黄眶离缘姬蜂卵巢和毒液器官的形态学及其超微结构

本文观察了内寄生蜂黄眶离缘姬蜂Trathala flavo-orbitalis Cameron卵巢和毒液器官的形态及超微结构,为了解黄眶离缘姬蜂与其寄主之间的协同进化关系奠定基础。在光学显微镜下解剖雌性生殖系统,并结合透射电镜进行形态学及超微结构观察,结果表明黄眶离缘姬蜂毒液器官主要由一个薄壁的毒囊、两条接于毒囊顶端的毒腺、通向产卵器的毒液导管组成。毒腺由分泌细胞层和环腔的内膜构成,分泌细胞层中存在大量的端器,主要功能是分泌毒液。毒囊由肌肉鞘层、扁平细胞层以及内膜层组成,肌肉鞘内肌纤丝规则排列,扁平细胞层内的细胞器极其稀少,内膜呈波浪状均匀加厚。黄眶离缘姬蜂的一对卵巢分别由8条卵巢小管组成,开口于两条侧输卵管,在卵巢与侧输卵管相接处略微膨大形成卵巢萼区,萼区内存在大量类似姬蜂病毒的颗粒。黄眶离缘姬蜂的毒液器官属于II型毒液器官,是较高等的进化模式,其毒液器官的形态结构与雌蜂的产卵行为相关。在萼区内发现的类似姬蜂病毒的颗粒,对研究寄生蜂调控寄主的免疫和发育生理具有重要意义。     

管氏肿腿蜂毒液器官超微结构观察

应用透射电镜技术,观察了管氏肿腿蜂Scleroderma guani毒液器官的超微结构.毒腺由基膜层、分泌细胞层、导管细胞层和内膜层构成,分泌细胞内含内质网、末端附器、分泌囊泡、分泌颗粒、液泡等细胞器,其内合成的毒液由末端附器输送至毒腺的腔体.毒囊由肌肉鞘层、上皮细胞层和内膜层组成,肌肉鞘内的肌纤丝规则排列不交错,上皮细胞层内细胞器稀少,内膜层呈波浪状均匀加厚.     

黄腹潜蝇茧蜂寄生因子的特性及其对寄主的生理效应

初步研究了黄腹潜蝇茧蜂Opius caricivorae Fischer寄生因子的特性及其对寄主美洲斑潜蝇Liriomyza sativae Blanchard幼虫的生理效应。黄腹潜蝇茧蜂携带的主要因子是毒液。毒液器官是由一个土黄色的锥形毒囊和7个透明的椭圆形的毒腺及导管构成的;毒液的电泳图谱显示约有12条蛋白带,其中绝大多数低于100 kD,含量最高的3条蛋白带为43.5、25.9和20.1 kD;杜氏腺约有15条左右蛋白质条带,其中有5条含量很高（121.4、77.0、51.5、42.7和36.5 kD）。通过透射电镜观察,在黄腹潜蝇茧蜂毒腺分泌细胞和卵巢表皮细胞中新发现存在一种类病毒颗粒,这些球状颗粒直径大约为50 nm。雌蜂经Co60辐射处理后再寄生（即假寄生）3龄寄主幼虫,被寄生后的寄主依然能正常化蛹,但不能羽化;7 h后寄生体壁开始出现红斑;脂肪体形态结构无显著变化;绝大多数的蜂卵没有被包囊。推测在正常寄生的情况下可能是毒液抑制了寄主的包囊作用,而新发现的类病毒颗粒是否参与了这一过程目前还不清楚。     

腰带长体茧蜂Macrocentrus cingulum毒液和卵巢蛋白的电泳分析

寄生蜂毒液和卵巢蛋白在寄生过程中起着重要作用,蛋白成分和性质的研究日益深入.本文主要通过SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)试验,分析了腰带长体茧蜂毒液和卵巢蛋白的分子量组成.结果表明,腰带长体茧蜂毒液蛋白图谱显示约有7条蛋白带介于43～100 kDa之间,含量较高的三条带为97kDa、64kDa、45kDa;卵巢蛋白图谱显示约有12条蛋白带,位于30～200 kDa之间,含量较高的两条带为39kDa、43kDa.并对毒液和卵巢的生物学功能进行了探讨.     

