青斑蝶雌成虫内生殖器官形态与发育

刘微芬，廖怀建，石 雷，杜 婷，周成理，邓 疆，马云强

(中国林业科学研究院资源昆虫研究所，国家林业局资源昆虫培育与利用重点实验室，云南 昆明 650224)

青斑蝶雌成虫内生殖器官形态与发育

刘微芬，廖怀建，石 雷*，杜 婷，周成理，邓 疆，马云强

(中国林业科学研究院资源昆虫研究所，国家林业局资源昆虫培育与利用重点实验室，云南 昆明 650224)

青斑蝶；内生殖器官；卵巢；发育分级

1 研究方法

1.1 虫源准备

1.2 内生殖器官解剖方法

上午9:30将成虫浸泡于40%乙醇中，使成虫晕厥后，置于0 ℃蒸馏水中备用。在自制的圆形蜡盘(直径：15 cm)中倒入0.7%的生理盐水进行虫体的解剖。解剖时将蜡盘置于体视显微镜(Leica MZ16A, Leica有限公司，德国)的载物台上，在10×目镜和1×物镜下进行。

1.3 内生殖器官图片采集

将雌成虫内生殖器官置于超景深三维显微镜(VHX-1 000，KEYENCE，日本)载物台下进行拍照采集图片。由于青斑蝶雌成虫内生殖器官尺寸较大，采集到的是内生殖器官的不同组成结构的局部图片。利用显微镜的实时2D图像合成功能，在放大20倍的情况下，将局部图片合成为内生殖器官的完整图片。然后，利用显微镜自带的XY测量系统，在合成的完整图片上测量各结构的物理参数(实验测定的各生殖器官大小的样本量与解剖的雌蝶样本量不一致是由于解剖过程部分器官会出现破损的情况，而对于有破损的器官没有进行测定)。

1.4 内生殖器官特征参数测定

表1 雌蝶内生殖器官特征参数测定的成虫样本量Table 1 The sample capacity for measuring the characteristic parameters of female adultTirumala limniace

图1 雌性青斑蝶卵巢内成熟(A)与未成熟卵(B)
Fig.1 The mature and immature oocyte in the ovaries of the female adult Tirumala limniace

1.5 数据分析

不同日龄间雌成虫卵巢小管长度与直径、精巢纵轴长与横轴长、以及侧输卵管直径的差异采用One-Way ANOVA进行检验，当差异达到显著时，则采用Duncan’s multiple-range test进行检验。不同日龄间雌成虫卵巢小管内未成熟卵和成熟卵数量的差异采用One-Way ANOVA进行检验，当差异达到显著时，则采用Duncan’s multiple-range test进行检验。同日龄雌成虫卵巢小管未成熟卵与成熟卵数量间差异，采用Student’s t test进行检验。所有统计分析均在SAS 9.4中进行。

2 结果分析

2.1 青斑蝶雌蝶内生殖器官

图2 雌性青斑蝶的内生殖器官结构
Fig.2 The female internal reproductive organs of the Tirumala limniace

2.2 雌性青斑蝶成虫的卵巢管长度、直径和侧输卵管直径随日龄的变化

注：不同小写字母表示不同日龄间数据达到显著差异(p<0.05)Note: Different letters indicate significant difference between ages. (p<0.05)图3 青斑蝶雌蝶卵巢管的长度和直径、侧输卵管的直径随日龄的变化Fig.3 Length and diameter of the ovariole, diameter of the oviduct of the female adult Tirumala limniace

图4 雌性青斑蝶卵巢中成熟卵、未成熟卵的数量以及卵的总量随日龄的变化Fig.4 Number of mature, immature, and total eggs in ovaries of one- to 12-day old female adult Tirumala limniace

2.4 青斑蝶雌蝶卵巢发育分级标准

注：图左上角数字表示日龄，比例尺：2 mmNote:Numbers on top left corner of figure mean age,Scale bars:2.0 mm.图5 不同日龄青斑蝶雌蝶的内生殖器官结构Fig.5 Developmental morphology of internal reproductive organs of 1- to 12-day old female adult Tirumala limniace

