浙江农业学报 ›› 2019, Vol. 31 ›› Issue (8): 1312-1320.DOI: 10.3969/j.issn.1004-1524.2019.08.12

• 园艺科学 • 上一篇    下一篇

双孢蘑菇内参基因的筛选与矫正

赵建霞1,2, 沈颖越1, 冯伟林1, 金群力1, 宋婷婷1, 范丽军1, 蔡为明1,*   

  1. 1.浙江省农业科学院 园艺研究所,浙江 杭州 310021;
    2.浙江师范大学 化学与生命科学学院,浙江 金华 321004
  • 收稿日期:2019-05-26 出版日期:2019-08-25 发布日期:2019-08-30
  • 通讯作者: 蔡为明,E-mail: caiwm527@126.com
  • 作者简介:赵建霞(1990-),女,河南安阳人,硕士研究生,研究方向为食用菌遗传育种。E-mail: 1713205733@qq.com
  • 基金资助:
    国家现代农业技术体系(CARS-20); 浙江省重大科技专项(2016C02057); (原)农业部公益性行业(农业)科研专项(201503137); 浙江省农业重大技术协同推广计划(2019XTTGSYJ03)

Screening of internal reference gene of Agaricus bisporus

ZHAO Jianxia1,2, SHEN Yingyue1, FENG Weilin1, JIN Qunli1, SONG Tingting1, FAN Lijun1, CAI Weiming1,*   

  1. 1. Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
    2. College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China
  • Received:2019-05-26 Online:2019-08-25 Published:2019-08-30

摘要: 根据特定试验材料与条件选择RT-qPCR分析中合适的内参基因,对于准确校正目的基因的表达至关重要。该研究采用RT-qPCR技术对8个内参基因在5个发育时期、3种不同组织器官和4个不同温度处理下的表达情况进行检测,通过ΔCt法、GeNorm和NormFiner对单基因、双基因组合和三基因组合的表达差异情况进行稳定性分析。结果表明:双孢蘑菇不同发育时期、不同组织,以及不同温度处理菌丝中最佳内参基因分别为40s+actin+Eif5基因组合、Eif5+ubiquitin+PRL14基因组合和EF1+ubiquitin+RPL14基因组合。该研究结果可为双孢蘑菇发育和温度相关基因的定量表达分析提供参考。

关键词: RT-qPCR, 内参基因, 表达稳定性, 双孢蘑菇

Abstract: According to specific experimental materials and conditions, the selection of appropriate internal reference genes in RT-qPCR analysis was crucial for accurate correction of target genes expression. In present study, eight internal genes were used to analyze expression stability between five development stages, three different tissues or organs, four different temperature by GeNorm, NormFiner softwares and Δ Ct methed. The results showed that the optimal internal reference genes of Agaricus bisporus at different developmental stages, in different tissues and at different temperatures were 40s+actin+Eif5 combination, Eif5+ubiquitin+PRL14 combination and EF1+ubiquitin+RPL14 combination, respectively. This study had important practical value for the quantitative expression analysis of key genes related with developmet and temperatural response in Agaricus bisporus.

Key words: real time quantitative PCR, internal reference genes, expression stability, Agaricus bisporus

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