Project description:Royal jelly has long been recognized as health food, with a high content of proteins. These proteins play important roles in honeybee caste and human health, but the proteomic analysis of those low-abundance proteins in royal jelly is always a challenge. Herein, we used the Osboren classification method to separate the royal jelly proteins of Xinjiang black bees, a sub-species of Apis mellifera mellifera, into various fractions. The globulin, ethanol-soluble protein and glutelin fractions were further separated by SDS-PAGE, and proteomic analysis was carried out by LC-MS/MS and searching against the NCBI database. A total of 63 proteins with definitive names were identified, in which 41 proteins were identified for the first time in royal jelly. The Osboren classification method combined with one-dimensional gel electrophoresis based proteomic analysis allows the identification of low-abundance proteins, and greatly extends the knowledge about the components and functions of royal jelly proteins.
Project description:The genome of the western honey bee (Apis mellifera) harbours ten different major royal jelly protein genes (mrjp1-10) which originate from a single-copy precursor via gene duplication. The evolutionary fate of duplicated genes is eventually determined over time as to result in loss due to pseudogenization, or in preservation due to neo- or sub-functionalization. Both fates were already observed in the mrjp gene cluster, as only mrjp1 - 9 are expressed, whereas mrjp10 was pseudogenized and represents an incomplete gene copy. In contrast, MRJP1 underwent neofunctionalization and developed an essential function within the food jelly of queen larvae, to guaranty the survival of the whole colony. We here show combining quantitative real time PCR with quantitative mass spectrometry that expression of most mrjps (mrjp1-5 and 7) shows an age dependent pattern in worker hypopharyngeal glands as well as in brains. Expression increases after hatching until the nurse bee period and is followed by a decrease in older workers that forage for different plant products. Mrjp6 expression deviates considerably from the expression profiles of the other mrjps and transcript abundance does not correlate with protein amount. Thus, either mrjp6 does fulfil a total different function or it might be on its way to pseudogenization. Furthermore, a tissue-specific function of the proteins MRJP8 and 9 in the hypopharyngeal glands and the brain can be excluded, suggesting a more general physiological than a nutritive function for both gene products.
Project description:Social caste determination in the honey bee is assumed to be determined by the dietary status of the young larvae and translated into physiological and epigenetic changes through nutrient-sensing pathways. We have employed microRNA gene-microarray, and observed that both worker jelly and royal jelly showed dynamic changes in miRNA content during the 4th to 6th day of larval development . Adding specific miRNAs to royal jelly elicited significant changes in queen larval mRNA expression and in morphological characters of the emerging adult queen bee. We propose that miRNAs in the nurse bee secretions constitute an additional element in the regulatory control of caste determination in the honey bee. We collected worker and royal jelly of the Italian honeybee (ZND No.1, Apis mellifera ligustica) at 73~90 hours (4th-day larvae), 97~114 hours (5th-day larvae), and 121~138 hours (6th-day larvae) after hatching. After total RNA was extractedM-BM- and quantified , equal amounts of total RNAs from each of the three sampling days were analyzed on the LC Science miRNA-array to observe the expression variation of miRNAs between worker jelly and royal jelly along with the development time points (4th-day, 5th-day and 6th-day).
Project description:Social caste determination in the honey bee is assumed to be determined by the dietary status of the young larvae and translated into physiological and epigenetic changes through nutrient-sensing pathways. We have employed Illumina/Solexa sequencing to examine the small RNA content in the bee larval food source, and show that worker jelly is enriched in miRNA complexity and abundance relative to royal jelly. The miRNA levels in worker jelly were 7-215 fold higher than in royal jelly, and both jellies showed dynamic changes in miRNA content during the 4th to 6th day of larval development. Adding specific miRNAs to royal jelly elicited significant changes in queen larval mRNA expression and in morphological characters of the emerging adult queen bee. We propose that miRNAs in the nurse bee secretions constitute an additional element in the regulatory control of caste determination in the honey bee. We collected worker and royal jelly of the Italian honeybee (ZND No.1, Apis mellifera ligustica) at 73~90 hours (4th-day larvae), 97~114 hours (5th-day larvae), and 121~138 hours (6th-day larvae) after hatching. After total RNA was extractedM-BM- and quantified , relative equal amounts of total RNAs from each of the three sampling days were pooled into respectively worker and royal jelly samples, and the fraction of small RNAs less than 30nt long was retained and sequenced on the Illumina/Solexa high-throughput platform (HTP).