Artificial Rearing Of The Diploid Honey Bee(Apis Cerana Cerana) Drones, Cannibalism Substance Identification And Sequencing Of Transcriptome

Using the Apis mellifera diploid drone cultivation technology and indoor artificiallybreeding larvae, this study has firstly nurtured Apis cerana cerana diploid drones.Morphological index of cultivated drones was measured and their cell ploidy wasdetermined using flow cytometry instrument. At the same time, the cannibalism substanceof larvae hatched within12h were analysed by gas chromatography/mass spectrometry(GC/MS) technology. Comparing the differences between experimental samples andcontrol samples, we preliminarily identified the composition of cannibalism pheromones.In addition, the transcriptome differences between diploid drones and haploid drones weredetected using high-throughput RNA-Seq.Queens mated with drones of sexual maturity by artificial insemination. Then the larvaehatched in12h were cultured by artificial technique. The results showed that there are tenpercent of drones survived. And then the cell ploidy of the drones were determined usingflow cytometry instrument. We found that92%of drones are diploid.Using gas chromatography/mass spectrometry (GC/MS) technology, the pheromonecomponents of the larvae hatched within12h were determined. There were5kinds ofcomposition only existing in all experimental samples, and they were not detected incontrol samples and blank samples. Therefore, the lack of these compositions may be oneof the causes of diploid drone larvae ingested.Transcriptome differences of diploid and haploid drones were detected using RNA-Seq.The results suggest that the Q20bases ratio of all the samples were more than97%and allthe reads were assembled into59,916uniques with a mean length of1357bp. Based onsequence similarity search against the three public databases (NR, GO, KEGG) with acut-off E-value of10-5using BLASTX, a total of24,625unigenes were annotated. Usingthe Apis cerana cerana transcriptome data as reference sequences, the gene expressiondifference was analysed between diploid and haploid drones by a tag-based digital geneexpression method. About7.50million tags were obtained from the haploid drones and4.44million tags from the diploid drones. A total of649genes were detected differentiallyexpressed between them, with507up-regulated and142down-regulated in the haploiddrones compared to the diploid drones.