RESUMO O melhoramento de plantas para resistência genética é um método importante para o manejo de doenças, pelos inúmeros benefícios e baixo custo de implementação. No presente estudo, progênies de 11 espécies de Coffea e 16 acessos selvagens de C. arabica foram testados quanto à resposta a Pseudomonas syringae pv. garcae, agente causal da mancha aureolada, doença disseminada nas principais regiões produtoras de café do Brasil e considerada fator limitante para o cultivo em áreas favoráveis a patógenos; e também para P. syringae pv. tabaci, agente causal da mancha foliar bacteriana, doença altamente agressiva detectada recentemente no Brasil. Experimentos separados para cada doença foram realizados em estufa, por meio da inoculação artificial dos patógenos em condições ideais de umidade para o desenvolvimento das doenças. Os resultados mostraram que as progênies Coffea canephora, C. congensis, C. eugenioides, C. stenophylla e C. salvatrix, além dos acessos selvagens de C. arabica Dilla & Alghe e Palido Viridis e da cultivar IPR 102, possuem níveis satisfatórios de resistência simultânea contra mancha aureolada e mancha foliar bacteriana. Os resultados descritos são úteis em programas de melhoramento para resistência duradoura a múltiplos agentes bióticos, fornecendo novas combinações de alelos de resistência por hibridização, bem como para estudos fitopatológicos, para identificar a variabilidade infraespecífica dos patógenos.

ABSTRACT Breeding for genetic resistance is an important method of crop disease management, due to the numerous benefits and low cost of establishment. In this study, progenies of 11 Coffea species and 16 wild C. arabica accessions were tested for their response to Pseudomonas syringae pv. garcae, the causal agent of bacterial halo blight, a widespread disease in the main coffee-producing regions of Brazil and considered a limiting factor for cultivation in pathogen-favorable areas; and also to P. syringae pv. tabaci, causal agent of bacterial leaf spot, a highly aggressive disease recently detected in Brazil. Separate experiments for each disease were carried out in a greenhouse, with artificial pathogen inoculations and ideal moisture conditions for disease development. The results showed that C. canephora, C. congensis, C. eugenioides, C. stenophylla, and C. salvatrix progenies, the wild C. arabica accessions Dilla & Alghe and Palido Viridis, and cultivar IPR 102 contain satisfactory levels of simultaneous resistance against bacterial halo blight and bacterial leaf spot. These results are useful in breeding programs for durable resistance to multiple biotic agents, providing new combinations of resistance alleles by hybridization, as well as for phytopathological studies, to identify infraspecific variability of the pathogens.

ABSTRACT Bacterial halo blight, caused by Pseudomonas syringae pv. garcae, is an important disease of coffee crop occurring in Brazil and other countries. In recent years, outbreaks of this disease have damaged several coffee crops in Brazil. Aggressiveness and genetic diversity of 25 strains of P. s. pv. garcae, obtained between the years 1958 and 2011, in 23 cities of São Paulo and Minas Gerais states, as well as three strains from Kenya were evaluated in this study. The strains were inoculated on coffee seedlings cultivar Mundo Novo, and their genetic diversity was evaluated by ERIC-PCR, REP-PCR, and their combination. All the strains were pathogenic to the coffee seedlings; the results of pathogenicity tests, in both experiments, could be divided in four aggressiviness classes (highly aggressive; aggressive; moderately aggressive and less aggressive). The Kenyan strains grouped separately from the Brazilian strains with ERIC-PCR and the combination of ERIC- and REP-PCR. The Brazilian strains could be grouped in two sub-clusters, the first including the older strains, obtained from 1958 to 1978, and the other comprising the remaining strains. With a few exceptions, strains isolated from 1997 to 2011, grouped mainly by their region of origin, were predominantly isolated from higher altitude regions, above 800 m. This probably occurred because the climatic conditions that prevail in these regions, characterized by milder temperatures and regular rainfall, are favorable for the coffee crop and for the production of high quality coffee beverage, but can be also favorable to bacterial halo blight.

ABSTRACT The bacterial-halo-blight (Pseudomonas syringae pv. garcae) is disseminated by the main coffee areas in the producing states of Brazil. On the other hand, the disease bacterial-leaf-spot (Pseudomonas syringae pv. tabaci) was reported only once in coffee seedlings in a sample collected in the State of São Paulo. In mid-2015, samples of coffee leaves with symptoms of foliar lesions surrounded by yellow halo, were collected in coffee plantations in the State of Paraná and fluorescent bacteria producing or not brown pigment in culture medium were isolated and determined as belonging to the Group I of P. syringae. Through biochemical, serological and pathogenicity tests, the pathogens were identified as P. syringae pv. garcae and P. syringae pv. tabaci, with prevalence of isolates belonging to pathovar tabaci and, as well as in certain samples, it was identified simultaneous infection by both etiological agents. Then, this is the first report of associated occurrence of garcae and tabaci pathovars of P. syringae and of the incidence of “bacterial-leaf-spot” under field conditions and in the State of Paraná.

