The psyllid fauna (Hemiptera: Psylloidea) of vegetable fields in Brazil

ABSTRACT Some psyllids transmit 'Candidatus Liberibacter solanacearum' (Lso), the causal agent of devastating plant diseases of cultivated Solanaceae and Apiaceae. The recent detection of Bactericera cockerelli and Lso in Ecuador seriously threatens these crops in South America. There, neither the role of native psyllids in the Lso epidemiology nor the psyllid fauna of vegetables are known. With the aim to identify potential vectors and risk scenarios for the spread of Lso in South America, a survey of the psyllid fauna of Solanaceae and Apiaceae crops and associated weeds was conducted in Brazil. Samples were taken at 29 localities in four states. A total of 2857 specimens were sampled, representing at least 37 species of 23 genera and seven families. The most frequent species on carrot, chilli pepper and potato were Russelliana solanicola, R. capsici and Isogonoceraia divergipennis, respectively. Immatures of R. capsici were found on chilli pepper and of R. solanicola on carrot and potato, confirming these plants as hosts. The two psyllid species have been suspected previously to transmit plant pathogens of unknown identity. Russelliana solanicola is one of the few polyphagous species. Here the species is reported for the first time from carrot. Recent collections in Rio Grande do Sul suggest that Solanum laxum represents the original host of R. capsici, which subsequently shifted to chilli pepper. Both, adaptation to agricultural crops and the possibility of ability to transmit pathogens, make the two Russelliana species dangerous potential vectors of Lso and other plant pathogens in South America.

First Report of a Mastrevirus (Geminiviridae) Transmitted by the Corn Leafhopper.

Maize striate mosaic virus (MSMV; genus Mastrevirus) was recently reported in maize plants in Brazil and also detected by metagenomic analyses in the corn leafhopper, Dalbulus maidis (DeLong & Wolcott). Although these findings suggested that D. maidis is a potential vector, no transmission studies have been performed. Here, we tested the transmission of MSMV by D. maidis from field-collected infected plants and plants infected with MSMV via leafhopper-mediated transmission in the laboratory; all plants were confirmed positive for MSMV by PCR. In each one of three transmission replicates, aviruliferous D. maidis nymphs and adults were confined together on a source plant during a 4-day acquisition access period (AAP) and subsequently transferred to healthy maize seedlings (10 individuals per test plant) in a series of 4-day inoculation access periods (IAPs). We also tested transmission by the corn aphid, Rhopalosiphum maidis (Fitch) and by mechanical inoculation of healthy maize seedlings. Only D. maidis transmitted MSMV, with overall transmission rates of 29.4 and 39.5% on field-collected infected plants and 18.5% on infected plants in laboratory. D. maidis transmitted MSMV until the third (8 to 12 days after the AAP) or fourth successive IAP (12 to 16 days), with gradual loss in transmission efficiency and rate of viruliferous insects over time, suggesting a persistent but nonpropagative mode of transmission. Infected test plants showed mottling symptoms with mild chlorotic streaks and height reduction. This is the first report of transmission of a mastrevirus by D. maidis, facilitating the completion of Koch's postulate for MSMV.

Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize.

Phytoplasmas are insect-transmitted bacterial pathogens that colonize a wide range of plant species, including vegetable and cereal crops, and herbaceous and woody ornamentals. Phytoplasma-infected plants often show dramatic symptoms, including proliferation of shoots (witch's brooms), changes in leaf shapes and production of green sterile flowers (phyllody). Aster Yellows phytoplasma Witches' Broom (AY-WB) infects dicots and its effector, secreted AYWB protein 11 (SAP11), was shown to be responsible for the induction of shoot proliferation and leaf shape changes of plants. SAP11 acts by destabilizing TEOSINTE BRANCHED 1-CYCLOIDEA-PROLIFERATING CELL FACTOR (TCP) transcription factors, particularly the class II TCPs of the CYCLOIDEA/TEOSINTE BRANCHED 1 (CYC/TB1) and CINCINNATA (CIN)-TCP clades. SAP11 homologs are also present in phytoplasmas that cause economic yield losses in monocot crops, such as maize, wheat and coconut. Here we show that a SAP11 homolog of Maize Bushy Stunt Phytoplasma (MBSP), which has a range primarily restricted to maize, destabilizes specifically TB1/CYC TCPs. SAP11MBSP and SAP11AYWB both induce axillary branching and SAP11AYWB also alters leaf development of Arabidopsis thaliana and maize. However, only in maize, SAP11MBSP prevents female inflorescence development, phenocopying maize tb1 lines, whereas SAP11AYWB prevents male inflorescence development and induces feminization of tassels. SAP11AYWB promotes fecundity of the AY-WB leafhopper vector on A. thaliana and modulates the expression of A. thaliana leaf defence response genes that are induced by this leafhopper, in contrast to SAP11MBSP. Neither of the SAP11 effectors promote fecundity of AY-WB and MBSP leafhopper vectors on maize. These data provide evidence that class II TCPs have overlapping but also distinct roles in regulating development and defence in a dicot and a monocot plant species that is likely to shape SAP11 effector evolution depending on the phytoplasma host range.

