{"title":"Sclerodermus guani (Hymenoptera: Bethylidae) RT-qPCR 分析参考基因的鉴定和验证。","authors":"Rina Zhao, Xiaomeng Guo, Ling Meng, Baoping Li","doi":"10.1017/S0007485324000427","DOIUrl":null,"url":null,"abstract":"<p><p>Gene expression studies in organisms are often conducted using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), and the accuracy of RT-qPCR results relies on the stability of reference genes. We examined ten candidate reference genes in <i>Sclerodermus guani</i>, a parasitoid wasp that is a natural enemy of long-horned beetle pests in forestry, including <i>ACT, EF1α, Hsc70, Hsp70, SRSF7, α-tubulin, RPL7A, 18S, 28S,</i> and <i>SOD1</i>, regarding variable biotic and abiotic factors such as body part, life stage, hormone, diet, and temperature. Data were analysed using four dedicated algorithms (ΔCt, BestKeeper, NormFinder, and geNorm) and one comparative tool (RefFinder). Our results showed that the most stable reference genes were <i>RPL7A</i> and <i>EF1α</i> regarding the body part, <i>SRSF7</i> and <i>Hsc70</i> regarding the diet, <i>RPL7A</i> and <i>α-tubulin</i> regarding the hormone, <i>SRSF7</i> and <i>RPL7A</i> regarding the life stage, and <i>SRSF7</i> and <i>α-tubulin</i> regarding temperature. To ascertain the applicability of specific reference genes, the expression level of the target gene (<i>ACPase</i>) was estimated regarding the body part using the most stable reference genes, <i>RPL7A</i> and <i>EF1α</i>, and the least stable one, <i>SOD1</i>. The highest expression level of <i>ACPase</i> was observed in the abdomen, and the validity of <i>RPL7A</i> and <i>EF1α</i> was confirmed. This study provides, for the first time, an extensive list of reliable reference genes for molecular biology studies in <i>S. guani</i>.</p>","PeriodicalId":9370,"journal":{"name":"Bulletin of Entomological Research","volume":" ","pages":"613-621"},"PeriodicalIF":1.6000,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Identification and validation of reference genes for RT-qPCR analysis in <i>Sclerodermus guani</i> (Hymenoptera: Bethylidae).\",\"authors\":\"Rina Zhao, Xiaomeng Guo, Ling Meng, Baoping Li\",\"doi\":\"10.1017/S0007485324000427\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Gene expression studies in organisms are often conducted using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), and the accuracy of RT-qPCR results relies on the stability of reference genes. We examined ten candidate reference genes in <i>Sclerodermus guani</i>, a parasitoid wasp that is a natural enemy of long-horned beetle pests in forestry, including <i>ACT, EF1α, Hsc70, Hsp70, SRSF7, α-tubulin, RPL7A, 18S, 28S,</i> and <i>SOD1</i>, regarding variable biotic and abiotic factors such as body part, life stage, hormone, diet, and temperature. Data were analysed using four dedicated algorithms (ΔCt, BestKeeper, NormFinder, and geNorm) and one comparative tool (RefFinder). Our results showed that the most stable reference genes were <i>RPL7A</i> and <i>EF1α</i> regarding the body part, <i>SRSF7</i> and <i>Hsc70</i> regarding the diet, <i>RPL7A</i> and <i>α-tubulin</i> regarding the hormone, <i>SRSF7</i> and <i>RPL7A</i> regarding the life stage, and <i>SRSF7</i> and <i>α-tubulin</i> regarding temperature. To ascertain the applicability of specific reference genes, the expression level of the target gene (<i>ACPase</i>) was estimated regarding the body part using the most stable reference genes, <i>RPL7A</i> and <i>EF1α</i>, and the least stable one, <i>SOD1</i>. The highest expression level of <i>ACPase</i> was observed in the abdomen, and the validity of <i>RPL7A</i> and <i>EF1α</i> was confirmed. This study provides, for the first time, an extensive list of reliable reference genes for molecular biology studies in <i>S. guani</i>.</p>\",\"PeriodicalId\":9370,\"journal\":{\"name\":\"Bulletin of Entomological Research\",\"volume\":\" \",\"pages\":\"613-621\"},\"PeriodicalIF\":1.6000,\"publicationDate\":\"2024-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Bulletin of Entomological Research\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1017/S0007485324000427\",\"RegionNum\":3,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/10/7 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"ENTOMOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bulletin of Entomological Research","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1017/S0007485324000427","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/10/7 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"ENTOMOLOGY","Score":null,"Total":0}
Identification and validation of reference genes for RT-qPCR analysis in Sclerodermus guani (Hymenoptera: Bethylidae).
Gene expression studies in organisms are often conducted using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), and the accuracy of RT-qPCR results relies on the stability of reference genes. We examined ten candidate reference genes in Sclerodermus guani, a parasitoid wasp that is a natural enemy of long-horned beetle pests in forestry, including ACT, EF1α, Hsc70, Hsp70, SRSF7, α-tubulin, RPL7A, 18S, 28S, and SOD1, regarding variable biotic and abiotic factors such as body part, life stage, hormone, diet, and temperature. Data were analysed using four dedicated algorithms (ΔCt, BestKeeper, NormFinder, and geNorm) and one comparative tool (RefFinder). Our results showed that the most stable reference genes were RPL7A and EF1α regarding the body part, SRSF7 and Hsc70 regarding the diet, RPL7A and α-tubulin regarding the hormone, SRSF7 and RPL7A regarding the life stage, and SRSF7 and α-tubulin regarding temperature. To ascertain the applicability of specific reference genes, the expression level of the target gene (ACPase) was estimated regarding the body part using the most stable reference genes, RPL7A and EF1α, and the least stable one, SOD1. The highest expression level of ACPase was observed in the abdomen, and the validity of RPL7A and EF1α was confirmed. This study provides, for the first time, an extensive list of reliable reference genes for molecular biology studies in S. guani.
期刊介绍：
Established in 1910, the internationally recognised Bulletin of Entomological Research aims to further global knowledge of entomology through the generalisation of research findings rather than providing more entomological exceptions. The Bulletin publishes high quality and original research papers, ''critiques'' and review articles concerning insects or other arthropods of economic importance in agriculture, forestry, stored products, biological control, medicine, animal health and natural resource management. The scope of papers addresses the biology, ecology, behaviour, physiology and systematics of individuals and populations, with a particular emphasis upon the major current and emerging pests of agriculture, horticulture and forestry, and vectors of human and animal diseases. This includes the interactions between species (plants, hosts for parasites, natural enemies and whole communities), novel methodological developments, including molecular biology, in an applied context. The Bulletin does not publish the results of pesticide testing or traditional taxonomic revisions.
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