两种寄生蜂的卵巢结构与卵成熟特点初步观察

对斜纹夜蛾的内寄生蜂双斑侧沟茧蜂MicroplitisbicoloratusChen和南美斑潜蝇的外寄生蜂潜蝇姬小蜂Diglyhusisaea(Walker)的卵巢结构进行了研究。双斑侧沟茧蜂的2个卵巢管底部膨大有大量的卵,通过凹陷的颈部与萼区相连接,在萼区里大量成熟待产的卵浸泡在萼液中。产卵量随产卵时间延长而降低,在日产卵实验中,日产卵量202(±114SE),从1000至1800每2h的产卵量分别占日产卵量的38,32,17和13%;在一生产卵量实验中,9d产卵期,每头雌峰一生平均产卵量为502(±78SE)粒,第1d产卵量占一生总产卵量的42%。潜蝇姬小蜂的每个卵巢由3条卵巢管组成,每条输卵管中仅有1粒成熟卵,整个卵巢中只有6粒成熟卵,每次产卵数不超过6粒。结果表明,M.bicoloratus是早熟卵(proovigenic)类昆虫;D.isaea是同步卵(synovigenic)类昆虫。     

腰带长体茧蜂寄生对亚洲玉米螟幼虫体内酚氧化酶活性的影响

通过对被腰带长体茧蜂Macrocentrus cingulum Brischke寄生的5龄亚洲玉米螟Ostrinia furnacalis Guenée幼虫体内不同组织中酚氧化酶活性的测定,采用体外注射腰带长体茧蜂雌性成蜂的萼液成分、毒液成分、萼液与毒液混合物的方法,研究了寄生蜂各种主要生理因子对寄主血清中酚氧化酶活性的影响。结果表明： 寄生蜂寄生可明显抑制寄主体内的酚氧化酶活性,减少黑色素产生;被寄生组FITC标记的血细胞阳性百分率低于未被寄生组,差异极显著( P<0.01）;萼液成分可明显地抑制亚洲玉米螟幼虫血清中酚氧化酶的活性 （P<0.01）;萼液与毒液混合物对酚氧化酶活性也有明显抑制作用（P<0.01）。研究认为寄生蜂产卵时注入的萼液、毒液可对寄主昆虫酚氧化酶活性产生明显的抑制作用,其中萼液是抑制寄主免疫能力的主要因素。     

