| 谭健俊,黄李珊,周熠玮,等.姜荷花苞片实时荧光定量PCR内参基因筛选[J].广东农业科学,2024,(12-2501):- |
| Screening of Reference Genes for Quantitative Real-time PCR in Curcuma alismatifolia Bracts |
| 投稿时间：2024-09-26 修订日期：2024-10-25 |
| DOI： |
|
| 摘要点击次数: 77 |
| 全文下载次数: 0 |
| Abstract: |
| 【Objective】 The quantitative real-time PCR is the most direct and efficient technique for quantifying gene expression levels in plant tissues, ensuring that the selection of stable reference genes is crucial for obtaining reliable and precise results. To date, the research of reference genes has not been documented in Curcuma alismatifolia. Identification of suitable reference genes for qRT-PCR analysis in C. alismatifolia bract will establishe the theoretical foundation for investigating the functional genes associated with bract color. 【Method】 Nine C. alismatifolia cultivars were employed in this experiment. Based on the genomic information, eight housekeeping genes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S rRNA, protein phosphatase 2A (PP2A), ubiquitin (UBQ), malic dehydrogenase (MDH), actin 1 (Actin), β-tubulin (TUB) and phenylalanine ammonia lyase (PAL) were selected, and four different calculation programs including ΔCt, geNorm, NormFinder and BestKeeper were used to evaluate the expression stability of 8 genes. The RefFinder program was utilized to comprehensively assess the optimal reference genes for C. alismatifolia bract tissue. 【Result】 Results indicated that all the eight reference genes amplified a single band, and the dissolution curves were all single peak with R2 ≥ 0.98 and amplification efficiency value of 99.5%~125.0%. The comprehensive ranking of candidate reference genes stability was obtained using the RefFinder program, with the order from high to low being MDH＞Actin＞TUB＞18S rRNA＞PP2A＞GAPDH＞UBQ＞PAL. The most suitable reference genes for different cultivars of C. alismatifolia bract were MDH, followed by Actin and TUB. The stability of PAL is the least favorable, rendering it unsuitable as a reference gene for C. alismatifolia bract tissue. 【Conclusion】 The stable expression of MDH in different cultivars of C. alismatifolia bract serves as a reliable reference gene for further analysis of the expression patterns of genes involved in color synthesis in C. alismatifolia. |
|
View Fulltext
查看/发表评论 下载PDF阅读器 |
|
|
|
