Evaluation of reference genes for gene expression analysis by real-time quantitative PCR (qPCR) in three stingless bee species (Hymenoptera: Apidae: Meliponini)

Stingless bees are generalist pollinators distributed through the pantropical region. There is growing evidence that their wild populations are experiencing substantial decline in response to habitat degradation and pesticides. Policies for conservation of endangered species will benefit from studie...

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Veröffentlicht in:Scientific reports 2019-11, Vol.9 (1), p.17692-13, Article 17692
Hauptverfasser: Freitas, Flávia C. P., Depintor, Thiago S., Agostini, Lucas T., Luna-Lucena, Danielle, Nunes, Francis M. F., Bitondi, Márcia M. G., Simões, Zilá L. P., Lourenço, Anete P.
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container_title Scientific reports
container_volume 9
creator Freitas, Flávia C. P.
Depintor, Thiago S.
Agostini, Lucas T.
Luna-Lucena, Danielle
Nunes, Francis M. F.
Bitondi, Márcia M. G.
Simões, Zilá L. P.
Lourenço, Anete P.
description Stingless bees are generalist pollinators distributed through the pantropical region. There is growing evidence that their wild populations are experiencing substantial decline in response to habitat degradation and pesticides. Policies for conservation of endangered species will benefit from studies focusing on genetic and molecular aspects of their development and behavior. The most common method for looking at gene expression is real-time quantitative polymerase chain reaction preceded by reverse transcription (RT-qPCR) of the mRNA of interest. This method requires the identification of reliable reference genes to correctly estimate fluctuations in transcript levels. To contribute to molecular studies on stingless bees, we used Frieseomelitta varia , Melipona quadrifasciata , and Scaptotrigona bipunctata species to test the expression stability of eight reference genes ( act , ef1-α , gapdh , rpl32 , rps5 , rps18 , tbp , and tbp-af ) in RT-qPCR procedures in five physiological and experimental conditions (development, sex, tissues, bacteria injection, and pesticide exposure). In general, the rpl32 , rps5 and rps18 ribosomal protein genes and tpb-af gene showed the highest stability, thus being identified as suitable reference genes for the three stingless bee species and defined conditions. Our results also emphasized the need to evaluate the stability of candidate genes for any designed experimental condition and stingless bee species.
doi_str_mv 10.1038/s41598-019-53544-0
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subjects 38/77
631/1647/2017
631/337/572
Animals
Bees
Bees - classification
Bees - genetics
Bees - growth & development
Bees - microbiology
Biodegradation
Endangered species
Environmental degradation
Escherichia coli
Escherichia coli Infections - genetics
Escherichia coli Infections - microbiology
Fat Body
Female
Gene expression
Gene Expression - drug effects
Genes, Essential
Glyceraldehyde-3-phosphate dehydrogenase
Head
Humanities and Social Sciences
Larva - genetics
Male
multidisciplinary
Ovary
Pesticides
Pesticides - pharmacology
Pollinators
Polymerase chain reaction
Pupa - genetics
Real-Time Polymerase Chain Reaction - methods
Reverse transcription
Science
Science (multidisciplinary)
Sex
title Evaluation of reference genes for gene expression analysis by real-time quantitative PCR (qPCR) in three stingless bee species (Hymenoptera: Apidae: Meliponini)
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