[1]王西成,吴伟民,巫建华,等.葡萄β-1,3-葡聚糖酶基因的克隆、序列分析及表达[J].江苏农业学报,2015,(04):899-903.[doi:10.3969/j.issn.1000-4440.2015.04.030]
　JP+WANG Xi-cheng,WU Wei-min,WU Jian-hua,et al.Cloning, characterization and expression analysis of beta-1,3-glucanase gene in grapevine[J].,2015,(04):899-903.[doi:10.3969/j.issn.1000-4440.2015.04.030]
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葡萄β-1,3-葡聚糖酶基因的克隆、序列分析及表达()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年04期
页码:
899-903
栏目:
园艺
出版日期:
2015-08-31

文章信息/Info

Title:
Cloning, characterization and expression analysis of beta-1,3-glucanase gene in grapevine
作者:
王西成12; 吴伟民12; 巫建华3; 赵密珍12; 王壮伟12; 钱亚明12; 解振强3
(1.江苏省农业科学院园艺研究所，江苏南京210014；2.江苏省高效园艺作物遗传改良重点实验室，江苏南京210014；3.江苏现代园艺工程技术中心，江苏句容212400)
Author(s):
〖JP+2〗WANG Xi-cheng12; WU Wei-min12; WU Jian-hua3; ZHAO Mi-zhen12; WANG Zhuang-wei12; QIAN Ya-ming12; XIE Zhen-qiang3
(1.Institute of Horticulture, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2.Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Nanjing 210014, China;3.Jiangsu Modern Horticultural Engineering Center, Jurong 212400, China)〖KH+1D〗
关键词:
葡萄; 霜霉病; β-1; 3-葡聚糖酶基因; 基因克隆; 基因表达
Keywords:
grapevine; downy mildew; beta-1; 3-glucanase gene; gene cloning;  gene expression
分类号:
S663.1
DOI:
10.3969/j.issn.1000-4440.2015.04.030
文献标志码:
A
摘要:
为验证葡萄β-1,3-葡聚糖酶基因在葡萄抗病防御机制中的作用，本研究以金星无核葡萄叶片为试验材料，采用RT-PCR技术克隆得到1个长度为1 244 bp的葡萄β-1,3-葡聚糖酶基因。采用生物信息学方法对其开放读码框和理化性质进行分析，结果显示，该基因包含1个长度为1 083 bp的完整开放阅读框，编码的蛋白质含360个氨基酸，分子量为89 440，理论等电点为4.83。系统进化分析结果显示，β-1,3-葡聚糖酶基因所编码的氨基酸序列与桃、豌豆和水稻的同源性分别为62.1%、60.8%和33.1%。定量PCR分析结果显示，在霜霉病菌侵染后的72 h内均可诱导该基因的表达，且表达量均高于对照，表明β-1, 3-葡聚糖酶基因在葡萄应答霜霉病菌侵染过程中可能发挥一定作用。
Abstract:
To identify the role of beta-1,3-glucanase gene in grape disease resistance, a beta-1,3-glucanase gene fragment with a length of 1 244 bp was cloned from grapevine Venus seedless leaves by RT-PCR method. Bioinformatics analysis revealed that the gene contained a complete open reading frame of1 083 bp in length encoding 360 amino acids. The molecular weight of deduced protein was 89 440, and the theoretical isoelectric point was 4.83. Phylogenetic analysis showed that amino acid sequence encoded by beta-1,3-glucanase gene shared 62.1%, 60.8% and 33.1% similarities with those in Prunus persica, Pisum sativum and Oryza sativa. Quantitative RT-PCR analysis revealed that the expressions of beta-1,3-glucanase gene was up-regulated within 72 h after the inoculation of Plasmopara viticola, and the level were much higher than that of control. It was suggested that beta-1,3-glucanase gene might play a certain role in response to grapevine downy mildew infection.

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备注/Memo

备注/Memo:
收稿日期：2015-01-28 基金项目：江苏省农业科技自主创新基金项目[CX(12)2013] 作者简介：王西成(1982-)，男，安徽亳州人，博士，助理研究员，主要从事葡萄种质资源收集、评价与利用以及葡萄分子生物学研究。(Tel)18652905096;(E-mail) wxcown@163.com 通讯作者：吴伟民, (E-mail) 5wm@163.com
更新日期/Last Update: 2015-08-31