Transmission of Spiroplasma citri by the aster leafhopper Macrosteles fascifrons (Homoptera: Cicadellidae)

The aster leafhopper (Macrosteles fascifrons), injected with an isolate of Spiroplasma citri obtained from brittle root-diseased horseradish (Armoracia rusticana), transmitted the spiroplasma to horseradish and China aster (Callistephus chinensis.) After feeding on plants infected with S. citri, M. fascifrons transmitted the spiroplasma from aster to aster and horseradish, from yellow rocket (Barbarea vulgaris) to aster, and from turnip (Brassica rapa) to turnip. Symptoms in infected horseradish were chlorosis and stunting of newly formed leaves, discoloration of root phloem, and reduced plant growth typical of brittle root disease. Chlorosis, stunting, and asymmetry of young leaves occurred in affected aster and turnip. Flowers of infected aster were small and pale in colour and occasionally showed other symptoms including asymmetry, petal distortion, or light green petals. Spiroplasmas were isolated from all plants showing symptoms. Transmission rates by M. fascifrons which acquired S. citri by feeding on infected plants were very low, but injected leafhoppers transmitted more frequently. This is the first report of the transmission of S. citri from diseased to healthy plants by M. fascifrons.     

Multiplication and morphology of Spiroplasma citri in the leafhopper Euscelis plebejus

Spiroplasma citri multiplied in all Euscelis plebejus leaf hoppers injected and sometimes reached titres of over 1 × 10⁷ colony forming units per insect. Spiroplasmas could be isolated from the haemolymph at all times although helices were only apparent for a few days after injection. The salivary glands of injected insects contained membrane bound pockets densely packed with mycoplasma-like bodies. These bodies were frequently infected with virus-like particles similar to those found in cultures of S. citri. Spiroplasmas had little effect on the longevity of the leafhoppers.     

Spiralin is not essential for helicity, motility, or pathogenicity but is required for efficient transmission of Spiroplasma citri by its leafhopper vector Circulifer haematoceps

Spiralin is the most abundant protein at the surface of the plant pathogenic mollicute Spiroplasma citri and hence might play a role in the interactions of the spiroplasma with its host plant and/or its insect vector. To study spiralin function, mutants were produced by inactivating the spiralin gene through homologous recombination. A spiralin-green fluorescent protein (GFP) translational fusion was engineered and introduced into S. citri by using an oriC-based targeting vector. According to the strategy used, integration of the plasmid by a single-crossover recombination at the spiralin gene resulted in the expression of the spiralin-GFP fusion protein. Two distinct mutants were isolated. Western and colony immunoblot analyses showed that one mutant (GII3-9a5) did produce the spiralin-GFP fusion protein, which was found not to fluoresce, whereas the other (GII3-9a2) produced neither the fusion protein nor the wild-type spiralin. Both mutants displayed helical morphology and motility, similarly to the wild-type strain GII-3. Genomic DNA analyses revealed that GII3-9a5 was unstable and that GII3-9a2 was probably derived from GII3-9a5 by a double-crossover recombination between plasmid sequences integrated into the GII3-9a5 chromosome and free plasmid. When injected into the leafhopper vector Circulifer haematoceps, the spiralinless mutant GII3-9a2 multiplied to high titers in the insects (1.1 x 10(6) to 2.8 x 10(6) CFU/insect) but was transmitted to the host plant 100 times less efficiently than the wild-type strain. As a result, not all plants were infected, and symptom production in these plants was delayed for 2 to 4 weeks compared to that in the wild-type strain. In the infected plants however, the mutant multiplied to high titers (1.2 x 10(6) to 1.4 x 10(7) CFU/g of midribs) and produced the typical symptoms of the disease. These results indicate that spiralin is not essential for pathogenicity but is required for efficient transmission of S. citri by its insect vector.     