杀虫蛋白Vip3Aa11对亚洲玉米螟及其寄生性天敌腰带长体茧蜂的影响

【目的】本文的研究目的是明确杀虫蛋白Vip3Aa11对亚洲玉米螟Ostrinia furnacalis及其寄生性天敌腰带长体茧蜂Macrocentrus cingulum的影响。【方法】首先利用生物测定的方法用含100μg/g Vip3Aa11蛋白的人工饲料饲喂亚洲玉米螟初孵幼虫,7 d后观察记录亚洲玉米螟的死亡情况和体重。然后用含100μg/g Vip3Aa11蛋白的人工饲料饲喂被腰带长体茧蜂寄生的亚洲玉米螟初孵幼虫,以含6μg/g印楝素的人工饲料作为阳性对照,10 d后记录亚洲玉米螟幼虫的死亡和被寄生情况,并在茧蜂化蛹结茧及羽化后分别记录茧重以及单个寄主的出蜂量,以此评价Vip3Aa11蛋白对腰带长体茧蜂的间接影响。再者,用含100μg/g Vip3Aa11蛋白的20%蜂蜜水饲喂腰带长体茧蜂成虫,以含100μg/g印楝素的20%蜂蜜水作为阳性对照,观察记录腰带长体茧蜂成蜂的死亡情况;不同处理的腰带长体茧蜂寄生亚洲玉米螟10 d后记录亚洲玉米螟的死亡和被寄生情况,并在子代茧蜂化蛹结茧及羽化后记录茧重及单个寄主的出蜂量,以此评价Vip3Aa11蛋白对腰带长体茧蜂的直接影响。【结果】生物测定结果显示,100μg/g的Vip3Aa11蛋白处理7 d后亚洲玉米螟幼虫的平均死亡率为50.7%,平均体重抑制率为77.1%。间接影响试验结果显示,被腰带长体茧蜂寄生后的亚洲玉米螟幼虫取食含有Vip3Aa11蛋白的人工饲料后死亡率明显升高,腰带长体茧蜂茧重和单头出蜂量大幅度下降,而寄生率及羽化后的成虫寿命没有受到不利影响。直接影响试验结果显示,腰带长体茧蜂取食含有100μg/g Vip3Aa11蛋白的蜂蜜水对腰带长体茧蜂成虫寿命和寄生率、亚洲玉米螟幼虫死亡率以及腰带长体茧蜂子代的茧重、单头出蜂量和成虫寿命均没有产生不利影响。【结论】本研究利用生测体系从直接和间接两方面评价了Vip3Aa11蛋白对腰带长体茧蜂的影响,结果表明腰带长体茧蜂对高于Bt作物中表达的Vip3Aa11蛋白浓度不敏感,Vip3Aa11蛋白不会对腰带长体茧蜂产生直接的不利影响;造成的间接影响可能主要是由于寄主自身质量的下降而引起。     

中国少棘蜈蚣（Scolopendra subspinipes mutillans）的毒腺结构

用免疫组化和光镜、透射电镜等观察了中国少棘蜈蚣毒腺的结构。结果显示,纵贯颚肢的弯月形毒腺为单管泡状腺,主要由柱状分泌细胞和介于其间的纤细表皮细胞组成。被肌肉束环绕的分泌细胞辐射状排列于几丁质的毒液导管外,其纤细的颈部由环状括约肌控制,分泌端以折叠回转的单向瓣膜经导管壁上的孔道直接伸入管腔,膨大的盲端直达毒腺底膜。高电子密度的分泌溶酶体向分泌口汇集时电子密度逐渐降低并降解为分泌小泡,其中的杆状结晶样毒蛋白也经无定型状态逐渐分散,经胞吐作用进入管腔并进一步疏散和均质化。免疫组化显示,分泌细胞颈部密集的分泌颗粒的主要成分为毒蛋白,毒蛋白在分泌细胞中呈明显的向心式梯度增强型分布。根据上述观察,提出了蜈蚣毒液以分泌溶酶体介导的非经典途径分泌的观点。     

中华蜜蜂咽下腺的电镜观察

王浆是由工蜂的咽下腺所分泌,咽下腺位于工蜂头腔内两侧,每侧腺体都有一条长而粗的分泌管,在分泌管的两侧平行排列有许多椭圆形小叶,每个小叶内有十多个细胞,每个细胞有一根胞外管与分泌管直接相通,细胞内有一条胞内管又与胞外管相连。王浆就是通过细胞内分泌块分泌而成,由胞内管把王浆输送到胞外管,最后由分泌管到咽喉部,在此处饲喂王浆给蜂王、幼蜂和雄蜂。通过咽下腺的超微结构观察,不仅了解到咽下腺各种细胞器的形态特征,同时也了解到输送王浆的途径。     

橙黄豆粉蝶生殖系统形态学研究

解剖并描述了橙黄豆粉蝶Colias fieldii Ménétriés的生殖系统结构。结果表明:橙黄豆粉蝶雄性内生殖器包括精巢(睾丸)、2个贮精囊、2条输精管、1对复射精管、2根附腺和单射精管。其中2个睾丸体彼此密接似单一器官包被在半透明的睾丸膜中,单射精管较长且分化为形态不同的4段。外生殖器包括抱器瓣、阳茎及其附属结构;雌性内生殖器由1对卵巢、2根侧输卵管、1根中输卵管、肾型受精囊、附腺、外生殖腔及产卵孔组成。卵巢左右对称,每个卵巢具4根多滋式卵巢管。外生殖器包括导精管、囊导管、交配囊及其附属结构、前后表皮突和肛突。     

