Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insect
Paederus fuscipes is a medically and agriculturally important insect all over the world. P. fuscipes cause not only a skin condition but is also an aggressive predator in different agro-ecosystems. Prior to performing RT-qPCR under a variety of conditions to investigate the function of target genes...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-01-01
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| Series: | Journal of King Saud University: Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1018364721003165 |
| _version_ | 1819175571923402752 |
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| author | Muhammad Musa Khan Chang-Fei Guo Jing Peng Ze-Yun Fan Muhammad Hafeez Daoud Ali Kai Wang Mohammed H.A. Almarzoug Bao-Li Qiu |
| author_facet | Muhammad Musa Khan Chang-Fei Guo Jing Peng Ze-Yun Fan Muhammad Hafeez Daoud Ali Kai Wang Mohammed H.A. Almarzoug Bao-Li Qiu |
| author_sort | Muhammad Musa Khan |
| collection | DOAJ |
| description | Paederus fuscipes is a medically and agriculturally important insect all over the world. P. fuscipes cause not only a skin condition but is also an aggressive predator in different agro-ecosystems. Prior to performing RT-qPCR under a variety of conditions to investigate the function of target genes in P. fuscipes, it is essential to screen reference genes. However, no P. fuscipes reference gene(s) has yet been discovered. Using the RefFinder software package, which includes ΔCt, geNorm, NormFinder, and BestKeeper, we evaluated the stability of seven housekeeping genes of P. fuscipes under five conditions (developmental stage, diet, temperature, tissue and sex). During the RT-qPCR analysis, geNorm software was used to evaluate pairwise variation in order to identify the most appropriate number of reference genes. The results revealed that for developmental stages RPL-13, EF1A and β-tubulin; for sex-related experiments β-tubulin and Actin, for tissue-related experiments PRS-18, 18S and RPL-13; for temperature-related experiments β-tubulin and PRL-13 and diet-related experiments, β-tubulin and PRS-18 are suitable reference genes. Furthermore, the associated HSP-70 (as the reporter gene) expression patterns differed significantly when the three most stable and three least stable reference genes were selected in different temperature treatments. This is the first time that a complete list of standardized reference genes has been given for P. fuscipes to use in an RT-qPCR study, which could be helpful for potential functional studies of target genes in P. fuscipes. |
| first_indexed | 2024-12-22T20:56:59Z |
| format | Article |
| id | doaj.art-0ab1087821ca45f2b83e6a3bdab76fdd |
| institution | Directory Open Access Journal |
| issn | 1018-3647 |
| language | English |
| last_indexed | 2024-12-22T20:56:59Z |
| publishDate | 2022-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of King Saud University: Science |
| spelling | doaj.art-0ab1087821ca45f2b83e6a3bdab76fdd2022-12-21T18:12:56ZengElsevierJournal of King Saud University: Science1018-36472022-01-01341101654Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insectMuhammad Musa Khan0Chang-Fei Guo1Jing Peng2Ze-Yun Fan3Muhammad Hafeez4Daoud Ali5Kai Wang6Mohammed H.A. Almarzoug7Bao-Li Qiu8Key Laboratory of Bio-Pesticide Innovation and Application, Guangdong Province, South China Agricultural University, Guangzhou 510642, China; Engineering Research Center of Biocontrol, Ministry of Education Guangdong Province, Guangzhou 510640, China; Maoming Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Maoming 525000, China; Corresponding authors at: Key Laboratory of Bio-Pesticide Innovation and Application, Guangdong Province, South China Agricultural University, Guangzhou 510642, China.Key Laboratory of Bio-Pesticide Innovation and Application, Guangdong Province, South China Agricultural University, Guangzhou 510642, China; Engineering Research Center of Biocontrol, Ministry of Education Guangdong Province, Guangzhou 510640, ChinaKey Laboratory of Bio-Pesticide Innovation and Application, Guangdong Province, South China Agricultural University, Guangzhou 510642, China; Engineering Research Center of Biocontrol, Ministry of Education Guangdong Province, Guangzhou 510640, ChinaKey Laboratory of Bio-Pesticide Innovation and Application, Guangdong Province, South China Agricultural University, Guangzhou 510642, China; Engineering Research Center of Biocontrol, Ministry of Education Guangdong Province, Guangzhou 510640, ChinaState Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences Hangzhou 310021, ChinaDepartment of Zoology, Collage of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi ArabiaDepartment of Computational Medicine and Bioinformatics, Medical School, University of Michigan, Ann Arbor, MI 48109, USADepartment of Zoology, Collage of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi ArabiaKey Laboratory of Bio-Pesticide Innovation and Application, Guangdong Province, South China Agricultural University, Guangzhou 510642, China; Engineering Research Center of Biocontrol, Ministry of Education Guangdong Province, Guangzhou 510640, China; Maoming Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Maoming 525000, China; Corresponding authors at: Key Laboratory of Bio-Pesticide Innovation and Application, Guangdong Province, South China Agricultural University, Guangzhou 510642, China.Paederus fuscipes is a medically and agriculturally important insect all over the world. P. fuscipes cause not only a skin condition but is also an aggressive predator in different agro-ecosystems. Prior to performing RT-qPCR under a variety of conditions to investigate the function of target genes in P. fuscipes, it is essential to screen reference genes. However, no P. fuscipes reference gene(s) has yet been discovered. Using the RefFinder software package, which includes ΔCt, geNorm, NormFinder, and BestKeeper, we evaluated the stability of seven housekeeping genes of P. fuscipes under five conditions (developmental stage, diet, temperature, tissue and sex). During the RT-qPCR analysis, geNorm software was used to evaluate pairwise variation in order to identify the most appropriate number of reference genes. The results revealed that for developmental stages RPL-13, EF1A and β-tubulin; for sex-related experiments β-tubulin and Actin, for tissue-related experiments PRS-18, 18S and RPL-13; for temperature-related experiments β-tubulin and PRL-13 and diet-related experiments, β-tubulin and PRS-18 are suitable reference genes. Furthermore, the associated HSP-70 (as the reporter gene) expression patterns differed significantly when the three most stable and three least stable reference genes were selected in different temperature treatments. This is the first time that a complete list of standardized reference genes has been given for P. fuscipes to use in an RT-qPCR study, which could be helpful for potential functional studies of target genes in P. fuscipes.http://www.sciencedirect.com/science/article/pii/S1018364721003165Reference genePaederus fuscipesRefFinderqRT-PCR analysisGene expression |
| spellingShingle | Muhammad Musa Khan Chang-Fei Guo Jing Peng Ze-Yun Fan Muhammad Hafeez Daoud Ali Kai Wang Mohammed H.A. Almarzoug Bao-Li Qiu Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insect Journal of King Saud University: Science Reference gene Paederus fuscipes RefFinder qRT-PCR analysis Gene expression |
| title | Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insect |
| title_full | Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insect |
| title_fullStr | Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insect |
| title_full_unstemmed | Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insect |
| title_short | Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insect |
| title_sort | screening and validation of reference genes using in rt qpcr for gene expression studies in paederus fuscipes a medically and agriculturally important insect |
| topic | Reference gene Paederus fuscipes RefFinder qRT-PCR analysis Gene expression |
| url | http://www.sciencedirect.com/science/article/pii/S1018364721003165 |
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