温度通过影响Wolbachia滴度调控赤眼蜂生殖方式

【目的】揭示温度对宿主体内Wolbachia滴度及其调控宿主生殖作用的影响。【方法】以感染Wolbachia且营孤雌产雌生殖的食胚赤眼蜂Trichogramma embryophagum为对象,在22,25,28和31℃4个梯度温度下连续培养5代,观察生殖方式、性比等生物学特性;采用实时荧光定量PCR以Wolbachia特异基因——外膜蛋白基因(wsp)、二磷酸果糖醛缩酶基因(fbp A)和酰胺转移酶基因(gat B)为靶标对内生菌进行定量分析。【结果】22和25℃条件下连续培养5代,食胚赤眼蜂生殖方式均未发生改变(无雄蜂出现),且5代赤眼蜂群体Wolbachia滴度没有明显差异;28℃下,食胚赤眼蜂在F3代开始有雄蜂出现,至F5代雄蜂比例明显增多,且体内Wolbachia滴度在F2代开始下降,至F5代显著下降;31℃下,F2代即有雄蜂出现,至F5代已经恢复成产雄孤雌生殖,体内Wolbachia滴度也在F2代开始下降,F3代开始显著降低,到F5代Wolbachia滴度极小甚至检测不到。【结论】高温可改变营孤雌产雌生殖赤眼蜂的生殖方式及体内Wolbachia滴度,且随着处理代数的增加以及温度的升高作用显著,即高温对营孤雌产雌生殖赤眼蜂生殖方式改变程度与其体内Wolbachia滴度呈负相关。     

短期高温对感染Wolbachia的松毛虫赤眼蜂发育和繁殖的影响

感染了沃尔巴克氏体Wolbachia的松毛虫赤眼蜂Trichogramma dendrolimi Matsumura营孤雌产雌生殖方式,但高温可能影响这一特性。为了模拟该赤眼蜂在田间遇到短期高温后的受影响情况,研究了柞蚕卵繁殖的Wolbachia感染了Wolbachia的松毛虫赤眼蜂孤雌产雌品系,于蛹中期和蛹后期经历6h32,35,38℃高温单次冲击处理,对当代羽化出蜂率、单卵出蜂数、单卵雌蜂率、寿命4个指标和处理子代羽化出蜂率、有效繁殖个体、单卵出蜂数、单卵雌蜂率4个指标的影响。结果表明,短期高温刺激会对孤雌产雌的松毛虫赤眼蜂产生不利影响,尤其是38℃高温不利影响明显,主要表现在羽化出蜂率和单卵出蜂数两指标明显降低;高温冲击对子代蜂各指标影响不明显。2代赤眼蜂均未有雄峰出现,表明短期高温冲击不能对Wolbachia调控其宿主生殖方式的作用产生影响。     

孤雌产雌生殖品系松毛虫赤眼蜂产卵强度对Wolbachia诱导的其生殖表型的影响

[目的]松毛虫赤眼蜂Trichogramma dendrolimi是一种用于鳞翅目害虫生物防治的重要卵期寄生蜂.本研究旨在明确孤雌产雌生殖品系松毛虫赤眼蜂的产卵强度对其体内Wolbachia滴度及Wolbachia诱导的孤雌产雌生殖表型的影响.[方法]在室内调查了3个处理组松毛虫赤眼蜂雌蜂不同产卵强度(每日仅1 h供寄...     

温度对感染沃尔巴克氏体的松毛虫赤眼蜂种群参数的影响

在16℃、20℃、24℃、28℃和32℃恒温下,观察了温度对感染沃尔巴克氏体的松毛虫赤眼蜂发育、存活和繁殖的影响,组建了相应温度下的实验种群生命表。结果表明,在实验温度范围内,感染沃尔巴克氏体的松毛虫赤眼蜂的发育历期随着温度的升高而逐渐缩短,5个温度下感染沃尔巴克氏体的松毛虫赤眼蜂世代的发育历期分别为23.2、17.8、13.2、11.1和7.8天,世代的发育起点温度和有效积温分别为7.40℃和212.69日·度,内禀增长率(r_m)和净生殖力(R₀)分别介于0.2448～0.4584和28.00～84.03之间。结果提示温度24℃～28℃为繁殖感染沃尔巴克氏体的松毛虫赤眼蜂的理想温度条件。     

Wolbachia在我国广赤眼蜂种群内的感染

Wolbachia是广泛分布于节肢动物生殖组织内的一类细胞内共生菌,它属于原细菌的α亚类,能够通过调控寄主的生殖活动而促进其在寄主种群中的扩散。通过对wsp 基因的克隆及PCR-RFLP分析确定了Wolbachia在我国广赤眼蜂种群内的存在,并发现有2种Wolbachia 菌系的感染,命名为wEvaA和wEvaB。经过克隆分离得到了这2种Wolbachia的wsp基因序列,在GenBank的登录号为AY390279和 AY390280 ,并由基于wsp基因的聚类树中发现,这两种Wolbachia菌系均属于A组。     

