Identification of Reference Genes for RT-qPCR Analysis in Gleditsia microphylla under Abiotic Stress and Hormone Treatment

Jiaqi Yang; Fengying Han; Li Yang; Jin Wang; Feng Jin; An Luo; Fuyong Zhao

doi:10.3390/genes13071227

Identification of Reference Genes for RT-qPCR Analysis in Gleditsia microphylla under Abiotic Stress and Hormone Treatment

Genes (Basel). 2022 Jul 10;13(7):1227. doi: 10.3390/genes13071227.

Authors

Jiaqi Yang¹, Fengying Han¹, Li Yang¹, Jin Wang¹, Feng Jin¹, An Luo¹, Fuyong Zhao¹

Affiliation

¹ College of Life Sciences, Yangtze University, Jingzhou 434025, China.

Abstract

Gleditsia microphylla is an important galactomannan gums source plant with characteristics of drought resistance, barren tolerance, and good adaptability. However, the underlying molecular mechanisms of the biological process are not yet fully understood. Real-time quantitative PCR (RT-qPCR) is an accurate and convenient method to quantify the gene expression level and transcription abundance of suitable reference genes. This study aimed to screen the best internal reference genes in G. microphylla under abiotic stresses, hormone treatments, and different tissues. Based on the transcriptome data, twelve candidate reference genes were selected, and ultimately, nine of them were further evaluated by the geNorm, NormFinder, BestKeeper, and RefFinder algorithms. These results show that TATA-binding protein 1 (TBP1)and Eukaryotic translation initiation factor 4A1 (EIF4A1)were the two most stable reference genes, and glyceraldehyde-3-phosphate dehydrogenase A subunit, chloroplastic (GAPA)and glyceraldehyde-3-phosphate dehydrogenase B subunit, chloroplastic (GAPB)were the two most unstable reference genes across all samples under the given experimental conditions. Meanwhile, the most stable reference genes varied among the different groups and tissues. Therefore, this study suggests that it is better to use a specific reference gene for a particular case rather than using a common reference gene.

Keywords: Gleditsia microphylla; abiotic stresses; hormone treatments; real-time quantitative PCR (RT-qPCR); reference genes.

Publication types

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

MeSH terms

Gene Expression Profiling / methods
Gleditsia* / genetics
Hormones
Real-Time Polymerase Chain Reaction / methods
Stress, Physiological / genetics

Substances

Hormones

Grants and funding

This research was funded by the National Natural Science Foundation of China (31700281) and Jingzhou Jiahua Technology Co., Ltd. (Jingzhou, Hubei Province, China). The sponsors had no role in the design, execution, interpretation, or writing of the study.