二种寄生蜂寄生抑制小菜蛾精巢生长和精子束形成的比较研究

我们曾发现菜蛾盘绒茧蜂Cotesia vestalis和半闭弯尾姬蜂Diadegma semiclausum寄生严重阻碍小菜蛾Plutella xylostella幼虫的精子发生。本研究着重比较2种蜂寄生对小菜蛾精巢生长和精子束形成的影响, 以探明寄生因子对昆虫生殖调控的作用途径。 采取过寄生和假寄生方法, 对2种蜂各自寄生后的小菜蛾精巢生长体积, 精子发生和形成过程中生精细胞、精子束的显微形态变化进行了比较。 结果表明: 茧蜂和姬蜂寄生均明显降低小菜蛾精子束的数量, 严重阻碍了寄主幼虫的精子发生和精子形成. 姬蜂寄生造成小菜蛾精巢畸形, 而茧蜂则造成小菜蛾精子束畸形, 且茧蜂对小菜蛾精巢生长的抑制程度明显强于姬蜂。过寄生造成寄主寄生性去势程度加剧, 茧蜂和姬蜂过寄生后的小菜蛾精巢体积分别为0.005 mm³和0.008 mm³, 仅为各自只寄生1次后精巢体积的33.1%和36.3%。假寄生后, 发现只有寄生蜂母代物质存在的前提下, 对小菜蛾精巢生长的抑制程度基本模拟了正常寄生时的状态, 说明多分DNA病毒（polydnavirus, PDV）和毒液发挥了主要作用。 由此推断分属姬蜂属PDV和茧蜂属PDV的2类PDV功能基因对小菜蛾精巢生长发育的调控机制可能存在较大差异。     

寄生蜂的秘密武器

寄生蜂以其他昆虫为宿主营寄生生活,是昆虫中进化程度最高的类群,更是控制农林害虫的重要物种。在长期的进化中,寄生蜂产生了许多克敌制胜的秘密武器,如多分DNA病毒、毒液和畸形细胞等。这些寄生因子在抑制宿主免疫系统的攻击、调节宿主的生长发育、保证寄生蜂成功寄生的过程中发挥重要作用。     

三种内寄生蜂寄生对小菜蛾幼虫精子发生的影响

内寄生蜂寄生可能会引起寄主的寄生性去势。对小菜蛾Plutella xylostella与菜蛾啮小蜂Oomyzus sokolowskii Kurdumov (膜翅目： 姬小蜂科)、半闭弯尾姬蜂Diadegma semiclausum Hellén (膜翅目： 姬蜂科)、菜蛾盘绒茧蜂Cotesia plutellae (Kurdj.) (膜翅目： 茧蜂科) 3个寄生体系,利用形态学方法和蛋白质技术,研究了寄生对小菜蛾幼虫精子发生的影响。结果表明：菜蛾啮小蜂寄生对寄主的精子发生过程没有影响。半闭弯尾姬蜂寄生造成寄主精母细胞的细胞核畸形,精细胞的染色质超浓缩并趋向核膜,但能形成少量的精子;半闭弯尾姬蜂寄生会导致寄主精巢总蛋白的含量显著下降。菜蛾盘绒茧蜂寄生对小菜蛾幼虫精子发生的抑制程度最强,被寄生寄主的精母细胞出现肿胀,核膜皱缩,胞质中的线粒体发生病变;精细胞的染色体也出现超浓缩并趋向核膜,大量的精子溶解,无正常的精子形成;其精巢总蛋白含量的下降程度比姬蜂寄生的更为明显,且导致分子量为63.4 kD的主蛋白缺失。     

寄生蜂多分DNA病毒对寄主昆虫的免疫抑制作用

多分DNA病毒（PDV）是在膜翅目姬蜂科和茧蜂科寄生蜂体内的一类很独特的病毒,寄生蜂产卵时,PDV随同卵和萼液一起被注射入寄主体内,能干扰寄主的细胞免疫和体液免疫,该病毒直接侵染或间接作用于血细胞,主要是浆细胞和颗粒细胞,导致血细胞变圆或凋亡,PDV也能抑制血淋巴酚氧化酶活性,诱导抗菌因子的大量合成,最近有关研究主要集中在病毒基因的表达和伴随寄主血细胞功能失常的分子事件上,一些寄主蜂的PDV与其他因子,如卵巢蛋白,畸形细胞或蜂毒等协同发挥作用。     

