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Open AccessArticle

Identification and Selection of Reference Genes for Quantitative Transcript Analysis in Corydalis yanhusuo

1
School of Pharmacy, Jiangsu Health Vocational College, Nanjing, Jiangsu 211800, China
2
School of Pharmacy, Jiangsu Health Vocational College, Nanjing, Jiangsu 211800, China
3
School of pharmacy, China Pharmaceutical University, Nanjing, Jiangsu 210009, China
4
Institute of Modern Chinese Medicine, School of Pharmacy, Wannan Medical College, Wuhu, Anhui 241002, China
*
Authors to whom correspondence should be addressed.
Genes 2020, 11(2), 130; https://doi.org/10.3390/genes11020130
Received: 11 December 2019 / Revised: 19 January 2020 / Accepted: 23 January 2020 / Published: 27 January 2020
(This article belongs to the Section Plant Genetics and Genomics)
Corydalis yanhusuo is a medicinal plant frequently used in traditional Chinese medicine, which has effective medical effects in many aspects. Real-time polymerase chain reaction (RT-PCR) has been one of the most widely used methods in biosynthesis research due to its high sensitivity and quantitative properties in gene expression analysis. To obtain accurate normalization, reference genes are often selected in advance; however, no reference genes are available in C. yanhusuo. Herein, 12 reference gene candidates, named cyclophilin 2 (CYP2), elongation factor 1-α (EF1-α), protein phosphatase 2 (PP2A), SAND protein family (SAND), polypyrimidine tract-binding protein (PTBP), TIP41-like protein (TIP41), lyceraldehyde-3-phosphate hydrogenase (GAPDH), ubiquitin-conjugating enzyme 9 (UBC9), cyclophilin 1 (CYP1), tubulin beta (TUBA), thioredoxin (YLS8), and polyubiquitin 10 (UBQ10), were selected for stability analysis. After being treated with hormone, UV, salt, metal, oxidative, drought, cold (4 °C), and hot stresses (40 °C), the qRT-PCR data of the selected genes was analyzed with NormFinder, geNorm, and BestKeeper. The result indicated that GAPDH, SNAD, and PP2A were the top three most stable reference genes under most treatments. This study selected and validated reliable reference genes in C. yanhusuo under various environmental conditions, which can provide great help for future research on gene expression normalization in C. yanhusuo.
Key word: qPCR; Corydalis yanhusuo; reference genes; geNorm; NormFinder; BestKeeper
Keywords: qPCR; Corydalis yanhusuo; reference genes; geNorm; NormFinder; BestKeeper qPCR; Corydalis yanhusuo; reference genes; geNorm; NormFinder; BestKeeper
MDPI and ACS Style

Bao, Z.; Zhang, K.; Lin, H.; Li, C.; Zhao, X.; Wu, J.; Nian, S. Identification and Selection of Reference Genes for Quantitative Transcript Analysis in Corydalis yanhusuo. Genes 2020, 11, 130.

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