| Honeybees are insects with complete metamorphosis, in which an individual passes through 4 developmental stages:egg, larva, pupa and adult. Pupa stage is quiescent white that superficially looks adult bees and the way it raised influence its adult future undertakings. There are some important organs and glands in the head such as hypopharyngeal gland, mandibular gland, salivary gland, antenna, brain and mushroom body.We investigated the protein profiling of pupae head from the higher royal jelly producing bees (RJB) (Apis mellifera L.) and the native Italian bees (ITB) (Apis mellifera L.) at day13, day15, day17, day19 and day20 using 2-DE and differential expressions by mass spectrometry. Aiming to illuminate the molecular mechanism of higher royal jelly producing and development regulation and provide the theoretic foundation for utilizing and breeding the higher royal jelly producing bees, we classify the proteins and carry out functional enrichment, proteins interaction net and cluster analysis.We tentatively concluded that royal jelly bee keep on the similar growth curve with its counterpart Italian honey bee even its head weight is significantly higher than Italian bees, that may be induced by the morphological changes of hypopharyngeal gland.Analysis on the proteome in the pupal head from ITB show that there are 57 proteins differentially expressed between five time points that were categorized into 11 biologically functional groups. Large numbers of proteins were upregulated (25 of 57) at early developmental stage of the pupae (13 days), five of them were from carbohydrate metabolism and energy production,3 from development related proteins,4 from protein folding,3 from cytoskeleton,4 from protein biosynthesis,1 from amino acid,3 from transporter,2 from nucleotide metabolizing.Eleven proteins were upregulated at middle developmental stage,2 of them were from carbohydrate metabolism and energy production,3 from development related proteins,2 from cytoskeleton,2 from protein biosynthesis,2 from amino acid. Only 21 proteins were upregulated at the late stage (19-20 days). Five of them were from carbohydrate metabolism and energy production,5 from development related proteins,3 from protein folding,4 from cytoskeleton,1 from protein biosynthesis,1 from transporter,1 from antioxidant,1 from fat acid metabolism. The results indicate that most of early stage to middle stage up regulated proteins participated in organogenesis and hence pupae head have shown significant growth, while late stage developmental stage expressed proteins contributed more to the development of neuron and glands functions. It show that the head of honeybee have been well prepared for behaviors of collecting pollen, secreting royal jelly and defense by identifying odor prior to full maturation.Analysis on the proteome in the pupal head from RJB show that 49 proteins differentially expressed between five time points. There are 8,17 and 24 proteins upregulated at early, middle and late stage respectively. Organogenesis and differentiation are at the early stage, cytoskeleton, protein folding, cell division, cell cycle, cell multiplication proteins were upregulated to meet the requirement of cell growth and increment. Proteins upregulated at middle stage participated in ontogenesis of neuron, nerve axon, differentiation of visual system cell and pigmentation of compound eyes and cuticle. The developmental proteins related to function improvement of nervous system, olfactory system, odor guidance behavior, visual system and glands were upregulated at the late stage. The developmental regulation in the head from RJB is consistent with that from ITB.The total number of proteins detected at every gel from five time points is almost 330 by comparing the proteome profiles of two strains. This number is more in pupal head of RJB than that of ITB at every different day that this experiment determinated.18,28,33,25 and 24 proteins differentially expressed at day13, day15,day17, day19 and day20, respectively. The results demonstrated that (1) the number of upregulated proteins which involve the metabolism of carbohydrate, energy, fat acid, amino acid, protein biosynthesis, antioxidant activities and transporter from the pupal head of RJB were more than that of ITB at the late stage. Most of upregulated proteins at late stage is in relation to the development of nerve system, olfactory system and gland function. The difference in royal jelly producing performance attributed to the proteins differentially expressed at late stage. This show that the sensitivity of nerve centre in the head of RJB is higher than that of RJB. when the sensitivity of the pheromone from larvae in the cell increase it can stimulate RJB to secrete royal jelly. This indicates that RJB is prepared for increasing the product of royal jelly. (2) The proteins related to metabolism and protein biosynthesis were upregulated in the pupal head of ITB, and development proteins were upregulated in that of ITB. This induce ITB to be good at foraging nectar and RJB to do well in producing royal jelly. |
