芒根组织在不同非生物胁迫下最适内参基因的筛选  

Optimum Reference Gene Selection in Miscanthus Sinensis Root Tissue with Various Abiotic Stress

作　　者：黄逸之 唐铭余 钟旻依 王芮嘉 陈嘉怡 叶云天 张新全[1] 聂刚[1] HUANG Yizhi;TANG Mingyu;ZHONG Minyi;WANG Ruijia;CHEN Jiayi;YE Yuntian;ZHANG Xinquan;NIE Gang(College of Animal Science and Technology,Sichuan Agricultural University,Chengdu 611130,China;College of Horticulture,Sichuan Agricultural University,Chengdu 611130,China)

机构地区：[1]四川农业大学动物科技学院,成都611130 [2]四川农业大学园艺学院,成都611130

出　　处：《四川农业大学学报》2020年第6期699-707,共9页Journal of Sichuan Agricultural University

基　　金：国家现代牧草产业技术体系(CARS-34);国家自然科学基金(31801432);教育部春晖计划(20191383-101)。

摘　　要：【目的】筛选不同非生物胁迫下芒根组织中稳定表达的内参基因,并为调控芒根组织中基因表达提供理论依据。【方法】以芒(Miscanthus sinensis)根组织为试验材料,通过实时荧光定量PCR技术检测12个候选内参基因在不同非生物胁迫环境(干旱、盐、镉、铬和砷)下的表达情况,并运用geNorm、BestKeeper、NormFinder和RefFinder软件分析它们在芒根组织中的稳定性。【结果】在不同的非生物环境胁迫条件下,最稳定的内参基因各不相同。12个内参基因表达稳定性由高到低排序为:Unigene26576,Unigene33024,Sb02g041180,eIF4A,ACTIN,GAPDH,Unigene33312,Sb09g019750,PP2A,SAMDC,18S rRNA,SAMS。其中在干旱胁迫条件下,Sb02g041180和ACTIN基因最稳定;在盐胁迫和砷胁迫处理下最稳定的内参基因分别是Sb09g019750和Unigene26576、Sb09g019750和PP2A;镉和铬胁迫下最稳定的内参基因分别为Unigene26576和PP2A、Sb09g019750和Unigene33312。【结论】Unigene26576和Unigene33024最适合作为研究芒根组织非生物胁迫的内参基因,而18S r RNA和SAMS最不适合作为内参基因。鉴定了候选基因在芒根组织中的稳定性,对进一步研究芒非生物胁迫下基因表达奠定了基础。【Objective】Screening of stable reference genes in Miscanthus sinensis root tissues under various abiotic stress provides a theoretical basis for the research on the regulation of gene expression in the roots of M.sinensis.【Method】The root tissue of M.sinensis was used as the material,and qRT-PCR technique was used to detect the expression of 12 candidate reference genes under different abiotic stresses(drought,salt,cadmium,chromium and arsenic),and their stability was analyzed by geNorm,BestKeeper,NormFinder and RefFinder software.【Result】The results showed that the most stable ref-erence genes were different under various abiotic stress.The stability of the expression of 12 reference genes was ranked from high to low as follows:Unigene26576,Unigene33024,Sb02g041180,eIF4A,ACTIN,GAPDH,Unigene33312,Sb09g019750,PP2A,SAMDC,18S rRNA,SAMS.Among Sb02g-041180 and ACTIN were found to be the two best reference genes under drought stress;Sb09g019750 and Unigene26576,Sb09g019750 and PP2A were the most stably expressed genes under salt stress and as stress;Unigene26576 and PP2A,Sb09g019750 and Unigene33312 were found to be the most suitable reference genes under Cd stress and Cr stress.【Conclusion】Unigene26576 and Unigene33024 were most suitable for the study of abiotic stress in M.sinensis root tissues,while 18S rRNA and SAMS are the least suitable reference genes.The stability of the candidate reference genes in the root tissue of the M.sinensis were identified,and has laid the foundation for further research on the expression of various genes under the abiotic stress of M.sinensis.

关 键 词：芒 实时荧光定量PCR 内参基因 非生物胁迫 根组织 

分 类 号：Q943.2[生物学—植物学]