Transmission Efficiency of Xylella fastidiosa subsp. pauca Sequence Types by Sharpshooter Vectors after In Vitro Acquisition.

Xylella fastidiosa subsp. pauca is genetically diverse and has many vector species. However, there is limited information on vector specificity and efficiency for different sequence types (STs) within the pathogen subspecies. Both STs of X. fastidiosa and vectors differ in their associations with plants; therefore, assessment of vector competence should include the standardized vector acquisition ability of bacteria from artificial diets. This work aimed to adapt and validate an in vitro acquisition system for strains of X. fastidiosa that cause citrus variegated chlorosis, and to compare the transmission efficiency of STs of subsp. pauca by different species of sharpshooter vector. First, acquisition and transmission of ST13 by Bucephalogonia xanthophis and Macugonalia leucomelas was tested using an artificial diet with bacteria grown on minimum defined medium (X. fastidiosa medium) with or without 1% galacturonic acid (GA). Subsequently, four sharpshooter species (B. xanthophis, M. leucomelas, M. cavifrons, and Sibovia sagata) were compared as vectors of ST13 acquired from artificial diets, and four STs of subsp. pauca (11, 13, 65, and 70) were tested for acquisition and transmission by M. leucomelas. The artificial system allowed efficient acquisition and transmission of ST13 to plants, with no differences between the media tested. ST13 was transmitted more efficiently by B. xanthophis and M. leucomelas when compared with M. cavifrons and S. sagata. Different STs influenced acquisition and transmission rates by M. leucomelas. The differences in vector competence, despite the standardized acquisition system, suggest that ST-vector foregut or vector-plant interactions may influence bacterial acquisition, retention and inoculation by the insect.

Distribution and Genetic Diversity of Xylella fastidiosa subsp. pauca Associated with Olive Quick Syndrome Symptoms in Southeastern Brazil.

In Brazil, the host expansion of Xylella fastidiosa subsp. pauca was recently demonstrated with the report of diseased olive trees (Olea europaea), whose symptoms were associated with olive quick decline syndrome previously described in southern Italy. We employed both polymerase chain reaction-based techniques and culture medium isolation to investigate the geographic distribution of X. fastidiosa as well as the genetic signatures of 21 strains isolated from 11 olive orchards in both São Paulo and Minas Gerais States in Brazil. X. fastidiosa subsp. pauca was detected in 83% of the orchards examined in the region, and was positively diagnosed in 43.7% of all sampled plants with typical scorching symptoms. Of the 21 strains characterized with fast-evolving microsatellite (single sequence repeat [SSR]) markers, 20 different multilocus microsatellite genotypes were observed with the overall allelic diversity of HNei = 0.38. Principal component analysis using the SSR markers clustered all strains, except for three, in one cluster demonstrating a limited range of genetic diversity. Multilocus sequence typing analysis showed the prevalence of a sequence type (ST16) in 75% of the samples; three other novel STs (84, 85, and 86), were detected, all belonging to the X. fastidiosa subsp. pauca cluster. These results show that genetically diverse strains of X. fastidiosa subsp. pauca are widely present in olive orchards in southeastern Brazil, which is consistent with the long history of this bacterium in that region.

Homologous Recombination and Xylella fastidiosa Host-Pathogen Associations in South America.

