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Reference genes selection for expression studies in Maconellicoccus hirsutus (Green) (Pseudococcidae: Hemiptera) under specific experimental conditions

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Abstract

Background

Maconellicoccus hirsutus is a destructive pest which causes severe losses of agricultural and horticultural crops. For the management of M. hirsutus, many insecticides have been used and it has been exposed to insufficient dosage or uneven spray coverage which resulted in the development of insecticide resistance. Xenobiotic metabolism can be better understood with the help of gene expression studies by unveiling the underlying molecular mechanisms. The qRT-PCR is the simplest method to analyse gene expression, however, it highly relies on suitable reference genes concerning the different experimental conditions.

Methods and results

We evaluated the stability of five reference genes in two sets of experimental conditions viz. developmental stages (nymphs and adults) and agrochemical stress (GA3 and Buprofezin sprayed) against M. hirsutus, using different softwares—NormFinder, geNorm, BestKeeper, and RefFinder. The study revealed that ATP51a and GAPDH can be used as reference genes for gene expression studies when exposed to Gibberellic acid. Additionally, the study revealed that the ideal pair of reference genes for data validation in M. hirsutus treated with Buprofezin was GAPDH and β-tubulin. The ideal reference gene combination for various developmental stages was found to be 28S and Actin.

Conclusion

According to the study, GAPDH can be utilized as a reliable reference gene in the agrochemical (GA3 and Buprofezin) exposure set. The genes can be utilized as a suitable reference for qRT-PCR gene expression studies of xenobiotic metabolism to understand the underlying molecular mechanism, which will help further to design suitable management strategies.

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Acknowledgements

We sincerely thank Dr Mei Zhang, Institute of Marine and Environmental Technology, University of Maryland Center for Environmental Science, China for providing the geNorm analysis software. KVNR acknowledges the Senior Research Fellowship from the Indian Council of Agricultural Research. The authors are grateful to the Director, ICAR-NBAIR, Bengaluru, India for the help and support & providing the required facilities for the research.

Funding

Funding information is not applicable. No funding was received.

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Authors and Affiliations

  1. Division of Genomic Resources, ICAR- National Bureau of Agricultural Insect Resources, Bengaluru, 560024, India

    K. V. Nagarjuna Reddy, Gandhi Gracy Ramasamy, Aditi Agrawal, Subhi Srivastava, Jyoti Pathak & Thiruvengadam Venkatesan

  2. Department of Entomology, College of Agriculture, IGKV, Raipur, 492012, India

    K. V. Nagarjuna Reddy & Dhanendra Kumar Rana

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  1. K. V. Nagarjuna Reddy
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Contributions

GRG, TV and DKR contributed to the study’s conception and design. AA and JP analysed the transcriptome data, provided the primer sequences and helped with primer designing. KVNR and SS performed material preparation, data collection, and analysis. The first draft of the manuscript was written by KVNR. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Gandhi Gracy Ramasamy.

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Reddy, K.V.N., Ramasamy, G.G., Agrawal, A. et al. Reference genes selection for expression studies in Maconellicoccus hirsutus (Green) (Pseudococcidae: Hemiptera) under specific experimental conditions. Mol Biol Rep 50, 1221–1230 (2023). https://doi.org/10.1007/s11033-022-08120-7

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