王浆高产蜜蜂(Apis mellifera L.)工蜂幼虫发育期蛋白质组分析

Analysis of the Proteome of the Larvae of the High Royal Jelly Producing Worker Bees (Apis mellifera L.)

【目的】20世纪90年代中国成功培育出的王浆高产蜜蜂(Apis mellifera L.)是当今世界最优良的蜂种,使中国的王浆年产量高达2600多吨,占世界总产量的90%以上,稳居世界第一,通过对该蜂种不同发育日龄工蜂幼虫的蛋白质组进行研究,以探明其发育机理。【方法】采用双向电泳法对王浆高产蜜蜂(Apis mellifera L.)不同发育期的工蜂幼虫进行蛋白质组研究。【结果】在幼虫期6d的发育过程中,2日龄、4日龄、6日龄幼虫中分别检测到了262、418、194个蛋白点。这些蛋白的分子量在12.2～88.2kD的较大范围之间,等电点在pH4.00～9.24之间。有84个蛋白在幼虫期整个发育过程中均有表达,其中33%呈上调趋势,21%呈下调趋势,46%蛋白的表达没有规律。2日龄、4日龄、6日龄幼虫中分别有88、209、63个特异表达的蛋白。除此之外,有84个蛋白在幼虫发育的6日龄表达关闭,而在2日龄和4日龄幼虫中表达;有41个蛋白在2日龄表达关闭,而在4日龄和6日龄中表达;仅有6个蛋白在4龄日表达关闭,而在2日龄和6日龄表达。【结论】幼虫的发育过程有大量基因参与表达和调控,是一个复杂而动态有序的过程,且发育到4日龄的幼虫蛋白表达最为活跃。幼虫整个发育过程中的共有蛋白是发育必需的保守蛋白。幼虫不同的发育阶段由不同的特异蛋白进行调控。

【Objective】The purpose was to investigate the protein composition of worker bees larvae of the high producing royal jelly bees （Apis mellifera L.）.【Method】Two-dimensional gel electrophoresis （2-DE） was employed to separate and quantify proteins of the larvae of the high producing royal jelly worker bees （Apis mellifera L.） during their six days development.【Result】 The results showed that 262, 418 and 194 proteins with a range of molecular weight 12.2-88.2 kD and relatively narrow scope of pI 4.00-9.24 could be detected on day 2, day 4 and day 6 during the developmental process of the larvae, respectively. Meanwhile 84 protein spots were constantly detected during the larvae development, among them 33 percent were expressed in up trend along with the larvae development, 21 percent were in down trend and 46 percent were irregular. In addition, the specific proteins expressed on day 2, day 4, and day 6 were 88, 209, and 63, respectively. Except the constant and specific expressed proteins, 84 proteins were expressed on day 2 and day 4 but silenced on day 6, 41 proteins were expressed on day 2 and day 4 but silenced on day 6, only 6 proteins were expressed on day 2 and day 6 but silenced on day 4.【Conclusion】The result indicates that the larvae development is a sequential and complex gene controlled process, while the proteins of larvae of day 4 express the most active. The eternal expressed proteins suggest that they are conservative and indispensable for this event. The specific proteins detected at different developmental stages suggesting it is indispensable to regulate the larvae’ growth.