摘要
用CodeHop方法设计简并引物并克隆得到了GAPDH、β-actin、α-tubulin、UBQ5、RPL4、eIF4A和eEF1α7个看家基因的部分序列,将这7个基因和GenBank中已公布的18SrRNA、NADHD2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-tubulin3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。
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.
出处
《作物学报》
CAS
CSCD
北大核心
2012年第3期471-478,共8页
Acta Agronomica Sinica
基金
国家现代农业产业技术体系建设项目(CARS-15-1-05)资助
