两种小菜蛾寄生蜂产卵器感器的形态和超微结构

半闭弯尾姬蜂和菜蛾盘绒茧蜂是两种重要的小菜蛾寄生蜂.采用扫描电镜对该两种寄生蜂雌蜂产卵器上的感受器的形态和超微结构进行了观察和研究,结果显示半闭弯尾姬蜂和菜蛾盘绒茧蜂的产卵器形态结构基本相同,都是由两个独立的腹产卵瓣和两个愈合在一起的背产卵瓣包围形成产卵道,与其它种类的寄生蜂相似.借背产卵瓣上的脊与腹产卵瓣的沟连锁在一起,腹产卵瓣可以沿背产卵瓣上的脊前后相对滑动.在半闭弯尾姬蜂产卵器上观察到1种钟形感器、2种腔锥感器、浅凹状感器和分泌毛孔.在菜蛾盘绒茧蜂产卵器上同样观察到了这几种感器,但菜蛾盘绒茧蜂产卵器端部的钟形感器突起较狭长,顶部开有气孔.此外菜蛾盘绒茧蜂产卵器上还存在第3种腔锥感器,其中央突起较低而长,表面有网孔.对各种感受器的形态和分布进行了描述,并对其功能进行了探讨,认为半闭弯尾姬蜂的产卵器较细长,且刺突部位有6个倒钩状突起,背、腹产卵瓣上分布更多的感受器,显示出更强的寄主搜寻定位和产卵寄生能力.     

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。     

闭弯尾姬蜂与菜蛾盘绒茧蜂寄生菜蛾幼虫时的种间竞争

在室内25℃下,以菜蛾3龄初幼虫作寄主,研究了菜蛾盘绒茧蜂Cotesia plutellae和半闭弯尾姬蜂Diadegma semiclausum的种间竞争。当寄主供2种蜂同时产卵寄生时,2种蜂各自的寄生率与其单独寄生时无显著差异,合计寄生率比一种蜂单独存在时有所提高,但差异不显著。2种蜂均能产卵寄生已被另一种蜂寄生了的寄主幼虫。当寄主被2种蜂寄生的间隔时间很短（少于10 h）时,所育出的蜂绝大部分（80%以上）为绒茧蜂;当寄主先被绒茧蜂寄生,并饲养2天以上再供弯尾姬蜂寄生时,所育出的全为绒茧蜂;当寄主先被弯尾姬蜂寄生,并饲养2天以上再供绒茧蜂寄生时,寄主幼虫绝大部分不能存活,只有少部分能育出寄生蜂,且多为弯尾姬蜂。当2种蜂的幼虫存在于同一寄主体内时,2种蜂的发育均受到另一种蜂的抑制;绒茧蜂1龄幼虫具有物理攻击能力,能将弯尾姬蜂卵或幼虫致死。这些结果表明,菜蛾盘绒茧蜂与半闭弯尾姬蜂在同一寄主中发育时,前者具有明显的竞争优势。     

二种寄生蜂寄生对不同虫龄小菜蛾精巢的影响

为探明寄生蜂引起寄主寄生性去势的机制,本文选取携带不同多分DNA病毒(Polydnavirus,PDV)的2种内寄生蜂与共同寄主小菜蛾Plutella xylostella(L.)为寄生体系,研究不同虫龄小菜蛾被寄生后雄性生精细胞、精子束形态和精巢发育体积变化,系统比较寄生性去势程度和分别拥有2类PDV的寄生蜂在寄主精子发生和形成过程中的作用。结果表明:携带Bracovirus PDV的菜蛾盘绒茧蜂Cotesiave stalis(Haliday)或拥有Ichvovirus PDV的半闭弯尾姬蜂Diadegma semiclausum Hellén寄生对不同虫龄小菜蛾的精子发生和形成过程均产生明显的抑制作用,表现为不能产生精子束或精子束数量减少,但抑制程度以寄生低龄寄主时最明显。2种蜂寄生均能抑制小菜蛾精巢体积的增长,但对低龄寄主的抑制程度明显强于高龄寄主,寄生性去势程度取决于寄生时寄主虫龄。相比而言,寄生不同虫龄小菜蛾时,茧蜂引起小菜蛾寄生性去势的程度均强于姬蜂。     

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。     

菜蛾盘绒茧蜂多分DNA病毒的特性及其对小菜蛾幼虫的生理效应

对菜蛾盘绒茧蜂Cotesia plutellae多分DNA病毒的特性及其对寄主小菜蛾Plutella xylostella幼虫的生理效应进行了研究。结果表明：菜蛾盘绒茧蜂雌蜂输卵管萼中含有大量的多分DNA病毒（polydnavirus, PDV）;一个PDV内含多个核衣壳,最多可达16个;核衣壳长40～168 nm,直径39～40 nm;PDV仅在输卵管萼细胞内复制;雌蜂产卵时,随蜂卵将PDV注入寄主血腔,并扩散到寄主的许多组织中;PDV可能先通过脱膜再侵染寄主组织。雌蜂经Co⁶⁰辐射处理后再寄生（即假寄生）小菜蛾2龄、3龄和4龄初期的幼虫,被寄生后的寄主幼虫几乎全部不能化蛹,但末龄（即4龄）幼虫期显著延长,并在寄生后期,幼虫胸部有褐色的短翅芽出现;即将化蛹的4龄末小菜蛾幼虫被假寄生后,即使每头寄主被过寄生9次,依然能正常化蛹,但不能羽化。假寄生与正常寄生后寄主的脂肪体数量和形态结构有明显的不同,推测在正常寄生的情况下蜂卵孵化时释放的畸形细胞及随后的幼蜂可能对脂肪体的结构产生了作用。     

