Abstract:

【Objective】 To investigation into the differentially expressed proteins between the higher royal jelly producing (HRJB) and the native Italian honeybee (NIB) (Apis mellifera L.), the proteome profilings of the worker pupal head of the two honeybee strains were compared on 3 time-points (day 13, 15, 17) for providing a theoretical principle of clarifing honeybee developmental biology. 【Method】 Two-dimensional gel electrophoresis (2-DE) was employed to establish the proteomic map of the worker pupal head of the two honeybee strains on 13, 15 and 17 days, then the protein expression abundance, molecular weight and pI was obtained. Part of differentially expressed proteins was identified by analysis of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/MS) and database research. 【Result】 A total of 279, 296 and 340 proteins were detected on 3 time-points in HRJB, and 268, 277 and 279 were observed in NIB, respectively. A significantly higher protein spot numbers were observed in HRJB than those in NIB. Two hundred and twelve proteins were presented in the worker pupal head of these two bees on day 13, 67 proteins were specific in the worker pupal head of HRJB and 56 proteins were exclusive to NIB. While on day 15, 203 shared proteins and 93, 74 unique proteins were detected in HRJB, NIB, respectively. Until day17, 246 proteins were resolved in the head of both bees strains in the pupal period, 94 and 33 proteins were found unique to the worker pupal head of the HRJB and NIB, respectively. During the determination of PMF, 11 proteins were unambiguously identified. Hierarchical cluster analysis divided the 11 proteins into two categories, i. g. tubulin, actin 88F, ATP synthase, glutathione S transferase S1, arginine kinase, heat shock protein, proteasome 2 subunit, vacuolar ATP synthase catalytic subunit A and eukaryotic translation initiation factor 5A were up-regulated their expression, while only peroxiredoxin 2 540 was down-regulated in the head during the pupal period in both strains. 【Conclusion】 The HRJB and NIB have significant differences in protein expression in the head during the pupal development. The increased numbers of total and specific proteins expressed in the HRJB compared with that in the NIB suggested the significant change in the pupal development in the two bee strains. Furthermore, altered expression of antioxidant, metabolism and cytoskeleton confirmed that the development and metabolism of salivary gland and hypopharyngeal gland are accelerated and the nevous system is restructured during the pupal period. Finally, the data of proteomic analysis have contributed to the well understanding of the function or roles of proteins towards the development of honeybee worker pupal head of each type.

Key words: native Italian honeybee')">native Italian honeybee, higher royal jelly producing honeybee, two-dimensional gel electrophoresis, proteome, pupae