| Hemolymph, flowing in the circulatory system of honeybees, not only transports ions, nutrients, hormones, but also acts as the body’s important immune defense barrier and monitors the body’s physiological states. The strain of royal jelly bees(RJB) which has high royal jelly yield, is selected from the Italian bee(ITB). However, researchs about high royal jelly producing mechanisms of physiological changes of RJB from hemolymph’ proteome and phosphoproteome have not been done till now.This experiment used mass-spectrometry-based proteome, phosphoproteome, bioinformatics method to analysis hemolymph proteome of larvae and adult bees of RJB and ITB. Of the 3395 nonredundant proteins, 2797 and 2470 were expressed in RJB and ITB respectively. Of the 1082 nonredundant phosphoproteins, 991 and 743 were expressed in RJB and ITB respectively. By analys ising the functions, metabolism pathways and interaction networks enrichment of proteins in larvae and adult bees of two species, we found that after breeding many years, despite the production of royal jelly is enhanced in RJB, proteinogram and phosphoproteinogram of RJB and ITB hemolymph from larvae to adult bees are matched. Larvae hemolymph proteins mainly enr iched in protein synthesis, folding, carbohydrate and energy metabolism, immunization and development related pathway,these made clear that it provided effective and efficient immunization, nutrition and energy supply to meet the need of larvae rapid body growth. The hemolymph phosphoproteins of larvae were mainly enriched in the functions of protein synthesis and protein phosphorylation, these indicated that phosphorylation modification play an important role in regulating the development of larvae. And larvae day 4 had a much more efficient translation function than other stages, it stated that development of larvae day 4 is crucial to the synthesis of hemolymph proteins. The proteins of adult bees were enriched in metabolism of energy and carbohydrate, signal transduction, immunity and metabolism of amino acids and nucleic acid, development of glands and motion system. And the phosphoproteins of adult bees were enriched in metabolism of energy, protein phosphorylation, immune defenses, glands, and motion system development process, these indicated that hemolymph proteins and phosphoproteins of adult bees supported the body development, but also met the need of bees feeding, collecting, cleaning and defense capabilities. Among these result, nurse hemolymph proteins were signif icantly enriched in more functions and activated more pathways showed that nurse bees had the most exuberant protein and energy metabolism to support the synthesis of royal jelly. Because of few overlap in hemolymph proteins and phosphoproteins and most of them expressed mutually independent, biological functions of hemolymph played different roles at the level of proteome and phosphoproteome.However, comparing RJB with ITB hemolymph proteome and phosphoproteome by different ages, we found that the larvae day 2 respectively had 128,50 differentally expressed proteins and phosphorproteins, including 49,79 up-regulated proteins and 41,9 up-regulated phosphorproteins in RJB and ITB separately. And at this age hemolymph proteins were enriched in protein synthesis and defense pathway, indicated that hemolymph provided proteins and immune protection for body development. During the larvae day 4, 358,51 differentally expressed proteins and phosphorproteins were identified, including 341,17 up-regulated proteins and 41,10 up-regulated phosphorproteins in RJB and ITB separately. In RJB hemolymph, up-regulated proteins enriched in translation function which suggested energy metabolism and protein synthesis were strengthened in RJB. While up-regulated proteins in ITB enriched in nucleotide metabolism, indicated ITB provided more nucleic acid materials for development. Two strains of bees in larvae day 4 have undergone signif icant difference in the hemolymph. During the larvae day 6, 232,52 differentally expressed proteins and phosphorproteins were identified, including 82,150 up-regulated proteins and 33,19 up-regulated phosphorproteins in RJ B and ITB separately. Up-regulated proteins and phosphorproteins in RJ B enriched in carbohydrates and energy metabolism indicated energy synthesis function were strengthen. While in ITB, up-regulated proteins and phosphorproteins enriched in protein synthesis and folding indicated protein biosynthesis function were strengthened. In newly emerged hemolymph, 173,41 differentally expressed proteins and phosphorproteins were identif ied, including 111,62 up-regulated proteins and 18,23 up-regulated phosphorproteins in RJB and ITB separately. RJB up-regulated proteins enriched in energy synthesis and ribosome metabolism pathway suggested RJB had stronger energy synthesis and protein synthesis ability. Meanwhile, ITB up-regulated proteins enriched in amino acid and sugar metabolism indicated amino acid and energy metabolism strengthen which met need of newly emerged development. In nurse hemolymph, 326,106 differentally expressed proteins and phosphorproteins were identified, including 297,29 up-regulated proteins and 99,7 up-regulated phosphorproteins in RJ B and ITB separately. RJB up-regulated proteins enriched in energy producing, phagosome, protein synthesis, phosphorylation pathway indicated energy metabolism, immune, protein synthesis, protein phosphorylation strengthen in RJB. Proteins of ITB hemolymph involved in nucleotide metabolism provided energy for nursing, but protein and energy metabolism were more vigorous in RJB. After breeding, RJB had more efficient protein synthesis system to ensure royal jelly high-yield. In forager, 77,42 differentally expressed proteins and phosphorproteins were identified, inc luding 31,46 up-regulated proteins and 14,28 up-regulated phosphorproteins in RJB and ITB separately. Proteins were involved in energy production in two strains of bees, but two strains of bees had different metabolism way. All above results showed that after long-term breeding of high yield royal jelly, the hemolymph proteome and phosphoproteome in RJ B and ITB have changed signif icantly. Two strains of bees have formed their own physiological characteristics of proteome and phosphoproteome features. Larvae day 4 of RJB significantly strengthened many metabolic pathways, especially in the hemolymph of nurse bees significantly strengthened hemolymph physiology in order to meet the physiological needs of high yield royal jelly. |
PDF Full Text Request