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国际药用蘑菇期刊
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ISSN 打印: 1521-9437
ISSN 在线: 1940-4344

国际药用蘑菇期刊

DOI: 10.1615/IntJMedMushrooms.2019030106
pages 301-309

Identification and Evaluation of Reference Genes for qRT-PCR Normalization in Sparassis latifolia (Agaricomycetes)

Chi Yang
Institute of Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China; National and Local Joint Engineering Research Center for Breeding and Cultivation of Featured Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China
Lu Ma
Institute of Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China; National and Local Joint Engineering Research Center for Breeding and Cultivation of Featured Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China
Donglai Xiao
Institute of Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China; National and Local Joint Engineering Research Center for Breeding and Cultivation of Featured Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China
Zhenghe Ying
Institute of Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China; National and Local Joint Engineering Research Center for Breeding and Cultivation of Featured Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China
Xiaoling Jiang
Institute of Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China; National and Local Joint Engineering Research Center for Breeding and Cultivation of Featured Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China
Yanquan Lin
Institute of Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China; National and Local Joint Engineering Research Center for Breeding and Cultivation of Featured Edible Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, China

ABSTRACT

Real-time quantitative polymerase chain reaction (qRT-PCR) has emerged as a powerful and popular tool for quantitating differences in transcriptional gene expression levels between samples. Validation of the stability of reference genes is a fundamental step before initiating qRT-PCR assays. Sparassis latifolia is an edible and medicinal fungus containing a remarkably high concentration of β-glucan, which has many biological and pharmacologic activities. S. latifolia may be a model species for studying fungal photobiology because its fruiting body formation requires more light than other fungi. However, suitable reference genes for qRT-PCR have not yet been determined. In the present study, 10 candidate reference genes in S. latifolia were evaluated and validated under different developmental stages and light conditions. To evaluate the suitability of candidate reference genes, three popular software programs (geNorm, NormFinder, and BestKeeper), along with the delta Ct method, were used to analyze these genes; the final ranking was determined using RefFinder. According to our results, Actin and GAPDH were expressed at the most stable levels under different developmental stages and light conditions.


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