Abstract
Lentinula edodes (shiitake mushroom) is a common edible mushroom with a number of potential therapeutic and nutritional applications. It contains various medically important molecules, such as polysaccharides, terpenoids, sterols, and lipids, were contained in this mushroom. Quantitative real-time polymerase chain reaction (qRT-PCR) is a powerful tool to analyze the mechanisms underlying the biosynthetic pathways of these substances. qRT-PCR is used for accurate analyses of transcript levels owing to its rapidity, sensitivity, and reliability. However, its accuracy and reliability for the quantification of transcripts rely on the expression stability of the reference genes used for data normalization. To ensure the reliability of gene expression analyses using qRT-PCR in L. edodes molecular biology research, it is necessary to systematically evaluate reference genes. In the current study, ten potential reference genes were selected from L. edodes genomic data and their expression levels were measured by qRT-PCR using various samples. The expression stability of each candidate gene was analyzed by three commonly used software packages: geNorm, NormFinder, and BestKeeper. Base on the results, Rpl4 was the most stable reference gene across all experimental conditions, and Atu was the most stable gene among strains. 18S was found to be the best reference gene for different development stages, and Rpl4 was the most stably expressed gene under various nutrient conditions. The present work will contribute to qRT-PCR studies in L. edodes.
Highlights
The basidiomycete Lentinula edodes (Berkeley) Pegler (Lentinus edodes), an edible mushroom, is the most economically important cultivated mushroom in East Asia and the second most popular in the world [1]
GeNorm, NormFinder, and BestKeeper, were used to identify and evaluate the expression stability of candidate reference genes. These results provide useful guidelines to select reference gene(s) for gene expression studies in L. edodes and provide a basis for further studies of Quantitative real-time polymerase chain reaction (qRT-PCR) data normalization in other fungi
A total of 10 candidate reference genes, including three genes (18S, 28S, and Pma) that have already been used as reference genes and seven new genes: Atu, Btu, adenine phosphoribosyl transferase (Apt), F-actin, Ribosomal protein L2 (Rpl2), Ribosomal protein L4 (Rpl4), and Tsb were chosen in this study
Summary
The basidiomycete Lentinula edodes (Berkeley) Pegler (Lentinus edodes), an edible mushroom (shiitake), is the most economically important cultivated mushroom in East Asia and the second most popular in the world [1]. L. edodes is valued for its nutritional and medicinal properties, and has wide culinary and industrial applications [2]. Various compounds with therapeutic properties have been isolated from the mycelium and fruiting body in the last few decades, among which the polysaccharide lentinan has been studied intensively.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
