模式植物狗尾草响应不同干旱及盐胁迫表达的内参基因actin及其应用

Internal Reference Gene Actin of Setaria viridis and Its Application in Response to Different Drought and Salt Stress

为了研究应用于狗尾草响应不同干旱及盐胁迫基因表达的内参基因,本研究从网站https://phytozome.jgi.doe.gov/pz/portal.html上获得狗尾草A10品系8个actin基因的cDNA序列,设计荧光定量PCR引物,以不同程度的干旱及不同浓度盐胁迫处理的狗尾草幼苗cDNA做模板,进行荧光定量PCR试验,通过实时荧光定量PCR扩增片段电泳分析,扩增曲线及溶解曲线综合分析,结果表明登录号Sevir.9G114100对应的actin基因是8个actin基因中最合适的内标基因。并对所选内标的实用性进行了验证,验证的结果与转录组数据的结果保持一致,证明所选内标基因可用于后续狗尾草响应不同干旱及盐胁迫狗尾草转录组基因表达的验证工作,为深入研究狗尾草功能基因奠定分子基础。

In order to study the internal reference genes applied to the expression of different drought and salt stress genes in S. viridis, the cDNA sequence of 8 actin genes of S. viridis(accession A10.1) was obtained from https://phytozome.jgi.doe. gov/pz/portal.html. The primers for real time PCR were designed, and the templates of cDNA for the seedlings of S. viridis treated with different concentrations of drought and salt stress were used. Through the amplification fragment electrophoresis analysis, amplification curve and dissolution curve comprehensive analysis of RT-PCR, the results indicated that the actin gene corresponding to accession number Sevir.9 G114100 was the most suitable internal standard gene among the eight actin genes. The practicability of the selected internal standard gene was verified. The results of the verification were consistent with the results of the transcriptome data, suggesting that the selected internal standard gene could be used in the verify experiments of S. viridis gene expression response to different drought and salt stress, giving a molecular base for further study of the functional gene of S. viridis.