Identification of Reliable Reference Genes under Different Stresses and in Different Tissues of Toxicodendron succedaneum

Dongxiao Ma; Qin Zhang; Jintao Zhou; Yu Lu; Xiaomeng Duan; Chengzhong He; Jinde Yu

doi:10.3390/genes13122396

Identification of Reliable Reference Genes under Different Stresses and in Different Tissues of Toxicodendron succedaneum

Genes (Basel). 2022 Dec 17;13(12):2396. doi: 10.3390/genes13122396.

Authors

Dongxiao Ma^{1

2}, Qin Zhang^{1

2}, Jintao Zhou^{1

2}, Yu Lu^{1

2}, Xiaomeng Duan^{1

3}, Chengzhong He^{1

2

3}, Jinde Yu¹

Affiliations

¹ College of Life Sciences, Southwest Forestry University, Kunming 650224, China.
² Key Laboratory for Forest Genetic and Tree Improvement and Propagation in Universities of Yunnan Province, Southwest Forestry University, Kunming 650224, China.
³ Key Laboratory of Biodiversity Conservation in Southwest China, State Forestry Administration, Southwest Forestry University, Kunming 650224, China.

Abstract

Toxicodendron succedaneum (L.) Kuntze (T. succedaneum) is an economic tree species that produces urushiol and urushi wax, and it is of great value in industry and medicine. However, the stability of reference genes (RGs) has not been systematically reported in T. succedaneum to date. In this study, the expression of 10 candidate RGs was analyzed by RT-qPCR in different tissues (roots, stems, leaves), stress treatments (high/low temperature, drought), and hormone stimulation (jasmonic acid, JA). Then, the stability ranking of 10 candidate genes was evaluated by ∆Ct analysis and three software programs: geNorm, NormFinder, and BestKeeper. Finally, RefFinder was used to comprehensively analyze the expression stability of 10 candidate genes. The comprehensive analysis showed that TsRG05/06, TsRG01/06, and TsRG03/ACT were stable under high/low-temperature stress, drought stress, and JA treatment, respectively. TsRG03 and ACT had stable expression in different tissues. While the TsRG03 and ACT were recommended as the suitable RGs for T. succedaneum in all samples. Meanwhile, UBQ was the least suitable as a reference gene for T. succedaneum. In addition, the results of geNorm showed that the combination of two stable RGs could make the results of gene expression more accurate. These results provide alternative RGs for the study of gene function, correction, and normalization of target gene expression and directed molecular breeding in T. succedaneum.

Keywords: RT-qPCR; Toxicodendron succedaneum; gene expression; reference gene.

Publication types

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

MeSH terms

Gene Expression Profiling* / methods
Real-Time Polymerase Chain Reaction / methods
Reference Standards
Software
Toxicodendron*

Grants and funding

This research was funded by the Yunnan Provincial Expert Workstation (202005AF150020), and the Yunnan Provincial “Ten-Thousand Program” for Leading Industry Innovation (YUWR-CYJS-2020-018).