Grapevine bacterial canker caused byXanthomonas citri pv. viticola (X. campestris pv. viticola) (Xcv), was detected in Brazil in 1998 and is currently regarded as a quarantine disease with limited distribution in the country. To improve sensitivity and speed in the detection of Xcv in asymptomatic grapevines, two pairs of primers were designed, targeting sequences of a pathogenicity gene (hrpB) and the xanthomonadin coding cluster. Both pairs were tested in conventional PCR (cPCR) and real-time PCR (qPCR) formats. Primers targeting the hrpB gene showed cross reactions with other Xanthomonas spp. but were effective for use in both cPCR and qPCR, whereas primers for the xanthomonadin gene were highly specific for Xcv but showed low efficiency in qPCR. Enrichment of plant extracts in semi-selective medium before qPCR allowed a significant increase in sensitivity when compared to total DNA extraction, making it possible to detect as low as 10 1 CFU ml -1 . Under natural infection conditions, symptomatic and asymptomatic grapevines were tested by qPCR with hrpB primers and cPCR with xanthomonadin primers. In both cases, plant extracts were enriched for 36-72h. Xcv was detected in all symptomatic samples by qPCR and the result was confirmed by cPCR. For the asymptomatic samples, Xcv was detected in 93.4% with qPCR and in 89.5% with cPCR. These two methods offer advantages in terms of sensitivity and specificity, and they could be useful in quarantine programs, certification of grapevine propagating material and detection of inoculum sources in alternative hosts, contributing to the prevention of pathogen spread to disease-free areas.Keywords Grapevine bacterial canker . Vitis vinifera . Xanthomonas campestris pv. viticola. PCR-based diagnosis . qPCR
TRATAMENTO QUÍMICO DE SEMENTES DE FEIJÃO (Phaseolus vulgaris L.) PARA O CONTROLE DE Dedico AGRADECIMENTOSA Deus, pela presença constante em minha vida, dando-me proteção, coragem e força para conquistar meus objetivos.Ao meu pai, Gilberto Mauro Villela, meu maior incentivador, pela valiosa ajuda na montagem e condução do experimento de campo.A minha mãe, Valdemira Alves da Silva, pelo amor incondicional e confiança.A minha irmã Nathalia Alves Villela, pelo companheirismo e ajuda na montagem do experimento de campo.Ao Prof. Carlos Hidemi Uesugi, pela orientação, estímulo, incentivo, confiança e, sobretudo, pela amizade.Ao técnico de laboratório de Bacteriologia Vegetal, Arenildo Soares, pelo conhecimento repassado.Ao Prof. Ernandes Rodrigues, pelo conhecimento repassado e disponibilização da estrutura para ozonização das sementes.Aos funcionários da Estação Experimental de Biologia, pela disposição e ajuda na condução dos experimentos.Ao Adalberto Gatto, pelo fornecimento das sementes usadas nos experimentos.Aos amigos que fiz durante o curso, Carina Lopes, Cleia Cabral, Débora Cervieri, Elenice Barbosa, Geane Fontes, Pedro Verlage, Rafaela Borges e Ricardo Nunes, os quais tornaram essa caminhada mais fácil e divertida.Aos amigos de Unaí, Bruno Monteiro, Fabrício Vilela, Luís Augusto Fontanelle, Rafael Frees, Hudson Teodoro, pelos momentos de alegria e por sempre torcerem por mim.Aos amigos da graduação, Antônio Nelson, Aureliano Dantas, Bernardo Coutinho, Bruno Costa, Thiago Peixoto, pessoas que sempre posso contar quando preciso. Ao CRUZEIRO ESPORTE CLUBE, pelas alegrias e emoções vividas.A Universidade de Brasília e ao Departamento de Fitopatologia pela oportunidade de realização do curso.Ao CNPq, pela concessão da bolsa de estudos.A todo corpo docente do Departamento de Fitopatologia, pelos ensinamentos e conselhos repassados.A todos os colegas e funcionários do Departamento de Fitopatologia, pelo apoio e companheirismo no decorrer desta caminhada. Aos professores Luiz Eduardo Bassay Blum e Reinaldo José de Miranda Filho quegentilmente aceitaram participar da banca avaliadora.A todos aqueles que, de alguma forma, contribuíram para a realização deste trabalho.Trabalho realizado junto ao Departamento de Fitopatologia do Instituto de Ciências Biológicas da Universidade de Brasília, sob orientação do Professor Carlos Hidemi Uesugi, com apoio do Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). kasugamycin, benzalkonium chlorides and peracetic acid, exposure to vapor ethanol and methanol, and ozonation. In the evaluation of treatments efficiency in eradicating of Cff, inoculated seeds were treated and subjected to extraction process, then checking the incidence of contaminated seeds and number of bacterial cells. Experiments were carried in field conditions and greenhouse conditions. In field experiment was evaluated the effects of treatments on seed physiological quality, and greenhouse was measured the effect of treatments on seed physiological quality, in vegetative plant development, in i...
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