Transovarial transmission of Rickettsia spp. and organ-specific infection of the whitefly Bemisia tabaci

The whitefly Bemisia tabaci is a cosmopolitan insect pest that harbors Portiera aleyrodidarum, the primary obligatory symbiotic bacterium, and several facultative secondary symbionts. Secondary symbionts in B. tabaci are generally associated with the bacteriome, ensuring their vertical transmission; however, Rickettsia is an exception and occupies most of the body cavity, except the bacteriome. The mode of Rickettsia transfer between generations and its subcellular localization in insect organs have not been investigated. Using electron and fluorescence microscopy, we show that Rickettsia infects the digestive, salivary, and reproductive organs of the insect; however, it was not observed in the bacteriome. Rickettsia invades the oocytes during early developmental stages and resides in follicular cells and cytoplasm; it is mostly excluded when the egg matures; however, some bacterial cells remain in the egg, ensuring their transfer to subsequent generations. Rickettsia was localized to testicles and the spermatheca, suggesting a horizontal transfer between males and females during mating. The bacterium was further observed at large amounts in midgut cells, concentrating in vacuole-like structures, and was located in the hemolymph, specifically at exceptionally large amounts around bacteriocytes and in fat bodies. Organs further infected by Rickettsia included the primary salivary glands and stylets, sites of possible secretion of the bacterium outside the whitefly body. The close association between Rickettsia and the B. tabaci digestive system might be important for digestive purposes. The vertical transmission of Rickettsia to subsequent generations occurs via the oocyte and not, like other secondary symbionts, the bacteriome.     

Morphology of the sweet potato whitefly,Bemisia tabaci (Homoptera,Aleyrodidae) relative to virus transmission

 The stylet bundle of the sweet potato whitefly, Bemesia tabaci, consists of paired mandibles and maxillae. The latter interlock to form the food and salivary canals. Its salivary system consists of paired primary and accessary glands in the thorax. Primary and accessory gland ducts on each side of the nerve cord fuse to form lateral ducts that course anteroventrally to the midline and continue in parallel down the hypopharynx to eventually fuse to form the single afferent duct of the salivary pump. Saliva exiting the pump via the efferent duct enters the salivary canal of the maxillae. Food from the maxillary food canal passes from the antecibarium to the postcibarium or sucking pump and, per os, to the pharynx and esophagus of the foregut. The esophagus extends from the head to the base of the abdomen where it and the anterior midgut intimately mingle with the anterior hindgut to form a filter chamber. The midgut then proceeds dorsocaudally before looping anteroventrally to join the hindgut. The latter gives off two fingerlike Malpighian tubules before entering the filter chamber, whence it proceeds dorsocaudally to the anus within the vasiform orifice. Where possible, the morphology of Bemisia is discussed in relation to plant virus transmission and the morphologies of more thoroughly studied homopteran vectors such as aphids and leafhoppers. Accepted: 9 July 1996     

Tissue-specific gene silencing by RNA interference in the whitefly Bemisia tabaci (Gennadius)

The hemipteran whitefly Bemisia tabaci (Gennadius) species complex and the plant viruses they transmit pose major constraints to vegetable and fiber production, worldwide. The whitefly tissue- and developmental-specific gene expression has not been exhaustively studied despite its economic importance. In 2002, a functional genomic project was initiated, which generated several thousands expressed sequence tags (ESTs) and their sequence. This project provides the basic information to design experiments aimed at understanding and manipulating whitefly gene expression. In this communication, for the first time we provide evidence that the RNA interference mechanism discovered in many organisms, including in Hemiptera, is active in B. tabaci. By injecting into the body cavity long double-stranded RNA (dsRNA) molecules, specifically directed against genes uniquely expressed in the midgut and salivary glands, we were able to significantly inhibit the expression of the targeted mRNA in the different organs. Gene expression levels in RNAi-silenced whiteflies were reduced up to 70% compared to whiteflies injected with buffer or with a green fluorescent protein (GFP)-specific dsRNA. Phenotypic effects were observed in B. tabaci ovaries following dsRNA targeting the whitefly Drosophila chickadee homologue. Disruption of whitefly gene expression opens the door to new strategies aimed at curbing down the deleterious effects of this insect pest to agriculture.     

