CO2麻醉对中华蜜蜂处女蜂王卵巢发育的影响

doi:10.16380/j.kcxb.2022.05.005

昆虫学报 ›› 2022, Vol. 65 ›› Issue (5): 577-586.doi: 10.16380/j.kcxb.2022.05.005

CO₂麻醉对中华蜜蜂处女蜂王卵巢发育的影响

周建阳^1,2,3, 宋华莉^1,2,3, 邓萍^1,2,3, 陶坤伶^1,2,3, 唐相友^1,2,3, 燕乐乐^1,2,3, 张小燕^1,2,3, 石鹏^1,2,3, 许金山^1,2,3,*

(1. 重庆师范大学生命科学学院, 重庆 401331; 2. 活性物质生物技术教育部工程研究中心, 重庆 401331; 3. 重庆市媒介昆虫重点实验室, 重庆 401331)

出版日期:2022-05-20 发布日期:2022-05-08

Effects of CO₂ narcosis on the ovarian development of virgin queens of Apis cerana cerana (Hymenoptera: Apidae)

ZHOU Jian-Yang^1,2,3, SONG Hua-Li^1,2,3, DENG Ping^1,2,3, TAO Kun-Ling^1,2,3, TANG Xiang-You^1,2,3, YAN Le-Le^1,2,3, ZHANG XiaoYan^1,2,3, SHI Peng^1,2,3, XU Jin-Shan^1,2,3,*

(1. College of Life Sciences, Chongqing Normal University, Chongqing 401331, China; 2. Engineering Research Center of Biotechnology for Active Substances, Ministry of Education, Chongqing 401331, China; 3. Chongqing Key Laboratory of Vector Insect, Chongqing 401331, China)

Online:2022-05-20 Published:2022-05-08

摘要/Abstract

摘要： 【目的】明确中华蜜蜂Apis cerana cerana蜂王羽化后生殖系统的动态变化，探索CO₂麻醉对中华蜜蜂处女蜂王卵巢发育的影响。【方法】通过人工育王技术和蜂王的储存获取不同日龄的中华蜜蜂处女蜂王，观察蜂王羽化后生殖系统的形态，测定生殖系统各项形态学指标(体重、卵巢重、受精囊直径和卵巢管数目)。中华蜜蜂处女蜂王连续2 d(分别于5和6日龄时)接受不同浓度(50%, 75%与90%)CO₂麻醉8 min, 以及不同日龄处女蜂王接受75% CO₂麻醉1~3次(每次8 min)，待蜂王发育至10日龄时，利用石蜡切片技术与HE染色法观察蜂王卵巢形态、测定蜂王卵巢重和蜂王卵巢最大卵母细胞长度，通过实时荧光定量PCR(RT-qPCR)测定10日龄蜂王头部卵黄原蛋白基因(Vg)的相对表达量。【结果】中华蜜蜂蜂王的卵巢管数目与受精囊直径不会随着日龄发生明显的波动，但蜂王卵巢重在6-12日龄明显增加，之后将趋于稳定。与对照组(未接受CO₂麻醉处理)相比，50%, 75%与90% CO₂麻醉后中华蜜蜂蜂王卵巢重与最大卵母细胞长度均增加，并且头部Vg基因的相对表达量上调。其中CO₂浓度为75%，麻醉次数2次显著促进了中华蜜蜂蜂王的卵巢发育。【结论】中华蜜蜂蜂王羽化后卵巢会发生明显的发育变化，而CO2麻醉能加速卵巢的发育。

关键词: 中华蜜蜂, 蜂王, CO₂麻醉, 生殖系统, 卵巢, 卵黄原蛋白

Abstract: 【Aim】 To clarify the dynamic changes of reproductive system of queens of the Chinese honeybee (Apis cerana cerana) after emergence, and to explore the effects of CO₂ narcosis on the ovarian development of their virgin queens. 【Methods】 Through artificial breeding technology and storage of the queens, we obtained different day-old virgin queens of A. cerana cerana, observed the morphological characteristics of their reproductive system after emergence, and determined the morphological indexes of their reproductive system including the body weight, ovarian weight, diameter of spermatheca, and number of ovarioles. After the virgin queens were narcotized with different concentrations (50%, 75% and 90%) of CO₂ in two consecutive days (at the 5- and 6-d-old, respectively) for 8 min, and different day-old virgin queens were respectively narcotized with 75% CO₂ for 1-3 times (each time for 8 min), the morphological characteristics of the ovaries of the 10-d-old queen were observed by paraffin section and HE staining, and their ovarian weight and the length of the largest oocyte of ovaries were detected. Meanwhile, the relative expression level of vitellogenin gene (Vg) in the head of the 10-d-old queens was measured by real-time fluorescence quantitative PCR (RT-qPCR). 【Results】 The number of ovarioles and the diameter of spermatheca of the queens of A. cerana cerana did not fluctuate obviously with the age increasing. However, the ovarian weight of queens increased obviously during the 6-12-d-old and then tended to be stable. As compared to the control (not subjected to CO₂ narcosis), narcosis with 50%, 75% and 90% CO₂ resulted in the increase of the ovarian weight and the length of the largest oocyte of queens of A. cerana cerana and the upregulation of the relative expression level of Vg gene in the queen head. Moreover, narcosis with 75% CO₂ twice significantly promoted the ovarian development of A. cerana cerana queens. 【Conclusion】 The ovaries of A. cerana cerana queens present obvious developmental changes after emergence, and CO₂ narcosis can accelerate their ovarian development.

Key words: Apis cerana cerana, queen, CO₂ narcosis, reproductive system, ovary; vitellogenin

周建阳, 宋华莉, 邓萍, 陶坤伶, 唐相友, 燕乐乐, 张小燕, 石鹏, 许金山. CO₂麻醉对中华蜜蜂处女蜂王卵巢发育的影响[J]. 昆虫学报, 2022, 65(5): 577-586.

ZHOU Jian-Yang, SONG Hua-Li, DENG Ping, TAO Kun-Ling, TANG Xiang-You, YAN Le-Le, ZHANG Xiao-Yan, SHI Peng, XU Jin-Shan. Effects of CO₂ narcosis on the ovarian development of virgin queens of Apis cerana cerana (Hymenoptera: Apidae)[J]. Acta Entomologica Sinica, 2022, 65(5): 577-586.

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CO₂麻醉对中华蜜蜂处女蜂王卵巢发育的影响

Effects of CO₂ narcosis on the ovarian development of virgin queens of Apis cerana cerana (Hymenoptera: Apidae)

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