Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata
Abstract Pinellia ternata (Thunb.) Breit. (Abbreviated as P. ternata). It is a commonly prescribed Chinese traditional medicinal herb for the treatment of phlegm, cough, and morning sick. Bulbil reproduction is one of the main reproductive methods of P. ternata. The accurate quantification of gene e...
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| Format: | Article |
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-12782-5 |
| _version_ | 1818213481247145984 |
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| author | Haoyu Fan Qiuling He Yiheng Dong Wenxin Xu Yanlin Lou Xuejun Hua Tao Xu |
| author_facet | Haoyu Fan Qiuling He Yiheng Dong Wenxin Xu Yanlin Lou Xuejun Hua Tao Xu |
| author_sort | Haoyu Fan |
| collection | DOAJ |
| description | Abstract Pinellia ternata (Thunb.) Breit. (Abbreviated as P. ternata). It is a commonly prescribed Chinese traditional medicinal herb for the treatment of phlegm, cough, and morning sick. Bulbil reproduction is one of the main reproductive methods of P. ternata. The accurate quantification of gene expression patterns associated with bulbil development might be helpful to explore the molecular mechanism involved in P. ternata reproduction. Quantitative real-time PCR was the most preferred method for expression profile and function analysis of mRNA. However, the reference genes in different tissues of P. ternata in different periods of bulbil development have not been studied in detail. In present study, the expression stability of eight candidate reference genes were determined with programs: geNorm, NormFinder, BestKeeper, and refFinder. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was identified as the top- rated reference gene in all samples of P. ternata, while different combinations of reference gene proved to be the most stable depending on development stage and tissue type. Furthermore, the reliability of GAPDH expression was verified by six P. ternata related genes in hormone and nutrient biosynthesis pathways, and the expression profiles of these genes were agreed with the results of RNA-seq digital gene expression analysis. These results can contribute to studies of gene expression patterns and functional analysis of P. ternata involved in bulbil development. |
| first_indexed | 2024-12-12T06:04:58Z |
| format | Article |
| id | doaj.art-02d33b0ba7754e6f99af1e43a71140ac |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-12T06:04:58Z |
| publishDate | 2022-05-01 |
| publisher | Nature Portfolio |
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| spelling | doaj.art-02d33b0ba7754e6f99af1e43a71140ac2022-12-22T00:35:18ZengNature PortfolioScientific Reports2045-23222022-05-0112111210.1038/s41598-022-12782-5Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternataHaoyu Fan0Qiuling He1Yiheng Dong2Wenxin Xu3Yanlin Lou4Xuejun Hua5Tao Xu6Key Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, Zhejiang Sci-Tech UniversityKey Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, Zhejiang Sci-Tech UniversityKey Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, Zhejiang Sci-Tech UniversityKey Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, Zhejiang Sci-Tech UniversityKey Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, Zhejiang Sci-Tech UniversityKey Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, Zhejiang Sci-Tech UniversityKey Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, Zhejiang Sci-Tech UniversityAbstract Pinellia ternata (Thunb.) Breit. (Abbreviated as P. ternata). It is a commonly prescribed Chinese traditional medicinal herb for the treatment of phlegm, cough, and morning sick. Bulbil reproduction is one of the main reproductive methods of P. ternata. The accurate quantification of gene expression patterns associated with bulbil development might be helpful to explore the molecular mechanism involved in P. ternata reproduction. Quantitative real-time PCR was the most preferred method for expression profile and function analysis of mRNA. However, the reference genes in different tissues of P. ternata in different periods of bulbil development have not been studied in detail. In present study, the expression stability of eight candidate reference genes were determined with programs: geNorm, NormFinder, BestKeeper, and refFinder. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was identified as the top- rated reference gene in all samples of P. ternata, while different combinations of reference gene proved to be the most stable depending on development stage and tissue type. Furthermore, the reliability of GAPDH expression was verified by six P. ternata related genes in hormone and nutrient biosynthesis pathways, and the expression profiles of these genes were agreed with the results of RNA-seq digital gene expression analysis. These results can contribute to studies of gene expression patterns and functional analysis of P. ternata involved in bulbil development.https://doi.org/10.1038/s41598-022-12782-5 |
| spellingShingle | Haoyu Fan Qiuling He Yiheng Dong Wenxin Xu Yanlin Lou Xuejun Hua Tao Xu Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata Scientific Reports |
| title | Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata |
| title_full | Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata |
| title_fullStr | Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata |
| title_full_unstemmed | Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata |
| title_short | Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata |
| title_sort | selection of suitable candidate genes for mrna expression normalization in bulbil development of pinellia ternata |
| url | https://doi.org/10.1038/s41598-022-12782-5 |
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