PM 7/24 (3) <i>Xylella fastidiosa</i>

doi:10.1111/epp.12469

Cited by 30 publications

(6 citation statements)

References 71 publications

(58 reference statements)

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“…The use of one of these tetraplex assays hence corresponds to the first requirement for Xf detection as reported in PM 7/98 (EPPO, 2014). So far, subspecies identification is done by sequencing two to seven housekeeping genes (Yuan et al, 2010;EPPO, 2018b). If one of the gene amplifications fails, or if sequencing is not feasible (in case of a too-low amount of DNA), then the subspecies cannot be assigned.…”

Section: Discussionmentioning

confidence: 99%

“…This bacterial species is genetically diverse as five subspecies including fastidiosa, morus, multiplex, pauca, and sandyi are currently described (EFSA, 2018b). Although this subspecies delineation was initially associated to Xf host range and places of occurrence, more and more observations report infection of a given host by various subspecies (Denancé et al, 2017;EPPO, 2018b;Denancé et al, 2019;Nunney et al, 2019). Homologous recombination events were detected in Xf and were suspected to be associated with host-shift, as documented for the subspecies morus (Nunney et al, 2014).…”

Section: Introductionmentioning

confidence: 98%

“…The subspecies identification is then routinely performed by MLST, but this method, while accurate and portable, is time consuming, labor intensive, and expensive. From 2018, sequences of only two housekeeping genes (rpoD and cysG or rpoD and malF) are required for subspecies identification in France, while other sets of gene pairs are recommended by EPPO (EPPO, 2018b).…”

Section: Introductionmentioning

confidence: 99%

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Novel Tetraplex Quantitative PCR Assays for Simultaneous Detection and Identification of Xylella fastidiosa Subspecies in Plant Tissues

et al. 2019

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Xylella fastidiosa (Xf) is an insect-borne bacterium confined to the xylem vessels of plants. This plant pathogen has a broad host range estimated to 560 plant species. Five subspecies of the pathogen with different but overlapping host ranges have been described, but only three subspecies are widely accepted, namely subspecies fastidiosa, multiplex, and pauca. Initially limited to the Americas, Xf has been detected in Europe since 2013. As management of X. fastidiosa outbreaks in Europe depends on the identification of the subspecies, accurate determination of the subspecies in infected plants as early as possible is of major interest. Thus, we developed various tetraplex and triplex quantitative PCR (qPCR) assays for X. fastidiosa detection and subspecies identification in planta in a single reaction. We designed primers and probes using SkIf, a bioinformatics tool based on k-mers, to detect specific signatures of the species and subspecies from a data set of 58 genome sequences representative of X. fastidiosa diversity. We tested the qPCR assays on 39 target and 30 non-target strains, as well as on 13 different plant species spiked with strains of the different subspecies of X. fastidiosa, and on samples from various environmental and inoculated host plants. Sensitivity of simplex assays was equal or slightly better than the reference protocol on purified DNA. Tetraplex qPCR assays had the same sensitivity than the reference protocol and allowed X. fastidiosa detection in all spiked matrices up to 10 3 cells.ml −1. Moreover, mix infections of two to three subspecies could be detected in the same sample with tetraplex assays. In environmental plant samples, the tetraplex qPCR assays allowed subspecies identification when the current method based on multilocus sequence typing failed. The qPCR assays described here are robust and modular tools that are efficient for differentiating X. fastidiosa subspecies directly in plant samples.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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Novel Tetraplex Quantitative PCR Assays for Simultaneous Detection and Identification of Xylella fastidiosa Subspecies in Plant Tissues

et al. 2019

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“…The main problem associated with these diseases is the obstruction of the xylem, with symptoms ranging from leaf marginal necrosis and leaf abscission to dieback, delayed growth, and death of plants through insufficient water flow [1,2]. The current forecast for the expansion and severity of these diseases, named as the grapevine Pierce's disease (PD) or the Olive Quick Decline Syndrome (OQDS), may increase shortly [1][2][3], but they are characterized by symptoms often similar to water stress [4]. The bacterium X. fastidiosa is known to colonize crops of different climatic zones worldwide.…”

Section: Introductionmentioning

confidence: 99%

Insecticidal Effect of Entomopathogenic Nematodes and the Cell-Free Supernatant from Their Symbiotic Bacteria against Philaenus spumarius (Hemiptera: Aphrophoridae) Nymphs

