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茎点枯病菌诱导下芝麻内参基因的筛选
引用本文:刘莉铭,刘红彦,田保明.茎点枯病菌诱导下芝麻内参基因的筛选[J].作物学报,2012,38(3):471-478.
作者姓名:刘莉铭  刘红彦  田保明
作者单位: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
基金项目:国家现代农业产业技术体系建设项目(CARS-15-1-05)资助
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
收稿时间:2011-09-19

Selection of Reference Genes from Sesame Infected by Macrophomina phaseo-lina
LIU Li-Ming,LIU Hong-Yan,and TIAN Bao-Ming.Selection of Reference Genes from Sesame Infected by Macrophomina phaseo-lina[J].Acta Agronomica Sinica,2012,38(3):471-478.
Authors:LIU Li-Ming  LIU Hong-Yan  and TIAN Bao-Ming
Institution: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
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, and qRT-PCR was an impor-tant 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.
Keywords:Sesame  Macrophomina phaseolina  qRT-PCR  Reference gene
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