Homologous recombination affects the evolution of bacteria such as Xylella fastidiosa, a naturally competent plant pathogen that requires insect vectors for dispersal. This bacterial species is taxonomically divided into subspecies, with phylogenetic clusters within subspecies that are host specific. One subspecies, pauca, is primarily limited to South America, with the exception of recently reported strains in Europe and Costa Rica. Despite the economic importance of X. fastidiosa subsp. pauca in South America, little is known about its genetic diversity. Multilocus sequence typing (MLST) has previously identified six sequence types (ST) among plant samples collected in Brazil (both subsp. pauca and multiplex). Here, we report on a survey of X. fastidiosa genetic diversity (MLST based) performed in six regions in Brazil and two in Argentina, by sampling five different plant species. In addition to the six previously reported ST, seven new subsp. pauca and two new subsp. multiplex ST were identified. The presence of subsp. multiplex in South America is considered to be the consequence of a single introduction from its native range in North America more than 80 years ago. Different phylogenetic approaches clustered the South American ST into four groups, with strains infecting citrus (subsp. pauca); coffee and olive (subsp. pauca); coffee, hibiscus, and plum (subsp. pauca); and plum (subsp. multiplex). In areas where these different genetic clusters occurred sympatrically, we found evidence of homologous recombination in the form of bidirectional allelic exchange between subspp. pauca and multiplex. In fact, the only strain of subsp. pauca isolated from a plum host had an allele that originated from subsp. multiplex. These signatures of bidirectional homologous recombination between endemic and introduced ST indicate that gene flow occurs in short evolutionary time frames in X. fastidiosa, despite the ecological isolation (i.e., host plant species) of genotypes.

Tomato Infection by Whitefly-Transmitted Circulative and Non-Circulative Viruses Induce Contrasting Changes in Plant Volatiles and Vector Behaviour.

Virus infection frequently modifies plant phenotypes, leading to changes in behaviour and performance of their insect vectors in a way that transmission is enhanced, although this may not always be the case. Here, we investigated Bemisia tabaci response to tomato plants infected by Tomato chlorosis virus (ToCV), a non-circulative-transmitted crinivirus, and Tomato severe rugose virus (ToSRV), a circulative-transmitted begomovirus. Moreover, we examined the role of visual and olfactory cues in host plant selection by both viruliferous and non-viruliferous B. tabaci. Visual cues alone were assessed as targets for whitefly landing by placing leaves underneath a Plexiglas plate. A dual-choice arena was used to assess whitefly response to virus-infected and mock-inoculated tomato leaves under light and dark conditions. Thereafter, we tested the whitefly response to volatiles using an active air-flow Y-tube olfactometer, and chemically characterized the blends using gas chromatography coupled to mass spectrometry. Visual stimuli tests showed that whiteflies, irrespective of their infectious status, always preferred to land on virus-infected rather than on mock-inoculated leaves. Furthermore, whiteflies had no preference for either virus-infected or mock-inoculated leaves under dark conditions, but preferred virus-infected leaves in the presence of light. ToSRV-infection promoted a sharp decline in the concentration of some tomato volatiles, while an increase in the emission of some terpenes after ToCV infection was found. ToSRV-viruliferous whiteflies preferred volatiles emitted from mock-inoculated plants, a conducive behaviour to enhance virus spread, while volatiles from ToCV-infected plants were avoided by non-viruliferous whiteflies, a behaviour that is likely detrimental to the secondary spread of the virus. In conclusion, the circulative persistent begomovirus, ToSRV, seems to have evolved together with its vector B. tabaci to optimise its own spread. However, this type of virus-induced manipulation of vector behaviour was not observed for the semi persistent crinivirus, ToCV, which is not specifically transmitted by B. tabaci and has a much less intimate virus-vector relationship.

Combined use of a new SNP-based assay and multilocus SSR markers to assess genetic diversity of Xylella fastidiosa subsp. pauca infecting citrus and coffee plants.

Two haplotypes of Xylella fastidiosa subsp. pauca (Xfp) that correlated with their host of origin were identified in a collection of 90 isolates infecting citrus and coffee plants in Brazil, based on a single-nucleotide polymorphism in the gyrB sequence. A new single-nucleotide primer extension (SNuPE) protocol was designed for rapid identification of Xfp according to the host source. The protocol proved to be robust for the prediction of the Xfp host source in blind tests using DNA from cultures of the bacterium, infected plants, and insect vectors allowed to feed on Xfp-infected citrus plants. AMOVA and STRUCTURE analyses of microsatellite data separated most Xfp populations on the basis of their host source, indicating that they were genetically distinct. The combined use of the SNaPshot protocol and three previously developed multilocus SSR markers showed that two haplotypes and distinct isolates of Xfp infect citrus and coffee in Brazil and that multiple, genetically different isolates can be present in a single orchard or infect a single tree. This combined approach will be very useful in studies of the epidemiology of Xfp-induced diseases, host specificity of bacterial genotypes, the occurrence of Xfp host jumping, vector feeding habits, etc., in economically important cultivated plants or weed host reservoirs of Xfp in Brazil and elsewhere.