Morphology of the Phytophagous bug Platyscytus decempunctatus (Carvalho) (Heteroptera: Miridae)

Platyscytus decempunctatus Carvalho is a small mirid living throughout its life cicle plant Solanum cernuum Vell. (Solanaceae). The internal morphology of P. decempunctatus was studied and found that the female has a telotrophic ovary with three ovarioles in the right ovary and five in the left one. In the male reproductive trait there is a well developed tubular accessory gland. The phytophagous feeding habitat of P. decempunctatus was confirmed by the observations of chlorophyll pigments in the gut of all dissected specimens. The tubular salivary glands and the Malpighian tubules were also discussed.     

Major differences in the larval anatomy of the digestive and excretory systems of three Oestridae species revealed by micro‐CT

Oestrid flies (Diptera: Oestridae) do not feed during the adult stage, so they depend on an efficient assimilation and storage of nutrients during their parasitic larval stage. We describe the general morphology and provide volumetric data for the digestive and excretory organs of the three larval instars of the nasal bot fly Oestrus ovis L., using micro‐computed tomography. The size of the digestive and excretory organs greatly increased across larval instars. In all instars, the two salivary glands were remarkably large and formed a ‘glandular band’ by coming together, but without lumina uniting, at their posterior ends. The distal region of the anterior Malpighian tubules was greatly enlarged and full of highly radio‐opaque concretions. Moreover, the anatomy of O. ovis third‐instar larva was compared to that of two species of, respectively, similar and different feeding habits: Cephenemyia stimulator (Clark) and Hypoderma actaeon Brauer. Whereas the general morphology and arrangement of the digestive and excretory systems of C. stimulator was similar to that of O. ovis, some differences were observed in H. actaeon: a swollen anterior region of the midgut, salivary glands shorter and not forming a ‘band’ and anterior Malpighian tubules narrowly uniform throughout their entire length.     

Alkaline phosphatase activity in whitefly salivary glands and saliva

Alkaline phosphatase activity was histochemically localized in adult whiteflies (Bemisia tabaci B biotype, syn. B. argentifolii) with a chromogenic substrate (5-bromo-4-chloro-3-indolylphosphate) and a fluorogenic substrate (ELF-97). The greatest amount of staining was in the basal regions of adult salivary glands with additional activity traced into the connecting salivary ducts. Other tissues that had alkaline phosphatase activity were the accessory salivary glands, the midgut, the portion of the ovariole surrounding the terminal oocyte, and the colleterial gland. Whitefly nymphs had activity in salivary ducts, whereas activity was not detected in two aphid species (Rhodobium porosum and Aphis gossypii). Whitefly diet (15% sucrose) was collected from whitefly feeding chambers and found to have alkaline phosphatase activity, indicating the enzyme was secreted in saliva. Further studies with salivary alkaline phosphatase collected from diet indicated that the enzyme had a pH optimum of 10.4 and was inhibited by 1 mM cysteine and to a lesser extent 1 mM histidine. Dithiothreitol, inorganic phosphate, and ethylenediaminetetraacetic acid (EDTA) also inhibited activity, whereas levamisole only partially inhibited salivary alkaline phosphatase. The enzyme was heat tolerant and retained approximately 50% activity after a 1-h treatment at 65 degrees C. The amount of alkaline phosphatase activity secreted by whiteflies increased under conditions that stimulate increased feeding. These observations indicate alkaline phosphatase may play a role during whitefly feeding.     

Studies on water and ion transport in homopteran insects: ultrastructure and cytochemistry of the cicadoid and cercopoid Malpighian tubules and filter chamber

The filter chamber is a complex junction of anterior and posterior extremities of the midgut and Malpighian tubules. The sac-like anterior extremity, or filter chamber proper, comprises two cell types. These are large cuboidal cells which secrete a mucoprotein, and extremely thin cells which have regular tubular invaginations of the basal plasma membrane. The posterior extremity of the midgut and the internal Malpighian tubules coil round the filter chamber proper. They consist of thin epithelial cells identical in ultrastructure. The basal plasma membrane in these cells is formed into leaflets. A thin cellular sheath and thick muscle layers surround the filter chamber. The filter chamber proper is lined by the mucoprotein secretion of the cuboidal cells. This secretion appears to bind potassium ions. ATPase and alkaline phosphatase cannot be detected in the filter chamber epithelia. The structure and cytochemistry of the filter chamber suggests that water flows from filter chamber proper to midgut and Malpighian tubules by passive osmosis. This may be facilitated by ion binding in the filter chamber proper and by hydrostatic pressure engendered by contraction of the muscular coat. The Malpighian tubules appear to be structurally and chemically adapted for ion secretion by active transport and possibly for reabsorption in the Malpighian duct segment.     

