Selection of Reference Genes for qRT-PCR Analysis in Lentinula edodes after Hot-Air Drying

Molecules. 2018 Dec 31;24(1):136. doi: 10.3390/molecules24010136.

Abstract

Volatile sulfur compounds gradually develop in Lentinula edodes after hot-air drying, and many genes are involved in the generation of these sulfur compounds. The expression stability of reference genes may vary in a particular experimental treatment when analyzing their expressions by quantitative real-time polymerase chain reaction (qRT-PCR). In this study, the expression profile of 17 candidate genes was assessed in L. edodes under treatment at 50 °C for 0, 1, 2, and 3 h, and the expression stability of each reference gene was analyzed by three statistical algorithms, including geNorm, NormFinder, and BestKeeper. Results indicated that the two optimal reference genes for mycelium and fruiting body were CAC and DAHP as well as CAC and NUP, respectively. Additionally, CAC and DAHP were found to be the two most stable reference genes across the mycelium and fruiting body set. Our results will provide a genetic foundation for further research on the metabolism genes of sulfur compounds in L. edodes.

Keywords: Lentinula edodes; hot-air drying; qRT-PCR; reference genes; volatile sulfur compounds.

MeSH terms

  • Algorithms
  • Desiccation / methods*
  • Fungal Proteins / genetics*
  • Gene Expression Profiling / standards
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Fungal
  • Hot Temperature
  • Real-Time Polymerase Chain Reaction / standards*
  • Reference Standards
  • Sequence Analysis, RNA / methods
  • Shiitake Mushrooms / genetics*
  • Shiitake Mushrooms / growth & development
  • Sulfur Compounds
  • Volatile Organic Compounds

Substances

  • Fungal Proteins
  • Sulfur Compounds
  • Volatile Organic Compounds