Complete Genome Sequence of "Candidatus Sulcia muelleri" ML, an Obligate Nutritional Symbiont of Maize Leafhopper (Dalbulus maidis).

"Candidatus Sulcia muelleri" is a symbiont of sap-feeding insects in the suborder Auchenorrhyncha. The strain "Ca. Sulcia muelleri" ML is associated with the maize leafhopper (Dalbulus maidis), collected in Brazil, which is a disease vector that affects corn production. Here, we report the complete genome sequence of this bacterium.

Temporal progression of 'Candidatus Liberibacter asiaticus' infection in citrus and acquisition efficiency by Diaphorina citri.

Over the last decade, the plant disease huanglongbing (HLB) has emerged as a primary threat to citrus production worldwide. HLB is associated with infection by phloem-limited bacteria ('Candidatus Liberibacter' spp.) that are transmitted by the Asian citrus psyllid, Diaphorina citri. Transmission efficiency varies with vector-related aspects (e.g., developmental stage and feeding periods) but there is no information on the effects of host-pathogen interactions. Here, acquisition efficiency of 'Candidatus Liberibacter asiaticus' by D. citri was evaluated in relation to temporal progression of infection and pathogen titer in citrus. We graft inoculated sweet orange trees with 'Ca. L. asiaticus'; then, at different times after inoculation, we inspected plants for HLB symptoms, measured bacterial infection levels (i.e., titer or concentration) in plants, and measured acquisition by psyllid adults that were confined on the trees. Plant infection levels increased rapidly over time, saturating at uniformly high levels (≈10(8) copy number of 16S ribosomal DNA/g of plant tissue) near 200 days after inoculation-the same time at which all infected trees first showed disease symptoms. Pathogen acquisition by vectors was positively associated with plant infection level and time since inoculation, with acquisition occurring as early as the first measurement, at 60 days after inoculation. These results suggest that there is ample potential for psyllids to acquire the pathogen from trees during the asymptomatic phase of infection. If so, this could limit the effectiveness of tree rouging as a disease management tool and would likely explain the rapid spread observed for this disease in the field.

Resistência de cultivares de batata a Myzus persicae (Sulz.)(Hemiptera: Aphididae) / Resistance of potato cultivars to Myzus persicae (Sulz.) (Hemiptera: Aphididae)

Aphids are the most important vectors of viruses infecting potato (Solanum tuberosum). We focused on the response of the aphid vector Myzus persicae (Sulzer) to five commercial potatocultivars: Ágata, Jaette Bintje, Mondial, Monalisa and Santè, by traditional antibiosis and antixenosis tests and by the EPG (Electrical Penetration Graph) technique, as a step forward to the design of effective management practices. Our aim was to identify plant factors involved in resistance of these cultivars against M. persicae, both at the surface and in deeper plant tissues. Results from the antixenosis test confirmed a strong preference of M. persicae for the Mondial cultivar. The antibiosis study indicated a lower population development of the aphid in 'Monalisa' when compared to 'Ágata' and 'Jaette Bintje'. EPG assays indicated that 'Santè' inhibited the initial feeding process of M. persicae, whereas 'Monalisa' showed a physical-type of resistance as demonstrated by a very high number of short probes. The cultivar Mondial showed average values for all EPG variables analyzed. The behavior in 'Jaette Bintje' indicated this cultivar was an ideal host for aphid feeding and reproduction. Together, the EPG data revealed the existence of pre and post-phloematics factors in the cultivars under study, which have important implications on the efficiency of transmission and spread of virus in potato by M. persicae.

[Faunistic analysis of leafhopper (Hemiptera: Cicadellidae) species in vineyards of Rio Grande do Sul State, Brazil]. / Análise faunística de cigarrinhas (Hemiptera: Cicadellidae) na cultura da videira no Rio Grande do Sul.

