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【目的】对比大豆Pre-miRNAs候选内参基因和传统内参基因的表达稳定性,筛选出适合盐、碱胁迫条件下RT-qPCR分析的最稳定内参基因。【方法】选用了6个保守的Pre-miRNAs基因和4个常规的内参基因作为候选内参基因,通过RT-qPCR的方法,利用GeNorm和NormFinder软件,评价了10个候选内参基因在大豆盐、碱胁迫条件下的稳定性。【结果】大豆盐胁迫下,叶中表达最稳定的基因组合是Pre-miR166和Pre-miR172,表达最稳定的单一基因是FboX;根中表达最稳定的基因组合是Act11和EF1A,表达最稳定的单一基因是Act11。大豆碱胁迫下,叶中表达最稳定的基因组合是Pre-miR393和Pre-miR172,表达最稳定的单一基因是Pre-miR393;根中表达最稳定的基因组合是Act11和60S,表达最稳定的单一基因是Pre-miR172。【结论】大豆Pre-miRNAs可以与传统内参基因一样作为内参定量目的基因。
【Objective】 The objective of this study was to compare the expression stability of candidate internal reference genes and traditional reference genes in soybean Pre-miRNAs and screen out the most stable reference genes for RT-qPCR analysis under salt and alkali stress conditions. 【Method】 Six conserved Pre-miRNAs and four conventional internal reference genes were selected as candidate reference genes. By RT-qPCR and GeNorm and NormFinder software, ten candidate reference genes were evaluated in soybean salt, alkali Stability under stress conditions. 【Result】 Under the salt stress of soybean, the most stable gene combination in leaves was Pre-miR166 and Pre-miR172, and the single gene with the most stable expression was FboX. The most stable gene combination in roots was Act11 and EF1A, and the expression was the most stable The single gene is Act11. The most stable gene combination in leaves was Pre-miR393 and Pre-miR172, and the single gene with the most stable expression was Pre-miR393. The most stable gene combination in roots was Act11 and 60S, and the expression was the most stable The single gene is Pre-miR172. 【Conclusion】 Soybean Pre-miRNAs can be the same as the traditional reference genes as internal reference quantitative target genes.