Disruption of a gene predicted to encode a solute binding protein of an ABC transporter reduces transmission of Spiroplasma citri by the leafhopper Circulifer haematoceps

Spiroplasma citri is transmitted from plant to plant by phloem-feeding leafhoppers. In an attempt to identify mechanisms involved in transmission, mutants of S. citri affected in their transmission must be available. For this purpose, transposon (Tn4001) mutagenesis was used to produce mutants which have been screened for their ability to be transmitted by the leafhopper vector Circulifer haematoceps to periwinkle plants. With one mutant (G76) which multiplied in leafhoppers as efficiently as S. citri wild-type (wt) strain GII-3, the plants showed symptoms 4 to 5 weeks later than those infected with wt GII-3. Thirty to fifty percent of plants exposed to leafhoppers injected with G76 remained symptomless, whereas for wt GII-3, all plants exposed to the transmission showed severe symptoms. This suggests that the mutant G76 was injected into plants by the leafhoppers less efficiently than wt GII-3. To check this possibility, the number of spiroplasma cells injected by a leafhopper through a Parafilm membrane into SP4 medium was determined. Thirty times less mutant G76 than wt GII-3 was transmitted through the membrane. These results suggest that mutant G76 was affected either in its capacity to penetrate the salivary glands and/or to multiply within them. In mutant G76, transposon Tn4001 was shown to be inserted into a gene encoding a putative lipoprotein (Sc76) In the ABCdb database Sc76 protein was noted as a solute binding protein of an ABC transporter of the family S1_b. Functional complementation of the G76 mutant with the Sc76 gene restored the wild phenotype, showing that Sc76 protein is involved in S. citri transmission by the leafhopper vector C. haematoceps.     

Comparative ultrastructure of the salivary glands of two phytopathogen vectors,the beet leafhopper,Circulifer tenellus (Baker), and the corn leafhopper,Dalbulus maidis DeLong and Wolcott (Homoptera : Cicadellidae)

The salivary glands of 2 leafhoppers, Circulifer tenellus and Dalbulus maidis (Homoptera : Cicadellidae) were examined by light and electron microscopy. Centrally located and occupying both the head and thorax, the salivary glands consist of 2 paired parts, the accessory glands and the principal glands. In C. tenellus and D. maidis, the accessory glands are large, multicelled lobes that lie anterior to the principal gland. They join the principal glands near the common salivary duct-gland junction via a thinner tubular duct. The principal glands of both species consist of large binucleate cells that differ in cytology and arrangement. These cells are easily distinguished by unique staining characteristics. Circulifer tenellus salivary gland cells are arranged in 2 lobes, the anterior lobe, made up of 3 concentric rings around the salivary duct and the posterior lobe, arranged in a loose pyramid extending above the foregut. Dalbulus maidis glands are similarly organized around the salivary duct.     

Searching and oviposition behavior of a mymarid egg parasitoid, Anagrus nigriventris, on five host plant species of its leafhopper host, Circulifer tenellus

Searching and oviposition behavior and parasitization ability of Anagrus nigriventris Girault (Hymenoptera: Mymaridae), an egg parasitoid of beet leafhopper, Circulifer tenellus (Baker) (Homoptera: Cicadellidae), were examined on five host plant species of beet leafhopper: sugar beet (Beta vulgaris L.), red stem filaree (Erodium cicutarium[L.]), peppergrass (Lepidium nitidum Nuttall), desert plantain (Plantago ovata Forsskal), and London rocket (Sisymbrium irio L.). Beet leafhopper embeds its eggs in the tissues of these plant species. For each plant species, A. nigriventris behavior was examined on plants with and without beet leafhopper eggs. Experimental design was a 5 (plant species) by 2 (host eggs present/absent) factorial. Additionally within each treatment, parasitoid behavior was observed over a 22-h period at five different observation periods: t=0, 3, 6, 9, and 22 h where t=0 h represents initial exposure of the insect with the plant. The behavioral events observed were: `fast walking' (general searching), `slow walking' (intensive searching), ovipositor probing, grooming, feeding, and resting. Significant differences (=0.05) among plant species in time spent on the plant, percentage of host eggs parasitized, and behavioral variables associated with intensive searching and oviposition all indicated that the plant species fell into two groups: `preferred' plants (sugar beet, London rocket, and peppergrass), and `unpreferred' plants (filaree and plantago). These variables also indicated that the parasitoids spent more time on, searched more, probed more, and oviposited more in plants with host eggs than plants without host eggs. Consistent effects of time (over the observation periods from t=0 to t=22 h) generally were detected only in the preferred plant species that had host eggs present. In these cases, intensive searching and probing decreased as time advanced, while variables related to general searching (`fast walking') and abandoning host egg patches (leaving the plant) tended to increase over time.     

