Avaliação de laranjeiras doces quanto à qualidade de frutos, períodos de maturação e resistência a Guignardia citricarpa

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mancha preta dos citros: expressão dos sintomas em frutos pela inoculação com conídios e controle do agente causal (Guignardia citricarpa)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção de estruturas reprodutivas e efeito do ambiente nos tipos de sintomas produzidos por Guignardia citricarpa em Citrus spp

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Período de incubação de Guignardia citricarpa em frutos de laranja ‘Valência’ e importância das pulverizações de cobre no controle da mancha preta

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Controle de Guignardia citricarpa e Penicillium digitatum em laranja com óleos essenciais e agentes de biocontrole

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Interferência da redução no volume de aplicação sobre o controle da mancha preta (Guignardia citricarpa Kiely) em frutos de laranja 'Valência'

The chemical control of the causal agent of citrus black spot (CBS) has been highlighted for the excessive required number of sprayings, considerably increasing citrus production costs. Improvements in the spray efficiency and reductions in the quantity of phytosanitary products have already been searched, but the results of that practice are not consistent yet for its use at commercial scale. Thus, the aim of the present study was to evaluate the interference of reduced spray volumes in black spot control in citrus fruits. The experiment was carried out in commercial citrus orchard with 16-year-old plants of 'Valencia' variety, during 2007 agricultural season. Treatments consisted of three spray volumes: 3.5; 4.5 and 8.5 liters.plant-1, applied with Arbus 2000/Export airblast sprayer with special manifold of hydraulic nozzles, using fungicides and periods recommended for the disease control, totaling four sprayings plus a control treatment (without spray). The disease incidence and severity were evaluated by visual diagrammatic scale of notes in two different periods (preharvest and harvest), at three plant heights (low, middle and top) and three horizontal sections (entrance, frontal and exit) in two sides of the plant. The fallen fruits were counted every fifteen days, for previously selected plants, from the beginning of maturation to harvest, and the production was quantified (kg.plant-1). The disease incidence and severity were significantly lower when sprayings were done with 8.5 liters.plant-1 in the first evaluation period (pre-harvest), but in the harvest period there were not differences between the same parameters when 4.5 or 8.5 liters.plant-1 were sprayed. None of those treatments reduced the disease on the top section of plants, compared to control. The plant sections with fruits more exposed to sun rays, top and right side of the plant, demonstrated higher disease incidence and severity. The volume reduction from 8.5 to 4.5 liters.plant-1 can be adopted for citrus orchards without damaging the CBS control level.

Estudo comparativo do genoma e do transcriptoma de Phyllosticta citricarpa (fase sexual Guignardia citricarpa) agente da Mancha Preta dos Citros e Phyllosticta capitalensis

Pós-graduação em Ciências Biológicas (Genética) - IBB

Período de incubação de Guignardia citricarpa em diferentes estádios fenológicos de frutos de laranjeira 'Valência'

A mancha preta dos citros (MPC), causada pelo fungo Guignardia citricarpa, produz lesões em frutos, os quais ficam depreciados para o mercado interno e os restringem para a exportação. O grande período de suscetibilidade dos frutos cítricos, em adição ao fato de G. citricarpa causar infecções latentes, dificulta o entendimento sobre o período de incubação da doença. O objetivo do trabalho foi determinar o período de incubação da MPC inoculando frutos de laranjeira 'Valência' em diferentes estádios fenológicos. Para a inoculação foram empregadas suspensões de conídios de G. citricarpa (10³, 10(4), 10(5) e 10(6) conídios mL-1) em diferentes diâmetros dos frutos (1,5; 2,0; 2,5; 3,0; 5,0 e 7,0 cm). O período de incubação da MPC para os diferentes diâmetros dos frutos inoculados apresentou uma relação polinomial negativa. Em frutos com até 3 cm de diâmetro o período de incubação médio foi superior a 200 dias, enquanto que em frutos com diâmetros superiores a 5 cm o período de incubação médio foi inferior a 84 dias. A MPC apresenta período de incubação variável dependente do estádio fenológico em que os frutos são infectados. A concentração de conídios de G. citricarpa, na infecção, não interfere no período de incubação da doença.

