[1]刘婷,杨若巍,王学敏,等.抗性候选基因GmMIOX4的表达分析、克隆及过表达载体的构建[J].大豆科学,2016,35(05):730-735.[doi:10.11861/j.issn.1000-9841.2016.05.0730]
　LIU Ting,YANG Ruo-wei,WANG Xue-min,et al.Resistance Candidate Genes GmMIOX4 Gene Expression Analysis,Cloning and Over Expression Vector Constructing[J].Soybean Science,2016,35(05):730-735.[doi:10.11861/j.issn.1000-9841.2016.05.0730]
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抗性候选基因GmMIOX4的表达分析、克隆及过表达载体的构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年05期 页码: 730-735 栏目: 出版日期: 2016-09-20
Title:
Resistance Candidate Genes GmMIOX4 Gene Expression Analysis,Cloning and Over Expression Vector Constructing
作者:
刘婷1; 杨若巍1; 王学敏1; 于佰双2; 王惠1; 段玉玺3
（1.沈阳农业大学 生物科学技术学院，辽宁 沈阳 110866； 2.黑龙江省农业科学院 大豆研究所，黑龙江 哈尔滨 150086； 3.沈阳农业大学 植物保护学院，辽宁 沈阳 110866）
Author(s):
LIU Ting1; YANG Ruo-wei1; WANG Xue-min1; YU Bai-shuang2; WANG Hui1; DUAN Yu-xi3
（1.Shenyang Agricultural University, College of Bioscience and Biotechnology, Shenyang 110866, China; 2.Soybean Research Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China; 3.Shenyang Agricultural University, College of Plant Protection, Shenyang 110866, China）
关键词:
GmMIOX4基因; 大豆胞囊线虫; 实时荧光定量PCR; 克隆; 过表达载体
Keywords:
:GmMIOX4 gene;  Soybean cyst nematode;  Real-time quantitative PCR;  Clone;  Over expression vector
DOI:
10.11861/j.issn.1000-9841.2016.05.0730
文献标志码:
A
摘要:
将两个不同抗性大豆品种灰皮支黑豆和辽豆15人工接种大豆胞囊线虫3号生理小种（SCN3），利用实时荧光定量PCR技术，对SCN3侵染后的早期和线虫完成一个整个生活史时的GmMIOX4基因相对表达量变化进行分析。结果表明：在SCN3侵染下，辽豆15根内的GmMIOX4基因在25 dpi时，相对表达量升高，而灰皮支黑豆根内的GmMIOX4基因在10 dpi时被诱导表达，相对表达量达到最高值，表明该基因在此时期发挥一定作用。线虫侵入大豆根内10 d是其通过合胞体吸取寄主营养从而维持正常生长发育的时期，在10 dpi该基因被诱导表达，使其合胞体结构发生改变，从而抑制线虫发育，最终导致死亡或使灰皮支黑豆根内成虫数量减少，起到抗病效果。随后以抗性品种灰皮支黑豆根系总RNA为模板，利用RT-PCR技术获得了939 bp的GmMIOX4基因的CDS序列，克隆得到质粒Pgem-MIOX4，再以得到的GmMIOX4序列为材料构建过表达载体pCAM-MIOX4，为进一步研究GmMIOX4基因功能奠定了基础。
Abstract:
Two kinds of different resistance soybean(Huipizhiheidou and Liaodou 15) were inoculated with SCN3 in the research.By real-time quantitative PCR method, GmMIOX4 gene expression changes were identified in early period and complete life cycle of SCN3.Real-time quantitative PCR results showed that: Under SCN3 infection, the relative expression of GmMIOX4 gene in roots of Liaodou 15 increased until 25 dpi after inoculation, the relative expression of GmMIOX4 gene in roots of Huipizhiheidou reached the highest level in 10 dpi, GmMIOX4 gene was induced and played an important role.Host’s nutrients was absorbed by SCN through syncytial to maintain normal growth and development period at 10 dpi, and this gene expression is induced, the gene can changes syncytial structure, inhibit nematode development, and thus played a disease-resistant effect. Total RNA of Huipizhi Heidou was used as template, 939 bp CDS of GmMIOX4 genezone was obtained by RT-PCR According to sequencing result, pGEM-MIOX4 over expression vector was cloned, it laid the foundation for further study of GmMIOX4 gene function.

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备注/Memo

基金项目：国家自然科学基金重点项目（31330063）。第一作者简介：刘婷（1990-），女，硕士，主要从事大豆胞囊线虫抗性基因相关研究。E-mail：360594542@qq.com。通讯作者：王惠（1972-），女，博士，副教授，主要从事大豆抗病、抗逆机制和分子生物学研究。E-mail：wanghuisyau@sina.com。段玉玺（1964-），男，教授，博导，主要从事植物线虫学研究。E-mail：duanyx6407@sina.com。

更新日期/Last Update: 2016-09-24