Reference Genes Screening and Gene Expression Patterns Analysis Involved in Gelsenicine Biosynthesis under Different Hormone Treatments in Gelsemium elegans
-
Published:2023-11-04
Issue:21
Volume:24
Page:15973
-
ISSN:1422-0067
-
Container-title:International Journal of Molecular Sciences
-
language:en
-
Short-container-title:IJMS
Author:
Zhang Yao1ORCID, Mu Detian1, Wang Liya1, Wang Xujun2, Wilson Iain W.3ORCID, Chen Wenqiang1ORCID, Wang Jinghan4ORCID, Liu Zhaoying5ORCID, Qiu Deyou6ORCID, Tang Qi1ORCID
Affiliation:
1. College of Horticulture, National Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, Hunan Agricultural University, Changsha 410128, China 2. Hunan Academy of Forestry, Changsha 410018, China 3. CSIRO Agriculture and Food, Canberra, ACT 2601, Australia 4. College of Forestry, Central South University of Forestry and Technology, Changsha 410004, China 5. College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China 6. State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China
Abstract
Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is an accurate method for quantifying gene expression levels. Choosing appropriate reference genes to normalize the data is essential for reducing errors. Gelsemium elegans is a highly poisonous but important medicinal plant used for analgesic and anti-swelling purposes. Gelsenicine is one of the vital active ingredients, and its biosynthesis pathway remains to be determined. In this study, G. elegans leaf tissue with and without the application of one of four hormones (SA, MeJA, ETH, and ABA) known to affect gelsenicine synthesis, was analyzed using ten candidate reference genes. The gene stability was evaluated using GeNorm, NormFinder, BestKeeper, ∆CT, and RefFinder. The results showed that the optimal stable reference genes varied among the different treatments and that at least two reference genes were required for accurate quantification. The expression patterns of 15 genes related to the gelsenicine upstream biosynthesis pathway was determined by RT-qPCR using the relevant reference genes identified. Three genes 8-HGO, LAMT, and STR, were found to have a strong correlation with the amount of gelsenicine measured in the different samples. This research is the first study to examine the reference genes of G. elegans under different hormone treatments and will be useful for future molecular analyses of this medically important plant species.
Funder
National Natural Science Foundation of China Postgraduate Scientific Research Innovation Project of Hunan Province Postgraduate Scientific Research Innovation Project of Hunan Agriculture University
Subject
Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis
Reference62 articles.
1. Zhao, X., Geng, Y., Hu, T., Zhao, Y., Yang, S., and Hao, D. (2022). Evaluation of Optimal Reference Genes for qRT-PCR Analysis in Hyphantria cunea (Drury). Insects, 13. 2. Freitas, B.L., Leach, L., Chaturvedi, V., and Chaturvedi, S. (2022). Reverse Transcription-Quantitative Real-Time PCR (RT-qPCR) Assay for the Rapid Enumeration of Live Candida auris Cells from the Health Care Environment. J. Clin. Microbiol., 60. 3. Zhao, J., Yang, J., Wang, X., Xiong, Y., Xiong, Y., Dong, Z., Lei, X., Yan, L., and Ma, X. (2022). Selection and Validation of Reference Genes for qRT-PCR Gene Expression Analysis in Kengyilia melanthera. Genes, 13. 4. Selection and Validation of Reference Genes for Quantitative Real-Time PCR Normalization in Athetis dissimilis (Lepidoptera: Noctuidae) Under Different Conditions;Tang;Front. Physiol.,2022 5. Deguchi, M., Potlakayala, S., Spuhler, Z., George, H., Sheri, V., Agili, R., Patel, A., and Rudrabhatla, S. (2021). Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp. PLoS ONE, 16.
Cited by
2 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献
|
|