Virus-like particles in venom of Meteorus pulchricornis induce host hemocyte apoptosis

Ultrastructural studies on the reproductive tract and venom apparatus of a female braconid, Meteorus pulchricornis, revealed that the parasitoid lacks the calyx region in its oviduct, but possesses a venom gland with two venom gland filaments and a venom reservoir filled with white and cloudy fluid. Its venom gland cell is concaved and has a lumen filled with numerous granules. Transmisson electron microscopic (TEM) observation revealed that virus-like particles (VLPs) were produced in venom gland cells. The virus-like particle observed in M. pulchricornis (MpVLP) is composed of membranous envelopes with two different parts: a high-density core and a whitish low-density part. The VLPs of M. pulchricornis is also found assembling ultimately in the lumen of venom gland cell. Microvilli were found thrusting into the lumen of the venom gland cell and seem to aid in driving the matured MpVLPs to the common duct of the venom gland filament. Injection of MpVLPs into non-parasitized Pseudaletia separata hosts induced apoptosis in hemocytes, particularly granulocytes (GRs). Rate of apoptosis induced in GRs peaked 48h after VLP injection. While a large part of the GR population collapsed due to apoptosis caused by MpVLPs, the plasmatocyte population was minimally affected. The capacity of MpVLPs to cause apoptosis in host's hemocytes was further demonstrated by a decrease ( approximately 10-fold) in ability of host hemocytes to encapsulate fluorescent latex beads when MpVLPs were present. Apparently, the reduced encapsulation ability was due to a decrease in the GR population resulting from MpVLP-induced apoptosis.     

The venom apparatus in stenogastrine wasps: Subcellular features of the convoluted gland

In the wasp venom apparatus, the convoluted gland is the tract of the thin secretory unit, i.e. filament, contained in the muscular reservoir. Previous transmission electron microscope investigation on Stenogastrinae disclosed that the free filaments consist of distal and proximal tracts, from/to the venom reservoir, characterized by class 3 and 2 gland patterns, respectively. This study aims to extend the ultrastructural analysis to the convoluted tract, in order to provide a thorough, subcellular representation of the venom gland in these Asian wasps. Our findings showed that the convoluted gland is a continuation of the proximal tract, with secretory cells provided with a peculiar apical invagination, the extracellular cavity, collecting their products. This compartment holds a simple end-apparatus lined by large and ramified microvilli that contribute to the processing of the secretory product. A comparison between previous and present findings reveals a noticeable regionalization of the stenogastrine venom filaments and suggests that the secretory product acquires its ultimate composition in the convoluted tract.     

Venom gland of the ichneumonid Diadromus collaris： morphology, ultrastructure and age-related changes

Females of the solitary parasitoid Diadromus collaris （Insecta： Hymenoptera： Ichneumonidae） lay eggs in the pupae of Plutella xylostella （Lepidoptera： Plutellidae）, and the venom is synchronously injected into hosts. The venom apparatus consists of two glandular tubules terminating in a common reservoir, A ductule connects the reservoir with the sting apparatus, by which the reservoir content enters the latter. Secretory units line the two glandular tubules. All secretory cells belong to dermal gland type Ⅲ. Dermal gland cells in glandular tubules are more abundant and developed than those in the reservoir. There are extensive rough endoplasmic reticulum and electrondense vesicles, and the microvilli are well developed. By the cuticle-lined central funnel secretion products of secretory units reach the reservoir. Moreover, the secretory apparatus undergoes age-related changes. The secretory units in the venom gland are better developed and more vigorous 7 days after eclosion than those 1 day after eclosion; autolytic processes occur 15 days after eclosion, and the tissue of the reservoir is more prostrate 15 day after eclosion than those 1 day after eclosion. The ovipostion peak of this parasitoid, about 3-7 days after eclosion, corresponds with the period when the venom gland is highly developed in the life span of the wasp.     

