少花蒺藜草荧光定量PCR内参基因筛选与验证
Screening and Verification of Internal Reference Gene of Cenchrus pauciflorus Benth.by Real-time PCR
徐礼莹 1田迅 1百灵 1吴佳丽 1张福霖 1黄菊 1马文静 1吕广超 1马红悦 1陈宇杰1
作者信息
- 1. 内蒙古民族大学生命科学与食品学院 内蒙古,通辽 028043
- 折叠
摘要
少花蒺藜草(Cenchrus pauciflorus Benth.)为禾本科蒺藜草属,是入侵我国北方地区的一年生恶性杂草,少花蒺藜草的入侵态势逐年扩增,因此对于少花蒺藜草的基因研究,以及在分子水平对于防治少花蒺藜草入侵的研究显得尤为重要.本研究以少花蒺藜草转录组数据为基础,候选了6个内参基因,荧光定量PCR检测候选内参基因的表达,通过geNorm、NormFinder、BestKeeper、比较△Ct法和RefFinder五种评价方式,综合评估筛选了最佳内参基因.结果表明,6个候选内参基因的综合排名为TCTP1>RH37>ACX4>TCTP2>GAPDH>TBP,TCTP1基因在少花蒺藜草不同时期不同组织样本中表达最高最稳定,最不稳定表达的是基因TBP,TCTP1为综合评估下少花蒺藜草的最佳内参基因,因此TCTP1可作为内参基因用于少花蒺藜草荧光定量PCR检测研究中.本研究为少花蒺藜草后续的基因研究以及在分子水平上为少花蒺藜草生物防治提供一定的理论基础.
Abstract
Cenchrus pauciflorus Benth.is a genus of Tribulus terrestris in the family Poaceae,which is an annual malignant weed that invades northern China,and the invasion trend of Cenchrus pauciflorus Benth.is expanding year by year,so it is particularly important to study the gene of Cenchrus pauciflorus Benth.and the prevention and control of Cenchrus pauciflorus Benth.at the molecular level.In this study,based on the transcriptome data of Cenchrus pauciflorus Benth.,6 Internal reference genes were selected,the expression of Internal reference genes was detected by Real-time PCR,and the best reference genes were comprehensively evaluated and screened through five evaluation methods:geNorm,NormFinder,BestKeeper,comparative △Ct method and RefFinder.The results showed that the comprehensive ranking of the six candidate reference genes was TCTP1>RH37>ACX4>TCTP2>GAPDH>TBP,TCTP1 gene was the highest and most stable expression in different tissue samples of Cenchrus pauciflorus Benth.at different stages,and the most unstable expression was the gene TBP,TCTP1 was the best internal reference gene of Cenchrus pauciflorus Benth.under comprehensive evaluation,so TCTP1 could be used as an internal reference gene for the detection of Cenchrus pauciflorus Benth.Real-time PCR.This study provides a theoretical basis for the follow-up genetic study of Cenchrus pauciflorus Benth.and the biological control of Cenchrus pauciflorus Benth.at the molecular level.
关键词
少花蒺藜草/荧光定量PCR/内参基因Key words
Cenchrus pauciflorus Benth./real-time PCR/internal reference genes引用本文复制引用
基金项目
国家自然科学基金(32260683)
国家自然科学基金(32360267)
内蒙古自治区硕士研究生科学创新项目(S20231211Z)
内蒙古自治区大学生创新创业训练计划(S202310136027)
出版年
2024
国家科技期刊平台
NSTL
万方数据