RESUMO: Há várias bactérias que causam problemas para o cafeeiro, incluindo Pseudomonas cichorii, P. syringae pv. garcae, P. syringae pv. tabaci, Burkholderia andropogonis e Xylella fastidiosa, todas elas já descritas no Brasil. Tentativas de diferenciar essas bactérias por testes serológicos de dupla difusão em ágar (dda), com antissoros produzidos contra células íntegras de P. s. pv. garcae, mostraram reações cruzadas, principalmente entre P. s. pv. garcae e P. s. pv. tabaci. Dessa forma, foram produzidos antissoros contra P. s. pv. garcae (linhagem patotipo IBSBF-248 - Coleção de Culturas de Fitobactérias do Instituto Biológico - IBSBF), obtidos por meio de imunizações de coelhos com antígenos de proteínas do complexo proteico da membrana (CPM). Esses antissoros foram testados por dupla difusão em agarose (dda) contra diversas formas de antígenos extraídos de P. cichorii, P. s. pv. garcae e P. s. pv. tabaci [células autoclavadas, células tratadas com formol, exopolissacarídeos (EPS), glicoproteínas (GP) da cápsula bacteriana, proteínas de membrana e suspensão bacteriana (SB) em NaCl 0,85%]. Os resultados mostraram que, dependendo do antígeno e do meio suporte da dupla difusão (com ou sem MgCl2 e/ou azul de tripano), os antissoros reagem somente com P. s. pv. garcae. Desse modo, esses antígenos podem ser usados para a rápida diagnose da mancha aureolada do cafeeiro nos testes de dda.

ABSTRACT: Some bacterial diseases have been described causing problems in coffee, whose causal agents are Pseudomonas cichorii, P. syringae pv. garcae, P. s. pv. tabaci and Burkholderia andropogonis, all of them also occurring in Brazil. Attempts to differentiate these bacteria by double diffusion agar (dda) technique with antisera produced against whole cells of P. s. pv. garcae showed cross-reactions, especially among P. s. pv. garcae and P. s. pv. tabaci. Thus, antisera were produced against P. s. pv. garcae (pathotype strain IBSBF-248 - Phytobacteria Culture Collection of Instituto Biológico - IBSBF), using rabbit immunizations with antigens of the protein complex of membranes. These antisera were tested against different kind of antigens (autoclaved cells, cells treated with formaldehyde, capsule polysaccharides, glycoproteins, bacterial membrane proteins and bacterial suspension in 0.85% NaCl) extracted from P. cichorii, P. s. pv. garcae and P. s. pv. tabaci. The results showed that depending on the kind of antigen and the medium used in the double diffusion test (with or without MgCl2 and/or trypan blue), the antiserum reacted only with P. s. pv. garcae. Thus, these antigens may be used as an auxiliary tool in the rapid diagnosis of bacterial halo blight of coffee in the double diffusion test.

O crestamento bacteriano comum do feijoeiro é uma doença amplamente distribuída no mundo e os sintomas comumente observados sáo manchas nas folhas, hastes, vagens e sementes. Em dezembro de 2009, plantas de feijoeiro cv. Uirapuru com sintomas de murcha similares aos produzidos por Curtobacterium flaccumfaciens pv. flaccumfaciens foram observadas em campos comerciais localizados no município de Itararé, estado de São Paulo, Brasil. As plantas encontravam-se no final do ciclo, com as vagens já formadas. Os sintomas foram notados em quase a totalidade da área cultivada. Observações ao microscópio óptico de fragmentos de tecido do sistema vascular das hastes de plantas doentes evidenciaram intenso fluxo bacteriano, confirmando tratar-se de doença bacteriana. Isolamentos realizados em meio nutriente ágar produziram colônias de coloração amarelada, brilhantes, convexas, lisas. A bactéria agente causal era Gram-negativa e produtora de pigmento marrom escuro em meio de cultura. Testes bioquímicos, culturais e fisiológicos confirmaram sua identidade como Xanthomonas fuscans subsp. fuscans (sin. Xanthomonas campestris pv. phaseoli “var. fuscans"). A patogenicidade dos isolados foi confirmada por inoculações artificiais em mudas de feijão cv. Carioca e os reisolamentos efetuados resultaram em colônias semelhantes às originais. Embora descrita na literatura, a infecção sistêmica não é usualmente observada nos plantios de feijoeiro em nosso país. Linhagens bacterianas encontram-se depositadas na Coleção de Culturas do Instituto Biológico (IBSBF) sob os números 2813 e 3028.