X-ray microanalysis of the filter chamber of the cicada,Cyclochila australasiae Don.

X-ray microanalysis of frozen-hydrated specimens of the cicada filter chamber shows that there are concentration changes in K+ and Cl- in the various compartments of the filter chamber from feeding to non-feeding states. These changes are consistent with an osmotic transfer of water between the anterior midgut and the posterior midgut and Malpighian tubules. K+ and Cl- account for a large proportion of the osmotically active solutes in these compartments, and it is suggested that the osmotic gradients are generated by secretion of K+ and Cl- into the posterior midgut and Malpighian tubule lumina. Analysis of freeze-substituted sections of filter chamber supports the analysis of frozen-hydrated specimens and indicates that Na+ may also be secreted into the posterior midgut and the Malpighian tubules. The system functions as a water-shunting complex.     

保护地间作芹菜对温室粉虱的防治作用

温室白粉虱Trialeurodes vaporariorum(Westwood)和烟粉虱Bemisia tabaci(Gennadius)是严重危害葫芦科、茄科和豆科等多种蔬菜的主要害虫,具有分布范围广、种群数量大、繁殖力强等特性。作者通过田间试验研究了蔬菜保护地内间作温室粉虱非嗜食植物芹菜(Apium graveliens L.)对其的防治效果。结果表明:与空白处理和常规化学防治相比,在番茄和黄瓜保护地内间作芹菜对温室粉虱均具有显著的防治效果,驱避效果分别达到98.0%和84.5%。这些结果是初步的,但其为进一步研究温室粉虱的寄主选择机制和非化学防治方法提供了依据。     

Ultrastructure and distribution of intracellular rickettsia-like microorganisms in various organs of the laboratory-reared adult tickArgas (Persicargas) arboreus (Ixodoidea: Argasidae)

Rod (RS) and coccoid (CS) rickettsia-like microorganisms were found in single and group forms in organs of the laboratory-reared adult ticksArgas (Persicarges) arboreus. RS are distributed in most organs but are mainly concentrated in the salivary glands, mid-gut, and testes. CS single forms were concentrated in the rectal sac, while the group forms were limited to Malpighian tubules and haemocytes of both sexes. The primary oocytes were heavily infected with both forms of CS. No RS or CS were detected in the muscles. Despite the structural differences between RS and CS, they are suggested to be different morphotypes of the same organism.     

烟粉虱和温室粉虱在甘蓝上的刺探取食行为比较

利用刺吸电波图技术研究B型、ZHJ_1型烟粉虱Bemisia tabaci(Gennadius)和温室粉虱Trialeurodes vaporariorum(Westwood)在甘蓝上的取食行为,将3种粉虱的电波图进行比较,其中,B型和ZHJ_1型烟粉虱记录到np,C,pd,E1,E2,F和G波7种波形,温室粉虱只记录到刺探波形,少有取食波形。B型的20个记录中只有1个没有持续吸食波形;ZHJ_1型的25个记录中有10个记录没有持续吸食波形;温室粉虱没有持续吸食记录。甘蓝叶片韧皮部的结构或汁液的化学成分与温室粉虱的抗性密切相关。     

四种粉虱种群超微形态特征多样性研究

运用扫描电子显微镜对烟粉虱Bemisia tabaci、非洲伯粉虱Bemisia afer、温室粉虱Trialeurodes vaporariorum和柑桔粉虱Dialeurodes citri4种粉虱4龄若虫的超微形态特征进行了初步研究。结果表明,扫描电子显微镜下的超微结构特征可很好地将4种粉虱区分开来,如烟粉虱圆锥状的舌状突,非洲伯粉虱尾沟两侧隆起上形成的横脊,温室粉虱形态典型的乳突和泌蜡孔,柑桔粉虱近圆形的皿状孔及其尾沟内的卵石状乳突等特异特征。并根据种间的超微结构特异特征,编制了不同种的简明检索表。超微形态特征为粉虱种群鉴定提供了可靠的理论参考。     

