Reference genes for gene expression studies by RT-qPCR in Brevipalpus yothersi (Acari: Tenuipalpidae), the mite vector of citrus leprosis virus

Rogerio, Luana Aparecida; Galdeano, Diogo Manzano; Arena, Gabriella Dias; Nunes, Maria Andréia; Machado, Marcos Antônio; Novelli, Valdenice M.

doi:10.1038/s41598-019-42993-2

Cited by 7 publications

(3 citation statements)

References 55 publications

(72 reference statements)

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“…Consistent with our findings, reference genes involving ribosomal protein genes have been selected frequently for expression analysis in various insects during the last 10 years ( Zhang et al, 2022 ; Shen et al, 2022 ). They are selected as reference genes in Coleopterans Ips sexdentatus ( RPS3 ) ( Sellamuthu et al, 2021 ), Tribolium castaneum ( RPS6 , RPL13a , RPS3 and RPL18 ) ( Toutges et al, 2010 ), L. decemlineata ( RP18 and RP4 ) ( Shi et al, 2013 ), H. vigintioctopunctata ( RPL13 and RPS18 ) ( Lü et al, 2018 ), H. vigintioctomaculata ( RPS18 and RPL13 ) ( Zhang et al, 2022 ) and P. brassicae ( RPL32 and RPL19 ) ( Ma et al, 2021 ), Lepidopteran species Helicoverpa armigera ( RPS15 and RPL27 ) ( Zhang et al, 2015 ), Plutella xylostella ( RPS13 and RPS23 ) ( Fu et al, 2013 ) and P. operculella ( RPL13 ) ( Shen et al, 2022 ), Thysanopterans Frankliniella occidentalis ( RPL32 ) ( Zheng et al, 2014 ), Dipterans Chlorops oryzae ( RPS15 ) ( Tian et al, 2019 ) and A. aphidimyza ( RPL8 and RPS3 ) ( Shen et al, 2023 ), Hymenopterans such as Apis mellifera ( RPS5 and RPS18 ) ( Jeon et al, 2020 ) and Anastatus japonicus Ashmead ( RPL13 and RPS6 ) ( Liu et al, 2022 ), Hemipterans N. lugens ( RPL5 , RPS8 and RPL14 ) ( Zhang et al, 2022 ), Aphis glycines ( RPS9 ) ( Bansal et al, 2012 ), Dichelops melacanthus ( RPL9 and RPS23 ) ( Pinheiro et al, 2020 ), Diaphorina citri ( RPL7 ) ( Bassan et al, 2017 ) and Rhopalosiphum padi ( RPL13 , RPS6 and RPS18 ) ( Li et al, 2021 ), and Orthopteran Locusta migratoria ( RPL32 ) ( Yang et al, 2014 ), as well as Acari Brevipalpus yothersi ( RPL13 and RPL32 ) ( Rogerio et al, 2019 ).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of reference genes for quantitative expression analysis in Mylabris sibirica (Coleoptera, Meloidae)

Shen,

Tang,

et al. 2024

Front. Physiol.

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Mylabris sibirica is a hypermetamorphic insect whose adults feed on oilseed rape. However, due to a shortage of effective and appropriate endogenous references, studies on molecular functional genes in Mylabris sibirica, have been tremendously limited. In this study, ten internal reference genes (ACT, ARF1, AK, EF1α, GAPDH, α-TUB, RPL6, RPL13, RPS3 and RPS18) were tested and assessed under four selected treatments including adult ages, adult tissues, temperatures, and sex by RT-qPCR based on five methods (Ct value, geNorm, NormFinder, BestKeeper and RefFinder). Our findings showed that RPL6 and RPL13 were the most optimal internal reference gene combination for gene expression during various adult ages and under diverse temperatures; The combination of RPL6 and RPS18 was recommended to test gene transcription levels under different adult tissues. AK and RPL6 were the best reference genes in male and female adults. RPL6 and RPL13 were the most appropriate reference gene pair to estimate gene expression levels under four different tested backgrounds. The relative transcript levels of a uridine diphosphate (UDP)-N-acetylglucosamine-pyrophosphorylase (MsUAP), varied greatly according to normalization with the two most- and least-suited reference genes. This study will lay the basis for further molecular physiology and biochemistry studies in M. sibirica, such as development, reproduction, sex differentiation, cold and heat resistance.

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Section: Discussionmentioning

confidence: 99%

Evaluation of reference genes for quantitative expression analysis in Mylabris sibirica (Coleoptera, Meloidae)

Shen,

Tang,

et al. 2024

Front. Physiol.

