Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress
Author:
Wang Xiaoyun1, Shu Xin1, Su Xiaoli1, Xiong Yanli1, Xiong Yi1ORCID, Chen Minli2, Tong Qi2, Ma Xiao1, Zhang Jianbo2, Zhao Junming1
Affiliation:
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.
Funder
Sichuan Forestry and Grassland Science and Technology Innovation Team Special Funding of China Sichuan Science and Technology Program Sichuan Forage Innovation Team Project National Natural Science Foundation of China
Subject
Genetics (clinical),Genetics
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