菜蛾盘绒茧蜂寄生对寄主小菜蛾幼虫丝腺的影响

菜蛾盘绒茧蜂Cotesia vestalis寄生对小菜蛾Plutella xylostella幼虫丝腺形态结构、丝腺内蛋白组分及含量有显著的影响。寄生后的小菜蛾幼虫的丝腺细胞不经历膨大期即开始凋亡,无法大量分泌蛋白,因此蛋白含量显著低于未寄生的小菜蛾丝腺内的蛋白,且蛋白组分也有一定程度的差异,主要是50 kDa以上的蛋白含量和种类的差别较大。     

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。     

菜蛾绒茧蜂和小菜蛾对杀虫剂的敏感性及酶学特性的比较研究

分别采用药膜法和浸叶法测定了菜蛾绒茧蜂Apanteles plutellae和小菜蛾Plutella xylostella对杀虫剂的敏感度。结果显示： 有机磷、氨基甲酸酯、拟除虫菊酯类杀虫剂、阿维菌素和锐劲特对菜蛾绒茧蜂高毒,而抑太保和Bt为低毒,然而,短时间(1 h)接触常规防治剂量的锐劲特、氰戊菊酯、氯氰菊酯和乙酰甲胺磷对菜蛾绒茧蜂低毒。增效剂胡椒基丁醚（PB）、磷酸三苯酯（TPP）和马来酸二乙酯（DEM）对菜蛾绒茧蜂的甲胺磷、克百威、氰戊菊酯、氯氰菊酯、阿维菌素和锐劲特敏感性增效显著,但对抑太保无增效作用。PB的增效作用显著高于TPP 和DEM。PB和TPP对菜蛾绒茧蜂羧酸酯酶(CarE),以及DEM对谷胱甘肽S转移酶(GST)具显著的活体抑制作用,但PB,TPP和 DEM对菜蛾绒茧蜂乙酰胆碱酯酶（AChE）无抑制作用。菜蛾绒茧蜂AChE的米氏常数(K_m)、最大反应速度(V_max)、CarE和GST活性分别为小菜蛾的0.22、2.08、4.60和0.45倍,甲胺磷、敌敌畏和克百威对菜蛾绒茧蜂AChE的双分子速度常数(Ki)分别为对小菜蛾的14.7、10.5 和26.0倍。酶与抑制剂反应温度增高将导致酶抑制率增高,尤其对菜蛾绒茧蜂AChE的抑制作用更为显著。上述结果表明,菜蛾绒茧蜂对有机磷和氨基甲酸酯类杀虫剂的高敏感性与其显著高的AChE敏感性有关,氧化代谢的解毒作用对菜蛾绒茧蜂耐药性的影响大于水解作用。此外,对小菜蛾和菜蛾绒茧蜂杀虫剂敏感性差异的毒理学原因进行了讨论。     

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.     

A comparison of the host-searching efficiency of two larval parasitoids of Plutella xylostella

1. A host specialist parasitoid is thought to have greater efficiency in locating hosts or greater ability to overcome host defence than a generalist species. This leads to the prediction that a specialist should locate and parasitise more hosts than a generalist in a given arena. The work reported here tested these predictions by comparing the host‐searching behaviour of Diadegma semiclausum (a specialist) and Cotesia plutellae (an oligophagous species), two parasitoids of larval Plutella xylostella. 2. Both parasitoids employed antennal search and ovipositor search when seeking hosts but D. semiclausum also seemed to use visual perception in the immediate vicinity of hosts. 3. Larvae of P. xylostella avoided detection by parasitoids by moving away from damaged plant parts after short feeding bouts. When they encountered parasitoids, the larvae wriggled vigorously as they retreated and often hung from silk threads after dropping from a plant. 4. These two parasitoids differed in their responses to host defences. Diadegma semiclausum displayed a wide‐area search around feeding damage and waited near the silk thread for a suspended host to climb up to the leaf, then attacked it again. Cotesia plutellae displayed an area‐restricted search and usually pursued the host down the silk thread onto the ground. 5. Diadegma semiclausum showed a relatively fixed behavioural pattern leading to oviposition but C. plutellae exhibited a more plastic behavioural pattern. 6. The time spent by the two parasitoids on different plants increased with increasing host density, but the time spent either on all plants or a single plant by D. semiclausum was longer than that of C. plutellae. Diadegma semiclausum visited individual plants more frequently than C. plutellae before it left the patch, and stung hosts at more than twice the rate of C. plutellae. 7. The results indicated that the host‐location strategies employed by D. semiclausum were adapted better to the host's defensive behaviour, and thus it was more effective at detecting and parasitising the host than was C. plutellae.     

