Dataset Information

Evaluation of reference genes for real-time quantitative PCR analysis in southern corn rootworm, Diabrotica undecimpunctata howardi (Barber).

ABSTRACT: Quantitative reverse transcription PCR (RT-qPCR) is one of the most efficient, reliable and widely used techniques to quantify gene expression. In this study, we evaluated the performance of six southern corn rootworm, Diabrotica undecimpunctata howardi (Barber), housekeeping genes (HKG), ?-actin (Actin), ?-tubulin (Tubulin), elongation factor 1 alpha (EF1?), glyceraldehyde-3 phosphate dehydrogenase (GAPDH), 40?S ribosomal protein S9 (RpS9) and ubiquitin-conjugating protein (Ubi), under different experimental conditions such as developmental stage, exposure of neonate and adults to dsRNA, exposure of adults to different temperatures, different 3^rd instar larva tissues, and neonate starvation. The HKGs were analyzed with four algorithms, including geNorm, NormFinder, BestKeeper, and delta-CT. Although the six HKGs showed a relatively stable expression pattern among different treatments, some variability was observed. Among the six genes, EF1? exhibited the lowest Ct values for all treatments while Ubi exhibited the highest. Among life stages and across treatments, Ubi exhibited the least stable expression pattern. GAPDH, Actin, and EF1? were among the most stable HKGs in the majority of the treatments. This research provides HKG for accurate normalization of RT-qPCR data in the southern corn rootworm. Furthermore, this information can contribute to future genomic and functional genomic research in Diabrotica species.

SUBMITTER: Basu S

PROVIDER: S-EPMC6656754 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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Publications

Evaluation of reference genes for real-time quantitative PCR analysis in southern corn rootworm, Diabrotica undecimpunctata howardi (Barber).

Basu Saumik S Pereira Adriano E AE Pinheiro Daniele H DH Wang Haichuan H Valencia-Jiménez Arnubio A Siegfried Blair D BD Louis Joe J Zhou Xuguo 'Joe' X' Vélez Ana Maria AM

Scientific reports 20190724 1

Quantitative reverse transcription PCR (RT-qPCR) is one of the most efficient, reliable and widely used techniques to quantify gene expression. In this study, we evaluated the performance of six southern corn rootworm, Diabrotica undecimpunctata howardi (Barber), housekeeping genes (HKG), β-actin (Actin), β-tubulin (Tubulin), elongation factor 1 alpha (EF1α), glyceraldehyde-3 phosphate dehydrogenase (GAPDH), 40 S ribosomal protein S9 (RpS9) and ubiquitin-conjugating protein (Ubi), under differen ...[more]

PMID: 31341190

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Project description:Abstract The EFSA Panel on Plant Health performed a pest categorisation of Diabrotica undecimpunctata howardi (Coleoptera: Chrysomelidae) for the EU. This subspecies occurs in North and Central America. Adults oviposit on annual plants in the families Asteraceae, Chenopodiaceae, Cucurbitaceae, Fabaceae, Poaceae, Polygonaceae and Solanaceae. Adults feed on tender plant parts in hosts in 40 additional botanical families. Preimaginal development takes place on the roots of the host plant, where larvae feed and pupate. D. undecimpunctata howardi is a multivoltine species. Overwintering adults, which may enter a facultative diapause, abandon crops in autumn and reinvade them in spring. D. undecimpunctata howardi is not known to occur in the EU and is regulated in Annex IIA of Commission Implementing Regulation 2019/2072. This species is a competent vector of Erwinia tracheiphila (Smith) Bergey et al., which can cause bacterial wilt, a serious disease of cucurbits. The bacterium, which is restricted to temperate midwestern and eastern North America, is not regulated in the EU. Within Commission Implementing Regulation 2019/2072, potential entry pathways for D. undecimpunctata howardi, such as Asteraceae, Poaceae and Solanaceae plants for planting with foliage and soil/growing medium, and soil/growing media by themselves can be considered as closed. However, plants for planting of the families Chenopodiaceae, Cucurbitaceae, Fabaceae and Polygonaceae are not specifically regulated. Should D. undecimpunctata howardi arrive in the EU, climatic conditions and availability of susceptible hosts provide conditions suitable for establishment and further spread. Economic impact is anticipated in maize and outdoor cucurbit production. D. undecimpunctata howardi satisfies the criteria that are within the remit of EFSA to assess for this species to be regarded as a potential Union quarantine pest. This species does not meet the criteria of being present in the EU, nor plants for planting being the main pathway for spread, for it to be regarded as a potential regulated non?quarantine pest.

Dataset Information

Evaluation of reference genes for real-time quantitative PCR analysis in southern corn rootworm, Diabrotica undecimpunctata howardi (Barber).

Publications

Evaluation of reference genes for real-time quantitative PCR analysis in southern corn rootworm, Diabrotica undecimpunctata howardi (Barber).

Similar Datasets

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