寄生蜂多分DNA病毒的基因表达产物及对寄主昆虫的生理作用

最近有关由多分DNA病毒介导的寄生蜂与寄主关系的研究主要集中在多分DNA病毒基因的表达和伴随寄主生理功能失常的分子机理上 ,本文介绍在寄生蜂作用于寄主时一些重要的PDV基因表达产物 ,包括CsPDV ,MdPDV ,CcPDV ,TrPDV ,HdPDV ,CrPDV和其它PDV的基因表达产物 ,并简叙这些基因表达产物对寄主的生理影响。     

多态DNA病毒对寄生蜂宿主生理功能的影响

多态DNA病毒(polydnavirus, PDV)由一组特异的病毒组成,其生活史与某些寄生蜂种类有着密不可分的联系.大量研究表明该种病毒是决定某些寄生蜂能否在寄主体内成功寄生的关键因子之一[1].本文将综述已有的研究报道,重点阐明PDV随寄生蜂产卵而传递到寄生蜂的寄主体内时所发挥的确切生理功能和作用方式.     

多态DNA病毒对寄生蜂宿主生理功能的影响

多态ＤＮＡ病毒（ｐｏｌｙｄｎａｖｉｒｕｓ，ＰＤＶ）由一组特异的病毒组成，其生活史与某些寄生蜂种类有着密不可分的联系。大量研究表明该种病毒是决定某些寄生蜂能否在寄主体内成功寄生的关键因子之一［１］。本文将综述已有的研究报道，重点阐明ＰＤＶ随寄生蜂产卵而...     

多分DNA病毒及其在寄生蜂与寄主关系中的作用

分DNA病毒(PDV)在协调寄生蜂与寄主的相互关系中起重要作用。该文就PDV 的特征、PDV与寄生蜂的相互关系、PDV对鳞翅目寄主的免疫抑制及发育调节等方面进行综述。     

半闭弯尾姬蜂寄生及其毒液对小菜蛾幼虫血细胞吞噬作用的影响

半闭弯尾姬蜂Diadegma semiclausum是小菜蛾Plutella xylostella的优势内寄生蜂, 拥有毒液、多分DNA病毒（PDV）等寄生因子,能有效调控寄主幼虫的营养生理和免疫系统, 但其毒液在这过程中的功能不明。本文利用SDS-PAGE方法分析了半闭弯尾姬蜂毒液的蛋白组分,利用寄主幼虫血细胞体外原代培养的方法,研究了小菜蛾幼虫血细胞噬菌能力在半闭弯尾姬蜂寄生后的变化情况。结果表明：半闭弯尾姬蜂毒液蛋白分子量主要集中在35~220 kDa之间,少数小于15 kDa,但分子量处于35~70 kDa之间的蛋白含量较高,与其他寄生蜂毒液蛋白相似。半闭弯尾姬蜂毒液单独对寄主小菜蛾幼虫功能血细胞（浆血细胞和颗粒血细胞）的延展能力和吞噬功能不产生破坏作用。但半闭弯尾姬蜂寄生后短时间内,寄主功能血细胞的延展受到抑制,然而功能血细胞仍然能识别外源异物, 却无法进一步吞噬外源物; 寄生后24 h,功能血细胞的延展力恢复,颗粒血细胞的吞噬作用可顺利完成。本研究证明了半闭弯尾姬蜂寄生能暂时性地抑制颗粒血细胞的延展性从而影响其噬菌过程。     

辽宁蚧虫寄生蜂二新种（膜翅目：跳小蜂科）

报道采自辽宁阜新的蚧虫寄生蜂2新种：微小玛丽跳小蜂Mayridia parva,Wu et Xu,sp.n和短棒多丽跳小蜂Doliphoceras brachyclavae Wu et Xu,sp.n.。本文也是玛丽跳小蜂属Mayridia Mecet和多丽跳小蜂属Doliphoceras Mercet在我国分布的首次记录,标本夏放于激光大学农业与生物技术学院应用昆虫研究所。     