Genetic Analysis of Bemisia (Hemiptera: Aleyrodidae) Populations by Isoelectric Focusing Electrophoresis

Twenty-one whitefly populations in the genus Bemisia were evaluated for genetic variation at 3 allozyme loci. Nine of the 22 populations that exhibited polymorphic loci were subjected to allozyme analysis using a minimum of 10 enzymes, representing 10 to 14 distinct loci. Among those nine variants examined, calculated genetic distances ranged between 0.03 and 0.52, with three main groups emerging from the analysis. One group comprised two closely related Western Hemisphere variants of B. tabaci: type A from California, United States and a geographically proximal population from Culiacan, Mexico. A second cluster contained five collections previously identified as B. tabaci type B and Bemisia argentifolii, while a third group contained a single population from Benin, Africa. The latter two groups were grouped separately from New World populations and are thought to have a recent origin in the Eastern Hemisphere.     

刺螫库蠓成虫内部器官系统组织学观察

【目的】刺螫库蠓Culicoides punctatus是一种重要媒介蠓虫,是施马伦贝格病毒(Schmallenberg virus, SBV)的主要传播载体。通过石蜡切片和苏木素 伊红染色技术观察刺螫库蠓成虫内部器官系统的组织结构。【方法】利用网扫法、灯诱法采集刺螫库蠓成虫,除足和翅外置于Duboscq-Brasil固定液中常温固定,然后经过逐级脱水、二甲苯透明、石蜡包埋、连续切片以及苏木素-伊红染色等过程制成玻片,利用光学显微镜进行观察和拍照。【结果】刺螫库蠓成虫消化系统不存在性别差异,由消化道及唾液腺组成。中枢神经系统由脑和腹神经索组成。脑可划分为前脑、中脑和后脑3个功能区。复眼和触角是刺螫库蠓主要的感觉器官。腹神经索可分为咽下神经节、胸神经节和腹神经节。呼吸系统主要由气管组成,气管遍布全身,无肺组织,胸部具有2对气门,分别位于中胸和后胸;腹支囊组织呈泡状,着色不明显。生殖系统包括内生殖器官和外生殖器官,其中雌性内生殖器官包括卵巢、输卵管、受精囊及生殖附腺,雄性内生殖器官包括精巢、输精管、射精管及生殖附腺。【结论】本研究明确了刺螫库蠓成虫的消化系统、神经系统、呼吸系统、生殖系统以及感觉器官的结构特征,为库蠓的发育研究提供了更为直接、准确的证据,有助于提高蠓媒监测、预报及防制的全面性和精确性。     

The effects of aposymbiosis and of an infection with Blastocrithidia triatomae (Trypanosomatidae) on the tracheal system of the reduviid bugs Rhodnius prolixus and Triatoma infestans

We have investigated the effects of the absence of symbionts and of infection with a trypanosome on the tracheal supply to different organs in larvae of the blood-sucking bugs, Rhodnius prolixus and Triatoma infestans. In bugs grown without symbionts there were extensive reductions in the tracheal supply to all the internal organs examined. These bugs also excreted less fluid after blood meals, perhaps because of a reduction in the oxygen supply to the Malpighian tubules. Inclusion of vitamin B in the diet of these affected insects reversed both the adverse effects on diuresis and on the extent of the respiratory supply to the different internal organs. The results of these studies suggest that vitamin B may play a key role in the development and maintenance of an adequate tracheal supply to the tissues in these insects. Infection with the trypanosome Blastocrithidia triatomae also greatly reduced the density of tracheoles supplying the rectum, small intestine and Malpighian tubules in infected T. infestans, but not in R. prolixus. It is possible, therefore, that the parasite exerts at least part of its pathogenic effect by causing a vitamin B deficiency, that in turn affects oxygen supply to the tissues.     