In some American countries, grapevines are affected by Pierce's disease (PD), which is caused by a particular strain of Xylella fastidiosa not yet reported in Brazil. In order to investigate the potential for PD spread in Brazil in case of pathogen introduction, we conducted a faunistic analysis of leafhoppers (Hemiptera: Cicadellidae) associated to vineyards in the State of Rio Grande do Sul, with emphasis in the subfamily Cicadellinae (sharpshooters), which includes the main potential vectors of X. fastidiosa. Leafhopper samplings were carried out fortnightly during two years (9/2004-9/2006) in four Vitis vinifera vineyards in the municipalities of Bento Gonçalves and Farroupilha (RS). Thirtyfour leafhopper and six spittlebug species were collected, but most (98.4%) of the 3,893 specimens trapped were leafhoppers, distributed in the subfamilies Cicadellinae (60.2%), Gyponinae (34.1%), Deltocephalinae (3.8%) and Coelidinae (0.3%). The sharpshooter specimens were divided in the tribes Cicadellini (68.5%; 12 species) and Proconiini (31.5%; 11 species). Based on the faunistic indices, five species of Cicadellini, Bucephalogonia xanthophis (Berg), Dilobopterus dispar (Germar), Macugonalia cavifrons Stal, Sibovia sagata (Signoret) and Spinagonalia rubrovittata Cavichioli, and three of Proconiini, Molomea consolida (Schöder), Oncometopia facialis (Signoret) and Oncometopia fusca Melichar were prevalent in the vineyards. The high diversity of native sharpshooters in Rio Grande do Sul indicates the existence of a high risk of PD spread if the pathogen is introduced in grapevines.

Análise faunística de cigarrinhas (Hemiptera: Cicadellidae) na cultura da videira no Rio Grande do Sul / Faunistic analysis of leafhopper (Hemiptera: Cicadellidae) species in vineyards of Rio Grande do Sul state, Brazil

In some American countries, grapevines are affected by Pierce's disease (PD), which is caused by a particular strain of Xylella fastidiosa not yet reported in Brazil. In order to investigate the potential for PD spread in Brazil in case of pathogen introduction, we conducted a faunistic analysis of leafhoppers (Hemiptera: Cicadellidae) associated to vineyards in the State of Rio Grande do Sul, with emphasis in the subfamily Cicadellinae (sharpshooters), which includes the main potential vectors of X. fastidiosa. Leafhopper samplings were carried out fortnightly during two years (9/2004-9/2006) in four Vitis vinifera vineyards in the municipalities of Bento Gonçalves and Farroupilha (RS). Thirtyfour leafhopper and six spittlebug species were collected, but most (98.4 percent) of the 3,893 specimens trapped were leafhoppers, distributed in the subfamilies Cicadellinae (60.2 percent), Gyponinae (34.1 percent), Deltocephalinae (3.8 percent) and Coelidinae (0.3 percent). The sharpshooter specimens were divided in the tribes Cicadellini (68.5 percent; 12 species) and Proconiini (31.5 percent; 11 species). Based on the faunistic indices, five species of Cicadellini, Bucephalogonia xanthophis (Berg), Dilobopterus dispar (Germar), Macugonalia cavifrons Stal, Sibovia sagata (Signoret) and Spinagonalia rubrovittata Cavichioli, and three of Proconiini, Molomea consolida (Schõder), Oncometopia facialis (Signoret) and Oncometopia fusca Melichar were prevalent in the vineyards. The high diversity of native sharpshooters in Rio Grande do Sul indicates the existence of a high risk of PD spread if the pathogen is introduced in grapevines.

[Resistance of potato cultivars to Myzus persicae (Sulz.) (Hemiptera: Aphididae)]. / Resistência de cultivares de batata a Myzus persicae (Sulz.)(Hemiptera: Aphididae).

Aphids are the most important vectors of viruses infecting potato (Solanum tuberosum). We focused on the response of the aphid vector Myzus persicae (Sulzer) to five commercial potatocultivars: Ágata, Jaette Bintje, Mondial, Monalisa and Santè, by traditional antibiosis and antixenosis tests and by the EPG (Electrical Penetration Graph) technique, as a step forward to the design of effective management practices. Our aim was to identify plant factors involved in resistance of these cultivars against M. persicae, both at the surface and in deeper plant tissues. Results from the antixenosis test confirmed a strong preference of M. persicae for the Mondial cultivar. The antibiosis study indicated a lower population development of the aphid in 'Monalisa' when compared to 'Ágata' and 'Jaette Bintje'. EPG assays indicated that 'Santè' inhibited the initial feeding process of M. persicae, whereas 'Monalisa' showed a physical-type of resistance as demonstrated by a very high number of short probes. The cultivar Mondial showed average values for all EPG variables analyzed. The behavior in 'Jaette Bintje' indicated this cultivar was an ideal host for aphid feeding and reproduction. Together, the EPG data revealed the existence of pre and post-phloematics factors in the cultivars under study, which have important implications on the efficiency of transmission and spread of virus in potato by M. persicae.