Common and fuscous blights of bean are diseases widely distributed in the world. The most commonly observed symptoms are spots on leaves, stems, pods and seeds. In December 2009, bean plants cv. Uirapuru showing symptoms of wilt similar to those induced by Curtobacterium flaccumfaciens pv. flaccumfaciens were observed in a commercial crop located in the county of Itararé, State of São Paulo, Brazil. The plants were at the last cycle stage with mature pods and these symptoms were noted in the majority of the growing area. Optical microscopic observations of discolored vascular tissue from diseased stems revealed the presence of bacterial masses oozed from infected tissue, indicating that the disease was caused by bacterial pathogen. Isolations on nutrient agar showed circular, convex, yellow colonies with smooth edges. The causal bacterium was Gramnegative and produced a dark brown pigment in culture medium. Biochemical, cultural and physiological tests confirmed its identity as Xanthomonas fuscans subsp. fuscans(syn. Xanthomonas campestris pv. phaseoli“var. fuscans"). The pathogenicity of the isolates was confirmed by artificial inoculations. Systemic infection has been reported in the literature but these kinds of symptoms are not currently observed in Brazilian fields. Bacterial strains were deposited on the Phytobacteria Culture Collection of Instituto Biológico (IBSBF - www.biologico.sp.gov.br/bacterias/php) under accession numbers 2813 and 3028.

Em 2005, foi constatada em dois campos comerciais de tomate no Estado de São Paulo, a ocorrência da queima bacteriana, causada por Pseudomonas cichorii. Em vista disso, foram desenvolvidos estudos visando a determinação da gama de hospedeiros de isolados de Pseudomonas cichorii (IBSBF 2309 e IBSBF 2323), obtidos de tomateiro, provenientes de campos comerciais localizados nos municípios de Bragança Paulista e Mogi Guaçú, SP. Plantas de abobrinha, alface, beldroega, berinjela, beterraba, cenoura, couvebrócolo, datura, fumo, girassol, jiló, melão, pepino, petúnia, pimentão, rabanete, repolho, rúcula, salsa e tomateiro foram inoculadas por pulverização, separadamente, com os dois isolados de P. cichorii de tomateiro e um isolado de girassol (GIR-1). Os isolados IBSBF 2309 e IBSBF 2323 foram patogênicos à beldroega, datura, girassol, pimentão e tomate; GIR-1 foi patogênico apenas à beldroega, datura e girassol, não sendo patogênico ao pimentão e ao tomateiro. No Brasil não se conhecem fontes de resistência dentro do gênero Lycopersicon ou a reação de cultivares de tomateiros a esta bactéria. Vinte e oito genótipos de tomateiro provenientes do Banco de Germoplasma da empresa Sakata Seed Sudamerica Ltda., foram avaliados quanto a reação aos isolados IBSBF 2309 e IBSBF 2323 de P. cichorii, pelo método de inoculação nas folhas. Os maiores níveis de resistência foram observados em AF 11768, AF 2521, AF 11766, AF 11772, AF 229, AF 5719-1 e AF 8162. O genótipo AF 5719-1, que possui o gene Pto, que confere resistência a P. syringae pv. tomato, apresentou um bom nível de resistência a P. cichorii. A identificação de genótipos que apresentem bons níveis de resistência a este patógeno é importante para utilização em programas de melhoramento genético do tomateiro, visando a incorporação de genes de resistência a P. cichorii.

The occurrence of the bacterial blight, caused by Pseudomonas cichorii, was observed in two commercial tomato fields in the State of São Paulo in 2005. In view of this, studies were carried out in order to determine the host range of Pseudomonas cichorii isolates (IBSBF 2309 and IBSBF 2323), obtained from tomato plants at commercial fields located in the cities of Bragança Paulista and Mogi Guaçú, SP, Brazil. Caserta pumpkin, lettuce, purslane, eggplant, beet, broccoli, carrot, Jimson weed, sunflower, tobacco, scarlet eggplant, melon, cucumber, petunia, green pepper, radish, cabbage, arugula, parsley, and tomato plants were spray-inoculated separately with two isolates of P. cichorii obtained from tomato and one from sunflower (GIR-1). The isolates IBSBF 2309 and IBSBF 2323 were pathogenic to purslane, Jimson weed, sunflower, green pepper, and tomato; GIR-1 was only pathogenic to purslane, Jimson weed, and sunflower, but not pathogenic to green pepper or tomato. In Brazil, no sources of resistance to this bacterium are known within the Lycopersicon genus. The reaction of tomato cultivars to the bacterium is also unknown. Twenty-eight tomato genotypes from the Sakata Seed Sudamerica Ltda. Germplasm Bank were evaluated for their reaction to P. cichorii isolates IBSBF 2309 and IBSBF 2323, using the leaf inoculation method. The highest resistance levels were observed in tomato genotypes AF 11768, AF 2521, AF 11766, AF 11772, AF 229, AF 5719-1, and AF 8162. The genotype AF 5719-1, wich has the Pto gene imparting resistance to P. syringae pv. tomato, showed a good level of resistance to P. cichorii. The identification of genotypes with good levels of resistance to this pathogen is important, since they represent potential resources to be used in tomato breeding programs for incorporation of resistance genes against P. cichorii.