世界性重要害虫B型烟粉虱的入侵机制

B型烟粉虱是近二十年来入侵世界各国并暴发成灾的一种世界性重要害虫。近年来, B型烟粉虱的入侵机制研究表明,其成功定殖、扩散、暴发与其对入侵地区的土著烟粉虱种群和其它昆虫种群的竞争取代有关;竞争取代包括生态位竞争,生殖干涉,其危害寄主对其它昆虫的影响等方面。B型烟粉虱抗药性的形成也是这种生物成功入侵的重要影响因素。同时,生态环境（包括寄主植物、气候因子、越冬场所等）的影响也不容忽视。最后本文还探讨了B型烟粉虱入侵机制研究动态及其意义。     

小地老虎成虫消化系统和马氏管的形态研究

本文研究了小地老虎Agrotis ypsilon Rott.成虫的消化系统和马氏管的形态,详细描述了消化道、唾腺和马氏管三个部分的构造。观察到有两种构造是前人有关文献中未描述过的:1.吸泵被区分为几丁质的底壁和极薄的背壁;2.唾窦背面存在有一对囊形小体,它们伸入到吸泵中。     

Morphology and ultrastructure of the alimentary canal of the cicada Platypleura kaempferi (Hemiptera: Cicadidae)

The alimentary canal of cicada Platypleura kaempferi is described. It comprises the oesophagus, filter chamber, external midgut section and hindgut. The elongate oesophagus expands posteriorly, with its posterior end constricting to become a bulb. The filter chamber consists of two parts: a very thin sheath and a filter organ. The filter organ is composed of the anterior and posterior ends of the midgut (internal midgut section), and the internal proximal ends of the Malpighian tubules. The external midgut section differentiates into a collapsed sac and a midgut loop. The latter is divided into three distinct segments. The hindgut contains a dilated rectum and a long narrow ileum. The distal portions of the four Malpighian tubules are enclosed in a peritoneal sheath together with the distal ileum before reaching to the rectum. Ultrastructurally, the oesophagus and the hindgut are lined with a cuticle. The filter chamber sheath consists of cells with large irregular nuclei. Filamentous substances coat the microvilli of the cells of the internal midgut section. The posterior end of the midgut comprises two types of cells, with the first type of cells containing many vesicles and scattered elements of rough endoplasmic reticulum. The anterior and posterior segments of the midgut loop cells have ferritin‐like granules. The ileum cells have well‐developed apical leaflets associated with mitochondria. Accumulations of virus‐like particles enclosed in the membrane are observed in the esophagus, conical segment, mid‐ and posterior segments of the midgut loop.     

合哑蝉成虫消化道形态、组织结构及超微结构（英文）

【目的】本研究通过合哑蝉Karenia caelatata成虫消化道的形态学、组织学和超微结构研究,进一步了解蝉科(Cicadidae)代表种类的消化道形态和功能分化。【方法】利用光学显微镜和透射电子显微镜技术,对合哑蝉雄成虫消化道的整体形态以及食道、滤室(中肠前端及后端、马氏管基部、后肠基部)、滤室外中肠(锥形体、中肠环)、后肠(回肠、直肠)的一般形态和超微结构进行了详细观察,同时对滤室的组织结构进行了研究。【结果】结果表明,合哑蝉消化道由食道、滤室、滤室外中肠及后肠组成。食道狭长,被有上表皮和内表皮。中肠前端、中肠后端、马氏管基部以及后肠基部被一肌肉鞘包围形成滤室构造。组成中肠前端和后端的细胞基膜高度内褶,顶端的微绒毛发达。中肠后端分布许多线粒体和高电子密度的分泌颗粒。滤室外的中肠包括膨大的锥形体、中肠环。其中,锥形体由两种细胞组成;中肠环分为前、中、后3个不同的区段。前中肠细胞包含大量的分泌颗粒、线粒体、粗面内质网和溶酶体;中中肠细胞含有分泌颗粒;后中肠细胞包括许多低电子密度的分泌颗粒和滑面内质网。类铁蛋白颗粒零星分布于中肠环的前、中区段。组成锥形体和中肠环前端的细胞顶端微绒毛被丝状物质覆盖。后肠被有一层表皮。食道、中肠环中段、直肠细胞中含有微生物。【结论】本研究获得的合哑蝉消化道形态、组织结构和超微结构方面的信息为其功能分化研究提供了重要信息。同时,相关微生物的发现为进一步探讨共生菌与蝉总科昆虫的协同进化提供了信息。     

THE DIGESTIVE AND MALE INTERNAL REPRODUCTIVE SYSTEMS OF SOME AUSTRALIAN ANTHOCORIDAE (HEMIPTERA)

Abstract
The digestive and male internal reproductive systems of 11 species of Australian Anthocoridae are described and figured. The alimentary canals and salivary glands show some specific differences; the reproductive systems are distinct at the tribal level. Results are discussed in relation to Carayon's system of classification.