草地早熟禾荧光定量PCR分析中内参基因的筛选

发布时间：2018-09-13 16:44

【摘要】：为筛选出草地早熟禾实时荧光定量中的最适内参基因,利用实时荧光定量PCR技术,分析检测6个传统内参基因ACT、GAPDH、UBQ、EF-1α、18s rRNA和β-tublin在草地早熟禾不同组织器官、叶片不同发育期、非生物胁迫和不同激素诱导下mRNA的表达差异情况。利用geNorm、NormFinder和BestKeeper软件综合评价6个候选内参基因的表达稳定性。结果表明,在不同组织、叶片不同发育期、激素诱导和非生物胁迫下,候选内参基因的表达稳定性存在差异。GAPDH在草地早熟禾不同组织器官中表达最为稳定;EF-1α在非生物胁迫下表达最为稳定;不同激素下首选β-tublin;在叶片不同发育期ACT表达最为稳定。综上所述,通过筛选出不同条件下适宜的内参基因不仅有助于提高草地早熟禾基因表达分析的准确性,也为早熟禾属植物其他内参基因的开发提供了理论参考。
[Abstract]:In order to screen the optimal internal reference gene in the real-time fluorescence quantitative analysis of Kentucky bluegrass, six traditional internal reference genes, ACT,GAPDH,UBQ,EF-1 伪 ~ (18 s) rRNA and 尾 -tublin, were detected by real-time fluorescence quantitative PCR technique in different tissues and organs and different developmental stages of leaves of Kentucky bluegrass. The difference of mRNA expression under abiotic stress and different hormone induction. The expression stability of 6 candidate internal reference genes was evaluated by geNorm,NormFinder and BestKeeper software. The results showed that in different tissues, leaves at different developmental stages, hormone induction and abiotic stress, There were differences in the expression stability of candidate endogenous reference genes. The expression of EF-1 伪 was the most stable in different tissues and organs of Kentucky bluegrass. The expression of EF-1 伪 was the most stable under abiotic stress; 尾 -tublinin was the first choice under different hormones; and the ACT expression was the most stable in different development stages of leaves. To sum up, screening suitable internal reference genes under different conditions can not only improve the accuracy of gene expression analysis of Kentucky bluegrass, but also provide a theoretical reference for the development of other internal reference genes in POA plants.
【作者单位】：北京林业大学林学院草坪研究所;
【基金】：国家自然基金项目(31302016) 深圳市科技计划项目(JSGG20160229155434792)资助
【分类号】：S688.4

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