Selection and Validation of Candidate Reference Genes for Quantitative Real-time PCR in Jasminum sambac Aiton

doi:10.7668/hbnxb.20191401

Abstract

Abstract: To screen suitable reference genes for jasmine, five tissues (root, stem, young leaf, matured leaf and flower) and four developmental stages of flowers (young floral bud, swollen floral bud, the longest floral bud and initial opening flower) of jasmine were used as experimental materials. The eight common candidate reference genes were selected for primer specificity analysis, and the results showed that all the eight genes amplified a single band, and the melting curve had only a single obvious peak. The expression levels of eight reference genes in different samples were analyzed by qRT-PCR. The results showed that the expression levels of eight candidate reference genes were different in the samples, and the expression level of each gene in flower was significantly lower than that of other samples. The stability of these genes was evaluated by software including geNorm, NormFinder and BestKeeper, and then the expression stability of them was comprehensively analyzed by software RefFinder. The results indicated that the best reference genes for different tissues with flowers were SAND and UPL7, for different tissues without flowers were UPL7 and GAPDH, and for different developmental stages of flowers were Actin and EF1α. Furthermore, the selected internal reference gene was verified by the target genes JsDXS and JsPAL2. The results showed that the expression levels of JsDXS and JsPAL2 presented the same trend at different stages when used Actin, EF1α and Actin+EF1α combination as references, however, the expression level trends were inconsistent when used the unstable PP2A as reference. In summary, this study we screened the reference genes for jasmine, and the optimal reference genes for different tissues including flowers were SAND and UPL7, but the most stable reference genes in different tissues without flowers were UPL7 and GAPDH. Simultaneously, Actin and EF1α were the most suitable genes for different developmental stages of flowers.

Key words: Jasmine, qRT-PCR, Reference gene, Expression stability

CLC Number:

S685.160.1

QI Xiangyu, CHEN Shuangshuang, FENG Jing, WANG Huadi, DENG Yanming. Selection and Validation of Candidate Reference Genes for Quantitative Real-time PCR in Jasminum sambac Aiton[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(6): 22-30. doi: 10.7668/hbnxb.20191401.

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