Feeding sites and food intake of Bucephalogonia xanthophis (Berg) (Hemiptera: Cicadellidae), a sharpshooter vector of Xylella fastidiosa, on citrus plants

A cigarrinha Bucephalogonia xanthophis (Berg) (Hemiptera: Cicadellidae) é um importante vetor da bactéria Xylella fastidiosa, agente causal da clorose variegada dos citros. Este trabalho teve como objetivo identificar o local preferido de alimentação e o período de maior atividade alimentar desta cigarrinha em citros, no sentido de elucidar o comportamento alimentar relacionado à transmissão da bactéria. O local de alimentação foi estudado em ensaio de escolha, no qual 30 insetos adultos foram liberados em gaiolas de observação (n = 10) contendo uma muda de laranja-doce [Citrus sinensis (L.) Osbeck]. Após 1; 15; 21; 25; 39; 45 e 49 h da liberação, contaram-se os insetos na parte superior (ramos com brotações) e inferior (haste principal, até H40 cm de altura) da muda. Nos ramos da parte superior, avaliou-se a preferência entre a haste, o pecíolo e o limbo foliar. Em um segundo ensaio, 20 machos e 20 fêmeas de B. xanthophis foram confinados individualmente sobre a haste de 'seedlings' de laranja-doce para determinar os períodos de alimentação, quantificando-se a excreção de 'honeydew' (medida indireta da ingestão) em períodos sucessivos de dia e noite, durante 48 h. A maioria dos indivíduos de B. xanthophis preferiu a haste dos ramos novos (62%), na parte superior da muda cítrica (91%). Nos 'seedlings', observou-se maior volume de excreção e proporção de indivíduos excretando durante a fotofase, independentemente do sexo. Portanto, em estudos de transmissão de X. fastidiosa, deve-se considerar a preferência de B. xanthophis pela haste de brotações cítricas e sua maior atividade alimentar durante a fotofase.The sharpshooter Bucephalogonia xanthophis (Berg) (Hemiptera: Cicadellidae) is an important vector of Xylella fastidiosa, a bacterium that causes citrus variegated chlorosis. This study aimed to identify the preferred feeding sites and periods of this vector on citrus, in order to improve our understanding of the feeding behavior related to the transmission of this pathogen. The feeding sites were determined in a choice experiment, in which 30 adults were released inside observation chambers (n = 10) containing a sweet orange seedling [Citrus sinensis (L.) Osbeck]. One, fifteen, twenty-one, twenty-five, thirty-nine, forty-five, and forty-nine hours after the release, the number of insects on the upper (stem with sprouts) and bottom (primary stem, up to 40 cm above soil) parts of the plant was recorded. To determine the feeding periods, 20 males and 20 females of B. xanthophis were individually confined on the stems of sweet orange seedlings, and the honeydew excretion (indirect measure of ingestion) was quantified for 48 hours, during periods of day and night. Most B. xanthophis individuals (91%) were observed on the upper part of the plant, mainly on the stems of the young shoots (62%). A larger volume of excretion and higher proportion of excreting individuals were observed during the photophase for both males and females, indicating that daytime is the preferred feeding period for this vector. Therefore, in transmission studies of X. fastidiosa one should consider the preference of B. xanthophis for stems of young shoots and its higher feeding activity during the photophase.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Identificação de uma planta não-hospedeira de Xylella fastidiosa para criação de insetos vetores sadios

