| 摘要: |
| 【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。 |
| 关键词: 大豆 盐胁迫 碱胁迫 荧光定量PCR Pre-miRNAs(前体miRNAs) 内参基因 |
| DOI： |
| 分类号: |
| 基金项目:国家转基因生物新品种培育重大专项(2014ZX08010-002)；国家自然科学基金项目(31271746，31201144)；吉林省发改委项目(JF2012C002-04) |
|
| Stability of Pre-miRNAs in soybean as reference genes for quantitative polymerase chain reaction under salt and alkali stresses |
|
ZHOU Yong-gang,WANG Qi,LIU Wei-can,et al
|
| Abstract: |
| 【Objective】This study selected the appropriate reference genes for RT-qPCR by comparing the stability of candidate precursor miRNAs and traditional housekeeping genes in different tissues of soybean under different stress treatments.【Method】Six conservative Pre-miRNAs genes and four regular housekeeping genes in soybean were selected as candidate reference genes to evaluate their expression stability under salt and alkali stresses during RT-qPCR process using GeNorm and NormFinder.【Result】In leaves,the most stable combination of genes under salt stress was Pre-miR166 and Pre-miR172 whereas the best single gene under this stress was FboX.While,in roots,the most stable combination of genes under salt stress was Act11 and EF1A whereas the best single gene was Act11.Under alkali stress conditions,the most stable combination of genes in leaves was Pre miR393 and Pre-miR172 whereas the best single gene was Pre-miR393.The most stable combination of genes in roots was Act11 and 60S whereas the best single gene was Pre-miR172.【Conclusion】Precursor miRNAs reference genes can be used as reference genes together with traditional housekeeping genes. |
| Key words: soybean salt stress alkali stress RT-qPCR Pre-miRNAs reference gene |
