藜科植物藜和灰绿藜实时荧光定量PCR内参基因的选择

发布时间：2018-05-02 13:34

本文选题：藜 + 灰绿藜　；参考：《广西植物》2016年12期

【摘要】：选择合适的内参基因是实时荧光定量PCR(qRT-PCR)研究的关键,目前对藜科耐盐(盐生)植物胁迫相关基因的表达分析中所用内参基因的报道较为有限。该研究利用Ge Norm、Norm Finder和Best Keeper 3个内参基因分析软件,对已选择过的β-TUBULIN、β-ACTIN、GAPDH 3个常用候选内参基因进行了比较分析,筛选出在Na Cl和PEG胁迫下藜和灰绿藜中表达相对稳定的内参基因。结果表明:Ge Norm、Norm Finder内参软件分析在Na Cl和PEG胁迫下,GAPDH是藜和灰绿藜中均共同稳定表达的内参基因,同时在藜和灰绿藜中也有各自表达较稳定的内参基因,β-ACTIN在藜中稳定表达,β-TUBULIN则在灰绿藜中稳定表达。对相同科不同种的植物内参基因表达差异进行比较,内参基因在相同科中具有相同稳定表达的内参;对相同胁迫下两种不同植物内参基因表达稳定性进行分析,内参基因的选择需根据实际的实验材料和实验条件而定。基于3个分析软件对以上3个常用内参基因的分析结果,初步确定了在藜科植物藜和灰绿藜中相对稳定的内参基因,为藜和灰绿藜胁迫相关基因的定量表达分析提供了参考依据。
[Abstract]:The selection of suitable internal reference genes is the key of real-time fluorescence quantitative PCRQ RT-PCR.At present, there are limited reports on the expression of genes related to salt tolerance (halophyte) stress in Chenopodiaceae. In this study, three common candidate genes of 尾 -TUBULIN, 尾 -ACTINGAPDH were compared and analyzed by using three internal reference gene analysis softwares GE Norm Finder and Best Keeper, and relatively stable internal reference genes were screened out under NaCl and PEG stress. The results showed that GAPDH was coexpressed stably in both Chenopodium and Atriplex under NaCl and PEG stress by using the software of 10 GE Norm Finder. 尾 -ACTIN and 尾 -TUBULIN were also expressed stably in Chenopodium and Atriplex, and 尾 -TUBULIN was also expressed stably in Chenopodium graminearum, 尾 -ACTIN and 尾 -TUBULIN were also expressed stably in Chenopodium hirsutum. The differences of gene expression in different plants of the same family and different species were compared, and the stability of gene expression in two different plants under the same stress was analyzed, the results showed that the internal ginseng gene had the same stable expression in the same family. The selection of internal reference genes depends on the actual experimental materials and experimental conditions. Based on the analysis results of the above three common internal reference genes, the relatively stable internal reference genes in Chenopodiaceae and Chenopodium graminis were preliminarily identified. It provides a reference for quantitative expression analysis of stress related genes in Chenopodium indica and Chenopodium graminis.
【作者单位】：新疆大学生命科学与技术学院/新疆生物资源基因工程重点实验室;
【基金】：国家自然科学基金(31260037,31460043,31660068) 新疆自治区优秀青年科技人才培养项目(2013721013)~~
【分类号】：Q943.2

【相似文献】