A obtenção de cigarrinhas (Hemiptera: Cicadellidae) livres de Xylella fastidiosa é importante para estudos de interação entre essa bactéria e seus vetores, sendo desejável a seleção de uma planta que permita a criação desses insetos, mas não a multiplicação da bactéria. Neste estudo, duas plantas hospedeiras de cigarrinhas, Vernonia condensata (boldo) e Aloysia virgata (lixeira), foram inoculadas por agulha com as estirpes de citros e de cafeeiro de X. fastidiosa, para avaliar a possibilidade deste patógeno colonizá-las. Não foram observados sintomas, nem se detectou a bactéria por isolamento em meio de cultura e/ou PCR em períodos curtos (7 e 14 dias) ou longos (1, 4, 6 e 12 meses) após a inoculação. Para obtenção de adultos sadios das cigarrinhas vetoras, Acrogonia citrina, Bucephalogonia xanthophis, Dilobopterus costalimai, Homalodisca ignorata e Oncometopia facialis, ninfas de primeiros ínstares foram criadas em plantas de boldo. Não foi detectada X. fastidiosa em nenhum de 175 adultos obtidos da criação. V. condensata e A. virgata não permitem a colonização de X. fastidiosa, possibilitando assim a obtenção de cigarrinhas sadias para estudos com vetores.Rearing leafhopper (Hemiptera: Cicadellidae) vectors free of Xylella fastidiosa is a requirement for studies of various aspects of vector-pathogen interactions. The selection of a plant that allows vector development but not bacterial multiplication is desirable to produce healthy vectors. In this study, two leafhopper hosts, Vernonia condensata ('boldo') and Aloysia virgata ('lixeira') were needle inoculated with citrus and coffee strains of X. fastidiosa to evaluate if these plants support pathogen colonization. The inoculated plants did not present symptoms and the pathogen was not detected by culture and PCR tests, neither soon after inoculation (7-14 days) nor later, at 1, 4, 6 and 12 months after inoculation. To obtain healthy adults of the leafhopper vectors Acrogonia citrina, Bucephalogonia xanthophis, Dilobopterus costalimai, Homalodisca ignorata and Oncometopia facialis, early-instar nymphs were reared on V. condensata. X. fastidiosa was not detected in any of 175 adults obtained. V. condensata and A. virgata are nonpropagative hosts of X. fastidiosa and enable the production of healthy leafhoppers for vector studies

Resistance of True Citrus species to Diaphorina citri

This is the peer reviewed version of the following article: Eduardo, W. I., Miranda, M. P., Volpe, H. X. L., Garcia, R. B., Girardi, E. A., Alquezar, B., ... & Peña, L. (2022). Resistance of True Citrus species to Diaphorina citri. Pest management science, 78(11), 4783-4792, which has been published in final form at [https://doi.org/10.1002/ps.7098]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.[EN] BACKGROUND Host genetic resistance is a promising strategy for the management of Diaphorina citri Kuwayama (Hemiptera: Psyllidae), and consequently Huanglongbing (HLB). To date, no study has investigated the resistance to D. citri in the clonal and vegetatively propagated plants of the Microcitrus, Eremocitrus, and Atalantia genera. This study assesses Near and True Citrus genotype antixenosis and antibiosis against D. citri, with trichome density and volatile emission as possible mechanisms of resistance. RESULTS All genotypes were oviposited by D. citri, however, 8 of 14 genotypes were less oviposited than Citrus x sinensis 'Valencia' (susceptible control). Diaphorina citri nymphs had lower nymphal viability in E. glauca (31%) and M. warburgiana (58%) than that in Citrus x sinensis (77%). The behavioral assay showed that 30% of D. citri nymphs in the last instars evaded E. glauca shoots, whereas no nymphs evaded Citrus x sinensis shoots. A higher trichome density was observed in E. glauca shoots compared to the other genotypes. Chemical analysis revealed differences in the volatile profiles of E. glauca and Citrus x sinensis. CONCLUSION Eremocitrus glauca and M. warburgiana genotypes were more resistant to D. citri than Citrus x sinensis. Higher trichome density in the shoots may negatively influence the development of D. citri nymphs. Eremocitrus glauca volatiles may also be involved in their resistance to D. citri.This work was funded by the Fund for Citrus Protection (Fundecitrus), the European Union H2020 Innovation Action Program (grant #817526), and the project PID2019-104569RB-I00 from AEI-Spain. WIE received a postdoctoral fellowship (Proc. 2019/19649-3) from SAo Paulo Research Foundation (FAPESP). MPM received a research fellowship (Proc. 301805/2018-0) from Council for Scientific and Technological Development (CNPq), Brazil. The authors thank the Metabolomic service from Instituto de Biologia Molecular y Celular de Plantas, CSIC/UPV (IBMCP), Valencia, Spain for their help with the volatile emission analysis. The authors also acknowledge Fundecitrus and EMBRAPA Cassava & Fruits for providing plant materials.Eduardo, WI.; Miranda, MP.; Linhares Volpe, HX.; Garcia, RB.; Alquézar-García, B.; Girardi, EA.; Espinosa Ruiz, A.... (2022). Resistance of True Citrus species to Diaphorina citri. Pest Management Science. 78(11):4783-4792. https://doi.org/10.1002/ps.709847834792781

