Ovary activation of worker honey beefs Apis mellifera L. and identification of genes involved

In an effort to uncover genes associated with ovary activation in honey bee workers, northern blot hybridization and quantitative real-time PCR were used to examine differential expression of the candidate genes in the workers with and without ovary development. From northern blot hybridization, the expression of vitellogenin and transferrin genes showed significantly different between wild-type workers and ovary activated anarchist workers. These two genes had more expressed in the abdomen of anarchist workers while they were less expression in the head part of those workers compared with wild-type workers. The quantitative real-time PCR was used to confirm the results and we found that vitellogenin was up-regulated about 4-fold in the abdomen of ovary activated anarchist workers when compared with ovary non-activated wild-type workers. Moreover, the honey bee workers that had ovary activation were constructed to use as the samples for examining the genes that regulate or involve in the network of ovary activation. Carbon dioxide and pheromone were the important keys for constructing these samples. Carbon dioxide affected the ovary retardation in the honey bee workers and also pheromone did the same effect. Vitellogenin and transferrin genes did show significantly differences in the abdominal tissue and head tissue part following the CO₂ narcosis. These two genes were up-regulated about 4-fold in the abdomen of ovary activated workers. In contrast, they were down-regulated about 3-fold in the head of those workers. These confirmed the previous results from northern blot analysis in the case of ovary activation. These vitellogenin and transferrin were up-regulated, about 14-fold and 9-fold respectively, in the abdomen of ovary activated queenless workers relative to ovary non-activated queenright workers. Moreover, gene encoding phosphoinositolglycan-peptide and tyramine receptor seem to be up-regulated in the abdomen of ovary activated queenless workers too.为探究与蜜蜂工蜂（honey bee workers）卵巢激活相关的基因，本研究采用Northern印迹杂交（Northern blot hybridization）与实时定量聚合酶链式反应（quantitative real-time PCR），检测候选基因在卵巢发育与未发育的工蜂中的差异表达水平。经Northern印迹杂交检测发现，卵黄原蛋白（vitellogenin）与转铁蛋白（transferrin）基因在野生型工蜂与卵巢激活型无政府工蜂（anarchist workers）中的表达水平存在显著差异：相较于野生型工蜂，这两种基因在无政府工蜂的腹部中表达量更高，而在其头部中表达量更低。随后采用实时定量聚合酶链式反应对上述结果进行验证，结果显示：与卵巢未激活的野生型工蜂相比，卵巢激活型无政府工蜂腹部的卵黄原蛋白表达量上调约4倍。此外，本研究构建了卵巢激活的工蜂作为实验样本，用以探究参与卵巢激活调控网络的基因。二氧化碳与信息素是构建此类实验样本的关键调控因子：二氧化碳可抑制蜜蜂工蜂的卵巢发育，信息素亦具有相同作用。经二氧化碳麻醉处理后，卵黄原蛋白与转铁蛋白在工蜂腹部组织与头部组织中的表达水平同样存在显著差异：这两种基因在卵巢激活工蜂的腹部中表达量上调约4倍，而在其头部中表达量下调约3倍，该结果与此前Northern印迹杂交得到的卵巢激活相关研究结果一致。与卵巢未激活的有蜂王工蜂相比，卵巢激活的无蜂王工蜂腹部的卵黄原蛋白与转铁蛋白表达量分别上调约14倍与9倍。此外，编码磷酸肌醇糖肽（phosphoinositolglycan-peptide）与酪胺受体（tyramine receptor）的基因在卵巢激活的无蜂王工蜂腹部中似乎亦呈现上调表达。