Venom gland of the digger wasp Liris niger: morphology, ultrastructure, age-related changes and biochemical aspects

Females of the parasitoid digger wasp species Liris niger hunt crickets as food for their future brood. The wasps paralyse the prey by injecting their venom directly into the CNS. The venom is produced in a gland consisting of two ramified glandular tubules terminating in a common reservoir. The reservoir contents enter the sting bulb via a ductus venatus. Secretory units of dermal gland type III line the two free gland tubules, the afferent ducts to the reservoir and the cap region within the reservoir. Secretion products of tubules reach the reservoir through the cuticle-lined central funnel. Secretory cells in the distal and middle parts of the tubules contain extensive rough endoplasmic reticulum and numerous electron-dense vesicles, whereas secretory cells of the afferent ducts and the cap region of the reservoir lack electron-dense vesicles and the endoplasmic reticulum is poorly developed. The secretory apparatus undergoes age-related changes. The secretory units in the venom gland tubules and inside the reservoir complete differentiation 1 day after imaginal ecdysis. After 30 days, massive autolytic processes occur in the secretory cells and in the epithelial cells of the reservoir. Analysis of the polypeptide composition demonstrates that the venom reservoir contains numerous proteins ranging from 3.4 to 200 kDa. A dominant component is a glycoprotein of about 90 kDa. In contrast the polypeptide composition of Dufour's gland is completely different and contains no glycoproteins. Comparison of the venom reservoir contents with the polypeptide pattern of venom droplets reveals that all of the major proteinaceous constituents become secreted. Thus the secreted venom contains exclusively proteins present in the soluble contents of the venom gland.     

Morphology and ultrastructure of a secretory region enclosed by the venom reservoir in social wasps (Insecta,Hymenoptera)

The morphology and ultrastructure of the convoluted gland inside the venom reservoir of four species of social Vespidae are described. The cells of the venom gland (including the convoluted gland) can be divided into six groups: (1) epithelial cells, (2) glandular cells with the end apparatus secreting into the tubule inside the convoluted gland (internal or embedded tubule), (3) a continuous arrangement of glandular cells with the end apparatus secreting directly into the venom reservoir, (4) glandular cells that are loosely dispersed along the tubule lumen between the free tubules and the embedded tubule of the convoluted gland, (5) secretory cells of the free tubules and (6) duct cells. One kind of secretory cell, hitherto unknown and described in this paper (group 3), is characterized by the presence of a well-developed end apparatus, usually with enlarged extracellular spaces, but lacking the normally associated duct cells. The secretory cells contain several stacks of granular endoplasmic reticulum, but these are mainly concentrated in the middle of the cell. The basal half of the cells contains many lipid droplets. Although the function of the convoluted gland is not yet understood, an hypothesis is related to what is known of the function of reservoir secretory cells in solitary wasps. All wasp species studied showed the same organization of the convoluted gland, which clearly distinguishes their venom gland from that of Sphecidae.     

Venom and Dufour's glands of the emerald cockroach wasp Ampulex compressa (Insecta,Hymenoptera, Sphecidae): Structural and biochemical aspects