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“…The result of refFinder demonstrated that GAPDH was the most reliable reference gene in different periods and different tissues of P. ternata (Table 2 ). GAPDH was commonly used RGs in many species and tissues 36 – 38 .…”

Section: Discussionmentioning

confidence: 99%

Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata

Fan

Dong

et al. 2022

Sci Rep

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Pinellia ternata (Thunb.) Breit. (Abbreviated as P. ternata). It is a commonly prescribed Chinese traditional medicinal herb for the treatment of phlegm, cough, and morning sick. Bulbil reproduction is one of the main reproductive methods of P. ternata. The accurate quantification of gene expression patterns associated with bulbil development might be helpful to explore the molecular mechanism involved in P. ternata reproduction. Quantitative real-time PCR was the most preferred method for expression profile and function analysis of mRNA. However, the reference genes in different tissues of P. ternata in different periods of bulbil development have not been studied in detail. In present study, the expression stability of eight candidate reference genes were determined with programs: geNorm, NormFinder, BestKeeper, and refFinder. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was identified as the top- rated reference gene in all samples of P. ternata, while different combinations of reference gene proved to be the most stable depending on development stage and tissue type. Furthermore, the reliability of GAPDH expression was verified by six P. ternata related genes in hormone and nutrient biosynthesis pathways, and the expression profiles of these genes were agreed with the results of RNA-seq digital gene expression analysis. These results can contribute to studies of gene expression patterns and functional analysis of P. ternata involved in bulbil development.

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“…It needs to be mentioned that this molecule, a component of the large ribosomal 60S unit, has been considered before as a reference gene for data normalization. Its high stability has been discussed for application to various tissues in, e.g., pig [30,31], rat [32], human [33] and chicken [34,35], and even in several invertebrates [36]. Notably, it was found to be stably expressed in several canine healthy and diseased, i.e., neoplastic, tissues [9,37,38], and was therefore proposed as a good reference gene.…”

Section: Discussionmentioning

confidence: 99%

Determination of novel reference genes for improving gene expression data normalization in selected canine reproductive tissues – a multistudy analysis

Nowak

Aslan²,

Kowalewski

2020

BMC Vet Res

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Background Real time RT-PCR (qPCR) is a useful and powerful tool for quantitative measurement of gene expression. The proper choice of internal standards such as reference genes is crucial for correct data evaluation. In female dogs, as in other species, the reproductive tract is continuously undergoing hormonal and cycle stage-dependent morphological changes, which are associated with altered gene expression. However, there have been few attempts published so far targeted to the dog aimed at determining optimal reference genes for the reproductive organs. Most of these approaches relied on genes previously described in other species. Large-scale transcriptome-based experiments are promising tools for defining potential candidate reference genes, but were never considered in this context in canine research. Results Here, using available microarray and RNA-seq datasets derived from reproductive organs (corpus luteum, placenta, healthy and diseased uteri) of dogs, we have performed multistudy analysis to identify the most stably expressed genes for expression studies, in each tissue separately and collectively for different tissues. The stability of newly identified reference genes (EIF4H, KDELR2, KDM4A and PTK2) has been determined and ranked relative to previously used reference genes, i.e., GAPDH, β-actin and cyclophillin A/PPIA, using RefFinder and NormFinder algorithms. Finally, expression of selected target genes (luteal IL-1b and MHCII, placental COX2 and VEGFA, and uterine IGF2 and LHR) was re-evaluated and normalized. All proposed candidate reference genes were more stable, ranked higher and introduced less variation than previously used genes. Conclusions Based on our analyses, we recommend applying KDM4A and PTK2 for normalization of gene expression in the canine CL and placenta. The inclusion of a third reference gene, EIF4H, is suggested for healthy uteri. With this, the interpretation of qPCR data will be more reliable, allowing better understanding of canine reproductive physiology.

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Reference genes for gene expression studies by RT-qPCR in Brevipalpus yothersi (Acari: Tenuipalpidae), the mite vector of citrus leprosis virus

Cited by 7 publications

References 55 publications

Evaluation of reference genes for quantitative expression analysis in Mylabris sibirica (Coleoptera, Meloidae)

Evaluation of reference genes for quantitative expression analysis in Mylabris sibirica (Coleoptera, Meloidae)

Selection of suitable candidate genes for mRNA expression normalization in bulbil development of Pinellia ternata

Determination of novel reference genes for improving gene expression data normalization in selected canine reproductive tissues – a multistudy analysis

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