3 讨论

昆虫雌性的内生殖器官通常由1对卵巢小管、附腺、一个或多个受精囊以及连接这些组织的导管组成[13]。青斑蝶雌性内生殖器官组成与之相同，包括1对卵巢、2根侧输卵管、1根中输卵管、交配囊腺、外生殖腔和产卵孔；不过，在青斑蝶雌性内生殖器官中没有发现附腺结构，这与黄粉蝶和橙黄豆粉蝶雌性内生殖器官结构存在差异[5,14]。

本研究对雌性青斑蝶的内生殖系统结构与发育的研究，均在恒温恒湿条件下进行。研究指出，适宜的温湿度及其昆虫的取食对昆虫的卵巢的发育起着至关重要的作用，温度太高而湿度太低会导致卵巢滞育，发育停滞不前；而当昆虫可取食的寄主食物由不可获取变为可以获取时，同等温湿度条件下滞育卵巢会变成熟[20]。因此，在实际工作中需要根据不同温湿度来判定具体的卵巢发育进度。

4 结论

雌性青斑蝶的内生殖器官由1对卵巢、1对侧输卵管、1根中输卵管、交配囊腺和受精囊组成；对卵巢小管长度、卵巢小管和侧输卵管直径测定发现随着日龄增大，其大小显著变大；随着日龄的增长，卵巢中成熟卵的数量先增大后减小，未成熟卵数量始终缓慢增加；根据卵巢的发育特征，将卵巢发育分为乳白透明期、卵黄沉积期、成熟待产期、产卵盛期、产卵末期。本研究将在青斑蝶的人工繁殖中为人为调控雌蝶产卵行为和有效提高其产卵量提供理论依据。

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(责任编辑：张 玲)

Morphology and Development of Reproductive System of Female AdultTirumalalimniace(Cramer)

LIUWei-fen,LIAOHuai-jian,SHILei,DUTing,ZHOUCheng-li,DENGJiang,MAYun-qiang

(Research Institute of Resources Insects of the Chinese Academy of Forestry/ Key Laboratory of Cultivation and Utilization of Resource Insects,State Forestry Administration, Kunming 650224, Yunnan, China)

[Objective]Tirumalalimniaceis a kind of important ornamental insects. To understand the structure and developmental state of internal reproductive organs can lay a theoretical basis for artificial reproduction and utilization ofT.limniace.[Method]The internal reproductive system of femaleT.limniacewith ages from one- to twelve-day old were dissected and observed, the characteristic parameters were measured, and the ovarian development was graded according to the developmental characteristics of ovary.[Result](1) The internal reproductive organs of femaleT.limniacecontain a couple of ovaries, a couple of lateral oviducts, a common oviduct, bursa copulatrix gland and spermatheca. (2) The development of internal reproductive organ was influenced significantly by age. The length of ovariole increased significantly with the age increasing and reached the maximum at the10th day, but the diameter of ovariole and lateral oviduct increased slowly with the increase of age. (3) With the increase of age, the amount of mature eggs in ovary increased at initial and then deduced, but the amount of immature eggs increased slowly all the time. (4) Based on the developmental characteristics, the ovarian development can be divided into 5 grades, i.e. milky and translucent stage, yolk deposition stage, egg maturation stage, peak stage of oviposition and final stage of oviposition. [Conclusion]The structure of internal reproductive system of femaleT.limniaceis the same as other butterflies, the development of internal reproductive organs is influenced significantly by age, and the eggs mature gradually with the increase of age. Ovarian grading ofT.limniacecan theoretically guide the artificial control of egg laying amount.

Tirumalalimniace(Cramer); reproductive system; ovary; development grading

10.13275/j.cnki.lykxyj.2017.01.017

2016-06-15

林业公益性行业科研专项经费(201504305); 云南省技术创新人才培养计划(2012HB054)。

刘微芬(1990—)，女，硕士研究生，主要从事蝴蝶行为生理生态学研究.
* 通讯作者：石 雷, 研究员.

S899

A

1001-1498(2017)01-0125-06