低温弱光胁迫下茄子幼苗实时荧光定量PCR内参基因的筛选与评价  

Screening and Evaluation of Reference Genes in Eggplant Seedlings by Real-time Fluorescence Quantitative PCR Under Low Temperature and Weak Light Stress

作　　者：林珲[1] 裘波音 朱海生[1] 温庆放[1] LIN Hui;QIU Bo-yin;ZHU Hai-sheng;WEN Qing-fang(Fujian Key Laboratory of Vegetable Genetics and Breeding/Fujian Engineering Research Center for Vegetables/Crops Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350013,China)

机构地区：[1]福建省农业科学院作物研究所/福建省蔬菜遗传育种重点实验室/福建省蔬菜工程技术研究中心,福建福州350013

出　　处：《福建农业科技》2024年第8期1-15,共15页Fujian Agricultural Science and Technology

基　　金：福建省科技计划省属公益类项目(2021R1031003);国家大宗蔬菜产业技术体系福州综合试验站项目(CARS-23-G-53);福建省种业创新与产业化工程项目(zycxny2021009);福建省农业科学院科技创新平台专项(CXPT202001);福建省农业科学院蔬菜遗传育种科技创新团队(CXTD2021038)。

摘　　要：为了探索茄子Solanum melongena L.的基因表达模式,利用qRT-PCR方法分析了茄子ACT、EF1α、TUA、TUB、GAPDH、eIF、UBQ、UBI3、PP2A、CYP、RPL28、L25、SAND、TBP、DNAJ、APRT、EXP、CAC、HSP20和CysPro 20个候选内参基因mRNA的表达差异情况,并运用GeNorm、NormFinder和BestKeeper 3种计算方法评价茄子20个候选内参基因在低温、弱光、低温弱光叶片样品中的表达稳定性。结果表明:茄子20个候选内参基因荧光定量引物扩增效率(E)数值在96.8%~117.6%,相关系数(R^(2))介于0.9688~0.9999,扩增效率良好,扩增反应具有高度的专一性,引物能特异性扩增,特异性好,均为单峰,样品间扩增曲线重复性强,可用于qRT-PCR扩增。茄子20个候选内参基因C_(T)值表达丰度分析,发现茄子20个候选内参基因平均CT值在19.76~31.20。3种计算方法的综合评价分析结果显示,TBP基因在所有样本中为最稳定的内参基因。研究可为茄子低温、弱光和低温弱光胁迫下基因特异性表达研究提供了合适的内参基因。In order to explore the gene expression pattern of Solanum melongena L.,the qRT-PCR method was used to analyze the expression differences of mRNA in the 20 candidate reference genes of eggplant,including ACT,EF1α,TUA,TUB,GAPDH,eIF,UBQ,UBI3,PP2A,CYP,RPL28,L25,SAND,TBP,DNAJ,APRT,EXP,CAC,HSP20 and CysPro.The three calculation methods,including GeNorm,NormFinder and BestKeeper,were used to evaluate the expression stability of 20 candidate reference genes in eggplant samples at low temperature,weak light and low temperature and weak light stress.The results showed that the fluorescence quantitative primer amplification efficiency(E)values of 20 candidate reference genes in eggplant ranged from 96.8%to 117.6%,and the correlation coefficient(R^(2))was between 0.9688−0.9999,indicating that the amplification efficiency was good,and the amplified reaction was highly specific.The primers could be specifically amplified,with good specificity,all of which were unimodal.The amplification curves among the samples had strong repeatability,which could be used for qRT-PCR amplification.By analyzing the expression abundance of C_(T)values of 20 candidate reference genes in eggplant,it was found that the average C_(T)values of 20 candidate reference genes in eggplant ranged from 19.76 to 31.20.The comprehensive evaluation results of the three calculation methods showed that the TBP gene was the most stable reference gene in all the samples.The study could provide a suitable reference gene for the study of gene-specific expression in eggplant under low temperature stress,weak light stress and low temperature and weak light stress.

关 键 词：茄子 内参基因 实时荧光定量PCR 标准化 基因表达 

分 类 号：S641.1[农业科学—蔬菜学]