Effect of contact and systemic insecticides on the sharpshooter Bucephalogonia xanthophis (Hemiptera: cicadellidae), a vector of Xylella fastidiosa in citrus

The knockdown and toxic effects of insecticides of different chemical groups and modes of action registered for citrus in Brazil were investigated for effective control of Bucephalogonia xanthophis, a sharpshooter vector of Xylella fastidiosa in citrus. The active ingredients dimethoate (1.2 mL/1.2L), imidacloprid (0.24 mL/1.2L) and lambda-cyhalothrin (0.24 mL/1.2L), as well as a control (water), were sprayed onto branches of potted-citrus nursery trees to evaluate the effect of residual contact. The insects were confined on sprayed branches by using sleeve cages, in groups of 10 per branch (5 branches/treatment). Lambdacyhalothrin showed a knockdown effect on B. xanthophis (>70% mortality within 2 h of exposure), and the residues were effective for approximately one wk. Imidacloprid, lambdacyhalothrin and dimethoate suppressed the vector populations for up to 3 wk after application, when the insects were exposed to sprayed plants for at least 24 h. In another experiment, 2 neonicotinoid insecticides (thiamethoxam and imidacloprid) were applied by soil drench to potted nursery trees, in order to study their systemic effect, i.e., mortality by ingestion on sharpshooter adults. Thiamethoxam and imidacloprid effectively controlled the vectors at all concentrations tested, when the insects were exposed to treated plants for 24 h (>80% mortality) or 48 h (near 100% mortality). The knockdown effect of thiamethoxam and lambda-cyhalothrin might be particularly important to prevent vector transmission of X. fastidiosa in citrus groves.Programa Nacional de Pos-Doutorado - PNPD/CAPES [23038.039426/2008-97

Engineered Orange Ectopically Expressing the Arabidopsis beta-Caryophyllene Synthase Is Not Attractive to Diaphorina citri, the Vector of the Bacterial Pathogen Associated to Huanglongbing

[EN] Huanglongbing (HLB) is a destructive disease, associated with psyllid-transmitted phloem-restricted pathogenic bacteria, which is seriously endangering citriculture worldwide. It affects all citrus species and cultivars regardless of the rootstock used, and despite intensive research in the last decades, there is no effective cure to control either the bacterial species (Candidatus Liberibacter spp.) or their insect vectors (Diaphorina citri and Trioza erytreae). Currently, the best attempts to manage HLB are based on three approaches: (i) reducing the psyllid population by intensive insecticide treatments; (ii) reducing inoculum sources by removing infected trees, and (iii) using nursery-certified healthy plants for replanting. The economic losses caused by HLB (decreased fruit quality, reduced yield, and tree destruction) and the huge environmental costs of disease management seriously threaten the sustainability of the citrus industry in affected regions. Here, we have generated genetically modified sweet orange lines to constitutively emit (E)-beta-caryophyllene, a sesquiterpene repellent to D. citri, the main HLB psyllid vector. We demonstrate that this alteration in volatile emission affects behavioral responses of the psyllid in olfactometric and no-choice assays, making them repellent/less attractant to the HLB vector, opening a new alternative for possible HLB control in the field.This work was funded by the Fundo de Defesa da Citricultura (Fundecitrus), Sao Paulo Research Foundation (FAPESP, grant #2015/07011-3), and EU H2020 Innovation Action Program (grant #817526). Consent for research and field trial of genetically modified organisms was granted by the National Technical Biosafety Commission from Brazil (CTNBio) to Fundecitrus.Alquézar-García, B.; Linhares Volpe, HX.; Facchini Magnani, R.; Pedreira De Miranda, M.; Almeida Santos, M.; Vieira Marques, V.; Rodrigues De Almeida, M.... (2021). Engineered Orange Ectopically Expressing the Arabidopsis beta-Caryophyllene Synthase Is Not Attractive to Diaphorina citri, the Vector of the Bacterial Pathogen Associated to Huanglongbing. Frontiers in Plant Science. 12:1-15. https://doi.org/10.3389/fpls.2021.6414571151

Survey of leafhopper (hemiptera: cicadellidae) vectors of Xylella fastidiosa in citrus groves of the north coast of Bahia state.

