Construction Of Genetic Linkage Map In Apis Cerana And Molecular Mechanism Of Development In Females Honey Bee

Honey bee is an importantly economic insect, who plays an important role inincreasing agricultural production and maintaining the ecological balance as the pollinator.In addition, honey bee has become a model organism for research on behavior plasticity,learning and memory.This study used Apis cerana cerana (ACC) as experimental materials to construct A.cerana family.126990single nucleotide polymorphic (SNP) locus were genotyped from103workers. After filtering low quality and those not passing the Mendel test, we obtained3000SNPs.1535of those were informative and used to construct the first A. ceranalinkage map. Results showed that this map contains1535markers spaning16linkagegroups. The total genetic distance is3942.7centimorgans (cM) and the largest linkagegroup (180locus) is574.5cM. The average marker interval of the entire16linkage groupsis2.6cM. A. cerana has high recombination rate (17.4cM/Mb) as well as A. mellifera.Furthermore, we analyzed the DNA methylomes of queen larvae (QL) and workerlarvae (WL) with different ages (2,4, and6days post hatching) from two single droneinseminated colonies (A. mellifera) by meDIP-sequencing. Our results showed that rates ofDNA methylation in WL increased with aging at all three time points, which increased firstand then decreased in QL. DNA methylation occurred mainly in intron. The number ofdifferentially methylated genes ranged from725to5049in QL and WL from2-to6-day-old larvae. We also identified and characterized the microRNAs (miRNAs) of4days old QL and WL from three single drone inseminated colonies (A. mellifera) byIllumina HiSeqTM2500. The results showed that the length of small RNAs in QL and WLwere clustered together in22nt-23nt.62,61known honey bee miRNAs were obtained inQL and WL, respectively. Compared with WL, there were20down-regulated and17up-regulated miRNAs in QL. These differentially methylated genes and differentiallyexpressed miRNAs were involved in development, metabolic regulation, reproduction,signal transduction and cellular structure.Moreover, we sequenced the miRNAs within royal jelly (RJ) from A. mellifera (RJM)and A. cerana (RJC) respectively. We then feed A. mellifera larvae with RJM and RJCseparately to compare the impact of the royal jelly origine on the lavae development byRNA-sequencing. We then determined the global transcriptomes of adult A. melliferadeveloped from larvae fed either with RJM (mRJM) or RJC (mRJC). Our results showedthat a high proportion (23.3%) of the affected genes was regulated by differentiallyexpressed miRNAs. Our results also demonstrated that4types of alternative splicing (exonskipping, intron retention, alternative5’splice site and alternative3’splice site) in adult A.mellifera were affected by the type of RJ (RJM and RJC). In the end, one-day-old larvae of A. mellifera ligustica and A. cerana cerana were fedwith RJM or RJC. The genome DNA was extracted from3-and6-day-old larvae to detectdynactin p62DNA methylation by Sequenom MassARRAY. Our results showed that thewhole DNA methylation of dynactin p62in female honey bee larvae decreasedsignificantly after feeding with heterospecific RJ. RJM and RJC had different effects onthe whole DNA methylated and methylated sites of dynactin p62in female honey beelarvae. These results indicate that different types of RJ have different biological effects.