Abstract

Juvenile hormones (JHs) play a crucial role in the development of honey bee (Apis mellifera) worker larvae. Juvenile hormone analogs (JHAs), insecticides widely used in pest control, have been reported to affect the health and survival of honey bee worker larvae. However, the molecular mechanisms of JHAs in the honey bee remain unclear. In this study, we treated honey bee worker larvae with pyriproxyfen, fenoxycarb, and methoprene, three different JHAs. We monitored the changes in the transcription of genes encoding major JH response enzymes (CYP15A1, CYP6AS5, JHAMT, and CHT1) using RT-qPCR and analyzed the transcriptome changes in worker larvae under JHA stress using RNA-seq. We found that the enrichment pathways differed among the treatment groups, but the classification of each pathway was generally the same, and fenoxycarb affected more genes and more pathways than did the other two JHAs. Notably, treatment with different JHAs in the honey bee changed the JH titers in the insect to various extents. These results represent the first assessment of the effects of three different JHAs on honey bee larvae and provide a new perspective and molecular basis for the research of JH regulation and JHA toxicity in the honey bee.

Highlights

  • The honey bee is widely distributed all over the world [1]

  • We selected four differentially expressed genes (DEGs) related to Juvenile hormone analogs (JHAs) stress, CHT1, CYP6AS5, CYP15A1, and juvenile hormone acid methyltransferase (JHAMT) genes, which were expressed in three different treatments (Table S2)

  • The results showed a similar trend of expression changes for RT-qPCR and RNA-Seq data, indicating that the RNA-Seq results were reliable (Figure 3), and the FPKM mean value of RNA-seq and RT-qPCR data had been shown in Tables 2 and 3

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Summary

Introduction

The honey bee is widely distributed all over the world [1]. It plays a crucial role in the pollination of crops [2,3], and a healthy and strong colony of honey bees well enhances the pollination area of local crops. Play a crucial role in the regulation of vitellogenesis, reproduction, and ovary development of insects [11,12,13] They function in insect migration, energy metabolism, and caste differentiation [14,15,16]. Other studies have suggested that JHA insecticides can affect the social activity of the honey bee and change its growth and development processes [25,26]. Genes such as cytochrome CYP6AS5 (CYP6AS5), chitinase (CHT1), cytochrome CYP15A1. With the wider use of JHA insecticides in pest control, it is becoming increasingly important to test the toxicity of pyriproxyfen, fenoxycarb, and methoprene in honey bees as well as their regulation and molecular mechanisms. The resulting transcriptome library may help to elucidate the regulation and molecular mechanisms of JHA on honey bees and screen the most effective JHA pesticide among these three JHAs in honey bees

Chemicals and Honey Bee Larvae Rearing
Extraction and Determination of JH Titers in Honey Bees
Transcriptomic Data Analysis
Functional Analyses of DEGs
Validation of Four DEGs
Statistics
Determination of JH Titers in Six-Day-Old Worker Larvae
Quality Analysis of Transcriptome Sequencing Results
Screening of DEGs
GO and KEGG
Validation of DEGs by Using RT-qPCR
Discussion
Findings
Methods
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