意大利蜜蜂(A.m.ligustica)雄蜂卵期发育蛋白质组分析

Analysis of Developmental Proteome at Egg Stage of Drone Honeybees (A. m. ligustica)

【目的】对原种意大利蜜蜂(Apis mellifera ligustica Spinola,1806)(意蜂)雄蜂的3d卵期发育进行蛋白质组研究,以探明该蜂种雄蜂卵期蛋白质表达调控方面的一些特点,从而揭示其发育的分子机理。【方法】采用双向电泳法对意蜂雄蜂卵期蛋白质组进行研究。【结果】3d的卵期发育中,按顺序分别检测到200、242、233个蛋白,这些蛋白的分子量在12.42～169.60kD,等电点为4.50～9.00。164个蛋白在3d卵期发育过程中均有表达,其中79个蛋白在3个日龄的表达量无显著差异(P>0.05),7个共有蛋白表达量随卵期发育呈显著上调(P<0.05)趋势,而4个共有蛋白呈显著下调(P<0.05)趋势。同时卵期3d分别有11、18和18个特异的蛋白表达。除共有及特有蛋白外,还有17个蛋白在第1日龄和第2日龄表达而在第3日龄关闭;43个蛋白在第2日龄和第3日龄表达而在第1日龄关闭;仅有8个蛋白在第2日龄关闭,而在第1日龄和第3日龄表达。【结论】结果初步表明意蜂雄蜂卵期发育过程中第2日龄蛋白表达最为活跃;卵期3d所表达的共有蛋白,可能是调控意蜂雄蜂卵期发育所必需的保守蛋白,这些蛋白的表达模式存在一定的差异;不同日龄表达的特异性蛋白,表明不同发育阶段的卵需要不同的特异性蛋白来调控。同时也表明雄蜂卵期发育较工蜂卵表达的保守蛋白更多,这将给蜜蜂的基因改良带来便利。

[Objective] An investigation on the proteome of drone egg development of native Italian bee （Apis mellifera ligustica Spinola, 1806） in order to prove the characteristics in protein expression and regulation at egg stage and understand the molecular mechanism of development. [Method] The experiment was carried out by two-dimensional gel electrophoresis. [ Result ] The results showed that there were 200, 242 and 233 proteins with wide range of molecular weight （12.42-169.60 kD） and relatively narrow scope of pI （4.50-9.00） were detected on day 1, day 2 and day 3, respectively, during the developmental process of the drone egg. Meanwhile, 164 protein spots were resolved at all the images （i.e. the protein was consistently expressed） along with the egg development, among which 7 were significantly up-expressed （P〈0.05） and 4 were significantly down-expressed （P〈0.05）, while seventy-nine had no significant difference （P〉0.05）. In addition, the specific proteins expressed proteins on day 1, day 2 and day 3 were 11, 18, 18, respectively. Besides, seventeen proteins expressed both on day 1 and day 2, but silenced on day 3, and forty -three proteins expressed both on day 2 and day 3, but silenced on day 1, while only eight proteins expressed both on day l and day 3, but silenced on day 2. [Conclusion] The results indicate that 2 day-old eggs is the most active expressional stage in the development of drone egg. The protein expressed at all images suggests that it should be indispensable for drone egg development,but their expression pattern is different. The proteins expressed at a specific age of egg suggest that different developmental stages need specific proteins to regulate. And there are more house-keeping proteins in the developmental process of the drone egg than that of worker egg, it will provide more target for gene improvement.