Research On The Effect Of The Instar Of The Grafted Larvae On Growth And Development And Ovary Proteomic Of Queen

Suitable instar of larvae is seen as key to queen artificial cultivation. Although the most focusing on cast differentiation and its molecular mechanism caused by different larvae about 3 days of age has been addressed, little is known about the efforts of grafting different instar of younger larva on the queen reproductive development.To reveal the molecular regulation mechanism of the effect of the instar of the grafted larvae on queen growth and development, the JHIII titer of 4d and Ecd titer of 9d were measured in different instar of the grafted larvae of the queen and the larvae in this paper; and the reproductive properties were measured of new queen developed from different instar of the grafted larvae; Meanwhile, iTRAQ method were used to analysis the influence of different instar of the grafted larvae on the queen ovary. The main results were as follows:1.The HPLC-MS was used to determine the JHIII titer of 4days larvae and the Ecd titer of 9d larvae. The results indicated that there were significant differences among the JHIII titer of 4days larvae and the Ecd titer of 9d larvae grafted in different instars. And the older the larvae grafted, the more significant differences showed.2. Methods of morphology and anatomy were used to measured correlated parameters on queen of different instar of the grafted larvae. The results indicated that birth weight, ovary weight, head weight and ovariole number were significantly of the virgin queen developed from different instar of the grafted larvae. The birth weight, ovary weight, head weight and ovariole number were significantly reduced with the increase of grafted instar. The ovariole number is linearly correlation with the birth weight(r=0.82). This study result shows that the earlier the larvae grafted, the earliethe better conditions the larvae get, the better development status of the queen. So, we should choose as early as possible of the worker larvae to foster the queen bee.3. iTRAQ method were used to analysis the ovarirs of queens developed from larvaes grafted in different instar. A total of 452 thousand spectrum were obtained. Total of 3542 proteins were mapped and identified.424 proteins were differentially expressed, of which 202 up-regulated and 222 down-regulated in 2 days age grafted larvae queen Compared with 1 day age.305 proteins were differentially expressed, of which 98 up-regulated and 207 down-regulated in 3 days age grafted larvae queen Compared with group 1 day age. Go enrichment analysis suggested that there were differences in metabolism, cell division and protein biosynthesis. The differential proteins of 1 day age and 2 days age group enriched in the carbohydrate metabolism, lipid metabolism, exogenous degradation, et al., And the result showed that this two groups queens have great difference in growth and metabolic rate. The differential proteins of 1 day age and 3 days age groupmain enriched in developing pathways(5.64%), ribosome pathways(5.24%) and lysosome metabolic, which indicated that the differentiation of organic development and the translation of genetic materials of this two groups was great.4. It was found that the expressions of MRJP1 and its precursor were significantly different among queens developed from larvaes grafted in different instar in the research of proteomics, which suggested that there were differences of MRJP1 between the royal jelly in worker cell and the royal jelly in queen cell.