出芽短梗霉菌株PA-2胁迫下藜qRT-PCR内参基因的筛选与验证  

Screening and Validation of qRT-PCR Reference Genes in Chenopodium album under the Stress of Aureobasidium pullulans PA-2

作　　者：刘晓芳 程亮 郭青云 LIU Xiaofang;CHENG Liang;GUO Qingyun(Institute of Plant Protection,Qinghai Academy of Agriculture and Forestry Science,Qinghai University/Key Laboratory of Integrated Management of Agricultural Pests in Qinghai Province/Xining Crop PestScientific Observation and Experimental Station,Ministry of Agriculture and Rural Affairs,Xining810016,China)

机构地区：[1]青海大学农林科学院植物保护研究所/青海省农业有害生物综合治理重点实验室/农业农村部西宁作物有害生物科学观测实验站,青海西宁810016

出　　处：《中国草地学报》2024年第2期48-56,共9页Chinese Journal of Grassland

基　　金：青海省自然科学基金面上项目(2022-ZJ-911);国家自然科学基金项目(31760539);青海省农业有害生物综合治理重点实验室项目(2022-ZJ-Y10)资助。

摘　　要：为筛选适合出芽短梗霉菌株PA-2胁迫下藜中稳定表达的内参基因,在转录组数据(NCBI登录号为:SRA:SRR12674257)分析的基础上,以PA-2不同时间胁迫的藜叶片为材料,分析藜中ACT-1、TUB-1、GAPDH、eEF1α、eIF3、UBQ、PTB、COX10、UPL6、TIP41L、RAN-B1和IDH-5等12个内参基因的表达水平,并利用ΔCt、GeNorm、NormFinder、BestKeeper及RefFinder分析候选内参基因的表达稳定性。结果表明,内参基因COX10是PA-2不同时间胁迫下藜中最稳定的内参基因,最优组合是COX10+eEF1α。通过分析藜ACC基因的相对表达水平,验证了内参基因的可靠性,为今后杂草藜胁迫响应基因的表达分析提供了参考基因和理论依据。In order to screen the stable expression of reference genes in Chenopodium album under the stress of Aureobasidium pullulans PA-2,based on the analysis of transcriptome data(NCBI access number:SRA:SRR12674257),the expression levels of 12 reference genes in Chenopodium album under different time stress of PA-2 were analyzed,including ACT-1,TUB-1,GAPDH,eEF1α,eIF3,UBQ,PTB,COX10,UPL6,TIP41L,RAN-B1 and IDH-5.The expression stability of candidate reference genes was analyzed byΔCt,GeNorm,NormFinder,BestKeeper and RefFinder.The results revealed that the reference gene COX10 was the most stable reference gene in Chenopodium album under different time stress of PA-2,and the optimal combination was COX10+eEF1α.The reliability of the reference genes was verified by ana⁃lyzing the relative expression levels of acetyl-CoA carboxylase gene(ACC)in C.album.The findings of the present study provide reference genes and theoretical basis for the expression analysis of stress response gene in C.album in the future.

关 键 词：出芽短梗霉菌株PA-2 QRT-PCR 藜 内参基因 表达稳定性 

分 类 号：S451[农业科学—植物保护]