高温胁迫和抗生素对携带Wolbachia短管赤眼蜂生殖的影响

发布时间：2018-03-26 17:48

  本文选题：短管赤眼蜂　切入点：Wolbachia　出处：《华南农业大学》2016年硕士论文

【摘要】：短管赤眼蜂(Trichogramma pretiosum)是膜翅目赤眼蜂科的重要天敌昆虫,主要寄生鳞翅目害虫的卵,在大田间有广泛的应用。受其体内共生菌Wolbachia的影响,短管赤眼蜂能够进行完全产雌孤雌生殖。本论文利用高温胁迫和抗生素对短管赤眼蜂的生殖生物学进行了研究,获得以下结果:1、利用Wolbachia的表面蛋白(Wolbachia Surface Protein,wsp)基因序列鉴定了所用短管赤眼蜂及其寄主米蛾(Corcyra cephalonica(Stainton))体内的Wolbachia,它们分别属于B超群的Kay亚群和Pip亚群。利用高温30℃和抗生素处理短管赤眼蜂,研究这些因素对短管赤眼蜂外部形态及卵巢管的影响。结果获得了雄性个体以及10种雌雄触角和外生殖器不同程度组合的雌雄嵌合体。同时发现,卵巢管变小及抱卵量减少的现象。论文还比较了含盐酸四环素、利福平、庆大霉素和青霉素四种抗生素三种不同浓度蜂蜜水对短管赤眼蜂及其体内Wolbachia的药效作用,结果表明,对于感染Wolbachia的短管赤眼蜂,盐酸四环素和利福平处理比庆大霉素处理和青霉素处理更早获得雌性后代及雌雄嵌合体,并且高浓度的抗生素(50 mg/mL、100 mg/m L)也会杀死赤眼蜂。此外,还比较了高温胁迫(30℃)和抗生素对携带Wolbachia的短管赤眼蜂后代性比的影响,结果表明,抗生素处理的短管赤眼蜂雄性百分率显著高于30℃高温处理的后代。利福平或庆大霉素处理的F0代短管赤眼蜂的产卵量显著低于青霉素处理的F0代产量量,到了F1、F2代,四种抗生素饲喂的短管赤眼蜂的产卵量并没有差异。随着浓度的升高,产卵量显著降低。2、利用实时荧光定量PCR技术对雌雄嵌合体内的Wolbachia进行拷贝数检测,发现不同表现型的短管赤眼蜂体内的Wolbachia拷贝数有显著的差异。同时,对饲喂10mg/mL盐酸四环素蜂蜜水获得的F1、F2、F3代雌性成虫进行Wolbachia进行拷贝数检测,发现这三个世代雌成虫体内的Wolbachia拷贝数没有显著差异,但呈现逐代降低的趋势。3、对短管赤眼蜂的转录组进行测序,共得到了31439条Unigenes。其中,29439个Unigenes比对上了KOG数据库的25个功能类别;10316条Unigenes被注释到了183个通路上;对参与的生物过程、细胞所处的位置和基因的分子功能中得到聚类数据,为进一步分析Wolbachia对短管赤眼蜂的生殖与行为调控机制提供基础。同时共得到9749个串联重复单元,为利用微卫星技术研究赤眼蜂的遗传结构或分化提供材料。
[Abstract]:Trichogramma pretiosum) is an important natural enemy insect of Hymenoptera Trichogramma, which mainly parasitizes the eggs of Lepidoptera pests and is widely used in large fields. The reproductive biology of Trichogramma brevis was studied by high temperature stress and antibiotics. The following results were obtained: 1. Wolbachia Surface protein wspsp. gene sequence of Wolbachia was used to identify Wolbachia brevis and its host, Corcyra cephalonicus Staintonia, which belong to B supergroup Kay subgroup and Pip subgroup, respectively. Treatment of Trichogramma brevis with biotin, The effects of these factors on the external morphology and ovarian duct of Trichogramma brevis were studied. Results male individuals and 10 different combinations of male and female antennae and external genitals were obtained. The effects of three different concentrations of honey water containing tetracycline hydrochloride rifampicin gentamicin and penicillin on Trichogramma brevis and its Wolbachia in vivo were compared. For Trichogramma brevis infected with Wolbachia, tetracycline hydrochloride and rifampicin treatments produced female offspring and female and male chimerism earlier than gentamicin and penicillin treatments, and a high concentration of 50 mg / mL 100 mg/m / L of antibiotics also killed Trichogramma. The effects of high temperature stress (30 鈩，

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