菜蛾盘绒茧蜂主要寄生因子导致的寄主小菜蛾幼虫脂肪体结构的变化

在不同的寄生状态下,菜蛾盘绒茧蜂Cotesia plutellae不同的寄生因子可引起寄主小菜蛾Plutella xylostella幼虫脂肪体结构发生相应的改变。显微和亚显微形态结构显示： 假寄生后多分DNA病毒和毒液对脂肪体结构的完整性没有显著影响,但细胞内脂质体变得小而密集,线粒体和内质网丰富,并有糖原积累; 正常寄生后,脂肪体结构被破坏,多数线粒体内嵴紊乱,脂质体也变得不规则,特别是当幼蜂完成在寄主体内发育时,寄主体内几乎无完整脂肪体存在。与此同时,同批未被寄生的小菜蛾幼虫发育到4龄末期时,体内脂肪体细胞发育正常,已开始向蛹期细胞形态转化,细胞内脂质体很大,细胞器数量较多、糖原积累丰富, 而且部分细胞已成为游离态细胞。由此证明,寄生蜂携带的寄生因子,如多分DNA病毒、毒液、畸形细胞和幼蜂等,均对寄主脂肪体结构的改变产生影响,但程度明显不同。     

小菜蛾颗粒体病毒PP31与两种宿主蛋白发生相互作用

在病毒侵染和复制过程中,病毒与宿主之间存在广泛的蛋白质-蛋白质相互作用。本研究建立了一个小菜蛾幼虫cDNA文库,用于筛查与小菜蛾颗粒体病毒(Plutella xylostella granulovirus,PlxyGV)蛋白相互作用的小菜蛾幼虫蛋白。pp31同源基因存在于所有鳞翅目昆虫杆状病毒。其编码产物是一种磷蛋白,与病毒基因表达调控相关。本研究通过酵母双杂交实验从小菜蛾幼虫cDNA文库中筛选出两个与PlxyGV PP31相互作用的蛋白质基因。序列分析结果显示这两个基因的预期编码产物分别是小菜蛾蛋白激酶C受体(RACK)和一种甲硫氨酰氨肽酶2(MetAP2)同源蛋白。原核表达和蛋白质纯化实验结果显示,rack与6-组氨酸编码序列的融合基因的表达产物是一个38kD多肽。在GST-pulldown实验中,RACK蛋白随同GST-PP31融合蛋白一起吸附于GST亲和树脂,进一步证实了小菜蛾RACK蛋白与PlxyGV PP31发生相互作用。     

Parasitism-induced effects on host growth and metabolic efficiency in Plutella xylostella larvae parasitized by Cotesia vestalis or Diadegma semiclausum

The nutritional physiology of the diamondback moth, Plutella xylostella, larvae was examined after parasitization by the solitary endoparasitoids Cotesia vestalis or Diadegma semiclausum. Examinations were performed in two phases, one was examined at the time point of 24 h post‐parasitization, and the other was done at the end of the 4th instar larvae of host. Rates of growth, food consumption, assimilation, excretion, and respiration were calculated as well as approximate digestibility and the rate ratios ECI (percent efficiency of conversion of ingested food to body substance), and ECD (percent efficiency of conversion of digested food to body substance). Parasitization by C. vestalis resulted in significant decrease in the rates of growth, feeding, excretion, assimilation, and respiration, but the final dry rate of respiration at the end of last larval stadium was elevated. The ECI and ECD were also reduced as the result of parasitization, but digestibility was increased. All these parameters in the larvae parasitized by D. semiclausum at 24 h post‐parasitization were also significantly changed compared to the control; however, these differences were quantitatively, but not qualitatively before pupation, similar to those resulted from parasitization by C. vestalis. In spite of the similarities of the parasitism‐induced effects caused by these endoparasitoids, the final metabolic rate, that is, the rate of intake of nutrients required to compensate for metabolism, was much lower in the larvae parasitized by C. vestalis than that of the larvae parasitized by D. semiclausum. All of the results discussed here will contribute toward explaining the different ways these two wasps regulate the parasitoid‐host relationship.     

