藜科植物藜和灰綠藜實(shí)時(shí)熒光定量PCR內(nèi)參基因的選擇
本文選題:藜 + 灰綠藜 ; 參考:《廣西植物》2016年12期
【摘要】:選擇合適的內(nèi)參基因是實(shí)時(shí)熒光定量PCR(qRT-PCR)研究的關(guān)鍵,目前對(duì)藜科耐鹽(鹽生)植物脅迫相關(guān)基因的表達(dá)分析中所用內(nèi)參基因的報(bào)道較為有限。該研究利用Ge Norm、Norm Finder和Best Keeper 3個(gè)內(nèi)參基因分析軟件,對(duì)已選擇過的β-TUBULIN、β-ACTIN、GAPDH 3個(gè)常用候選內(nèi)參基因進(jìn)行了比較分析,篩選出在Na Cl和PEG脅迫下藜和灰綠藜中表達(dá)相對(duì)穩(wěn)定的內(nèi)參基因。結(jié)果表明:Ge Norm、Norm Finder內(nèi)參軟件分析在Na Cl和PEG脅迫下,GAPDH是藜和灰綠藜中均共同穩(wěn)定表達(dá)的內(nèi)參基因,同時(shí)在藜和灰綠藜中也有各自表達(dá)較穩(wěn)定的內(nèi)參基因,β-ACTIN在藜中穩(wěn)定表達(dá),β-TUBULIN則在灰綠藜中穩(wěn)定表達(dá)。對(duì)相同科不同種的植物內(nèi)參基因表達(dá)差異進(jìn)行比較,內(nèi)參基因在相同科中具有相同穩(wěn)定表達(dá)的內(nèi)參;對(duì)相同脅迫下兩種不同植物內(nèi)參基因表達(dá)穩(wěn)定性進(jìn)行分析,內(nèi)參基因的選擇需根據(jù)實(shí)際的實(shí)驗(yàn)材料和實(shí)驗(yàn)條件而定;3個(gè)分析軟件對(duì)以上3個(gè)常用內(nèi)參基因的分析結(jié)果,初步確定了在藜科植物藜和灰綠藜中相對(duì)穩(wěn)定的內(nèi)參基因,為藜和灰綠藜脅迫相關(guān)基因的定量表達(dá)分析提供了參考依據(jù)。
[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.
【作者單位】: 新疆大學(xué)生命科學(xué)與技術(shù)學(xué)院/新疆生物資源基因工程重點(diǎn)實(shí)驗(yàn)室;
【基金】:國(guó)家自然科學(xué)基金(31260037,31460043,31660068) 新疆自治區(qū)優(yōu)秀青年科技人才培養(yǎng)項(xiàng)目(2013721013)~~
【分類號(hào)】:Q943.2
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