Levantamento populacional de cigarrinhas (Hemiptera: Cicadellidae) associadas à transmissão de Xylella fastidiosa em pomares cítricos do Litoral Norte da Bahia / Survey of sharpshooters (Hemiptera: Cicadellidae) associated with Xylella fastidiosa transmission in citrus groves of the North Coast of Bahia State

The causal agent of citrus variegated clorosis, Xylella fastidiosa, is transmitted by leafhoppers of the subfamily Cicadellinae, whose species vary regionally. The goal of this study was to identify potential vectors of this pathogen in citrus groves of Bahia North Coast, Brazil. The survey was done from March/2002 to February/2003 in three seven- to nine-year-old sweet orange (Citrus sinensis, Pêra variety) groves located in Rio Real, BA. Fifteen yellow sticky cards (8.5x11.5 cm) were installed 40 m apart in each grove, hanged at 1.5 m high on the upper north side of citrus canopies, and replaced fortnightly. A sweep net was periodically used to sample leafhoppers on herbaceous weeds inside the groves, by selecting five points at random and performing 30 sweeps in each point. A total of 1,360 specimens of 49 Auchenorrhyncha species were collected in this study, mostly in the family Cicadellidae (90.2 percent). The subfamily Cicadellinae, which includes the sharpshooter vectors of X. fastidiosa, showed the largest number of species (14) and specimens (84.8 percent). Acrogonia flagellata Young, A. citrina Marucci & Cavichioli, Homalodisca spottii Takiya, Cavichioli & McKamey and an unidentified Cicadellini (species 1) were the dominant species trapped on citrus canopies, while Hortensia similis (Walker) and Erythrogonia dubia (Medler) were dominant in the weedy vegetation. Among the Cicadellinae species already known as vectors of X. fastidiosa in citrus, only A. citrina, Bucephalogonia xanthophis (Berg) e Ferrariana trivittata (Signoret) were found. The two latter species were accidentally trapped by sweep net in the weedy vegetation.

O agente causal da clorose variegada dos citros, Xylella fastidiosa, é transmitido por cigarrinhas da subfamília Cicadellinae, cujas espécies variam regionalmente. O objetivo deste trabalho foi identificar potenciais vetores do patógeno em pomares cítricos do Litoral Norte da Bahia. Os levantamentos foram conduzidos de mar/2002 a fev/2003, em três propriedades no município de Rio Real, em pomares de laranjeira doce (Citrus sinensis) variedade Pêra, com sete a nove anos. Em cada pomar, 15 cartões adesivos amarelos foram dispostos a 40 m entre si, instalados a 1,5 m de altura em ramos da face norte, na periferia da copa, sendo substituídos quinzenalmente. Também foram realizadas amostragens com rede de varredura na vegetação rasteira dos pomares, em cinco pontos escolhidos aleatoriamente, efetuando-se 30 redadas por ponto. No total, foram coletados 1360 espécimes pertencentes a 49 espécies de Auchenorrhyncha, a maioria da família Cicadellidae (90,2 por cento). A subfamília Cicadellinae, que inclui os vetores de X. fastidiosa, apresentou maior número de espécies (14) e de indivíduos coletados (84,8 por cento). Entre as espécies de Cicadellinae, Acrogonia flagellata Young, A citrina Marucci & Cavichioli, Homalodisca spottii Takiya, Cavichioli & McKamey e uma espécie não-identificada de Cicadellini (espécie 1) foram dominantes na copa das plantas cítricas, enquanto que Hortensia similis (Walker) e Erythrogonia dubia (Medler) foram dominantes na vegetação rasteira. Entre os cicadelíneos já conhecidos como vetores de X. fastidiosa em citros, apenas A. citrina, Bucephalogonia xanthophis (Berg) e Ferrariana trivittata (Signoret) foram observados, sendo que as duas últimas espécies tiveram ocorrência acidental na vegetação rasteira.

Diversidade de Hemiptera Auchenorrhyncha em citros, café e fragmento de floresta nativa do Estado de São Paulo / Diversity of Hemiptera Auchenorrhyncha in citrus, coffee and a fragment of native forest of the State of São Paulo

