Ribosomal protein L7 as a suitable reference gene for quantifying gene expression in gastropod Bellamya aeruginosa

Liu, Qing; Lei, Kun; Ma, Qingqing; Qiao, Fei; Li, Zicheng; An, Lihui

doi:10.1016/j.etap.2016.03.007

Cited by 8 publications

(4 citation statements)

References 24 publications

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“…It is also one of the genes that displayed stable expression across seasons. However it did not behave ideally in all data sets, a result that agrees with differences in BACT expression between tissues and treatments in other molluscs [ 20 , 21 , 22 , 23 , 24 ]. Our results also identified the traditional housekeeping gene UBI as a good candidate reference gene in multiple data sets in both spring, autumn and spring + autumn (e.g.…”

Section: Discussionsupporting

confidence: 59%

“…Because the literature concerning the identification of stable reference genes in molluscs is extremely small we can compare our results to these previous efforts. In a study testing eight reference genes in the pond snail ( Bellamya aeruginosa ) the highly expressed ribosomal protein L7 gene was identified as the most stably expressed [ 24 ]. However several studies in diverse animals have demonstrated that ribosomal genes are not reliable candidates for stable expression [ 20 , 21 , 38 ], therefore we did not consider them for investigation here.…”

Section: Discussionmentioning

confidence: 99%

“…To date, only a handful of molluscan species have had reference genes tested for stable expression [ 20 , 21 , 22 , 23 , 24 ], and in C . nemoralis this has never been performed.…”

Section: Introductionmentioning

confidence: 99%

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Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

Affenzeller

Cerveau

2018

PLoS ONE

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Identifying and understanding mechanisms that generate phenotypic diversity is a fundamental goal of evolutionary biology. With a diversity of pigmented shell morphotypes governed by Mendelian patterns of inheritance, the common grove snail Cepaea nemoralis (Linnaeus, 1758) has been a model for evolutionary biologists and population geneticists for decades. However, the genetic mechanisms by which C. nemoralis generates this pigmented shell diversity remain unknown. An important first step in investigating this pigmentation pattern is to establish a set of validated reference genes for differential gene expression assays. Here we have evaluated eleven candidate genes for reverse transcription quantitative polymerase chain reaction (qPCR) in C. nemoralis. Five of these were housekeeping genes traditionally employed as qPCR reference genes in other species, while six alternative genes were selected de novo from C. nemoralis transcriptome data based on the stability of their expression levels. We tested all eleven candidates for expression stability in four sub-adult tissues of C. nemoralis: pigmented mantle, unpigmented mantle, head and foot. We find that two commonly employed housekeeping genes (alpha tubulin, glyceraldehyde 3-phosphate dehydrogenase) are unsuitable for use as qPCR reference genes in C. nemoralis. The traditional housekeeping gene UBIquitin on the other hand performed very well. Additionally, an RNA-directed DNA polymerase (RNAP), a Potassium Channel Protein (KCHP) and a Prenylated Rab acceptor protein 1 (PRAP), identified de novo from transcriptomic data, were the most stably expressed genes in different tissue combinations. We also tested expression stability over two seasons and found that, although other genes are more stable within a single season, beta actin (BACT) and elongation factor 1 alpha (EF1α) were the most reliable reference genes across seasons.

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Section: Discussionsupporting

confidence: 59%

Section: Discussionmentioning

confidence: 99%

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Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

Affenzeller

Cerveau

2018

PLoS ONE

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“…Previous research has shown that genes encoding ribosomal proteins consistently display stable expression and are widely used as reference genes in molecular studies of insects in the past decade [47]. The ribosomal protein family, including RPS23 and RPL32, is also identified as the most suitable reference genes in insects [30,46] and other organisms [48][49][50], and it was ranked sixth by the ICG website (https://ngdc.cncb.ac.cn/icg/, accessed on 12 October 2023). In the present study, RPS23 was assessed as the most stable gene in both nymph age and temperature treatments.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in the Bean Bug, Riptortus pedestris (Hemiptera: Alydidae)

Wang,

Liu,

Guo

et al. 2023

Insects

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Quantitative real-time PCR (qRT-PCR) is widely accepted as a precise and convenient method for quantitatively analyzing the expression of functional genes. The data normalization strongly depends upon stable reference genes. The bean bug, Riptortus pedestris (Hemiptera: Alydidae), is a significant pest of leguminous crops and broadly distributed across Southeast Asia. In this study, a total of 16 candidate reference genes (RPL32, RPS23, SDHA, UBQ, UCCR, GST, TATA−box, HSP70, GAPDH, RPL7A, SOD, RPS3, Actin, α−tubulin, AK, and EF1) were carefully chosen in R. pedestris, and their expression levels were assessed across various conditions, including different developmental stages, diverse tissues, temperature treatments, adult age, molting time, and mating status. Following this, the stability of these reference genes was evaluated using four algorithms (ΔCt, GeNorm, NormFinder, and BestKeeper). Ultimately, the comprehensive rankings were determined using the online tool RefFinder. Our results demonstrate that the reference gene for qRT-PCR analysis in R. pedestris is contingent upon the specific experimental conditions. RPL7A and EF1 are optimal reference genes for developmental stages. Furthermore, α−tubulin and EF1 exhibit the most stable expression across various adult tissues. RPL32 and RPL7A exhibit the most stable expression for adult age. For nymph age, RPL32 and SOD display the most stable expression. For temperature conditions, RPS23 and RPL7A were identified as the most suitable for monitoring gene expression. Lastly, we verified the practicability of evaluating expression levels of odorant-binding protein 37 (RpedOBP37) and cytochrome P450 6a2 (RpedCYP6) throughout developmental stages, tissues, and temperature conditions. These findings are a significant addition to the qRT-PCR analysis studies on R. pedestris, serving as a fundamental groundwork for future investigations on stable reference genes in R. pedestris as well as other organisms.

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Genome-Wide Identification and Evaluation of New Reference Genes in Pineapple (Ananas comosus L.) during Stamen and Ovule Development

Jin

Hou

Zhao

et al. 2020

Tropical Plant Biol.

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Ribosomal protein L7 as a suitable reference gene for quantifying gene expression in gastropod Bellamya aeruginosa

Cited by 8 publications

References 24 publications

Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in the Bean Bug, Riptortus pedestris (Hemiptera: Alydidae)

Genome-Wide Identification and Evaluation of New Reference Genes in Pineapple (Ananas comosus L.) during Stamen and Ovule Development

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