阳桃(Averrhoa carambola)实时荧光定量PCR内参基因的筛选与验证  

Selection and Validation of Reference Genes of Averrhoa carambola by Quantitative Real-time PCR

作　　者：李笑平 朱雅婷 赵亚梅 陈蕾 任惠[2] 吴沙沙[1] 翟俊文[1] Li Xiaoping;Zhu Yating;Zhao Yamei;Chen Lei;Ren Hui;Wu Shasha;Zhai Junwen(Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization,College of Landscape Architecture,Fujian Agriculture and Forestry University,Fuzhou,350002;Horticultural Research Institute,Guangxi Academy of Agricultural Sciences,Nanning,530007)

机构地区：[1]福建农林大学园林学院,兰科植物保护与利用国家林业和草原局重点实验室,福州350002 [2]广西农业科学院园艺研究所,南宁530007

出　　处：《分子植物育种》2024年第3期789-799,共11页Molecular Plant Breeding

基　　金：福建农林大学艺术学院园林学院博士生导师培育项目(YSYL-bdpy-2);广西科技重大专项(桂科AA17204-045-4);广西农业科学院基本科研业务专项(桂农科2021YT046)共同资助。

摘　　要：阳桃(Averrhoa carambola)具有很高的营养、药用及观赏价值,但功能基因组方面的研究基础较为薄弱。为使阳桃相关功能基因表达分析的定量实验结果更加准确,需要选择稳定性好的内参基因。然而目前还没有关于阳桃内参基因筛选的报道。本研究基于阳桃基因组及转录组数据,选取了α-TUB、β-TUB、Actin、UBQ、UBC4、EF1α和H2B共7个管家基因作为候选内参基因;利用实时荧光定量PCR(RT-qPCR)技术检测各候选内参基因在茎、叶、果、花序芽、叶芽及不同时期花序中的表达情况。使用软件geNorm、Normfinder和BestKeeper综合评估了候选内参基因的稳定性。结果表明,α-TUB和β-TUB在阳桃不同器官中稳定性最好,β-TUB和UBC4在阳桃不同时期花序中稳定性最好。此外,通过分析成花基因AP1(APETALA1)和开花抑制因子TFL1(TERMINAL FLOWER 1)的表达模式,进一步验证了内参基因筛选结果的可靠性。本研究筛选得到的稳定内参基因为后期开展阳桃相关功能基因表达分析的研究提供了基础。The starfruit(Averrhoa carambola)has high nutritional,medicinal and ornamental value,but the basic research on functional genomics is still lacking.To carry out research on the expression analysis of related functional genes in starfruit,it is necessary to select suitable reference genes to reduce the deviation of experimental results.However,there is no report on the screening of reference genes of starfruit.In this study,seven housekeeping genes,α-TUB,β-TUB,Actin,UBQ,UBC4,EF1α and H2B,were selected as candidate reference genes based on the starfruit genome and transcriptome data of starfruit leaves,inflorescence buds and leaf buds.The expression of seven candidate reference genes were assessed by quantitative real-time PCR in different tissues such as mature leaves,stems,fruits,inflorescence buds,leaf buds,and inflorescence of different stages of starfruit,and the stability of them were analyzed by geNorm,NormFinder,and BestKeeper programs.The results showed that α-TUB and β-TUB had the best stability in different organs of starfruit,β-TUB and UBC4 had the best stability in inflorescence of different developmental stages of starfruit.In addition,by analyzing the expression patterns of the flowering gene AP1(APETALA1)and the flowering inhibitor TFL1(TERMINAL FLOWER 1),the reliability of the screening results of reference genes was further verified.The stable reference genes screened in this study laid the foundation for future research on the related functional gene expression of starfruit.

关 键 词：阳桃 内参基因 实时荧光定量PCR 表达稳定性 

分 类 号：S667.9[农业科学—果树学]