Wolbachia属共生细菌及其对节肢动物生殖活动的调控作用

Wolbachia属是广泛分布于节肢动物生殖组织内的一类共生细菌。这些共生菌通过卵的细胞质传播并参与多种调控其宿主生殖活动的机制,包括:诱导生殖不亲和、诱导孤雌生殖、雌性化、雄性致死和调节繁殖力。Wolbachia被认为与性别决定、共生关系和物种形成等重要生物学问题密切相关,是探索这些研究领域的新线索。而且Wolbachia可作为特定的载体对其宿主种群进行遗传调控,如增强寄生蜂在害虫生物防治中的作用,控制线虫引起的疾病传播。该文综述了Wolbachia的形态学及存在部位、基因组结构、系统发育、种的命名、水平传递和Wolbachia对其宿主生殖活动的调控作用,并分析了Wolbachia研究的科学意义和发展趋势,以期引起我国生物学家对Wolbachia研究的注意和快速切入。     

首次发现沃尔巴克氏体Wolbachia对我国南亚果实蝇的感染现象

沃尔巴克氏体Wolbachia是广泛分布于节肢动物体生殖组织内的一类共生细菌,泰国等已发现Wolbachia感染南亚果实蝇Bactrocera(Zeugodacus)tau(Walker)的现象。利用沃尔巴克氏体wsp基因的1对通用引物(81F,691R)从我国南亚果实蝇的总DNA中扩增到1段500 bp左右的wsp基因片段,并克隆得到了Wolbachia相关序列,首次发现了沃尔巴克氏体对我国南亚果实蝇的感染现象。     

哺乳动物性别分化的调控

哺乳动物性别分化调控的分子机制的研究特别是性别分化的层次调控、剂量补偿和性染色体进化这三个领域,已取得快速进展。已经发现Ｙ染色体性别决定区基因（ＳＲＹ）、Ｘ染色体ＤＳＳ－ＡＨＣ决定区基因１（ＤＡＸ－１）、甾类生成因子１基因（ＳＦ１）和Ｗｉｌｍｓ瘤抑制基因（ＷＴ－１）等与哺乳动物性别决定有关。ＳＲＹ启动睾丸分化,但胚胎发育成雄性的其余步骤由睾丸分泌的激素控制。ＤＡＸ－１编码一种女性特异功能的蛋白质,     

我国麦蚜体内的沃尔巴克氏体(Wolbachia)的检测

沃尔巴克氏体是广泛分布于节肢动物体内的一类共生细菌 ,它们参与多种调控其寄主生殖活动的机制。本研究通过wsp基因的特异性扩增证实了沃尔巴克氏体在麦长管蚜Sitobionavenae(Fabricius)体内的共生。而且在麦长管蚜的两种体色型 (绿色型和红色型 )中都检测到了沃尔巴克氏体的共生。     

Wolbachia在玉米螟赤眼蜂内的三重感染

Wolbachia是一类广泛存在于节肢动物体内的共生菌。玉米螟赤眼蜂Trichogramma ostriniae是我国玉米田间的优势赤眼蜂种, 据报道, 赤眼蜂种内有Wolbachia感染。本文利用Wolbachia的16s rDNA和wsp基因引物通过PCR方法对玉米螟赤眼蜂的野生种群进行了调查, 发现以wsp基因为鉴定依据, 检测的所有个体都感染了3种Wolbachia ［wOstGDAa (GenBank accession no. EU157103), wOstGDAb (GenBank accession no. EU157104) 和 wOstGDB (GenBank accession no. EU157105)］。本文首次报道了野生赤眼蜂种群内Wolbachia的三重感染率几乎为100%。根据本研究的结果, 可以推测当不同种赤眼蜂寄生同一寄主时, Wolbachia可能会在不同赤眼蜂种间进行横向传播。     