Vicente-Díez

Blanco-Pérez

González-Trujillo

et al. 2021

Insects

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The meadow spittlebug Philaenus spumarius (Hemiptera: Aphrophoridae) is the primary vector of Xylella fastidiosa (Proteobacteria: Xanthomonadaceae) in Europe, a pest–disease complex of economically relevant crops such as olives, almonds, and grapevine, managed mainly through the use of broad-spectrum pesticides. Providing environmentally sound alternatives to reduce the reliance on chemical control is a primary challenge in the control of P. spumarius and, hence, in the protection of crops against the expansion of its associated bacterial pathogen. Entomopathogenic nematodes (EPNs) are well-known biocontrol agents of soil-dwelling arthropods. Recent technological advances in field applications, including improvements in obtaining cell-free supernatant from their symbiotic bacteria, allow their successful implementation against aerial pests. Thus, this study aimed to evaluate, for the first time, the efficacy of EPN applications against nymphal instars of P. spumarius. We tested four EPN species and the cell-free supernatant of their corresponding symbiotic bacteria: Steinernema feltiae–Xenorhabdus bovienii, S. carpocapsae–X. nematophila, S. riojaense–X. kozodoii, and Heterorhabditis bacteriophora–Photorhabdus laumondii subsp. laumondii. First, we showed that 24 and 72 h exposure to the foam produced by P. spumarius nymphs did not affect S. feltiae virulence. The direct application of steinernematid EPNs provided promising results, reaching 90, 78, and 53% nymphal mortality rates after five days of exposure for S. carpocapsae, S. feltiae, and S. riojaense, respectively. Conversely, the application of the cell-free supernatant from P. laumondii resulted in nymphal mortalities of 64%, significantly higher than observed for Xenorhabdus species after five days of exposure. Overall, we demonstrated the great potential of the application of specific EPNs and cell-free supernatant of their symbiont bacteria against P. spumarius nymphs, introducing new opportunities to develop them as biopesticides for integrated management practices or organic vineyard production.

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“…A equação de regressão Para detecção da bactéria nos insetos foi realizada a extração total do DNA seguida da análise de qPCR. As cigarrinhas provenientes dos experimentos tiveram a cabeça separada para a extração, a fim de minimizar a presença de inibidores presentes no restante do corpo do inseto(EPPO 2018). A extração de DNA total dos insetos seguiu a metodologia porRogers & Bendich (1988) com protocolo a base de CTAB (Cetyl Trimethyl Ammonium Bromide).…”

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N-acetilcisteína em estudos de transmissão de >i<Xylella fastidiosa>/i< por cigarrinhas

Kudlawiec¹

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N-actilcisteína em estudos de transmissão de Xylella fastidiosa por cigarrinhas / Karla Kudlawiec --versão revisada de acordo com a resolução CoPGr 6018 de 2011. --Piracicaba , 2021. 75 p. Dissertação (Mestrado) --USP / Escola Superior de Agricultura "Luiz de Queiroz". 1. Clorose variegada dos citros 2. Vetor 3. NAC 4. Controle I. Título AGRADECIMENTOSAgradeço a Deus por nortear meus caminhos... Agradeço aos meus pais por todo apoio, amor e dedicação e a toda minha família que eu amo e admiro muito.À minha irmã Carol por ser minha parceira de vida e por estar sempre ao meu lado em todos os momentos.Ao professor João Roberto Spotti Lopes pela orientação e apoio nas decisões ao longo do mestrado.Aos pesquisadores do IAC-Cordeirópolis, Alessandra Alves de Souza e Helvécio Dela-Coletta Filho pela ajuda com o projeto de pesquisa e pela orientação no desenvolvimento do projeto.Ao Dr Alberto Fereres, que me recebeu em seu laboratório durante o estágio no exterior e as pesquisadoras Aranzazu Moreno e Elisa Garzo por toda ajuda. Estendo meu agradecimento à técnica María Plaza e ao pós -doutorando Daniele Cornara por todo auxilio. Ao prof Dr. Walmes Zeviani -UFPR pela ajuda com as análises estatísticas.Aos colegas e companheiros que tornaram meus 6 meses em Madri uma experiência enriquecedora profissional e pessoalmente. Alan, Marina, Clara, Elena, Carolina, Cristina, Pierpaolo e Ana, vocês são 10! Os echo mucho de menos! Aos colegas do laboratório de insetos vetores, em especial a Maria Fernanda e a Mariana pela ajuda no decorrer do experimentos. Aos meus amigos, que mesmo com a distância levo comigo Mayara, Gaúcho, Marília, Bianca e Fernanda.Aos colegas da pós-graduação com os quais compartilhei momentos de estudos, apoio e de descontração, em especial à Sabrina, Thais, Matheus e Fernando, que tornaram-se amigos do coração. Incluo aqui a amiga Kelly com quem compartilhei muitos momentos felizes nesta etapa final e a pessoa que foi fundamental para minha evolução pessoal.

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PM 7/24 (3) Xylella fastidiosa

Abstract: Specific scope This Standard describes a diagnostic protocol for Xylella fastidiosa. It should be used in conjunction with PM 7/76 Use of EPPO diagnostic protocols. Specific approval and amendment First approved in 2004‐09. Revised in 2016‐09 and 2018‐04.

Cited by 30 publications

References 71 publications

Novel Tetraplex Quantitative PCR Assays for Simultaneous Detection and Identification of Xylella fastidiosa Subspecies in Plant Tissues

Novel Tetraplex Quantitative PCR Assays for Simultaneous Detection and Identification of Xylella fastidiosa Subspecies in Plant Tissues

Insecticidal Effect of Entomopathogenic Nematodes and the Cell-Free Supernatant from Their Symbiotic Bacteria against Philaenus spumarius (Hemiptera: Aphrophoridae) Nymphs

N-acetilcisteína em estudos de transmissão de >i<Xylella fastidiosa>/i< por cigarrinhas

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