Screening and verification of reference genes for analysis of gene expression in winter rapeseed (Brassica rapa L.) under abiotic stress

PLoS One. 2020 Sep 17;15(9):e0236577. doi: 10.1371/journal.pone.0236577. eCollection 2020.

Abstract

Winter rapeseed (Brassica rapa L.) is the main oilseed crop in northern China and can safely overwinter at 35 (i.e., Tianshui, China) to 48 degrees north latitude (i.e., Altai, Heilongjiang, Raohe, and Xinjiang, China). In order to identify stable reference genes to understand the molecular mechanisms of stress tolerance in winter rapeseed, internal reference genes of winter rapeseed under four abiotic stresses were analyzed using GeNorm, NormFinder, BestKeeper, and RefFinder software. The most stable combinations of internal reference genes were β-actin and SAND in cold-stressed leaves, β-actin and EF1a in cold-stressed roots, F-box and SAND in high temperature-stressed leaves, and PP2A and RPL in high temperature-stressed roots, SAND and PP2A in NaCl-stressed leaves, RPL and UBC in NaCl-stressed roots, RPL and PP2A in PEG-stressed leaves, and PP2A and RPL in PEG-stressed roots. Expression profiles of PXG3 were used to verify these results. The stable reference genes identified in this study are useful tools for identifying stress-responsive genes to understand the molecular mechanisms of stress tolerance in winter rapeseed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brassica rapa / genetics*
  • Brassica rapa / metabolism
  • Cold-Shock Response*
  • Gene Expression Profiling / methods
  • Gene Expression Profiling / standards*
  • Gene Expression Regulation, Plant
  • Osmotic Pressure*
  • Plant Leaves / metabolism
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Plant Roots / metabolism
  • Reference Standards
  • Seasons

Substances

  • Plant Proteins

Grants and funding

This study was financially supported by the Scientific research start-up funds for openly-recuited doctors of Gansu Agricultural University (GAU-KYQD-2019-17), the Utilization Technology of Rapeseed Heterosis and Creation of Strong Heterosis of China (2016YFD0101300), Agriculture Research System of China (CARS-12), and the Agriculture Research System of Gansu Province (GARS-TSZ-1), and the National Natural Science Foundation of China (31860388). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.