Screening and validating of endogenous reference genes in Chlorella sp. TLD 6B under abiotic stress
Abstract Chlorella sp. TLD 6B, a microalgae growing in the Taklamakan Desert, Xinjiang of China, is a good model material for studying the physiological and environmental adaptation mechanisms of plants in their arid habitats, as its adaptation to the harsh desert environment has led to its strong r...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-28311-x |
| _version_ | 1828051785481191424 |
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| author | Yongshun Zhou Fanze Meng Kai Han Kaiyue Zhang Jianfeng Gao Fulong Chen |
| author_facet | Yongshun Zhou Fanze Meng Kai Han Kaiyue Zhang Jianfeng Gao Fulong Chen |
| author_sort | Yongshun Zhou |
| collection | DOAJ |
| description | Abstract Chlorella sp. TLD 6B, a microalgae growing in the Taklamakan Desert, Xinjiang of China, is a good model material for studying the physiological and environmental adaptation mechanisms of plants in their arid habitats, as its adaptation to the harsh desert environment has led to its strong resistance. However, when using real-time quantitative polymerase chain reaction (RT-qPCR) to analyze the gene expression of this algae under abiotic stress, it is essential to find the suitable endogenous reference genes so to obtain reliable results. This study assessed the expression stability of 9 endogenous reference genes of Chlorella sp. TLD 6B under four abiotic stresses (drought, salt, cold and heat). These genes were selected based on the analysis results calculated by the three algorithmic procedures of geNorm, NormFinder, and BestKeeper, which were ranked by refinder. Our research showed that 18S and GTP under drought stress, 18S and IDH under salt stress, CYP and 18S under cold stress, GTP and IDH under heat stress were the most stable endogenous reference genes. Moreover, UBC and 18S were the most suitable endogenous reference gene combinations for all samples. In contrast, GAPDH and α-TUB were the two least stable endogenous reference genes in all experimental samples. Additionally, the selected genes have been verified to be durable and reliable by detecting POD and PXG3 genes using above endogenous reference genes. The identification of reliable endogenous reference genes guarantees more accurate RT-qPCR quantification for Chlorella sp. TLD 6B, facilitating functional genomics studies of deserts Chlorella as well as the mining of resistance genes. |
| first_indexed | 2024-04-10T19:43:37Z |
| format | Article |
| id | doaj.art-ef1c5c2273c24157a539fa1788ebb401 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-10T19:43:37Z |
| publishDate | 2023-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-ef1c5c2273c24157a539fa1788ebb4012023-01-29T12:13:10ZengNature PortfolioScientific Reports2045-23222023-01-0113111210.1038/s41598-023-28311-xScreening and validating of endogenous reference genes in Chlorella sp. TLD 6B under abiotic stressYongshun Zhou0Fanze Meng1Kai Han2Kaiyue Zhang3Jianfeng Gao4Fulong Chen5College of Life Sciences, Shihezi UniversityCollege of Life Sciences, Shihezi UniversityCollege of Life Sciences, Shihezi UniversityCollege of Life Sciences, Shihezi UniversityCollege of Life Sciences, Shihezi UniversityCollege of Life Sciences, Shihezi UniversityAbstract Chlorella sp. TLD 6B, a microalgae growing in the Taklamakan Desert, Xinjiang of China, is a good model material for studying the physiological and environmental adaptation mechanisms of plants in their arid habitats, as its adaptation to the harsh desert environment has led to its strong resistance. However, when using real-time quantitative polymerase chain reaction (RT-qPCR) to analyze the gene expression of this algae under abiotic stress, it is essential to find the suitable endogenous reference genes so to obtain reliable results. This study assessed the expression stability of 9 endogenous reference genes of Chlorella sp. TLD 6B under four abiotic stresses (drought, salt, cold and heat). These genes were selected based on the analysis results calculated by the three algorithmic procedures of geNorm, NormFinder, and BestKeeper, which were ranked by refinder. Our research showed that 18S and GTP under drought stress, 18S and IDH under salt stress, CYP and 18S under cold stress, GTP and IDH under heat stress were the most stable endogenous reference genes. Moreover, UBC and 18S were the most suitable endogenous reference gene combinations for all samples. In contrast, GAPDH and α-TUB were the two least stable endogenous reference genes in all experimental samples. Additionally, the selected genes have been verified to be durable and reliable by detecting POD and PXG3 genes using above endogenous reference genes. The identification of reliable endogenous reference genes guarantees more accurate RT-qPCR quantification for Chlorella sp. TLD 6B, facilitating functional genomics studies of deserts Chlorella as well as the mining of resistance genes.https://doi.org/10.1038/s41598-023-28311-x |
| spellingShingle | Yongshun Zhou Fanze Meng Kai Han Kaiyue Zhang Jianfeng Gao Fulong Chen Screening and validating of endogenous reference genes in Chlorella sp. TLD 6B under abiotic stress Scientific Reports |
| title | Screening and validating of endogenous reference genes in Chlorella sp. TLD 6B under abiotic stress |
| title_full | Screening and validating of endogenous reference genes in Chlorella sp. TLD 6B under abiotic stress |
| title_fullStr | Screening and validating of endogenous reference genes in Chlorella sp. TLD 6B under abiotic stress |
| title_full_unstemmed | Screening and validating of endogenous reference genes in Chlorella sp. TLD 6B under abiotic stress |
| title_short | Screening and validating of endogenous reference genes in Chlorella sp. TLD 6B under abiotic stress |
| title_sort | screening and validating of endogenous reference genes in chlorella sp tld 6b under abiotic stress |
| url | https://doi.org/10.1038/s41598-023-28311-x |
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