Wolbachia和蝇蛹数量对蝇蛹金小蜂适合度的影响

发布时间：2018-04-02 11:12

本文选题：蝇蛹金小蜂　切入点：Wolbachia　出处：《山东农业大学》2015年硕士论文

【摘要】：Wolbachia是广泛存在于节肢动物及线虫体内的胞内共生菌,可以通过寄主卵的细胞质母系遗传给后代。Wolbachia不仅可以调控寄主的生殖方式(主要为胞质不相容、孤雌生殖、雌性化和杀雄作用),参与昆虫物种的形成,而且还可以影响昆虫寄主的适合度。蝇蛹金小蜂Nasonia vitripennis隶属于膜翅目金小蜂科,是一种双翅目蝇类蛹期的寄生蜂。因为其分布范围广、世代周期短、种群数量大、雌雄易于区分、易于饲养等特点,加上其基因组测序已经完成,遗传背景清晰,已经逐步成为实验室内的研究昆虫,尤其是膜翅目的“模式昆虫”。实验室前人应用不同抗生素浓度处理自然感染Wolbachia的蝇蛹金小蜂,得到不感染Wolbachia的株系并稳定遗传,初步探讨了内共生菌Wolbachia对蝇蛹金小蜂适合度的影响。本实验是在前人的研究基础上,深入探究了Wolbachia对蝇蛹金小蜂的生殖调控以及蝇蛹数量和内共生菌二者对寄主适合度的影响。首先,明确了我们所养蝇蛹金小蜂种群体内Wolbachia对寄主的生殖调控作用是胞质不相容;然后,我们通过对5种利福平浓度(0.1mg/ml、0.3mg/ml、0.5mg/ml、3mg/ml、10mg/ml)处理的和对照(自然状态下感染单株系Wolbachia)蝇蛹金小蜂连续饲养多代,统计其后代个体在每天不同蝇蛹量(1、3、5粒)下每天的出蜂数量及总出蜂量;并分别统计其性比。最后研究了Wolbachia对蝇蛹金小蜂寿命的影响。主要结果如下:(1)Wolbachia对蝇蛹金小蜂的生殖调控表现为胞质不相容。针对Wolbachia不同感染状态的雌雄虫,我们设计四组交配实验:感染的雌虫和感染的雄虫(♀B×♂B)、感染的雌虫和不感染的雄虫(♀B×♂D)、不感染的雌虫和不感染的雄虫(♀D×♂D)和不感染的雌虫和感染的雄虫(♀D×♂B),分别统计其后代性比,前三组子代性比分别为0.83、0.83、0.71,而♀D×♂B杂交子代的性比仅为0.11,显著低于其他三种杂交组合,表现出强烈的胞质不相容。结合实验室分子检测数据,证明我们单株系Wolbachia对蝇蛹金小蜂的生殖调控表现是胞质不相容。(2)每天给交配后的蝇蛹金小蜂放置1粒蝇蛹,感染Wolbachia的蝇蛹金小蜂的出蜂量约219头,而去除Wolbachia的蝇蛹金小蜂的出蜂量约189头;每天3粒蝇蛹时,感染Wolbachia的蝇蛹金小蜂的出蜂量约392头,而去除Wolbachia的蝇蛹金小蜂的出蜂量约305头;在每天5粒蝇蛹时,感染Wolbachia的蝇蛹金小蜂的出蜂量约444头,而去除Wolbachia的蝇蛹金小蜂的出蜂量约407头。从结果可以看出,一方面,Wolbachia能够提高蝇蛹金小蜂的出蜂量;另一方面,随着每天放置蝇蛹量的增加,出蜂量明显增加。(3)实验结果表明感染Wolbachia的蝇蛹金小蜂的子代性比在0.83以上,而去除Wolbachia的蝇蛹金小蜂的子代性比在0.80以下。说明Wolbachia能够提高蝇蛹金小蜂的子代性比。每天放置1粒、3粒和5粒蝇蛹时,蝇蛹金小蜂的性比分别为0.80、0.82和0.83。蝇蛹数量对蝇蛹金小蜂子代性比的影响不显著。(4)Wolbachia对蝇蛹金小蜂寿命的影响。我们对感染Wolbachia与去除Wolbachia的雌雄蝇蛹金小蜂的寿命分别进行了统计,结果表明Wolbachia虽然对蝇蛹金小蜂的平均寿命有一定影响,但差异不显著。Wolbachia和蝇蛹数量对蝇蛹金小蜂的繁殖力产生积极影响,并且Wolbachia可以显著提高蝇蛹金小蜂的性比,在以后的实验中,可以利用Wolbachia和蝇蛹数量来增加蝇蛹金小蜂的数量,为科研和生物防治提供便利。
[Abstract]:Wolbachia is widely present in animal and arthropod elegans intracellular symbiotic bacteria, can host egg cytoplasm maternal inherited.Wolbachia can not only control the host reproduction (mainly cytoplasmic incompatibility, parthenogenesis, feminization and male killing), involved in the formation of insect species, but also can be suitable for influence of host insects. The pupae Nasonia Nasonia vitripennis belonging to Hymenoptera Pteromalidae, is a parasitoid of Diptera flies in the pupal stage. Because of its wide distribution, short generation cycle, large population, and easy to distinguish, easy feeding and other characteristics, and its genome sequencing has been completed, genetic background clearly, it has gradually become the research of insects in the laboratory, especially the Hymenoptera "model insects." previous application of different concentration of antibiotic treatment of Wolbachia natural infection from gold Bee, get Wolbachia infection strains and genetic stability, discusses the endosymbiont Wolbachia effects on the fitness of the pupae p.puparum. This experiment is on the basis of previous research, in-depth study of the Wolbachia of the pupae Nasonia reproductive regulation and the pupae number and endosymbionts two effects on the fitness of the host. First of all, we have defined the pupal Nasonia species groups within Wolbachia reproductive regulation effects on host is cytoplasmic incompatibility; then, we passed on the 5 concentration of rifampicin (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 3mg/ml, 10mg/ml (Wolbachia) treatment and control the natural state of plant infection) from p.puparum continuous multi generation, statistics of its offspring in different pupae per day (1,3,5 grain) daily and total number of adult bee; we evaluated the sex ratio. Finally, the effects of Wolbachia on pupal p.puparum life The influence. The main results are as follows: (1) on reproductive regulation Nasonia Wolbachia pupae showed cytoplasmic incompatibility. According to different Wolbachia infection status of the male and female worms, we designed four groups of mating experiments: infection of female and male infection (male B, female B *) infection and infection of male female (male D, female B *) no infection and no infection in female male (female and male D * D) infection of female and male infection (male B, female D *) evaluated the sex ratio of the offspring, the three groups before the offspring sex ratio was 0.83,0.83,0.71, while the female D *. The B hybrids of the sex ratio is only 0.11, significantly lower than the other three hybrid combinations showed strong cytoplasmic incompatibility. Combined with the laboratory testing data prove that Wolbachia molecules, we plant Department of reproductive regulation is the pupal Nasonia cytoplasmic incompatibility. (2) to the day after mating from Jin Xiaofeng place 1 grains of fly pupae, Wo infection Lbachia gold wasp bee pupae amount of about 219, and the removal of Wolbachia from bee pupae p.puparum about 189; 3 capsules per day pupae, Wolbachia infected pupae p.puparum a bee quantity about 392, and the removal of Wolbachia from bee pupae p.puparum about 305 in the head; every day 5 grains of pupae infected with Wolbachia, the bee pupae p.puparum amount of about 444, and the removal of Wolbachia from bee pupae p.puparum about 407 head. It can be seen from the results, on the one hand, Wolbachia can improve the quantity of bee pupae p.puparum; on the other hand, with the increase of the pupae placed every day the bee was significantly increased (3). The experimental results show that Wolbachia infection of pupae p.puparum offspring sex ratio is above 0.83, and the removal of Wolbachia from p.puparum offspring sex ratio below 0.80. The results showed that Wolbachia can improve the pupal p.puparum offspring sex ratio. Every place 1 grain 3, and 5 seeds per pupae, pupal Nasonia sex ratios were 0.80,0.82 and 0.83. pupae had no significant effect on pupal p.puparum offspring sex ratio. (4) the effect of Wolbachia on pupal Nasonia life. Our male and female pupae infected with Wolbachia Jin Xiaofeng and the removal of Wolbachia statistics for life the results show that, although Wolbachia has a certain effect on the average life expectancy of p.puparum pupae, but have a positive impact on the fecundity of.Wolbachia and no significant differences in the numbers of pupae of p.puparum pupae, and Wolbachia can significantly improve the ratio of the pupae of Jin Xiaofeng, after the experiment, you can use the Wolbachia number to increase the number of pupae and pupae of p.puparum and provide convenience for scientific research and biological control.

【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S476.3

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