Este trabalho foi realizado com o objetivo de identificar cigarrinhas vetoras ou potenciais vetoras de Xylella fastidiosa Wells et al. em pomares cítricos do litoral norte da Bahia, determinando-se as espécies predominantes com ênfase na subfamília Cicadellinae. Os levantamentos foram conduzidos no período de março/2002 a fevereiro/2003, em cinco propriedades do litoral norte do Estado da Bahia, nos municípios de Alagoinhas, Inhambupe e Rio Real, em pomares de laranja doce [Citrus sinensis (L.) Osbeck] cv. Pêra, enxertada sobre limão cravo (Citrus limonia Osbeck) com idade de 7 a 9 anos. Em cada pomar, foram instalados 15 cartões adesivos amarelos de 8,5 X 11,5 cm, dispostos em um espaçamento de 40x40 m. Os cartões foram amarrados a 1,5 m de altura em ramos da face norte, na periferia da copa, sendo substituídos periodicamente. Também foram realizadas avaliações periódicas na vegetação rasteira presente nas entrelinhas dos pomares através de rede de varredura, amostrando-se, em cada avaliação, cinco pontos escolhidos aleatoriamente e efetuando-se 30 redadas por ponto. Os dados de coleta foram submetidos a uma análise faunística, calculando-se os índices de constância, freqüência, abundância, dominância, diversidade, eqüitabilidade e similaridade. Coletaram-se 1860 espécimes pertencentes a 54 espécies de sete famílias da subordem Auchenorrhyncha, sendo a maioria deles das famílias Cicadellidae (84,3%) e Membracidae (14,2%). Um maior número de espécies de Auchenorrhynca foi observado em pomar vizinho a mata nativa pouco degradada. A subfamília Cicadellinae, que inclui os vetores de X. fastidiosa, foi a mais representativa em número de espécies (15) e de indivíduos coletados (63,92%). Observou-se variação na composição de espécies de Cicadellinae dependendo do método e local de amostragem, evidenciando-se a existência de dois grupos de potenciais vetores habitando a copa das laranjeiras e a vegetação rasteira. Acrogonia flagellata Young, A. citrina Marucci & Cavichioli, Homalodica sp. e Cicadellini sp.1 predominaram nos cartões adesivos amarelos (árvores cítricas), enquanto que Hortensia similis (Walker) e Erythrogonia dubia (Medler) foram dominantes na rede de varredura (vegetação rasteira). Entre as espécies de Cicadellinae já conhecidas como vetoras de X. fastidiosa em citros, apenas A. citrina, Bucephalogonia xanthophis (Berg) e Ferrariana trivittata (Signoret) foram observadas, sendo que as duas últimas foram de ocorrência acidental na vegetação rasteira. Entre os novos potenciais vetores, A. flagellata, Homalodisca sp. e Cicadellini sp.1 são espécies predominantes nos pomares cítricos do litoral norte da Bahia, devendo ser investigadas quanto à capacidade de transmissão de X. fastidiosa para citros.This study was carried out to identify vectors or potential vectors of Xylella fastidiosa Wells et al. in citrus groves of the north coast of Bahia State, Brazil, based on a faunistic survey and determination of predominant sharpshooter leafhoppers (subfamily Cicadellinae). The survey was carried out from March/2002 to February/2003 in five old sweet orange [Citrus sinensis (L.) Osbeck, cv. Pêra, grafted on rangpur lime (Citrus limonia Osbeck); 7-9 years old] groves of northeastern Bahia State, located in Alagoinhas, Inhambupe and Rio Real. Fifteen yellow sticky cards (8,5 X 11,5 cm) were installed 40x40 m apart in each grove, hanged at the height of 1.5 m on the upper north side of citrus canopies. Cards were replaced monthly or fortnightly. A sweep net was periodically used to sample leafhoppers in the herbaceous weeds inside the groves, by selecting five points at random and performing 30 sweeps in each point (sample unit). Data was uses to calculate indices of constancy, frequency, abundance, dominance, diversity, equitability and similarity. A total of 1860 specimens of 54 species belonging to 7 families of suborder Auchenorrhyncha were collected, most of them in the families Cicadellidae (84,3%) and Membracidae (14,2%). A larger number of species was observed in a grove located nearby a non-degraded native wood. The subfamily Cicadellinae, which includes the sharphooter vectors of X. fastidiosa, showed the largest number of species (15) and specimens (63,92%). There was a variation in Cicadellinae species composition depending upon the sampling method, which indicates that two distinct groups of potential sharpshooter vectors are inhabiting the citrus canopy and the weedy vegetation. Acrogonia flagellata Young, A. citrina Marucci & Cavichioli, Homalodica sp. e Cicadellini sp.1 predominate in the yellow sticky cards hanged on the citrus canopy, whereas Hortensia similis (Walker) e Erythrogonia dubia (Medler) were dominant species on weeds. 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. Among the new potential vectors, A. flagellata, Homalodisca sp. e Cicadellini sp.1 are predominant species in citrus groves of the north coast of Bahia and should be tested with respect to their ability to transmit X. fastidiosa to citrus