Changes in hemocytes of Plutella xylostella after parasitism by Diadegma semiclausum

We examined the changes of hemocytes in the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae), due to parasitism by the endoparasitoid Diadegma semiclausum (Hymenoptera: Ichneumonidae). Necrosis of prohemocytes in different stages was observed while cell death was absent in the mature hemocytes in the parasitized larvae, which was related to the declined total hemocyte count per microliter (THC). THC in the host hemolymph declined sharply by 12 h post-parasitization and then remained at a low level. When hemocytes of the parasitized larvae were cultured in vitro, encapsulation ability was suppressed coincidently with the inhibited spreading ability; however, such effects were transient. Simultaneously, activation of the prophenoloxidae from the hemocytes was inhibited. Unlike the results of previous studies, the decrease in hemocytes, which was due to the necrosis of the prohemocytes instead of the mature hemocytes in our study, was not responsible for the impaired encapsulation. Our studies suggest that parasitism by D. semiclausum have some effects on hematopoietic regulation and on hemocyte immune reaction of P. xylostella larvae.     

Interspecific competition between Diadegma semiclausum Hellen (Hym., Ichneumonidae) and Cotesia plutellae (Kurdjumov) (Hym., Braconidae) in parasitizing Plutella xylostella (L.) (Lep., Plutellidea)

Abstract:  Interspecific competition between Diadegma semiclausum and Cotesia plutellae was investigated at 25°C in the laboratory, by exposing the third instar larvae of the diamondback moth, Plutella xylostella to both species together, either species alone or by exposing the host larvae already parasitized by one species, at different intervals, to the other. When host larvae were exposed simultaneously to two species in one arena, parasitism rates of the host by each species were not reduced by the presence of the other species; joint parasitism rate by two species was not significantly higher than that by either parasitoid alone. Both parasitoids could lay eggs into the host larvae which had previously been parasitized by the other species, leading to the occurrence of multiparasitized hosts. When host larvae were parasitized first by D. semiclausum and then being followed within 1–2 h by exposing to C. plutellae , or vice versa, ensuing parasitoid cocoons from the multiparasitized host larvae were nearly all C. plutellae . When host larvae were parasitized initially by C. plutellae and then being followed by D. semiclausum two or more days later, all parasitoids ensued from the multiparasitized hosts were C. plutellae . In contrast, when host larvae were parasitized initially by D. semiclausum and then being followed by C. plutellae two or more days later, most host larvae could not survive to prepupae and most of the ensuing parasitoid adults from the surviving hosts were D. semiclausum . Dissections of host larvae at various time intervals after parasitization by the two parasitoids showed that development of both parasitoids in multiparasitized hosts were somewhat arrested, and that the first instar larvae of C. plutellae could initiate a physical attack on the larvae of D. semiclausum and remove the latter.     

Effects of insecticides on the foraging behaviour and survival of Cotesia vestalis,a larval parasitoid of the diamondback moth,Plutella xylostella

The effects of the insecticides etofenprox, malathion, diazinon, methomyl, alanycarb, clothianidin and emamectin benzoate on the foraging behaviour and survival of Cotesia vestalis (Hymenoptera: Braconidae), a larval parasitoid of the diamondback moth Plutella xylostella (Lepidoptera: Plutellidae) were investigated in the laboratory. The period required for C. vestalis females to reach P. xylostella‐infested Komatsuna (Brassica rapa) plants treated with etofenprox, methomyl or malathion was significantly longer than that to infested plants treated with emamectin benzoate or clothianidin; and the period to reach alanycarb or diazinon‐treated plants was intermediate between them. The period to reach emamectin benzoate‐ or clothianidin‐treated plants was not significantly different from that to distilled water‐sprayed plants, suggesting that etofenprox, methomyl and malathion have strong inhibitory effects on the flight response, whereas the inhibitory effects of alanycarb and diazinon are relatively weak. By contrast, emamectin benzoate and clothianidin showed no inhibitory effect. The time of residency on the clothianidin‐treated plants was significantly longer than that on other insecticide‐treated plants (etofenprox, methomyl, alanycarb, malathion or diazinon), with the time of residency on emamectin benzoate‐treated plants being intermediate between them. These results suggest that etofenprox, methomyl, alanycarb, malathion, and diazinon have a strong inhibitory effect on host‐searching behaviour, while emamectin benzoate and clothianidin have a relatively weak effect. Furthermore, the mortality of wasps after foraging on the clothianidin‐treated plants was significantly higher than after foraging on other insecticide‐treated plants (etofenprox, methomyl or malathion), with subsequent mortality on alanycarb‐, diazinon‐ and emamectin benzoate‐treated plants being intermediate between them. Our results suggest that subsequent mortality increases with decreased inhibitory effect on the foraging behaviour of C. vestalis.