粳稻品种‘云引’谷胱甘肽S-转移酶基因OsGST的克隆及序列分析

毛小辉; 魏毅东; 张建福; 谢华安

doi:10.19303/j.issn.1008-0384.2014.03.001

粳稻品种‘云引’谷胱甘肽S-转移酶基因OsGST的克隆及序列分析

doi: 10.19303/j.issn.1008-0384.2014.03.001

毛小辉^1,2,3,4,5,6,
魏毅东^2,3,4,5,6,
张建福^2,3,4,5,6,
谢华安^2,3,4,5,6

1. 福州大学生物科学与工程学院，福建福州 350108;
2. 农业部华南杂交水稻种质创新与分子育种重点实验室/福州国家水稻改良分中心/福建省作物分子育种工程实验室/福建省水稻分子育种重点实验室，福建福州 350003;
3. 福建省农业科学院水稻研究所，福建福州 350019;
4. 福建省作物种质创新与分子育种省部共建国家重点实验室培育基地，福建福州 350003;
5. 水稻国家工程实验室，福建福州 350003;
6. 杂交水稻国家重点实验室华南基地，福建福州 350003

基金项目:

福建省财政专项——福建省农业科学院科技创新团队建设项目(CXTD-1-1301)

国家“973”计划项目(2010CB723003)

国家转基因农作物新品种培育重大专项(2013ZX08001-002)

详细信息

作者简介:
毛小辉(1989-),女,硕士研究生,研究方向:遗传学(E-mail:814709913@qq.com);张建福(1971-),男,博士,研究员,主要从事水稻分子生物学与分子育种研究(E-mail:jianfzhang@163.com);谢华安(1941-),男,研究员,中国科学院院士,主要从事水稻遗传育种研究(E-mail:huaanxie@163.com)

中图分类号: S511.22
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出版历程
- 收稿日期: 2013-11-05
- 刊出日期: 2014-03-18

Cloning and Sequence Analysis of Glutathione S-transferase(GST)Gene of Japonica Variety‘Yunyin'

MAO Xiao-hui^1,2,3,4,5,6,
WEI Yi-dong^2,3,4,5,6,
ZHANG Jian-fu^2,3,4,5,6,
XIE Hua-an^2,3,4,5,6

1. College of Biological Science and Technology,Fuzhou University;
2. Key Laboratory of Germplasm Innovation and Molecular Breeding of Hybrid Rice for South China,Ministryof Agriculture,P.R China/Fuzhou branch,National Rice Improvement Center of China/Fujian EngineeringLaboratory of Crop Molecular Breeding/Fujian Key Laboratory of Rice Molecular Breeding;
3. Rice Research Institute,Fujian Academy of Agricultural Sciences;
4. Incubator of National Key Laboratory of Fujian Germplasm Innovation and Molecular Breeding between Fujian and Ministry of Sciences & Technology,P.R.China;
5. National Rice Engineering Laboratory of China;
6. South China Bases of National Key Laboratory of Hybrid Rice for China

摘要

摘要: 在对粳稻‘云引’受稻瘟病菌诱导的近等基因系蛋白质组学分析的基础上,通过实时定量PCR等方法,以普感稻瘟病品种丽江新团黑谷(LTH)为遗传背景,研究含有云引抗稻瘟病菌株“四川-43”基因的近等基因系的抗病和感病材料,预测基因谷胱甘肽转移酶OsGST(Genbank序列号:Os05g0116100)对稻瘟病菌诱导的响应及响应程度的差别。利用RACE技术克隆目的基因cDNA全长序列,并在近等基因系抗病材料的抗病株系和感病株系中进行序列比对,寻找差异位点。结果表明,该基因与短柄二叶草中编码谷胱甘肽转移酶的基因亲缘关系最接近,结合其编码的蛋白质序列分析进一步论证该基因可能作为一个相对保守的抗病防卫相关基因、编码谷胱甘肽转移酶,将基因命名为OsGST。同时构建了该基因的过表达载体、干扰载体,为后续揭示其在感抗材料中响应稻瘟病菌程度差异的原因、研究其功能及抗病机制奠定基础。
- 云引 /
- 近等基因系 /
- 谷胱甘肽S-转移酶 /
- 序列分析
Abstract: Based on the proteomic analysis of blast-resistant near-isogenic lines derived from japonica rice ‘Yunyin',infected with M.oryzae,real-time quantitative PCR and other methods were used to explore the blast resistant and susceptible materials of near-isogenic lines using LTH as genetic background and containing blast ‘Sichuan-43'resistance strain to forecast for rice blast fungus-induced response and the different degrees between the responses of Glutathione S-transferase gene OsGST(Genbank sequence number:Os05g0116100).Using RACE technology to get the full cDNA Sequence,and comparing the sequence between the blast resistant lines and the blast susceptible lines.The phylogenetic tree showed the gene had the closest genetic relationship with glutathione transferase encoding gene of brachypodium distachyon.Combined with the analysis of the protein sequence,OsGST was suggested to be a conserved gene that encoding glutathione transferase.The overexpression and RNAi expression vector were constructed for subsequent study of its functional identification and resistant mechanisms.
- ‘Yunyin' /
- near-isogenic lines /
- glutathione S-transferase /
- sequence analysis