The population of Hemiptera Auchenorrhyncha was studied in sweet citrus groves (Citrus sinensis), coffee plantations (Coffea arabica) and a semi-deciduous seasonal forest with shrub physiognomy in Bebedouro, SP, to evaluate the influence of the natural ecosystem on the species composition of the agroecosystems. Monitoring was carried out by using yellow stick cards, which were replaced every 15 days and all Auchenorrhyncha collected were counted and identified. Seven families, 11 subfamilies and 98 species were collected, with Cicadellidae being the most abundant. The native forest presented larger wealth, diversity and equitability of Auchenorrhyncha species, demonstrating to be more stable than the other habitats. The high values of similarities obtained between the agroecosystems and the forest demonstrated that great part of Auchenorrhyncha species occurring in the agricultural habitats was also occurring at the forest, indicating that the last may serve as reservoir of species. The abundance of the taxonomic groups of Auchenorrhyncha collected varied with the evaluated habitats, with Proconiini being the most abundant in the coffee plantation next to the forest, Athysanini, Scaphytopiini, Neocoelidiinae and Coelidiinae in the orange orchard and coffee plantation distant from the forest; Cicadellinae and Agalliinae were not related to any of the habitats. The presence of vector insects and possible vectors of plant diseases in the appraised habitats indicate the need of the implementation of strategies for landscape management.

A população de Hemiptera Auchenorrhyncha foi estudada em pomares de laranja doce (Citrus sinensis), cafezais (Coffea arabica) e floresta estacional semidecidual com fisionomia arbustiva, em Bebedouro (SP), com o objetivo de avaliar a influencia do ecossistema natural na composição de espécies do agroecossistemas. O monitoramento foi feito com cartões adesivos amarelos, os quais foram trocados a cada 15 dias, efetuando-se a contagem e identificação dos Auchenorrhyncha coletados. Capturou-se o total de sete famílias, 11 subfamílias e 98 espécies, sendo Cicadellidae o grupo mais abundante. A floresta nativa apresentou a maior riqueza, diversidade e equitabilidade de espécies de Auchenorrhyncha, demonstrando ser mais estável que os demais habitats. Os altos valores de similaridades obtidos entre os agroecossistemas e a floresta demonstram que grande parte das espécies de cigarrinhas que está ocorrendo nos habitats agrícolas também está ocorrendo na floresta, indicando que esta pode estar funcionando como reservatório de espécies. A abundância de grupos taxonômicos de Auchenorrhyncha coletados variou nos habitats avaliados, sendo Proconiini o mais abundante no cafezal próximo à floresta, Athysanini, Scaphytopiini, Neocoelidiinae e Coelidiinae no pomar de laranja e cafezal distante da floresta; Cicadellinae e Agalliinae não se relacionaram a nenhum dos habitats. A presença de insetos vetores e possíveis vetores de doenças de plantas cultivadas nos habitats avaliados indicam a necessidade da implantação do manejo de pragas nessa área.

[Diversity of Hemiptera Auchenorrhyncha in citrus, coffee and a fragment of native forest of the state of São Paulo]. / Diversidade de Hemiptera Auchenorrhyncha em citros, café e fragmento de floresta nativa do Estado de São Paulo.

The population of Hemiptera Auchenorrhyncha was studied in sweet citrus groves (Citrus sinensis), coffee plantations (Coffea arabica) and a semi-deciduous seasonal forest with shrub physiognomy in Bebedouro, SP, to evaluate the influence of the natural ecosystem on the species composition of the agroecosystems. Monitoring was carried out by using yellow stick cards, which were replaced every 15 days and all Auchenorrhyncha collected were counted and identified. Seven families, 11 subfamilies and 98 species were collected, with Cicadellidae being the most abundant. The native forest presented larger wealth, diversity and equitability of Auchenorrhyncha species, demonstrating to be more stable than the other habitats. The high values of similarities obtained between the agroecosystems and the forest demonstrated that great part of Auchenorrhyncha species occurring in the agricultural habitats was also occurring at the forest, indicating that the last may serve as reservoir of species. The abundance of the taxonomic groups of Auchenorrhyncha collected varied with the evaluated habitats, with Proconiini being the most abundant in the coffee plantation next to the forest, Athysanini, Scaphytopiini, Neocoelidiinae and Coelidiinae in the orange orchard and coffee plantation distant from the forest; Cicadellinae and Agalliinae were not related to any of the habitats. The presence of vector insects and possible vectors of plant diseases in the appraised habitats indicate the need of the implementation of strategies for landscape management.

[Survey of sharpshooters (Hemiptera: Cicadellidae) associated with Xylella fastidiosa transmission in citrus groves of the North Coast of Bahia State]. / Levantamento populacional de cigarrinhas (Hemiptera: Cicadellidae) associadas à transmissão de Xylella fastidiosa em pomares cítricos do Litoral Norte da Bahia.

