een 1.8 and two.0 for RNA) along with the concentration of each and every RNA sample had been measured according to our prior study [56]. RNA samples were standardized for reverse transcription. cDNA was synthesized applying MLV reverse transcriptase (Takara, Japan), in accordance with the manufacturer’s directions. The -action gene was made use of as an internal housekeeping gene. A total of ten genes from RNA-Seq final results have been randomly chosen as target genes for qRT-PCR analysis (Fig. S2 and S3). Primers for these genes have been designed utilizing HSP90 site Primer five.0 application (Table S12). qRT-PCR cycling conditions have been as follows: 94 for 2 min, 40 cycles, followed by 94 for 15 s, 60 for 30 s, and 72 for 30 s. The specificity with the PCR products was verified by melt curve analysis for every single sample. For every gene, 3 biological replicates (with 5 technical replicates for every single biological replicate) had been performed. Control and target genes for each and every sample have been run on the similar plateAcknowledgements We thank Dr. Freddie Partridge and Prof. Qiang Huang for reviewing the paper.Liu et al. BMC Genomics(2021) 22:Page 11 ofAuthors’ contributions XJH, ZZJ and YBL conceived and created the experiments. YBL performed the experiments. YY and AA helped in experiments. YBL and YLZ analyzed the information. YBL and XJH wrote the paper. All authors study and approved the final manuscript. Funding This work was supported by the National All-natural Science Foundation of China (31702193), All-natural Science Foundation of cIAP Purity & Documentation Jiangxi province (20171BAB214018) and Key Research and Improvement Project of Jiangxi province (20181BBF60019). The funding bodies played no part in the style from the study and collection, evaluation, and interpretation of information and in writing the manuscript. Availability of information and components We confirm that all relevant information are integrated within the report and its supplementary information files. The raw information of honeybee larval meals and morphological information are uploaded on Dryad database: 5061/dryad.v9s4mw6v1. The transcriptome information of 18 samples are uploaded on SRA with accession numbers as comply with. RNA-Seq raw information of 3rd instar drone: WCs: NCBI SRA: SRR12031955; SRR12031954; SRR12031956. QCs: NCBI SRA: SRR12031970; SRR12031969; SRR12031960. DCs: NCBI SRA: SRR12031959; SRR12031958; SRR12031957. RNA-Seq raw data of adult drones: WCs: NCBI SRA: SRR12031953; SRR12031968; SRR12031967. QCs: NCBI SRA: SRR12031966; SRR12031965; SRR12031964. DCs: NCBI SRA: SRR12031963; SRR12031962; SRR12031961.DeclarationsEthics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare that they’ve no competing interests. Author particulars 1 Honeybee Research Institute, Jiangxi Agricultural University, 330045 Nanchang, China. 2Jiangxi Essential Laboratory of Honeybee Biology and Bee Maintaining, 330045 Nanchang, Jiangxi, China. 3Jiangxi University of Traditional Chinese Medicine, Jiangxi 330004 Nanchang, P. R. China. Received: 12 July 2020 Accepted: 15 SeptemberReferences 1. Skoko J. Environmental regulation of animal improvement. Theological Research. 2011;70(two):4370. 2. Burggren W, Dubansky B. Improvement and Atmosphere. Springer-Verlag press; 2018. three. Fusco G, Minelli A. Phenotypic plasticity in improvement and evolution: facts and concepts. Phil Trans R Soc B. 2010;365:5476. 4. Gilbert SF, Bosch TCG, Ledon-Rettig C. Eco-Evo-Devo: developmental symbiosis and developmental plasticity as evolutionary agents. Nat Rev Genet. 2015;16:6112. 5. Miyaz