Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress

Xiaoyun Wang; Xin Shu; Xiaoli Su; Yanli Xiong; Yi Xiong; Minli Chen; Qi Tong; Xiao Ma; Jianbo Zhang; Junming Zhao

doi:10.3390/genes14101874

Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress

Genes (Basel). 2023 Sep 26;14(10):1874. doi: 10.3390/genes14101874.

Authors

Xiaoyun Wang¹, Xin Shu¹, Xiaoli Su¹, Yanli Xiong¹, Yi Xiong¹, Minli Chen², Qi Tong², Xiao Ma¹, Jianbo Zhang², Junming Zhao¹

Affiliations

¹ College of Grassland Science and Technology, Sichuan Agricultural University, Wenjiang, Chengdu 611130, China.
² Sichuan Academy of Grassland Sciences, Pidu, Chengdu 611731, China.

Abstract

As a C4 warm-season turfgrass, centipedegrass (Eremochloa ophiuroides (Munro) Hack.) is known for its exceptional resilience to intensive maintenance practices. In this research, the most stably expressed reference genes in the leaves of centipedegrass under different stress treatments, including salt, cold, drought, aluminum (Al), and herbicide, were screened by the quantitative real-time PCR (RT-qPCR) technique. The stability of 13 candidate reference genes was evaluated by software GeNorm V3.4, NormFinder V20, BestKeeper V1.0, and ReFinder V1.0. The results of this experiment demonstrated that the expression of the UBC (ubiquitin-conjugating enzyme) remained the most stable under cold and Al stress conditions. On the other hand, the MD (malate dehydrogenase) gene exhibited the best performance in leaf tissues subjected to salt and drought stresses. Under herbicide stress, the expression level of the RIP (60S ribosomal protein L2) gene ranked the highest. The expression levels of abiotic stress-associated genes such as PIP1, PAL, COR413, ALMT9, and BAR were assessed to validate the reliability of the selected reference genes. This study provides valuable information and reference points for gene expression under abiotic stress conditions in centipedegrass.

Keywords: RT-qPCR; abiotic stress; centipedegrass; reference genes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Gene Expression Profiling
Gene Expression Regulation, Plant
Genes, Plant*
Herbicides*
Real-Time Polymerase Chain Reaction / methods
Reproducibility of Results
Sodium Chloride
Stress, Physiological / genetics

Substances

Sodium Chloride
Herbicides

Grants and funding

This research was funded by the Sichuan Forestry and Grassland Science and Technology Innovation Team Special Funding of China (Grant number: CXTD2023005), the Sichuan Science and Technology Program (Grant number: 2022YFN0035), the Sichuan Forage Innovation Team Project (Grant number: sccxtd-2020-16), and the National Natural Science Foundation of China (Grant number: 32071885).