Secretome analysis of Trichoderma atroviride T17 biocontrol of Guignardia citricarpa

The fungal species Guignardia citricarpa is an important pathogen in citriculture. Members of the fungal genus Trichoderma are recognized as biocontrol agents but studies on the interactions between both fungi are scarce. This study aimed to identify extracellular proteins secreted by Trichoderma atroviride T17 that are related to the control of G. citricarpa. Two-dimensional gel electrophoresis (2D) was used to study the patterns of proteins secreted by T. atroviride T17 in medium containing glucose (control) and in medium containing G. citricarpa GC3 inactivated mycelium. We identified 59 of the 116 spots differentially expressed (50.86%) by LC–MS/MS. Of these, we highlight the presence of glycoside hydrolases (CAZy families 3, 43, 54, 76 and 93), chitinase, mutanase, a-1,3-glucanase, a-1,2-mannosidase, carboxylic hydrolase ester, carbohydrate-binding module family 13, glucan 1,3-b-glucosidase, a-galactosidase and Neutral protease 2. These proteins are related to mycoparasitism processes, stimuli and therefore to the biological control of pathogens. The results obtained are in agreement with reports describing an increase in the secretion of proteins related to mycoparasitism and biological control and a reduction in the secretion of proteins related to the metabolism of Trichoderma species grown in the presence of the pathogen. Moreover, these results are pioneer in understanding T. atroviride interaction with G. citricarpa. For the first time, we identified potential candidate proteins that may have a role in the antagonism mechanism of G. citricarpa by T. atroviride T17. Thus our results shed a light into the molecular mechanisms that T. atroviride use to control G. citricarpa.

Control of citrus black spot (Guignardia citricarpa) by biological control agents and other alternative products.

2007

Análise filogenética entre Citrus spp. e Guignardia spp

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Postharvest harpin or Bacillus thuringiensis treatments suppress citrus black spot in `Valencia` oranges

Citrus black spot (CBS) caused by Guignardia citricarpa represents an important threat to citriculture in Brazil. Limited information is available regarding potential biological control agents and new alternative compounds that may provide protection of orange fruits against the disease. In this study, the effects of commercial products based on Bacillus thuringiensis var. kurstaki (Bt) bacterium, Bt pure isolates and Harpin protein (Messenger (R)) on the postharvest control of CBS, were evaluated in `Valencia` sweet orange fruits harvested for three consecutive years in a citrus grove. The fruits were sprayed with the following products: DiPel (R) WP (Bt, subspecies, kurstaki strain HD-1,16,000 International Units mg(-1), 32 g active ingredient kg(-1)) (1, 20 and 50 mg ml(-1)), Dimy Pel (R) WP (Bt, subspecies, kurstaki, strain HD-1, 17,600 IU mg(-1), 26 g active ingredient l(-1)) (2, 20 and 50 mg ml(-1)), Messenger (R) (3% harpin protein) (1 and 2 mg ml(-1)) and fungicide Tecto (R) Flowable SC (thiabendazole, 485 gl(-1)) (0.8g active ingredient l(-1)), besides the Bt isolates, Bt- HD-567, Bt- DiPel and Bt- Dimy (9 x 10(8) CFU ml(-1)). Ten days after treatment, the number of newly developed CBS lesions and pycnidia produced were evaluated using fifty fruits per treatment. The Dimy Pel (R) and Messenger (R) reduced the number of new developed CBS lesions on fruits in up to 67% and 62%, respectively. All applied treatments drastically decreased the number of pycnidia produced in the CBS lesions on orange fruits with 85% to 96% reductions compared to the untreated control. Volatile compounds produced by the isolates Bt- HD-567, Bt- Dimy and Bt- DiPel, reduced the number of lesions on treated fruits by 70%, 65% and 71% compared to the control, respectively. In addition, the survival of Bt isolates on orange fruit surfaces were evaluated by recovering and quantifying the number of CFU every seven days for up to 28 days. The declines in survival rates on orange fruit surfaces were drastic for the three strains of Bt in the first week. The CFU numbers of all applied isolates declined by 4 to 5 orders of magnitude after storage at room temperature for 28 days. In vitro assays revealed that the Bt isolates significantly reduced the mycelial growth of the pathogen, ranging from 32% to 51%, compared to the control, whereas no inhibitory effect was observed in the presence of Messenger (R). (C) 2010 Elsevier Ltd. All rights reserved.