Characterization of feeding behavior of Bucephalogonia xanthophis (Berg) (Hemiptera: Cicadellidae) in citrus and its implications for transmission of Xylella fastidiosa

Xylella fastidiosa é uma bactéria limitada ao xilema de plantas, sendo transmitida principalmente por cigarrinhas da subfamília Cicadellinae (Hemiptera: Cicadellidae). No Brasil, é o agente causal da Clorose variegada dos citros (CVC), doença que afeta laranja-doce [Citrus sinensis (L.) Osbeck]. O objetivo deste trabalho foi caracterizar o comportamento alimentar da cicadelíneo vetor Bucephalogonia xanthophis (Berg) em citros e correlacionar suas atividades estiletares com a transmissão de X. fastidiosa. Inicialmente, testes de escolha e análises de excreção de honeydew foram realizadas para determinar os locais e períodos preferidos para alimentação em mudas de citros. B. xanthophis preferiu a haste dos ramos novos, na parte superior da muda. Esta cigarrinha ingeriu seiva do xilema e apresentou maior volume médio de excreção e maior percentual de indivíduos que excretaram durante a fotofase. Assim, estudou-se a penetração estiletar do vetor na haste de brotações cítricas, durante a fotofase, pela técnica de \"Electrical Penetration Graph\" (EPG, sistema DC). Os principais padrões de EPG foram correlacionados com análises histológicas e de \"honeydew\" para determinação da posição exata dos estiletes no tecido vegetal e atividades envolvidas. Seis padrões foram descritos: (S) secreção de bainha salivar e caminhamento dos estiletes através de células da epiderme ou parênquima; (R) estiletes inseridos na planta, porém sem nenhuma atividade aparente; (X) contato dos estiletes com os vasos do xilema; (Xi) ingestão ativa no xilema; (I) breve interrupção durante X ou Xi; (W) retirada dos estiletes da planta. Durante uma prova, a seqüência de eventos com maior probabilidade de ocorrência foi penetração estiletar através da epiderme e parênquima (S) (100% dos insetos), seguida de contato com o xilema (X) (67,6%). Entre os indivíduos que exibem o padrão X, 88,3% passam para Xi. Os vasos do xilema foram localizados pelo inseto após uma média de 2,2 provas. O tempo médio para atingir o xilema (X) e iniciar ingestão (Xi) após o início da primeira prova foi 27.8 min e 34,2 min, respectivamente. Entretanto, verificou-se ingestão prolongada no xilema (Xi > 5 min) somente após 39,8 min, em média. Em um outro estudo, investigou-se a relação dos padrões S, X e Xi com os processos de aquisição e inoculação de X. fastidiosa em citros. B. xanthophis adquiriu X. fastidiosa somente no padrão Xi. Após 1 h de ingestão no xilema, esta cigarrinha transmitiu a bactéria para plantas-teste com uma eficiência de 7,7%. A inoculação ocorreu durante os padrões S, S+X e S+X+Xi, com eficiência de 3,5; 7,1 e 7,4%, respectivamente. Um fato intrigante foi a ocorrência de inoculação de X. fastidiosa pelo inseto antes de atingir os vasos do xilema, durante o padrão S. Contudo, as maiores taxas de transmissão ocorreram após o contato com o xilema (S+X ou S+X+Xi). Por fim, estudou-se o efeito da infecção sintomática e assintomática por X. fastidiosa em plantas cítricas sobre a penetração estiletar de B. xanthophis. O comportamento alimentar foi semelhante em plantas sadias e infectadas sem sintomas. A infecção sintomática não afetou a capacidade de B. xanthophis localizar os vasos do xilema, mas reduziu o tempo gasto por este inseto ingerindo seiva dos mesmos. Estes resultados sugerem que a aquisição de X. fastidiosa pode ser mais eficiente em plantas infectadas assintomáticas do que em plantas com sintomas severos de CVC. As informações sobre penetração estiletar do vetor B. xanthophis em citros são importantes para estudos mais avançados de mecanismos de transmissão de X. fastidiosa, bem