Immunosuppression induced by expression of a viral RNase enhances susceptibility of Plutella xylostella to microbial pesticides

Abstract Polydnaviruses are a group of insect DNA viruses and are characterized in their segmented genome that is located in the chromosome(s) of host wasps. A polydnavirus, Cotesia plutellae bracovirus (CpBV), encodes a viral ribonuclease (RNase) T2 in a specific segment #3 (CpBV‐S3). This study tested its effect on gene expression associated with host immune responses in the diamondback moth, Plutella xylostella. Micro‐injection of CpBV‐S3 into nonparasitized larvae induced expression of its two encoded genes, CpBV‐ORF301 (=CpBV‐RNase T2) and CpBV‐ORF302. In response to a bacterial challenge, four antimicrobial peptide genes (hemolin, gloverin, cecropin and lysozyme) and six phenoloxidase (PO)–associated genes (proPO‐activating proteinase, PO, serine proteinase homolog and serpins 1–3) were up‐regulated in their expressions. However, the transient expression of CpBV‐S3 suppressed the expressions of cecropin, PO and serpin 1. Double‐stranded RNA specific to the viral RNase T2 could specifically knockdown the viral gene expression and restored the three gene expressions suppressed in the larvae injected with CpBV‐S3. The inhibitory activity of the viral RNase T2 on the target genes was further proven by the suppression of PO activation in response to bacterial challenge in the larvae injected with CpBV‐S3. This immunosuppression by the expression of the viral RNase T2 resulted in significant increase of pathogen susceptibility of P. xylostella against Bacillus thuringiensis or baculovirus infection.     

小菜蛾幼虫空间格局的地统计学分析

应用地统计学方法分析了小菜蛾Plutellaxylostella (L .)幼虫空间格局 ,建立了东西和南北 2个方向上多个半变异函数曲线模型。结果表明 ,在菜花 (Brassicaolenaceavar.botrylis)和早甘蓝 (Brassisaoleracea)地中小菜蛾幼虫种群的半变异函数曲线皆为球形 ,其空间格局为聚集型 ;小菜蛾幼虫在菜花上和在早甘蓝上的空间变程分别为 0 62～ 1 5 9m和 2 47～ 5 98m ,空间不连续性强度分别为 0 8779～0 9999和 0 862 1～ 0 91 2 4。     

卵磷脂促进小菜蛾颗粒体病毒感染的研究

采用有机溶剂从鸡蛋中提取卵磷脂,与小菜蛾颗粒体病毒混合添食感染2—3龄小菜蛾幼虫,所得数据经统计学方法处理,得到三组死亡率与时间的回归直线方程为y=18.1x-59.8(GV对照)、y=27.23x-99.85(GV+30mg卵磷脂/ml)、y=13.6x-18(GV+70mg卵磷脂/ml)。GV+70mg卵磷脂/ml与GV对照组比较,感染性提高了17％,LT_(50)提前了1.066天。结果表明,卵磷脂对小菜蛾颗粒体病毒感染具有促进作用。     

CLP gene family,a new gene family of Cotesia vestalis bracovirus inhibits melanization of Plutella xylostella hemolymph

Polydnaviruses (PDVs) are obligatory symbionts of parasitoid wasps and play an important role in suppressing host immune defenses. Although PDV genes that inhibit host melanization are known in Microplitis bracovirus, the functional homologs in Cotesia bracoviruses remain unknown. Here, we find that Cotesia vestalis bracovirus (CvBV) can inhibit hemolymph melanization of its host, Plutella xylostella larvae, during the early stages of parasitization, and that overexpression of highly expressed CvBV genes reduced host phenoloxidase activity. Furthermore, CvBV-7-1 in particular reduced host phenoloxidase activity within 12 h, and the injection of anti-CvBV-7-1 antibody increased the melanization of parasitized host larvae. Further analyses showed that CvBV-7-1 and three homologs from other Cotesia bracoviruses possessed a C-terminal leucine/isoleucine-rich region and had a similar function in inhibiting melanization. Therefore, a new family of bracovirus genes was proposed and named as C -terminal L eucine/isoleucine-rich P rotein (CLP). Ectopic expression of CvBV-7-1 in Drosophila hemocytes increased susceptibility to bacterial repression of melanization and reduced the melanotic encapsulation of parasitized D. melanogaster by the parasitoid Leptopilina boulardi. The formation rate of wasp pupae and the eclosion rate of C. vestalis were affected when the function of CvBV-7-1 was blocked. Our findings suggest that CLP genes from Cotesia bracoviruses encoded proteins that contain a C-terminal leucine/isoleucine-rich region and function as melanization inhibitors during the early stage of parasitization, which is important for successful parasitization.