Incidence of black spot in citrus fruits from different processing stages in packinghouses and from Sao Paulo`s wholesale market

The purposes of this work were a) to evaluate citrus black spot (CBS) incidence in `Valencia` oranges and `Murcott` tangors aimed at the export market, and in Pera`, `Lima` and `Natal` oranges, and `Murcott` tangors, aimed at the domestic market after different processing stages in packinghouses in 2004/05 and 2005/06; b) to evaluate CBS incidence in Pera` and `Lima` oranges and `Murcott` tangors sold at Ceagesp-SP, the biggest wholesale market in the State of Sao Paulo, in 2006. Citrus fruits were collected at the packinghouse, on their arrival, after pre-washing and de-greening, from the packing table, from the pallet and at Ceagesp. They were stored for 14 to 21 days at 25 degrees C and 85-90% RH. The incidence of CBS was visually evaluated after one day and at the end of the storage period. CBS incidence in fruits aimed at the export market decreased, with values under 2.0% on arrival and no CBS symptoms observed on fruits from the pallet. The average incidence of CBS in `Pera`, `Lima` and `Natal` oranges, and `Murcott` tangors in the packinghouse aimed at the domestic market were 64.1, 39.0, 32.1 and 19.3%, respectively, after one day of storage, then remaining constant in all processing stages. The incidence of CBS in Ceagesp fruits was low in winter months and increased in the spring. The increase in disease incidence during the storage period (21 days) was not significant in collected fruits.

Spatial pattern of black spot incidence within citrus trees related to disease severity and pathogen dispersal

Guignardia citricarpa, the causal agent of citrus black spot, forms airborne ascospores on decomposing citrus leaves and water-spread conidia on fruits, leaves and twigs. The spatial pattern of diseased fruit in citrus tree canopies was used to assess the importance of ascospores and conidia in citrus black spot epidemics in Sao Paulo State, Brazil. The aggregation of diseased fruit in the citrus tree canopy was quantified by the binomial dispersion index (D) and the binary form of Taylor`s Power Law for 303 trees in six groves. D was significantly greater than 1 in 251 trees. The intercept of the regression line of Taylor`s Power Law was significantly greater than 0 and the slope was not different from 1, implying that diseased fruit was aggregated in the canopy independent of disease incidence. Disease incidence (p) and severity (S) were assessed in 2875 citrus trees. The incidence-severity relationship was described (R-2 = 88.7%) by the model ln(S) = ln(a) + bCLL(p) where CLL = complementary log-log transformation. The high severity at low incidence observed in many cases is also indicative of low distance spread of G. citricarpa spores. For the same level of disease incidence, some trees had most of the diseased fruit with many lesions and high disease severity, whereas other trees had most of the fruit with few lesions and low disease severity. Aggregation of diseased fruit in the trees suggests that splash-dispersed conidia have an important role in increasing the disease in citrus trees in Brazil.

Influência do alinhamento de plantio na severidade da mancha-preta-dos-citros, produção e qualidade do suco

Os níveis de incidência e severidade de sintomas das doenças resultam da interação hospedeiro-patógeno-ambiente. Nesse contexto, a favorabilidade do ambiente, em particular, depende do nível de umidade, intensidade de radiação solar, luminosidade e molhamento foliar, dentre outros fatores. Por outro lado, a magnitude de expressão desses fatores é dependente do ciclo e porte das plantas, e possivelmente, também, do alinhamento de plantio. No presente estudo, foi avaliada a influência dos alinhamentos de plantio N-S, L-O, NE-SO E NO-SE no desempenho agronômico de plantas de laranjas 'Natal'e 'Valência', assim como na severidade de sintomas da mancha-preta-dos-citros (MPC), causada por Guignardia citricarpa. Para tal, foram selecionadas cinco propriedades distribuídas ao longo do cinturão citrícola paulista, de talhões semelhantes, a partir das quais foram coletadas amostras de 50 frutos, em dez plantas de mesma linha, da região central dos mesmos. Tais frutos foram levados para laboratórios e analisados quanto aos níveis de severidade de sintomas da MPC, produção e qualidade do suco. Constatouse que os frutos oriundos de pomares implantados no alinhamento N-S apresentavam menores níveis de severidade da doença, enquanto no alinhamento NE-SO, os mais elevados. Propriedades localizadas mais ao sul do Estado de São Paulo apresentaram menores níveis de doença, provavelmente em consequência da ocorrência recente da doença em tais áreas. Em relação à produção, os alinhamentos de plantio N-S e L-O proporcionaram maior número de frutos por planta, assim como quanto ao número de caixas por planta. Por outro lado, a menor produção foi verificada nos alinhamentos NO-SE. Em relação à qualidade do suco, não foi possível estabelecer um padrão definido em termos de ratio e ºBrix e os respectivos alinhamentos de plantio. Foi verificado que os valores de ratio foram menores nos frutos de pomares localizados mais ao sul do Estado, de maiores latitudes.