幼虫信息素酯类混合物对中华蜜蜂育王质量的影响

摘要

[目的]基于免移虫育王技术,探究幼虫信息素酯类混合物在育王中对中华蜜蜂Apis cerana cerana 蜂王质量的影响,为优质蜂王的培育奠定基础.[方法]利用中华蜜蜂免移虫育王生产器培育蜂王,在幼虫60 - 64 h 时向王台内注入1 μL 不同酯类组成的混合物(酯类混和物的浓度梯度分别为0,0. 1％,1. 0％和10. 0％),待蜂王出房后测定蜂王初生重、胸重、胸宽以及单侧卵巢管数量;并利用荧光定量PCR 测定蜂王卵巢卵黄原蛋白基因Vitellogenin (Vg),昆虫储存蛋白基因 hexamerin70b (hex70b) 和hexamerin110 (hex110) 的表达量.[结果]免移虫育王过程中添加3 种信息素酯类混合物3E(1. 0％甲基亚油酸酯、16. 0％ 甲基亚麻酸酯和35. 0％ 甲基油酸酯),不同浓度处理时蜂王的初生重较对照(石蜡油) 均显著增加,但蜂王胸宽和胸重均无显著变化; 0. 1％ 和 1. 0％浓度处理组蜂王单侧卵巢管数显著增加; 不同浓度处理组Vg 和hex70b 的表达量均无显著变化,但1. 0％和10. 0％浓度处理组蜂王卵巢的hex110 表达量显著升高.4 种信息素酯类混合物4E(4. 5％甲基棕榈酸酯、1. 0％ 甲基亚油酸酯、16. 0％ 甲基亚麻酸酯和35. 0％ 甲基油酸酯) 试验组只有10. 0％浓度处理组蜂王的初生重和单侧卵巢管数较对照显著上升(P ＜ 0. 05),0. 1％和1. 0％浓度处理对蜂王的个体发育指标均无显著影响(P ＞ 0. 05); 0. 1％和1. 0％浓度处理组Vg 的表达量、 10. 0％浓度处理组hex70b 的表达量以及1. 0％和10. 0％ 浓度处理组hex110 的表达量均显著上升(P ＜ 0. 05).10 种信息素酯类混合物10E(4. 5％ 甲基棕榈酸酯、2. 5％ 甲基硬脂酸、35. 0％ 甲基油酸酯、1. 0％甲基亚油酸酯、16. 0％甲基亚麻酸酯、3. 5％乙基棕榈酸酯、1. 5％乙基硬脂酸酯、18. 0％乙基油酸酯、0. 5％ 乙基亚油酸酯和17. 5％ 乙基亚麻酸酯) 试验组,各个浓度处理蜂王的初生重以及Vg 和hex110 的表达量均显著下降(P ＜ 0. 05),但蜂王的单侧卵巢管数和胸部指标无显著变化(P ＞ 0. 05),10. 0％浓度处理时hex70b 的表达量也显著降低(P ＜ 0. 05).[结论]中华蜜蜂育王过程中添加1. 0％的3E 和10. 0％的4E 幼虫信息素酯类可以在一定程度上提高蜂王质量.%[Aim]The objective of this study is to investigate the effects of mixtures of brood pheromone esters on the queen quality of the Chinese honeybee,Apis cerana cerana,reared by queen-rearing technology without larvae-grafting,and to find a way to rear high-quality queen. [Methods]Queenrearing technology without larvae-grafting was employed to rear queen of A. cerana cerana. Using a micro sampling syringe injector,1 μL of mixture of brood pheromone esters with the concentration gradients of 0,0. 1％,1. 0％ and 10. 0％ was added into queen cells,respectively,when the queen larvae were 60 - 64 h old. The body weight,the weight and width of thorax,the number of ovarioles in one ovary of the newly-emerged queens were tested. And the gene expression levels of vitellogenin (Vg ), hexamerin70b (hex70b) and hexamerin110 (hex110) were quantified by qPCR. [Results]The results showed that the body weight of the newly-emerged queens of A. cerana cerana was increased significantly(P ＜ 0. 05) in the treatments with the mixture of three brood pheromone esters (3E) (1. 0％ methyl linoleate,16. 0％ methyl linolenate,and 35. 0％ methyl oleate) at all the three concentrations,while the width and weight of thorax of reared queens had no significant change as compared with the control(paraffin oil) (P ＞ 0. 05). The number of ovarioles in one ovary was increased significantly (P ＜ 0. 05) in 0. 1％ and 1. 0％ treatment groups. The expression levels of Vg and hex70b in all 3E treatment groups had no significant change,while that of hex110 in 1. 0％ and 10. 0％ 3E treatment groups increased significantly (P ＜ 0. 05) as compared with the control. When the queens were reared with the mixture of four brood pheromone esters (4E) (4. 5％ methyl palmitate,1. 0％ methyl linoleate,16. 0％ methyl linolenate,and 35. 0％ methyl oleate),the body weight of the newly-emerged queens and the number of ovarioles in one ovary significantly increased in 10. 0％ 4E treatment group (P ＜ 0. 05 ),while no significant difference of developmental indexes existed in 0. 1％ and 1. 0％ 4E treatment groups (P ＞ 0. 05). The expression levels of Vg in ovaries in 0. 1％ and 1. 0％ 4E treatment groups,hex70b in 10. 0％ 4E treatment group and hex110 in 1. 0％ and 10. 0％ 4E treatment groups all significantly increased as compared with the control (P ＜ 0. 05). When the queens were reared with the mixture of 10 brood pheromone esters (10E) (4. 5％ methyl palmitate,2. 5％ methyl stearate,35. 0％ methyl oleate, 1. 0％ methyl linoleate,16. 0％ methyl linolenate,3. 5％ ethyl palmitate,1. 5％ ethyl stearate,18. 0％ ethyl oleate,0. 5％ ethyl linoleate,and 17. 5％ ethyl linolenate),the body weight of the newly-emerged queens,and the expression levels of Vg and hex110 in ovaries in all treatment groups significantly decreased as compared with the control (P ＜ 0. 05),while the number of ovarioles in one ovary and the thorax indexes showed no significant change (P ＞ 0. 05). The expression level of hex70b also reduced significantly in 10％ 10E treatment group (P ＜ 0. 05). [Conclusion]To a certain extent,the 1. 0％ 3E and 10. 0％ 4E added in queen-rearing process of A. cerana cerana could improve the queen quality.