{"title":"Identification and validation of stage-specific reference genes for gene expression analysis in Callosobruchus maculatus (Coleoptera: Bruchidae)","authors":"Gagandeep Singh Brar , Gurmeet Kaur , Satnam Singh , Jayendra Nath Shukla , Suneet Pandher","doi":"10.1016/j.gep.2022.119233","DOIUrl":null,"url":null,"abstract":"<div><p>In light of a number of recent studies highlighting the increasing research interest in bruchids, it is crucial to validate suitable reference genes that could be used in quantitative gene expression studies. <span><em>Callosobruchus maculatus</em></span> is a serious pest of stored grains and field legumes in which reference genes have not been assessed and validated to date. The present study aimed to identify and validate reference genes in different developmental stages of <em>C. maculatus</em> shortlisted from commonly used reference genes such as <span><span><span><em>VATPase</em><em>, TRIP12, </em></span><em>TBP</em><em>, TF11D, ACTIN, </em></span><em>GST</em><span><em>, ANNEXIN, PTCD3, </em><em>RPL32</em><em>,</em></span></span> and <em>β -Tub</em> in various insects. Dedicated algorithms like GeNorm, NormFinder, and BestKeeper were used to analyze the stability of these candidate genes, which revealed <em>GST</em> for third instar, <em>ANNEXIN</em> and <em>PTCD3</em> for the fourth instar, <em>TF11D</em> and <em>VATPase</em> for male pupa, <em>RPL32</em> and <em>β-tub</em> for female pupa, <em>β-tub</em> and <em>TBP</em> for adult male and <em>VATPase</em> and <em>GST</em> for adult females as suitable reference genes for expression studies in <em>C. maculatus.</em> The final comprehensive ranking using RefFinder identified <em>GST</em> and <em>TBP</em> as the best reference genes for all the developmental stages of <em>C. maculatus</em>. To the best of our knowledge, this is the first report which evaluates and validates stable reference genes in <em>C. maculatus</em>. The information of stage-specific gene expression, generated in this study will be useful for future molecular, physiological, and biochemical studies on <em>C. maculatus</em> and other closely related bruchids.</p></div>","PeriodicalId":55598,"journal":{"name":"Gene Expression Patterns","volume":null,"pages":null},"PeriodicalIF":1.0000,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Gene Expression Patterns","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1567133X22000035","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"DEVELOPMENTAL BIOLOGY","Score":null,"Total":0}
引用次数: 2
Abstract
In light of a number of recent studies highlighting the increasing research interest in bruchids, it is crucial to validate suitable reference genes that could be used in quantitative gene expression studies. Callosobruchus maculatus is a serious pest of stored grains and field legumes in which reference genes have not been assessed and validated to date. The present study aimed to identify and validate reference genes in different developmental stages of C. maculatus shortlisted from commonly used reference genes such as VATPase, TRIP12, TBP, TF11D, ACTIN, GST, ANNEXIN, PTCD3, RPL32, and β -Tub in various insects. Dedicated algorithms like GeNorm, NormFinder, and BestKeeper were used to analyze the stability of these candidate genes, which revealed GST for third instar, ANNEXIN and PTCD3 for the fourth instar, TF11D and VATPase for male pupa, RPL32 and β-tub for female pupa, β-tub and TBP for adult male and VATPase and GST for adult females as suitable reference genes for expression studies in C. maculatus. The final comprehensive ranking using RefFinder identified GST and TBP as the best reference genes for all the developmental stages of C. maculatus. To the best of our knowledge, this is the first report which evaluates and validates stable reference genes in C. maculatus. The information of stage-specific gene expression, generated in this study will be useful for future molecular, physiological, and biochemical studies on C. maculatus and other closely related bruchids.
期刊介绍:
Gene Expression Patterns is devoted to the rapid publication of high quality studies of gene expression in development. Studies using cell culture are also suitable if clearly relevant to development, e.g., analysis of key regulatory genes or of gene sets in the maintenance or differentiation of stem cells. Key areas of interest include:
-In-situ studies such as expression patterns of important or interesting genes at all levels, including transcription and protein expression
-Temporal studies of large gene sets during development
-Transgenic studies to study cell lineage in tissue formation