The causal agent of citrus variegated clorosis, Xylella fastidiosa, is transmitted by leafhoppers of the subfamily Cicadellinae, whose species vary regionally. The goal of this study was to identify potential vectors of this pathogen in citrus groves of Bahia North Coast, Brazil. The survey was done from March/2002 to February/2003 in three seven- to nine-year-old sweet orange (Citrus sinensis, Pêra variety) groves located in Rio Real, BA. Fifteen yellow sticky cards (8.5x11.5 cm) were installed 40 m apart in each grove, hanged at 1.5 m high on the upper north side of citrus canopies, and replaced fortnightly. A sweep net was periodically used to sample leafhoppers on herbaceous weeds inside the groves, by selecting five points at random and performing 30 sweeps in each point. A total of 1,360 specimens of 49 Auchenorrhyncha species were collected in this study, mostly in the family Cicadellidae (90.2%). The subfamily Cicadellinae, which includes the sharpshooter vectors of X. fastidiosa, showed the largest number of species (14) and specimens (84.8%). Acrogonia flagellata Young, A. citrina Marucci & Cavichioli, Homalodisca spottii Takiya, Cavichioli & McKamey and an unidentified Cicadellini (species 1) were the dominant species trapped on citrus canopies, while Hortensia similis (Walker) and Erythrogonia dubia (Medler) were dominant in the weedy vegetation. Among the Cicadellinae species already known as vectors of X. fastidiosa in citrus, only A. citrina, Bucephalogonia xanthophis (Berg) e Ferrariana trivittata (Signoret) were found. The two latter species were accidentally trapped by sweep net in the weedy vegetation.

Transmission efficiency of Xylella fastidiosa by sharpshooters (Hemiptera: Cicadellidae) in coffee and citrus.

Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner) is a bacterial pathogen transmitted by several sharpshooters in two tribes of Cicadellinae (Proconiini and Cicadellini). Here, we compared the transmission efficiency of X. fastidiosa in coffee (Coffea arabica L.) and citrus [Citrus sinensis (L.) Osbeck] by Cicadellini [Bucephalogonia xanthophis (Berg) and Dilobopterus costalimai Young] and Proconiini [Homalodisca ignorata Melichar and Oncometopia facialis (Signoret)] sharpshooters that occur in both crops. At different seasons, healthy adults of each species were submitted to a 48-h acquisition access period on citrus or coffee source plants infected with X. fastidiosa isolates that cause Citrus variegated chlorosis (CVC) and Coffee leaf scorch (CLS), respectively, and then confined on healthy seedlings of the corresponding host plant for a 48-h inoculation access period. No significant effect of inoculation season was observed when comparing infection rates of citrus or coffee plants inoculated by vectors at different times of the year. In citrus, the transmission rate by single insects was significantly higher for H. ignorata (30%) in relation to B. xanthophis (5%) and O. facialis (1.1%), but there was no difference among vector species in coffee, whose transmission rates ranged from 1.2 to 7.2%. Comparing host plants, H. ignorata was more effective in transmitting X. fastidiosa to citrus (30%) in relation to coffee (2.2%), whereas the other vectors transmitted the bacterium to both hosts with similar efficiencies. Despite these variations, vector efficiency in coffee and citrus is lower than that reported in other hosts.

Genetic structure and biology of Xylella fastidiosa strains causing disease in citrus and coffee in Brazil.

Xylella fastidiosa is a vector-borne, plant-pathogenic bacterium that causes disease in citrus (citrus variegated chlorosis [CVC]) and coffee (coffee leaf scorch [CLS]) plants in Brazil. CVC and CLS occur sympatrically and share leafhopper vectors; thus, determining whether X. fastidiosa isolates can be dispersed from one crop to another and cause disease is of epidemiological importance. We sought to clarify the genetic and biological relationships between CVC- and CLS-causing X. fastidiosa isolates. We used cross-inoculation bioassays and microsatellite and multilocus sequence typing (MLST) approaches to determine the host range and genetic structure of 26 CVC and 20 CLS isolates collected from different regions in Brazil. Our results show that citrus and coffee X. fastidiosa isolates are biologically distinct. Cross-inoculation tests showed that isolates causing CVC and CLS in the field were able to colonize citrus and coffee plants, respectively, but not the other host, indicating biological isolation between the strains. The microsatellite analysis separated most X. fastidiosa populations tested on the basis of the host plant from which they were isolated. However, recombination among isolates was detected and a lack of congruency among phylogenetic trees was observed for the loci used in the MLST scheme. Altogether, our study indicates that CVC and CLS are caused by two biologically distinct strains of X. fastidiosa that have diverged but are genetically homogenized by frequent recombination.