[1]马辉,韩永涛,高玉荣,等. 葡萄中RPW8.2同源基因克隆与表达分析[J].西北林学院学报,2015,30(01):60-68.[doi:doi:10.3969/j.issn.1001-7461.2015.01.10]
　MA Hui,HAN Yong-tao,GAO Yu-rong,et al. Molecular Cloning and Expression Analysis of RPW8.2 Homologous Genes in Grapevine[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2015,30(01):60-68.[doi:doi:10.3969/j.issn.1001-7461.2015.01.10]
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 葡萄中RPW8.2同源基因克隆与表达分析()
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《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]

卷:
第30卷
期数:
2015年01期
页码:
60-68
栏目:
出版日期:
2015-01-31

文章信息/Info

Title:
 Molecular Cloning and Expression Analysis of RPW8.2 Homologous Genes in Grapevine
文章编号:
1001-7461(2015)01-0060-09
作者:
 马辉1; 4; 韩永涛1; 2; 3; 高玉荣1; 2; 3; 徐虹1; 4; 王跃进1; 2; 3; 文颖强1; 2; 3*
 （1.旱区作物逆境生物学国家重点实验室，陕西 杨陵 712100；2.农业部西北地区园艺作物生物学与种质创制重点实验室，陕西 杨陵 712100；3.西北农林科技大学 园艺学院，陕西 杨陵 712100；4.西北农林科技大学 生命科学学院，陕西 杨陵712100）
Author(s):
 MA Hui1; 4;  HAN Yong-tao1; 2; 3;  GAO Yu-rong1; 2; 3;  XU Hong1; 4;  WANG Yue-jin1; 2; 3;  WEN Ying-qiang1; 2; 3*
 (1.State Key Laboratory of Crop Stress Biology in Arid Areas, Yangling, Shaanxi 712100, China; 2. Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Yangling, Shaanxi 712100, China; 3. College of Horticulture, Northwest A&F University, Yangling, Shaanxi 712100, China)
关键词:
 ; RPW8.2; 葡萄; 同源克隆; 表达分析
Keywords:
 ; RPW8.2;  Vitis;  homology cloning;  expression analysis
分类号:
S663.1
DOI:
doi:10.3969/j.issn.1001-7461.2015.01.10
文献标志码:
A
摘要:
 利用同源克隆技术分别从白河35-2、广西-1、黑比诺、五月紫4种葡萄中克隆得到了拟南芥广谱抗病基因RPW8.2的同源基因，分别命名为VpR8H-BH2、VpR8H-GX1、VvR8H-PN和VvR8H-MP，氨基酸多序列比对和系统发育分析显示，该4条序列均属于RPW8.2同源基因。4种葡萄R82H基因cDNA开放阅读框分别为2 457 bp、2 445 bp、2 445 bp和2 448 bp，分别编码819、815、815和816个氨基酸。采用半定量RT-PCR技术分析了R82H基因在4种葡萄根、茎、成熟叶、幼嫩叶、卷须不同组织中的表达模式，R82H基因在不同葡萄组织中均有表达，但是表达丰度不尽相同，其中，在白河35-2和五月紫葡萄成熟叶中表达量最高，而在广西-1和黑比诺中幼嫩叶片中表达量高于其他组织。
Abstract:
 Four genes designated VpR8H-BH2, VpR8H-GX1, VvR8H-PN and VvR8H-MP were cloned and sequenced by using homology-based cloning method from Chinese wild grapevine Vitis pseudoreticulata accession Baihe-35-2, accession Guangxi-1, and Vitis vinifera cv. Pinot Noir, and Vitis vinifera cv. May Purple, respectively. Amino acid sequences alignment and phylogenesis results showed that four genes were Arabidopsis broad-spectrum disease resistance gene RPW8.2 homologous genes. The full-length ORF of VpR8H-BH2, VpR8H-GX1, VvR8H-PN and VvR8H-MP were 2 457 bp, 2 445 bp, 2 445 bp and 2 448 bp, respectively, and encoded polypeptides of 819, 815, 815 and 816 amino acids, respectively. Semi-quantitative RT-PCR results indicated that four genes were expressed in all of the grapevine tissue types, including root, stem, mature leaves, young leaves and tendrils, but the expressions were different. The VpR8H-BH2 expression level of accession Baihe35-2 and the VvR8H-MP expression level of Vitis vinifera cv. May Purple were the most in mature leaves, but the expression level of young leaves of accession Guangxi-1 and Vitis vinifera cv. Pinot Noir were higher than other tissues.

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备注/Memo

备注/Memo:
 收稿日期：2014-03-19 修回日期：2014-04-14
基金项目：国家自然科学基金项目（31071772）；高校基本科研业务费专项资金项目（QN2011005）。
作者简介：马辉，男，硕士研究生，研究方向：果树育种与生物技术。E-mail: tianma1203@126.com
*通信作者：文颖强，男，博士，副教授，研究方向：葡萄种质资源与生物技术育种。E-mail: wenyq@nwsuaf.edu.cn
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