中国小麦花叶病毒(CWMV)侵染条件下小麦内参基因的选择

doi:10.3969/j.issn.1004-1524.2018.07.11

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (7): 1182-1187.DOI: 10.3969/j.issn.1004-1524.2018.07.11

中国小麦花叶病毒(CWMV)侵染条件下小麦内参基因的选择

吴讷^{1, 2}, 陈炫^{2, 3}, 姜瑶瑶^{2, 3}, 张天烨^{2, 3}, 羊健², 朱统泉⁴, 张恒木^{2, *}, 陈剑平^{1, 2, *}

1.浙江农林大学农业与食品科学学院,浙江杭州 311300;
2.浙江省农业科学院病毒学与生物技术研究所,浙江杭州310021;
3.浙江农林大学林业与生物技术学院,浙江杭州 311300;
4.驻马店农业科学院,河南驻马店 463000

收稿日期:2017-09-13 出版日期:2018-07-20 发布日期:2018-08-02
通讯作者: 张恒木,E-mail: zhhengmu@tsinghua.org.cn;陈剑平,E-mail: jpchen2001@126.com
作者简介:吴讷(1992—),女,浙江绍兴人,硕士研究生,主要从事植物病理学研究。E-mail: wune0330@126.com
基金资助:
国家自然科学基金(3150160); 国家农业产业体系(CARS-3-1); 国家小麦转基因专项(2016ZX08002001)

Selection of reference genes in wheat infected by Chinese wheat mosaic virus (CWMV)

WU Ne^{1, 2}, CHEN Xuan^{2, 3}, JIANG Yaoyao^{2, 3}, ZHANG Tianye^{2, 3}, YANG Jian², ZHU Tongquan⁴, ZHANG Hengmu^{2, *}, CHEN Jianping^{1, 2, *}

1. Collge of Agriculture and Food Science, Zhejiang A&F University, Hangzhou 311300, China;
2. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
3. College of Forestry and Biotechnology, Zhejiang A&F University, Hangzhou 311300, China;
4. Zhumadian Academy of Agriculture Sciences, Zhumadian 463000, China

Received:2017-09-13 Online:2018-07-20 Published:2018-08-02

摘要/Abstract

摘要： 实时定量PCR(qRT-PCR)是分析功能基因表达水平的常用方法之一,qRT-PCR数据统计分析离不开合适内参基因的选择。为了选择中国小麦花叶病毒(Chinese wheat mosaic virus, CWMV)侵染条件下的小麦内参基因,本实验通过PCR扩增效率和扩增特异性分析,从10个持家基因中选择8个候选基因,然后以接种CWMV的小麦样品为材料,利用qRT-PCR技术进一步检测上述8个基因在CWMV侵染条件下小麦样品中的时空表达特性。基于这些数据,通过geNorm、NormFinder程序分析,结果表明,2个小麦基因(即26S和CDC)在CWMV侵染前后以及不同组织中表达最为稳定,26S和CDC组合可选作CWMV与小麦互作过程中功能基因表达分析的内参基因。

关键词: 内参基因, 小麦, 中国小麦花叶病毒, qRT-PCR

Abstract: Quantitative PCR (qRT-PCR) has been widely applied to the expression analysis of functional genes. The selection of appropriate reference genes is required for the statistical analysis of qRT-PCR data. To select the suitable reference genes in wheat infected by Chinese wheat mosaic virus(CWMV), 10 housekeeping genes were screened by calculations of PCR efficiencies and melting curve analysis, suggesting that 8 out of 10 genes could be used as candidate reference genes. And then their temporal and spatial expression patterns were investigated by qRT-PCR in the different tissues and at the different stages of wheat inoculated with CWMV. These quantitative data were further evaluated by the computer programs geNorm and NormFinder in this study. The analysis results consistently revealed that two genes, 26S and CDC, were stably expressed in all tested tissues and CWMV-infected wheat samples, indicating that the combination of 26S and CDC could be selected as reference genes for expression analysis of functional gene during the interaction between CWMV and wheat.

Key words: reference gene, wheat, Chinese wheat mosaic virus, qRT-PCR

中图分类号:

S435.121

吴讷, 陈炫, 姜瑶瑶, 张天烨, 羊健, 朱统泉, 张恒木, 陈剑平. 中国小麦花叶病毒(CWMV)侵染条件下小麦内参基因的选择[J]. 浙江农业学报, 2018, 30(7): 1182-1187.

WU Ne, CHEN Xuan, JIANG Yaoyao, ZHANG Tianye, YANG Jian, ZHU Tongquan, ZHANG Hengmu, CHEN Jianping. Selection of reference genes in wheat infected by Chinese wheat mosaic virus (CWMV)[J]. , 2018, 30(7): 1182-1187.

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中国小麦花叶病毒(CWMV)侵染条件下小麦内参基因的选择

Selection of reference genes in wheat infected by Chinese wheat mosaic virus (CWMV)

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