Disruption of a Gene Predicted To Encode a Solute Binding Protein of an ABC Transporter Reduces Transmission of Spiroplasma citri by the Leafhopper Circulifer haematoceps

Spiroplasma citri is transmitted from plant to plant by phloem-feeding leafhoppers. In an attempt to identify mechanisms involved in transmission, mutants of S. citri affected in their transmission must be available. For this purpose, transposon (Tn4001) mutagenesis was used to produce mutants which have been screened for their ability to be transmitted by the leafhopper vector Circulifer haematoceps to periwinkle plants. With one mutant (G76) which multiplied in leafhoppers as efficiently as S. citri wild-type (wt) strain GII-3, the plants showed symptoms 4 to 5 weeks later than those infected with wt GII-3. Thirty to fifty percent of plants exposed to leafhoppers injected with G76 remained symptomless, whereas for wt GII-3, all plants exposed to the transmission showed severe symptoms. This suggests that the mutant G76 was injected into plants by the leafhoppers less efficiently than wt GII-3. To check this possibility, the number of spiroplasma cells injected by a leafhopper through a Parafilm membrane into SP4 medium was determined. Thirty times less mutant G76 than wt GII-3 was transmitted through the membrane. These results suggest that mutant G76 was affected either in its capacity to penetrate the salivary glands and/or to multiply within them. In mutant G76, transposon Tn4001 was shown to be inserted into a gene encoding a putative lipoprotein (Sc76) In the ABCdb database Sc76 protein was noted as a solute binding protein of an ABC transporter of the family S1_b. Functional complementation of the G76 mutant with the Sc76 gene restored the wild phenotype, showing that Sc76 protein is involved in S. citri transmission by the leafhopper vector C. haematoceps.     

Production and characteristics of antisera to Spiroplasma citri and clover phyllody-associated antigens derived from plants

Antisera were produced to clover phyllody- and Spiroplasma citri-associated antigens partially purified from infected Vinca rosea plants. Separate antisera were made to ‘membrane fraction’ (MF) preparations comprising the resuspended pellet obtained by high speed centrifugation, and to ‘soluble fraction’ (SA) preparations, comprising the supernatant from high speed centrifugation concentrated by freeze-drying. All antisera showed considerable activity against normal plant antigens but after cross-absorption with extracts of healthy plants the MF antisera were used in F(ab')₂based ELISA tests to detect S. citri- or clover phyllody-associated antigens in infected plants. The ‘clover phyllody’ antiserum also reacted specifically with extracts of clover plants with phyllody, and with naturally-infected strawberry plants showing symptoms of green petal disease. Both the ‘clover phyllody’ and S. citri antisera were specific for their respective homologous antigens. No cross-reactions were observed in heterologous tests or between either antiserum and extracts of V. rosea infected with various MLOs obtained from different host plants.     

Infection and replication sites of Spiroplasma kunkelii (Class: Mollicutes) in midgut and Malpighian tubules of the leafhopper Dalbulus maidis

Spiroplasma kunkelii distribution and infection mechanisms in the intestines and Malpighian tubules of Dalbulus maidis were investigated by transmission electron microscopy. Spiroplasmas were found between microvilli and in endocytic vesicles of the midgut epithelium. At the basal part, cytoplasmic vesicles contained multiple spiroplasmas with tube-like extensions and spiroplasmas accumulated between the laminae rara and densa of the basal lamina. Tip structures of flask-shaped spiroplasmas pierced the lamina densa that was discontinuous in close proximity to spiroplasmas. Spiroplasmas were found in hemolymph, crossed the basal lamina of Malpighian tubule epithelium and accumulated at high numbers in muscle cells that had cytopathogenic changes. S. kunkelii had perithrochous approximately 8nm diameter structures determined to be fimbriae protruding from the cell surface, and similar structures were adhering to the basal lamina of midgut epithelium and to external lamina of muscle cells. Further, spiroplasmas had pili-like appendages at one or both cell poles and appeared to conjugate. This is the first time that fimbriae and pili have been observed in a mollicutes.     