两种抗生素对携带Wolbachia的短管赤眼蜂生殖模式的影响

携带有Wolbachia的短管赤眼蜂为完全的产雌孤雌生殖。前期的实验表明,利用四环素对携带有Wolbachia的短管赤眼蜂进行除菌处理,但结果并不能获得100%恢复孤雌产雄。本实验拟用环丙沙星与磺胺嘧啶这两种抗生素对携带有Wolbachia的短管赤眼蜂进行除菌效果筛选。实验分3组药剂处理,分别为使用环丙沙星、磺胺嘧啶及环丙沙星与磺胺嘧啶1:1混配液进行除菌处理。各组实验分别进行浓度为0.1 mg/mL、1.0 mg/mL和10.0 mg/mL 3个浓度处理。以无抗生素蜂蜜水处理作为实验对照组。分别检测、记录各处理的F0 - F1代的寄生卵量,F1 - F2代羽化个体数、雌雄个体数,雌雄相嵌体的个体数。结果表明：磺胺嘧啶对短管赤眼蜂的F0代产卵量降低,但F1代恢复产卵量;仅有0.1 mg/mL环丙沙星与磺胺嘧啶1:1混配液对短管赤眼蜂F2代的卵羽化率具有抑制作用,其它的处理与对照组结果无差异。环丙沙星处理后的携带Wolbachia的短管赤眼蜂没出现雄性后代,浓度为0.1 mg/mL环丙沙星与磺胺嘧啶1:1混配液处理在F2代出现了雌雄相嵌体。浓度为1.0 mg/mL和10 mg/mL的环丙沙星与磺胺嘧啶1:1混配液处理和磺胺嘧啶处理在F2代出现了完全雄性后代。使用磺胺嘧啶处理携带Wolbachia的短管赤眼蜂,可更方便快捷获得100%雄性后代。     

Biological Control Potential of Wolbachia -infected Versus Uninfected Wasps: Laboratory and Greenhouse Evaluation of Trichogramma cordubensis and T. deion Strains

The effects of thelytoky-inducing Wolbachia ( &#102 -proteobacteria) on Trichogramma cordubensis and T. deion (Hymenoptera, Trichogrammatidae) were studied in laboratory and greenhouse conditions. One infected (thelytokous, all female) line of each wasp species was compared with its conspecific uninfected (arrhenotokous, sexual) counterpart for several fecundity and dispersal traits. Arrhenotokous lines had a higher fecundity than their thelytokous counterparts, which suggests that Wolbachia negatively affect the fecundity of Trichogramma females. The arrhenotokous females dispersed more in the laboratory than their thelytokous counterparts. In the greenhouse, the opposite effect or no difference between lines was found, indicating that the laboratory set-up used to measure dispersal is not useful to predict relative dispersal of the females in the greenhouse. Calculations show that by releasing 100 adult wasps of both lines, thus including arrhenotokous males in the sexual line, more eggs are parasitized by the thelytokous wasps. Therefore, in spite of their lower individual female fecundity, thelytokous lines have a better potential for biological control than their arrhenotokous counterparts.     

Quantification of Wolbachia copy number in Trichogramma eggs (Hymenoptera: Trichogrammatidae): Lysozyme treatment significantly improves total gene yield from the Gram-negative bacterium

The Wolbachia bacterium is one of the most prevalent endosymbionts of arthropods. The bacterium is known to induce four distinct reproductive manipulations in its various hosts; namely, cytoplasmic incompatibility, feminization, male killing and parthenogenesis induction. The bacterial titer has been thought to have an important role in expression of these phenotypes. Although host eggs of the bacterium are the place where all of the manipulations take place, in most cases, they are not big enough to be subjected to mechanical disruption to extract genetic materials from them. Furthermore, DNA extraction protocols generally involve washing steps in which part of the DNA is lost. With the simple three step digestion protocol we developed, we were able to preserve the whole genome and measure the titer of Wolbachia in individual Trichogramma eggs by real-time quantitative polymerase chain reaction (PCR), which was comparable to the titer by visual observation.     

Reduced competitive ability due to Wolbachia infection in the parasitoid wasp Trichogramma kaykai

Several hymenopteran parasitoids are infected with parthenogenesis‐inducing (PI) Wolbachia. Infected wasps produce daughters instead of sons from unfertilized eggs. Thus far, little is known about the direct effects of PI Wolbachia on their host's fitness. Here, we report reduced competitive ability due to Wolbachia infection in a minute parasitoid wasp, Trichogramma kaykai Pinto and Stouthamer (Hymenoptera: Trichogrammatidae). Immature survival of infected individuals in a host parasitized by a single infected female, laying a normal clutch of eggs, was lower than those parasitized by a single uninfected individual. When the offspring of infected and uninfected females shared the same host, the infected immatures had significantly lower survival rates than their uninfected counterparts. The survival rate of infected immatures was higher when they competed with other infected immatures from a different infected parent than in competition with uninfected immatures of conspecific wasps. Thus, the host Trichogramma can suffer a substantial reduction in fitness when it is infected with the PI Wolbachia. We discuss why such a reduction is to be expected when populations of infected and uninfected individuals co‐occur, and how the reduced competitive ability of PI Wolbachia influences the spread of the bacteria in the field.