Selection of Reliable Reference Genes for Gene Expression Studies of a Promising Oilseed Crop, Plukenetia volubilis, by Real-Time Quantitative PCR
文献类型：期刊论文
| 作者 | Xu, Zeng-Fu
|
| 刊名 | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
 |
| 出版日期 | 2015 |
| 卷号 | 16期号:6页码:12513-12530 |
| ISSN号 | 1422-0067 |
| 中文摘要 | Real-time quantitative PCR (RT-qPCR) is a reliable and widely used method for gene expression analysis. The accuracy of the determination of a target gene expression level by RT-qPCR demands the use of appropriate reference genes to normalize the mRNA levels among different samples. However, suitable reference genes for RT-qPCR have not been identified in Sacha inchi (Plukenetia volubilis), a promising oilseed crop known for its polyunsaturated fatty acid (PUFA)-rich seeds. In this study, using RT-qPCR, twelve candidate reference genes were examined in seedlings and adult plants, during flower and seed development and for the entire growth cycle of Sacha inchi. Four statistical algorithms (delta cycle threshold (Delta C-t), BestKeeper, geNorm, and NormFinder) were used to assess the expression stabilities of the candidate genes. The results showed that ubiquitin-conjugating enzyme (UCE), actin (ACT) and phospholipase A22 (PLA) were the most stable genes in Sacha inchi seedlings. For roots, stems, leaves, flowers, and seeds from adult plants, 30S ribosomal protein S13 (RPS13), cyclophilin (CYC) and elongation factor-1alpha (EF1 alpha) were recommended as reference genes for RT-qPCR. During the development of reproductive organs, PLA, ACT and UCE were the optimal reference genes for flower development, whereas UCE, RPS13 and RNA polymerase II subunit (RPII) were optimal for seed development. Considering the entire growth cycle of Sacha inchi, UCE, ACT and EF1 alpha were sufficient for the purpose of normalization. Our results provide useful guidelines for the selection of reliable reference genes for the normalization of RT-qPCR data for seedlings and adult plants, for reproductive organs, and for the entire growth cycle of Sacha inchi. |
| 原文出处 | 10.3390/ijms160612513 |
| 语种 | 英语 |
| 公开日期 | 2015-08-11 |
| 源URL | [http://ir.xtbg.org.cn/handle/353005/9223]  |
| 专题 | 西双版纳热带植物园_能源植物分子育种研究组 |
| 通讯作者 | Xu, Zeng-Fu |
| 推荐引用方式 GB/T 7714 | Xu, Zeng-Fu. Selection of Reliable Reference Genes for Gene Expression Studies of a Promising Oilseed Crop, Plukenetia volubilis, by Real-Time Quantitative PCR[J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,2015,16(6):12513-12530. |
| APA | Xu, Zeng-Fu.(2015).Selection of Reliable Reference Genes for Gene Expression Studies of a Promising Oilseed Crop, Plukenetia volubilis, by Real-Time Quantitative PCR.INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,16(6),12513-12530. |
| MLA | Xu, Zeng-Fu."Selection of Reliable Reference Genes for Gene Expression Studies of a Promising Oilseed Crop, Plukenetia volubilis, by Real-Time Quantitative PCR".INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 16.6(2015):12513-12530. |
入库方式： OAI收割
来源：西双版纳热带植物园
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