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| Selection and Characterization of Reliable Reference Genes in Chinese Cherry (Prunus pseudocerasus) Using Quantitative Real-time PCR (qRT-PCR) |
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Abstract Chinese cherry (Prunus pseudocerasus) is one of the most important deciduous fruit tree in China. In order to enrich stable and reliable reference genes and analyze the gene expression more accurately in Chinese cherry by quantitative Real-time PCR (qRT-PCR), the present research selected 7 housekeeping genes of tubulin (TUB), β-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor 1β (EF1B), ubiquitin-conjugating enzyme (UBCE), 18S rRNA and translation initiation factor 5A (TIF5A) based on Chinese cherry flower bud transcriptome data. The expression stability of 7 candidate reference genes were analyzed by Genorm, Bestkeeper, NormFinder and Delta Ct, respectively, under 4 ℃, NaCl (300 mmol/L) and abscisic acid (ABA) (100 mmol/L) treamtent as well as during endodormancy release. This result indicated that the expression of the above 7 genes was not significantly different. GAPDH could expressed stablly under 4 ℃ and NaCl stress, and ACTB and UBCE should be selected as internal controls during ABA treatment and dormancy transition, respectively. Identifying stably expressed housekeeping genes for candidate reference genes from transcriptome database was found to be reliable and efficient. The reference genes selected in this study will be helpful to improve the quality of gene expression studies under various stresses in Chinese cherry.
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Received: 19 November 2014
Published: 17 March 2015
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