como para estabelecer estratégias que visem interferir neste processo.Xylella fastidiosa is a xylem-limited bacterium transmitted mainly by leafhoppers (Hemiptera: Cicadellidae) in the subfamily Cicadellinae. In Brazil, it is the causal agent of Citrus variegated chlorosis, a disease that affects sweet orange [Citrus sinensis (L.) Osbeck]. The goal of this research was to characterize the feeding behavior of the cicadeline vector Bucephalogonia xanthophis (Berg) in citrus and correlate its feeding activities with transmission of X. fastidiosa. Initially, choice tests and honeydew excretion analyses were carried out to determine preferred feeding sites and periods on citrus nursery trees. B. xanthophis preferred the stems of young shoots, in the upper part of the plant. This species ingested sap from the xylem vessels and showed larger excretion volume and higher proportion of excreting individuals during the day. Thus, vector stylet penetration was studied on the stem of citrus shoots in the photophase, by using the Electrical Penetration Graph (EPG, DC system) technique. The main EPG waveforms were correlated with histological and honeydew excretion analyses to determine the precise stylet position in the plant tissue and feeding activities. Six waveforms and proposed activities are described: (S) secretion of salivary sheath and intracellular pathway; (R) stylets inserted into the plant, without any apparent activity; (X) contact of stylets with xylem vessels; (Xi) active xylem ingestion; (I) interruption between X and Xi; and (W) stylet withdrawal from the plant. During a probe, the most likely sequence of events is stylet pathway (S) through epidermal and parenchymal cells (all individuals), followed by contact with xylem (X) (67.6% of all individuals) and then active ingestion (Xi) (88.3% of those that exhibit waveform X). The xylem was reached after an average of 2.2 probes. The mean time to contact the xylem (X) and initiate ingestion (Xi) after onset of the first probe was 27.8 and 34.2 min, respectively. However, sustained xylem ingestion (Xi > 5 min) was established only after 39.8 min, on average. In a second study, the waveforms S, X and Xi were correlated with X. fastidiosa acquisition and inoculation in citrus. Acquisition of X. fastidiosa from infected plants occurred only after onset of the pathogen to 7.7% of test plants. In healthy plants, inoculation took place during waveform Xi (xylem ingestion); but with just 1 h in Xi, B. xanthophis subsequently transmitted waveforms S (salivary sheath formation and stylet pathway), S+X (X= first xylem contact by stylets) and S+X+X1, with efficiencies of 3.5, 7.1 and 7.4%, respectively. Although higher transmission rates were recorded after the first contact with xylem (S+X and S+X+Xi), it is intriguing the fact that inoculation of this xylem-limited bacterium also occurred before that (during S). Finally, the effect of X. fastidiosa infection on the feeding behavior of B. xanthophis was studied by comparing stylet penetration on: a) healthy citrus; b) symptomless infected citrus; c) infected citrus with CVC symptoms. Based on the analysis of 26 EPG parameters, no significant differences were found in stylet penetration on healthy versus asymptomatic infected citrus. Symptomatic infection did not affect the ability of B. xanthophis to locate xylem vessels, but reduced the time spend by this vector ingesting xylem sap. These results suggest that X. fastidiosa acquisition may be more efficient on symptomless infected plants than on citrus with severe CVC symptoms. Information on vector feeding behavior is basic for future studies on transmission mechanisms of X. fastidiosa and to establish control strategies aimed to interfere with this process

