短命植物新疆小拟南芥高盐胁迫处理下内参基因的筛选

The Screening of Internal Reference Genes in Xinjiang Ephemeral Plants Arabidopsis pumila under High Salt Stress Conditions

新疆小拟南芥(Arabidopsis pumila)是早春短命植物,广泛分布于新疆极端环境,是研究生物和环境适应机制较好的模式材料.实时荧光定量PCR(qRT-PCR)是一种常用的可以快速获取基因表达的分析方法,而选择合适的内参基因是实时荧光定量PCR的前提.本研究从小拟南芥全长转录组数据库中选取了14个候选内参基因,利用qRT-PCR分析内参基因在250 mmol/L NaCl胁迫处理0、3、6、12、24和48 h共6个不同时间点样品中的基因表达,利用geNorm、Norm Finder和Bestkeeper三种软件进行数据分析.结果显示:在250 mmol/L NaCl胁迫处理下,较为稳定表达的内参基因是GAPDH和ACT1.本研究筛选出高盐胁迫处理下的稳定表达的GAPDH和ACT1基因作为内参基因,对后续验证基因的表达和揭示小拟南芥耐逆机制具有重要意义.

Arabidopsis pumila is an early springephemeral plant that is widely distributed in Xinjiang extreme environment.It is a good model material for studying biological and environmental adaptation mechanisms.Quantitativereal-time PCR（q RT-PCR）is a commonly used analytical method that can quickly obtain gene expression profiles.Selection of the appropriate internal reference gene is a prerequisite for q RT-PCR analysis.In this study,14candidate internal reference genes were selected from the Arabidopsis pumila full-length transcriptome database,and q RT-PCR was used to analyze the gene expression profiles of internal reference genes at six different time points（0,3,6,12,24 and 48 h）,time points treated with 250 mmol/L Na Cl.Three application tools,geNorm,Norm Finder,and Bestkeeper,were usedfor data analysis.The results showed that under high salt stress conditions,GAPDH and ACT1 were relatively stable expression of the reference genes.This study screened stable genes under high salt stress as an internal reference,which is important for subsequent verification of gene expression profiles and understanding the mechanism of salt tolerancein Arabidopsis pumila.