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作物学报 ›› 2012, Vol. 38 ›› Issue (03): 471-478.doi: 10.3724/SP.J.1006.2012.00471

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

茎点枯病菌诱导下芝麻内参基因的筛选

刘莉铭1,2,3,刘红彦1,2,3,*,田保明1   

  1. 1 郑州大学生物工程系，河南郑州 450001；2 河南省农业科学院植物保护研究所，河南郑州 450002；3 河南省农作物病虫害防治重点实验室，河南郑州 450002
  • 收稿日期:2011-09-19 修回日期:2011-12-15 出版日期:2012-03-12 网络出版日期:2012-01-04
  • 通讯作者: 刘红彦, E-mail: liuhy1219@163.com, Tel: 0371-65730166
  • 基金资助:

    本研究由现代农业产业技术体系建设专项资金(CARS-15-1-05)资助。

Selection of Reference Genes from Sesame Infected by Macrophomina phaseolina

LIU Li-Ming1,2,3,LIU Hong-Yan1,2,3,*,TIAN Bao-Ming1   

  1. 1 Bioengineering Department, Zhengzhou University, Zhengzhou 450001, China; 2 Institute of Plant Protection, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; 3 Henan Key Laboratory for Control of Crop Diseases and Insect Pests, Zhengzhou 450002, China
  • Received:2011-09-19 Revised:2011-12-15 Published:2012-03-12 Published online:2012-01-04
  • Contact: 刘红彦, E-mail: liuhy1219@163.com, Tel: 0371-65730166

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1296

被引次数

15

摘要： 用CodeHop方法设计简并引物克隆得到了GAPDH、β-actin、α-tubulin、UBQ5、RPL4、eIF4A和eEF1α 7个看家基因的部分序列，将这7个基因和GenBank中已公布的18S rRNA、NADHD 2个基因共9个基因作为候选内参基因，利用实时荧光定量PCR技术，分析其在茎点枯病菌诱导下芝麻中的表达稳定性。经BestKeeper、NormFinder和GeNorm软件分析可知，UBQ5、eIF4A、α-tubulin 3个基因表达均较稳定，eEF1α变化最大。当使用多基因作为内参基因时，选择这3个最稳定的候选内参基因即可准确矫正定量结果。

关键词: 芝麻, 茎点枯病菌, qRT-PCR, 内参基因

Abstract: Stem rot caused by Macrophomina phaseolina (Tassi) Goid. is the most important disease in sesame, worldwide, the use of sesame cultivars resistant to stem rot provides an effective, economical, and environmentally friendly method to control the disease. The study on sesame defense response plays important roles in breeding resistant cultivars, andqRT-PCRwas an important method in studying plant defense response. In order to select the proper reference genes which can normalize expression of the target gene in sesame induced by Macrophomina phaseolina, nine genes 18S rRNA, NADHD, GAPDH, β-actin, α-tubulin, UBQ5, RPL4, eIF4A, and eEF1α, was analyzed by qRT-PCR. Among the nine genes, last seven were designed using CodeHop. BestKeeper, NormFinder and GeNorm analyses showed that UBQ5, eIF4A and α-tubulin were the top three optimum choices while eEF1α varied greatly. The analysis revealed that the three most stable genes UBQ5, eIF4A, α-tubulin were enough to obtain an accurate result when using multiple genes as control.

Key words: Sesame, Macrophomina phaseolina, qRT-PCR, Reference gene

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