Selection of Sclerodermus pupariae Reference Genes for Quantitative Real-Time PCR

被引：0
作者：
Zhou, Ting [1 ]
Feng, Huahua [1 ]
Zhang, Jie [2 ]
Tang, Yanlong [2 ]
Dong, Xiaoling [1 ]
Kang, Kui [2 ,3 ]
机构：
[1] Yangtze Univ, Coll Agr, MARA Key Lab Sustainable Crop Prod Middle Reaches, Jingzhou 434025, Peoples R China
[2] Zunyi Normal Univ, Coll Biol & Agr, Zunyi 563006, Peoples R China
[3] Hubei Univ Med, Biomed Res Inst, Shiyan 442000, Peoples R China
来源：
INSECTS | 2025年 / 16卷 / 03期
基金：
中国国家自然科学基金;
关键词：
S; pupariae; reference gene screening; qRT-PCR; developmental stages; AGRILUS-PLANIPENNIS COLEOPTERA; EXPRESSION; NORMALIZATION; HYMENOPTERA; BUPRESTIDAE; BETHYLIDAE; BIOLOGY;
D O I：
10.3390/insects16030268
中图分类号：
Q96 [昆虫学];
学科分类号：
摘要：
S. pupariae is a newly discovered species of parasitoid wasps. Research into its development, behavioral genetics, and molecular mechanisms provides valuable insights for improving integrated pest management strategies. Quantitative real-time PCR (qRT-PCR) is the most commonly used method for analyzing gene expression. This method requires the identification of stable reference genes to accurately evaluate transcriptional level variations. In this study, eight candidate reference genes (TUB, TBP, RPS18, GAPDH, 18S rRNA, RPL32, Actin, and EF1-alpha) were identified and evaluated for their suitability as reference genes. Gene expression levels across different developmental stages were analyzed using three software tools, GeNorm, NormFinder, and BestKeeper, and the online tool RefFinder. The overall ranking of reference gene stability was as follows: RPS18 > 18S rRNA > RPL32 > GAPDH > Actin > TUB > TPB > EF1-alpha. Ultimately, RPS18 was determined to be the most stable reference gene.
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页数：12
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共 52 条
[1]   Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets [J].
Andersen, CL ;
Jensen, JL ;
Orntoft, TF .
CANCER RESEARCH, 2004, 64 (15) :5245-5250
[2]   Venom Proteins from Endoparasitoid Wasps and Their Role in Host-Parasite Interactions [J].
Asgari, Sassan ;
Rivers, David B. .
ANNUAL REVIEW OF ENTOMOLOGY, VOL 56, 2011, 56 :313-335
[3]   Evaluation of reference genes for expression studies in the broad mite, Polyphagotarsonemus latus (Acari: Tarsonemidae) [J].
Augustine, Neenu ;
Selvapandian, Upasna ;
Venkatesan, Thiruvengadam ;
Srinivasa, Nagappa ;
Rao, Annabathula Mohan ;
Saraswathy, Benherlal Palayyan ;
Mohan, Muthugounder .
APPLIED ENTOMOLOGY AND ZOOLOGY, 2024, 59 (01) :31-40
[4]   Regulation of Translation Initiation by RNA Binding Proteins [J].
Babitzke, Paul ;
Baker, Carol S. ;
Romeo, Tony .
ANNUAL REVIEW OF MICROBIOLOGY, 2009, 63 :27-44
[5]   Protein-poor diet reduces host-specific immune gene expression in Bombus terrestris [J].
Brunner, Franziska S. ;
Schmid-Hempel, Paul ;
Barribeau, Seth M. .
PROCEEDINGS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES, 2014, 281 (1786)
[6]   Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays [J].
Bustin, SA .
JOURNAL OF MOLECULAR ENDOCRINOLOGY, 2000, 25 (02) :169-193
[7]   Validation of Reference Genes in Solenopsis invicta in Different Developmental Stages, Castes and Tissues [J].
Cheng, Daifeng ;
Zhang, Zhiling ;
He, Xiaofang ;
Liang, Guangwen .
PLOS ONE, 2013, 8 (02)
[8]   The molecular biology toolbox and its use in basic and applied insect science [J].
Cusson, Michel .
BIOSCIENCE, 2008, 58 (08) :691-700
[9]   Reference genes for Eucalyptus spp. under Beauveria bassiana inoculation and subsequently infestation by the galling wasp Leptocybe invasa [J].
Daude, Matheus Martins ;
Sagio, Solange Aparecida ;
Rodrigues, Jovielly Neves ;
Lima, Nivea Maria Pereira ;
Lima, Andre Almeida ;
Sarmento, Maira Ignacio ;
Sarmento, Renato Almeida ;
Barreto, Horllys Gomes .
SCIENTIFIC REPORTS, 2024, 14 (01)
[10]   RNA-seq validation: software for selection of reference and variable candidate genes for RT-qPCR [J].
de Brito, Marcio Wilson Dias ;
de Carvalho, Stephanie Serafim ;
Mota, Maria Beatriz dos Santos ;
Mesquita, Rafael Dias .
BMC GENOMICS, 2024, 25 (01)
123456