巴基斯坦角顶叶蝉亚科两新纪录属（半翅目：叶蝉科）

报道巴基斯坦角顶叶蝉亚科1新纪录属Changwhania Kwon和1新纪录亚属Limotettix(Scleroracus)Van Duzee,并记述2新纪录种Changwhania ceylonensis Baker,1925 n.rec.和Limotettix(Scleroracus)cacheolus n.rec.。     

A remarkable new opsiine leafhopper genus (Hemiptera: Cicadellidae: Deltocephalinae) from Southern China,with notes on its phylogenetic position

A remarkable new genus of tribe Opsiini (Deltocephalinae), Yinformibus gen. nov. including Yinformibus menglaensis sp. nov. is described from Yunnan of China. The new genus is placed in the tribe Opsiini based on male adults having a pair of aedeagal shafts, each with its own gonopore. Partial 28S rDNA and Histone H3 sequences are provided for the new species, and a phylogenetic analysis based on these markers for Deltocephalinae retrieved from GenBank suggests Opsiini is monophyletic and supports the placement of Yinformibus gen. nov. in Opsiini. The molecular results placed the new genus in a clade comprising Hishimonus Ishihara, Opsius Fieber, and an undescribed genus from Zambia. Additionally, we also analysed the relationships of Yinformibus gen. nov. with other genera based on morphology.

http://www.zoobank.org/urn:lsid:zoobank.org:pub:85C23AC3-8B19-4F29-9F55-BFC392242B51     

Three new species of the leafhopper genus Tambocerus Zhang & Webb (Hemiptera,Cicadellidae) from southern China

Three new species, Tambocerus dentatus, T. longicaudatus and T. robustispinus spp. n. from southern China, are described and illustrated. A checklist and distribution to the Tambocerus species from China is provided together with a key for their separation.     

A new species and new records of the leafhopper genus Taperus Li & Wang, 1994 (Hemiptera,Cicadellidae, Evacanthinae) from China

The paper deals with the species of the Oriental leafhopper genus Taperus Li & Wang. A new species, Taperus daozhenensissp. n., from Guizhou Province, China is described and new records for other Chinese species are given together with a key for their separation. The type specimens of the new species are deposited in the Institute of Entomology, Guizhou University (GUGC).     

Description of two new species of the leafhopper subgenus Pediopsoides (Pediopsoides) (Hemiptera,Cicadellidae, Macropsinae) from Guangxi Province,Southern China

Two new species of the Macropsinae leafhopper subgenus Pediopsoides (Pediopsoides) Matsumura, 1912, Pediopsoides (Pediopsoides) damingshanensis Li, Dai & Li, sp. n. and Pediopsoides (Pediopsoides) tishetshkini Li, Dai & Li sp. n., are described and illustrated from Guangxi Province of southern China. A key to males is provided to distinguish the species of the subgenus along with a map showing the distribution of the new species.     

Two new leafhopper genera of the Alebroides genus group (Hemiptera: Cicadellidae: Typhlocybinae) from China,with a key to genera of the group

Two new microleafhopper genera in the Alebroides genus group, Nulliata Lu, Xu & Qin, gen. nov. , based on the type species N. rubrostriata Lu, Xu & Qin, sp. nov. , and Inflatopina Lu, Dietrich & Qin, gen. nov. , based on the type species I. intonsa Lu, Dietrich & Qin, sp. nov. , are described from southwest China. Five known species in the Alebroides sohii species group are transferred to Inflatopina as new combinations. Keys to genera of the Alebroides genus group and species of Inflatopina are given.     

A new species of the leafhopper genus Diomma Motschulsky (Hemiptera, Cidadellidae, Typhlocybinae) from China

In the present paper, a new species is added to the genus Diomma Motschulsky (Hemiptera: Cicadellidae: Typhlocybinae) from Southwest China, Diomma pincersasp. n. At the same time, a key can distinguish all Chinese species of the genus is provided.     

甘肃武威地区酿酒葡萄园叶蝉种群动态及其与气象因子的关系研究

为明确甘肃武威地区酿酒葡萄园叶蝉种群发生动态及其与气象因子之间的关系.连续2年应用黄板诱集法对酿酒葡萄园叶蝉种群动态进行定时定点监测;采用相关分析、回归分析、通径分析、主成分分析、灰色关联度等方法分析其与气象因子的关系.结果表明,酿酒葡萄园叶蝉一年有4个发生高峰,分别为5月底6月初、7月中旬、8月上中旬和9月中旬,5月...