盐胁迫下蚕豆不同组织实时荧光定量PCR内参基因的筛选  

Selection of reference genes in the different tissues of faba bean under the salt stress via real-time quantitative PCR

作　　者：樊有存 张红岩[1,2] 韩芊 杨旭升 武学霞 刘玉皎 FAN Youcun;ZHANG Hongyan;HAN Qian;YANG Xusheng;WU Xuexia;LIU Yujiao(Qinghai University,Xining 810016,China;Academy of Agricultural and Forestry,Qinghai University,Xining 810016,China;State Key Laboratory of Plateau Ecology and Agriculture,Qinghai University,Xining 810016,China;College of Eco-Environmental Engineering,Qinghai University,Xining 810016,China)

机构地区：[1]青海大学,青海西宁810016 [2]青海大学农林科学院,青海西宁810016 [3]省部共建三江源生态与高原农牧业国家重点实验室,青海大学,青海西宁810016 [4]青海大学生态环境工程学院,青海西宁810016

出　　处：《青海大学学报》2021年第1期16-23,共8页Journal of Qinghai University

基　　金：青海省科学技术厅自然科学基金项目(2018-ZJ-940Q);省部共建三江源生态与高原农牧业国家重点实验室自主课题项目(2017-ZZ-12,2016-ZZ-04);国家食用豆产业技术体系建设专项(CARS-08)。

摘　　要：为筛选出蚕豆盐胁迫下不同组织中表达稳定的内参基因,本研究以不同浓度氯化钠胁迫下的蚕豆叶片、茎、根为材料,利用实时荧光定量PCR技术检测ELF1A、ACT2、GAPA、18S rRNA、TUA和CYP2六个候选内参基因的表达情况;通过Ct值分析、及GeNorm、NormFinder、BestKeeper软件综合分析候选内参基因表达稳定性。结果表明:在不同浓度氯化钠处理下,蚕豆叶片中候选内参基因表达稳定性由高到低排序为ACT2>18S rRNA>CYP2>ELF1A>GAPA>TUA;蚕豆茎中候选内参基因表达稳定性由高到低排序为CYP2>ACT2>ELF1A>GAPA>18S rRNA>TUA;蚕豆根中候选内参基因表达稳定性由高到低排序为CYP2>ELF1A>18S rRNA>ACT2>TUA>GAPA。本研究确定的蚕豆在盐胁迫下不同组织中适宜的内参基因,可为后续研究与盐胁迫相关功能基因提供理论依据。In order to select the stable reference genes in the different tissues of faba bean under the salt stress,the expression of these six candidate reference genes including ELF1A,ACT2,GAPA,18S rRNA,TUA and CYP2 is detected in this paper via real-time quantitative PCR in leaves,stems and roots of faba bean under the different concentrations of NaCl stress;the expression stability of candidate reference genes is analyzed by Ct value analysis and three softwares including GeNorm,NormFinder and BestKeeper.The results show that under the different concentrations of NaCl,the order of expression stability of candidate reference genes in the leaves is ACT2>18S rRNA>CYP2>ELF1A>GAPA>TUA;the order of expression stability of candidate reference genes in the stems is CYP2>ACT2>ELF1A>GAPA>18S rRNA>TUA;the order of expression stability of candidate reference genes in the roots is CYP2>ELF1A>18S rRNA>ACT2>TUA>GAPA.The suitable reference genes in the different tissues of faba bean under the salt stress can provide a theoretical basis for further research on the functional genes of salt stress.

关 键 词：蚕豆 盐胁迫 实时荧光定量PCR 内参基因 组织 

分 类 号：Q786[生物学—分子生物学] S529[农业科学—作物学]