Comparative Analysis Of Differentially Expressed Proteins And Phosphoproteins Of Drone Embryos Between Apis Mellifera Ligustica And Apis Cerana Cerana

Apis mellifera ligustica and Apis cerana cerana are two main bee species for beekeeping industry in China. Honeybee drone has an important contribution to the genetic makeup of the honeybee colony. To investigate into the characteristic of embryonic development and gain an indepth understanding of the developmental biology of the drone embryos of Apis mellifera ligustica and Apis cerana cerana, the proteome and phosphoproteomic during the three days development were compared. This will lay foundation for the future genetic improvement on honeybees. Two-dimensional gel electrophoresis (2-DE) was employed to compare the proteomic and phosphoproteomic differences between the drone embryogenesis of the two bee species at three time-points of24,48and72h, respectively. Level of protein expression, pI and molecular weight were determined. Furthermore, some of differentially expressed proteins were analyzed by mass spectrometry and bioinformatics.In total proteome,304,316and309protein spots were detected in drone embryos of Apis mellifera ligustica from day1to day3. Of these,103,110and106proteins were phosphorylated, respectively, In the drone embryos of Apis cerana cerana,289,311and306proteins were detected, in which92,98and96proteins were phosphorylated. Meanwhile,243,251and245proteins were commonly expressed during the three days of drone embryonic development. A larger number of proteins and phosphoproteins were expressed in Apis mellifera ligustica than those in Apis cerana cerana during the drone embryogenesis, indicating Apis mellifera ligustica needs more protein to regulate its drone embryonic development. About1/3protein phosphorylated in both honeybee embryos might be associated with signal transduction, cell proliferation and differentiation during drone embryogenesis.The results of analysing the differentially expressed proteins showed that103,110,106were highly regulated in Apis mellifera ligustica’s drone embryonic development and83,97,89in Apis cerana cerana. From those proteins,30,50,42 were identified in Apis mellifera ligustica and24,40,34in Apis cerana cerana. For the differentially expressed phosphoproteins,33,39,39were highly regulated in Apis mellifera ligustica’s drone embryonic development and27,34,30in Apis cerana cerana. From those proteins,12,15,14were identified in Apis mellifera ligustica and9,13,12in Apis cerana cerana. The identified proteins were involved in development, carbohydrate metabolism and energy production, amino acid, fatty acid and nucleotide metabolism, protein biosynthesis, Antioxidant system, protein folding and cytoskeletal.Function analysis indicates that in the long-term evolutionary process has shaped the two honeybee species perusing their own embryonic development ways to fit their biological characteristics. Some of important biological characteristics, such as Apis mellifera ligustica has higher production of honey and royal jelly, and Apis cerana cerana is good at collecting sporadic nectar and pollen and defending wasps have been partially unraveled by this proteome dataset. Furthermore, the network analysis of identified protein show that development, carbohydrate metabolism and energy production were highly connected in the network, and followed by proteins which related to the metabolism of amino acids, fatty acids and nucleotides, indicating that they are key node proteins that play a central role in drone embryonic development. These differentially expressed key proteins in two species may be associated with some of their specific traits, which provide the protein in two species for genetic modification.