Drench Application of Systemic Insecticides Disrupts Probing Behavior of Diaphorina citri (Hemiptera: Liviidae) and Inoculation of Candidatus Liberibacter asiaticus

© 2020 by the authorsCandidatus Liberibacter asiaticus (CLas) is a phloem-limited bacterium that is associated with the Huanglongbing (HLB) disease of citrus and transmitted by the psyllid, Diaphorina citri. There are no curative methods to control HLB and the prevention of new infections is essential for HLB management. Therefore, the objective of our study was to determine the effects of systemic insecticides, such as the neonicotinoids imidacloprid, thiamethoxam, and a mixture of thiamethoxam and chlorantraniliprole (diamide) on the probing behavior of CLas-infected D. citri and their effect on CLas transmission. The electrical penetration graph (EPG-DC) technique was used to monitor the stylet penetration activities of CLas-infected D. citri on sweet orange [Citrus sinensis (L.) Osbeck] ‘Valencia’ treated with systemic insecticides. Systemic insecticides disrupted the probing behavior of CLas-infected D. citri, in a way that affected CLas transmission efficiency, particularly by negatively affecting the stylet activities related to the phloem phase. All insecticides reduced (by 57–73%) the proportion of psyllids that exhibited sustainable phloem ingestion (waveform E2 > 10 min), with significant differences observed on plants treated with thiamethoxam and thiamethoxam + chlorantraniliprole. The transmission rate of CLas with high inoculum pressure (five CLas-infected D. citri per plant and a seven-day inoculation access period) to untreated control plants was 93%. In contrast, CLas transmission was reduced to 38.8% when test plants were protected by systemic insecticides. Our results indicated that all insecticides tested presented a potential to reduce CLas inoculation by an average of 59%; therefore, these insecticides can be used to reduce the spread of HLB.Financial support was received from National Council for Scientific and Technological Development (CNPq) and supported by Fund for Citrus Protection (Fundecitrus). First author received a Posdoc scholarship from CNPq/Brazil (300581/2016-5); the last author received a fellowship CNPq/Brazil (Proc. 301805/2018-0).Peer reviewe

Push-pull and kill strategy for Diaphorina citri control in citrus orchards

The development of sustainable methods to control Diaphorina citri Kuwayama (Hemiptera: Liviidae) is crucial to improve management of huanglongbing (HLB), the most important citrus disease worldwide. In a flushing Citrus × sinensis L. Osbeck (Rutaceae) orchard, this study assessed the combined effects of insecticide-treated Bergera koenigii L. (Rutaceae) trees in the orchard border, as a lethal trap crop, and the application of processed kaolin to the citrus crop, as a repellent (push-pull and kill strategy), in reducing D. citri infestation and dispersal. Additionally, the volatile organic compound (VOC) emission profile and attractiveness of B. koenigii shoots were investigated in indoor experiments. The push-pull and kill strategy reduced the number of D. citri adults on citrus trees by 95, 84, and 63% compared to the untreated control, trap crop only, and kaolin only treatments, respectively. These high reductions can be attributed to the combination of attractive stimuli (VOCs) and repellent stimuli by the trap crop and kaolin, respectively. The number of psyllids observed on citrus trees subjected to treatments with trap crop and kaolin separately was 69 and 86% lower than on untreated control trees, respectively. In addition, all treatments reduced and delayed D. citri dispersal inside the orchard, especially the push-pull and kill strategy. In olfactometric assays, D. citri females showed a significant preference for B. koenigii odors over C. × sinensis cv. ‘Valencia’ flush odors, likely related with the different VOC emission profiles of the two plants. This study shows the effectiveness of push-pull and kill as a new management strategy to control D. citri and consequently HLB.Peer reviewe