Identification and validation of reference genes for qRT-PCR analysis in mulberry (Morus alba L.)

PLoS One. 2018 Mar 15;13(3):e0194129. doi: 10.1371/journal.pone.0194129. eCollection 2018.

Abstract

Mulberry (Morus alba L.) is an important economic tree species in many countries. Quantitative real time PCR (qRT-PCR) has become a widely used method for gene expression studies in plants. A suitable reference gene is essential to ensure accurate and reliable results for qRT-PCR analyses. However, no reports describing the selection of reference genes have been published for mulberry. In this work, we evaluated the stability of twenty candidate reference genes in different plant tissues and under different stress conditions by qRT-PCR in mulberry using algorithms in two programs-geNorm and NormFinder. The results revealed that TUB2, UBI4, ACTIN3 and RPL4 were ranked as the most stable reference genes in the samples subsets, whereas EF1α4 and TUB3showed the least stability with both algorithms. To further validate the stability of the reference genes, the expression patterns of six genes of mulberry were analyzed by normalization with the selected reference genes. Our study will benefit future analyses of gene expression in mulberry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression Profiling / methods
  • Gene Expression Regulation, Plant
  • Genes, Plant*
  • Morus / genetics*
  • Morus / physiology
  • Real-Time Polymerase Chain Reaction / methods
  • Reference Standards
  • Stress, Physiological

Grants and funding

This work was supported by grants from the National Nature Science Foundation of China (No. 31500527) (FD), the science and technology project of Guangzhou city, China (No. 201710010129 (FD); No. 2014Y2-00514 (CT)), the science and technology project of Guangdong province, China (No. 2016A020210033 (FD)). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.