The digger wasp species Ampulex compressa produces its venom in two branched gland tubules. They terminate in a short common duct, which is bifurcated at its proximal end. One leg is linked with the venom reservoir, the other one extends to the ductus venatus. Each venom gland tubule possesses, over its entire length, a cuticle-lined central duct. Around this duct densely packed class 3 gland units each composed of a secretory cell and a canal cell are arranged. The position of their nuclei was demonstrated by DAPI staining. The brush border of the secretory cells surrounds the coiled end-apparatus. Venom is stored in a bladder like reservoir, which is surrounded by a thin reticulated layer of muscle fibres. The reservoir as a whole is lined with class 3 gland units. The tubiform Dufour's gland has a length of about 350 μm (∅ 125 μm) only and is surrounded by a network of pronounced striated muscle fibres. The glandular epithelium is mono-layered belonging to the class 1 type of insect epidermal glands. The gland cells are characterized by conspicuous lipid vesicles. Secretion of material via the gland cuticle into the gland lumen is apparent. Analysis of the polypeptide composition demonstrated that the free gland tubules and the venom reservoir contain numerous proteins ranging from 3.4 to 200 kDa. The polypeptide composition of the Dufour's gland is completely different and contains no lectin-binding glycoproteins, whereas a dominant component of the venom droplets is a glycoprotein of about 80 kDa. Comparison of the venom reservoir contents with the polypeptide pattern of venom droplets revealed that all of the major proteinaceous constituents are secreted. The secreted venom contains exclusively proteins present in the soluble contents of the venom gland. The most abundant compound class in the Dufour's gland consisted of n-alkanes followed by monomethyl-branched alkanes and alkadienes. Heptacosane was the most abundant n-alkane. Furthermore, a single volatile compound, 2-methylpentan-3-one, was identified in various concentrations in the lipid extract of the Dufour's gland.     

Development of female accessory glands of Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae) during oogenesis

Females of Chrysomya putoria (Diptera: Calliphoridae) have two sexual accessory glands, which are tubular and more dilated at the distal extremity. The glands open independently into the common oviduct. Two morpho-physiological regions were distinguished in the longitudinal semi-thin sections of the glands. The secretory region is constituted by three layers: a cuticular intima, lining the lumen, followed by a layer of small cells, and then a layer of very large secretory cells. The ductal region of the gland presents only two layers: the cuticular intima and a cellular layer. In both regions a basement membrane is present. Each secretory cell has in its apical region a reservoir, which enlarges throughout oogenesis; in its basal region there is a large nucleus. The ductal cells are cylindrical and smaller than the secretory cells. The glandular secretion is synthesized in the cytoplasm of the secretory cells, stored and/or modified in the reservoir, then drained to the lumen through an end apparatus seen in the apical region of the secretory cell. Histochemical tests indicate that this secretion is a glycoprotein. Measurements of the glands from females at different physiological conditions and fed on different diets correlate with the results obtained for changes in the ovary during oogenesis. Cell number averaged 561.2 ± 77.54 per gland. There was no increase in cell number during oogenesis.     

Ultrastructure of the venom gland of the scorpion,Centruroides sculpturatus (Ewing)

The venom apparatus of the scorpion, C. sculpturatus (Ewing) was studied with light and electron microscopy. Each of the paired glands is lined by secretory epithelium made up of a single layer of columnar cells. Extensive folding in the epithelial layer creates a primitive acinar gland. The secretory products are either membrane-bound or unbound vesicles with discrete morphologies and are observed in the extruded venom, within the lumen of the gland, and within single secretory cells. The venom apparatus, including connective tissues, nerve cells, and muscle tunic is described and correlations are made with observations in other Athropods.     

Morphology and ultrastructure of the convoluted gland in the ant Dinoponera australis (Hymenoptera: Formicidae)

The morphology and fine structure of the convoluted gland inside the venom reservoir of the ponerine ant Dinoponera australis (Hymenoptera: Formicidae) are described. The cells of the convoluted gland can be divided into 3 major groups: (1) epithelial cells, (2) glandular cells with end apparatus secreting into the tubule inside the convoluted gland, and (3) glandular cells with end apparatus secreting directly into the venom reservoir. A fourth group of cells belonging to the venom gland of this ant is also discussed as (4) secretory cells of the free tubule (not a part of the convoluted gland). The epithelial cells in the convoluted gland do not have many organelles. Most cells of group 3 are characterized by numerous mitochondria. In some of these cells, the mitochondria possess tubular cristae. Tubule cells of group 2 inside the convoluted gland, possess little rough endoplasmic reticulum when compared with cells of group 4, situated in the free tubule.