Dataset Information

Identification and validation of reference genes for quantitative real-time PCR in Drosophila suzukii (Diptera: Drosophilidae).

ABSTRACT: To accurately evaluate gene expression levels and obtain more accurate quantitative real-time RT-PCR (qRT-PCR) data, normalization relative to reliable reference gene(s) is required. Drosophila suzukii, is an invasive fruit pest native to East Asia, and recently invaded Europe and North America, the stability of its reference genes have not been previously investigated. In this study, ten candidate reference genes (RPL18, RPS3, AK, EF-1?, TBP, NADH, HSP22, GAPDH, Actin, ?-Tubulin), were evaluated for their suitability as normalization genes under different biotic (developmental stage, tissue and population), and abiotic (photoperiod, temperature) conditions. The three statistical approaches (geNorm, NormFinder and BestKeeper) and one web-based comprehensive tool (RefFinder) were used to normalize analysis of the ten candidate reference genes identified ?-Tubulin, TBP and AK as the most stable candidates, while HSP22 and Actin showed the lowest expression stability. We used three most stable genes (?-Tubulin, TBP and AK) and one unstably expressed gene to analyze the expression of P-glycoprotein in abamectin-resistant and sensitive strains, and the results were similar to reference genes ?-Tubulin, TBP and AK, which show good stability, while the result of HSP22 has a certain bias. The three validated reference genes can be widely used for quantification of target gene expression with qRT-PCR technology in D.suzukii.

SUBMITTER: Zhai Y

PROVIDER: S-EPMC4157791 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Publications

Identification and validation of reference genes for quantitative real-time PCR in Drosophila suzukii (Diptera: Drosophilidae).

Zhai Yifan Y Lin Qingcai Q Zhou Xianhong X Zhang Xiaoyan X Liu Tingli T Yu Yi Y

PloS one 20140908 9

To accurately evaluate gene expression levels and obtain more accurate quantitative real-time RT-PCR (qRT-PCR) data, normalization relative to reliable reference gene(s) is required. Drosophila suzukii, is an invasive fruit pest native to East Asia, and recently invaded Europe and North America, the stability of its reference genes have not been previously investigated. In this study, ten candidate reference genes (RPL18, RPS3, AK, EF-1β, TBP, NADH, HSP22, GAPDH, Actin, α-Tubulin), were evaluate ...[more]

PMID: 25198611

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Project description:The spotted wing drosophila (SWD), Drosophila suzukii Matsumura (Diptera: Drosophilidae), is an invasive polyphagous pest of soft-skinned fruit that has started to threaten small fruit production in Europe since 2008. High reproductive capacity, short generation time, and difficulties in visualizing early damage contribute to its rapid spread. Currently, the control strategy against D. suzukii mostly relies on treatment with synthetic insecticides. Keeping in mind that this pest causes the greatest damage during the harvesting period, control using chemicals is not recommended due to the increased risk of high pesticide residue levels in the fruit. With the aim of reducing the use of insecticides, there is a need for developing an environmentally safer way of control. Alternative solutions could rely on the use of essential oils (EOs), which can be used in conventional and organic production systems. Four essential oils, geranium (Pelargonium graveolens), dill (Anethum graveolens), Scots pine (Pinus sylvestris), and bergamot (Citrus bergamia), were assessed for their insecticidal effects using four different tests (contact toxicity, fumigant-contact toxicity, repellent effect, and multiple-choice tests). The EOs applied were dissolved in acetone at three different concentrations. The most promising one was the geranium essential oil, which had the best effect in all conducted tests, even at the lowest applied concentration. Geranium oil caused very high mortality (95%), even at the lowest concentration applied, after 24 h in the fumigant-contact test, and it caused a mortality of over 85% in the contact test. It was also noticed that geranium demonstrated a deterrent effect by repelling females from laying eggs for four days after at the lowest applied concentration. Scots pine and dill EOs have moderate to strong effects in most tests. The mortality of 100% was achieved for the highest applied concentration in the fumigant-contact and contact tests. Bergamot EO did not have any significant insecticidal activity. Geranium, Scots pine, and dill have great potential to be used as an environmentally friendly way of controlling D. suzukii as they exhibit deterrent, repellent, and insecticidal effects.

Dataset Information

Identification and validation of reference genes for quantitative real-time PCR in Drosophila suzukii (Diptera: Drosophilidae).

Publications

Identification and validation of reference genes for quantitative real-time PCR in Drosophila suzukii (Diptera: Drosophilidae).

Similar Datasets

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