Journal of Nuclear Agricultural Sciences ›› 2019, Vol. 33 ›› Issue (3): 473-481.DOI: 10.11869/j.issn.100-8551.2019.03.0473

• Induced Mutations for Plant Breeding·Agricultural Biotechnology • Previous Articles     Next Articles

Screening and Evaluation of Reference Genes for RT-qPCR in Wax Gourd (Benincasa hispida Cogn.)

YE Xinru, ZHU Haisheng*, LIN Hui, LIU Jiangting, WANG Bin, CHEN Mindong, WEN Qingfang*   

  1. Fujian Engineering Research Center for Vegetables/Vegetable Research Center, Fujian Academy of Agricultural Sciences/Crops Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013
  • Received:2018-07-31 Revised:2018-10-13 Online:2019-03-10 Published:2019-01-23

冬瓜实时荧光定量PCR内参基因的筛选与评价

叶新如, 朱海生*, 林珲, 刘建汀, 王彬, 陈敏氡, 温庆放*   

  1. 福建省农业科学院作物研究所/福建省农业科学院蔬菜研究中心/福建省蔬菜工程技术研究中心,福建 福州 350013
  • 通讯作者: 朱海生,男,研究员,主要从事分子生物学研究。E-mail:541585978@qq.com;温庆放,男,研究员,主要从事蔬菜栽培研究。E-mail:fjvrc@163.com。同为通讯作者。
  • 作者简介:叶新如,女,研究实习员,主要从事园艺植物分子生物学研究。E-mail:731122298@qq.com
  • 基金资助:
    福建省属公益类科研院所基本科研专项(2018R1026-2), 福建省农业科学院蔬菜科技创新团队(STI2017-1-2),福建省农业科学院“青年科技英才百人计划”(YC2017-5),中央引导地方科技发展专项(2018L3005)

Abstract: The present study aimed at selecting the stable reference genes to ensure the reliability and accuracy in gene expression analysis of wax gourd (Benincasa hispida Cogn.). By RT-PCR technology, combined with GeNorm, NormFinder, BestKeeper and ReFinder analysis, we investigated the expression stability of seven candidate reference genes(28SrRNAUBQRP ⅡGAPDHEF-1αUBC21、TUA). The results showed that, in different tissues and with different stress treatment,the expression abundance and stability of these seven reference genes were different. The expression stability of EF-1α was the best under low temperature stress., UBQ and TUA showed the best expression stability under high temperature and drought stress, EF-1α was the best reference gene for different tissues. The results of this study can be used as a reference for the selection of internal reference genes in wax gourd.

Key words: wax gourd, reference gene, abiotic stress

摘要: 为筛选适合冬瓜稳定表达的内参基因,以黑皮冬瓜低温、高温、干旱胁迫处理不同时间的叶片和正常处理不同组织为试验材料,利用RT-qPCR技术,结合GeNorm、NormFinder和BestKeeper软件评价7个候选内参基因(28SrRNAUBQRP ⅡGAPDHEF-1αUBC21和TUA)稳定性。结果表明,7个候选内参基因在不同组织和不同胁迫处理下表达丰度及稳定性存在差异; EF-1α在低温胁迫处理下表达稳定性最好;UBQTUA在高温和干旱胁迫处理下表达稳定性最好; EF-1α 在不同组织中表达稳定性最好。本研究结果为冬瓜内参基因的选择提供了一定的理论参考。

关键词: 冬瓜, 内参基因, 非生物胁迫