分析[J].江苏农业学报,2015,(06):1296-1303.[doi:doi:10.3969/j.issn.1000-4440.2015.06.016]
　YANG Xia,WANG Xiao,GUAN Rong-zhan,et al.Cloning and characterization of two glutathione S-transferases genes from herbicide-resistant Echinochloa crusgalli[J].,2015,(06):1296-1303.[doi:doi:10.3969/j.issn.1000-4440.2015.06.016]
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多抗性稗草中 ２ 个谷胱甘肽转移酶基因的克隆与分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年06期
页码:
1296-1303
栏目:
植物保护
出版日期:
2015-12-31

文章信息/Info

Title:
Cloning and characterization of two glutathione S-transferases genes from herbicide-resistant Echinochloa crusgalli
作者:
杨霞1; 王笑12; 管荣展2; 李永丰1; 董明超12; 张自常1
(1.江苏省农业科学院植物保护研究所,江苏南京210014;2.南京农业大学农学院,江苏南京210095)
Author(s):
YANG Xia1; WANG Xiao12; GUAN Rong-zhan2; LI Yong-feng1; DONG Ming-chao12; ZHANG Zi-chang1
(1.Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;2.College of Agriculture,Nanjing Agricultural University,Nanjing 210095,China)
关键词:
稗草; 谷胱甘肽转移酶（GST）; 基因克隆; 原核表达; 实时荧光定量PCR
Keywords:
Echinochloa crus-galli; glutathione S-transferases; gene cloning; prokaryotic expression; quantitative real-time PCR
分类号:
S451.21
DOI:
doi:10.3969/j.issn.1000-4440.2015.06.016
文献标志码:
A
摘要:
为深入揭示稗草中谷胱甘肽转移酶(GST)对除草剂的解毒机理,本研究通过PCR方法从多抗性稗草中获得了phi类EcGSTF1和zeta类EcGSTZ1基因,并进行生物信息学、基因表达研究及酶活性测定。系统进化树分析结果显示稗草GST家族成员与禾本科植物的亲缘关系较近;将EcGSTF1和EcGSTZ1分别插入pET28b载体中进行0.4 mmol／L IPTG诱导表达,蛋白质分子量大小为26 000;实时荧光定量PCR结果显示EcGST在多抗性稗草中的表达量高于在敏感性稗草,其中EcGSTF1基因表达量比EcGSTZ1高;GST酶比活力分析显示表明未施药处理的多抗性稗草GST酶比活力高于敏感性稗草,施药7 d后多抗性稗草GST酶比活力高于敏感性稗草。综上所述,稗草中GST酶对除草剂有解毒作用,其中phi类GST更为突出。
Abstract:
To achieve a better understanding of the detoxification mechanism of plant glutathione S-transferases (GST),phi EcGSTF1 and zeta EcGSTZ1 genes were identified from multiple herbicide-resistant and susceptible barnyardgrass (Echinochloa crus-galli) using PCR method and analyzed using bioinformatics. Gene expression and enzyme activity were measured as well Phylogenetic analysis revealed that EcGSTs was highly homologous to GST from Gramineae plants. The molecular weight of HIS::EcGST fusion proteins induced by 0.4 mmol／L IPTG was 26 000 in weight. Quantitative real-time PCR showed that the expression level of EcGSTF1 was higher than that of EcGSTZ1,and EcGST gene expression levels in resistant populations was higher than that in susceptible populations.The specific activity of GST was higher in resistant biotypes than that in susceptible biotypes seven days after herbicides application.

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备注/Memo

备注/Memo:
收稿日期：2015-07-01 基金项目：国家自然科学基金项目（31371953、31200334）；江苏省农业科技自主创新基金项目[SCX(13)3063]；江苏省面上基金项目（SBK2014020595） 作者简介：杨霞（1981-），女，江苏姜堰人，博士，副研究员，主要从事杂草抗药性机制研究。（E-mail）yangxia@jaas.ac.cn 通讯作者：李永丰,（E-mail）liyongfeng_2010@163.com
更新日期/Last Update: 2015-12-31