Celso Dornelas Fernandes

Genetic Diversity in Colletotrichum gloeosporioides from Stylosanthes spp. at Centers of Origin and Utilization.

ABSTRACT Using molecular markers, this work compares the genetic diversity in Colletotrichum gloeosporioides infecting species of the tropical forage legume Stylosanthes at the center of origin in Brazil and Colombia with that of Australia, China, and India, where Stylosanthes spp. have been introduced for commercial use. There was extensive diversity in the pathogen population from Brazil, Colombia, China, and India. The Australian pathogen population was least diverse probably due to its geographical isolation and effective quarantine. The extensive diversity in China and India means that threats from exotic pathogen races to Stylosanthes pastures can potentially come from countries outside the South American center of origin. In Brazil and India, both with native Stylosanthes populations, a high level of genetic differentiation in the pathogen population was associated with sites where native or naturalized host population was widely distributed. There was limited genetic diversity at germplasm evaluation sites, with a large proportion of isolates having identical haplotypes. This contrasts recent pathogenicity results for 78 of the Brazilian isolates that show hot spots of complex races are more common around research stations where host germplasm are tested, but few are found at sites containing wild host populations. For a pathogen in which the same races arise convergently from different genetic backgrounds, this study highlights the importance of using both virulence and selectively neutral markers to understand pathogen population structure.

Genetic diversity in South American Colletotrichum gloeosporioides isolates from Stylosanthes guianensis, a tropical forage legume

The degree of genetic diversity of 127 Colletotrichum gloeosporioides isolates from Stylosanthes guianensis genotypes in South America was measured at the molecular level by random amplified polymorphic DNA (RAPD) with nine arbitrary primers of 10 bases, and by restriction fragment length polymorphism (RFLP) with a non-LTR (long terminal repeats) retrotransposon DNA sequence. The RAPD products revealed scorable polymorphism among the isolates, and a total of 80 band positions were scored. Sixty-three of the 127 isolates were clustered into 13 distinct lineages usually correlating with geographic origin. Where isolates from various regions were clustered together, most had identical host genotype origin. The pathogen population sampled from Carimagua, Colombia, a long-time Stylosanthes breeding and selection site, with a savanna ecosystem, was highly diverse. A set of 12 S. guianensis genotype differentials was used to characterize pathogenic variability of 104 isolates and their virulence patterns were grouped into 57 pathotypes. However, when they were tested on four Australian differentials, they grouped into 11 pathotypes. As shown in previous studies, no strict correlations existed between genetic diversity measured by RAPD or RFLP, and pathotype defined by pathogenicity pattern on the differentials. Southern blot analysis of the 127 isolates revealed 23 hybridizing fragments, resulting in 41 fingerprint patterns among the 127 isolates. Relationships between RFLP and RAPD variables were examined using Spearmans Rank Correlation Coefficient, which showed that the two measures of genotypic variation are in agreement.

Formação de mudas de mamão em ambientes de cultivo protegido em diferentes substratos

The cultivation of fruits in the State of Mato Grosso do Sul, Brazil, lacks information on the potential of seedlingproduction. The aim of this study was to evaluate the production of papaya cultivar Sunrise Solo, in industrializedsubstrates and soil. The experiment was carried out in the State University of Mato Grosso do Sul (UEMS), Aquidauana,from 18/11/2008 to 05/01/2009. Three protected cultivation environments, three sizes of polyethylene bags and sixsubstrates were used in the treatments. The experimental design was a complete randomized in a split-plot scheme. Theresults showed that the 15.0 x 21.5 cm bag (1539.8 cm

Nematofauna fitopatogênica de sementes comerciais de forrageiras tropicais

The objective of this work was to identify and quantify the phytopathogenic nematofauna of forage seeds destined for internal and external markets. Seeds of Brachiaria brizantha (cvs. Marandu and Xaraes), B. decumbens cv. Basilisk, Panicum maximum (cvs. Massai, Mombaca and Tanzania), and Stylosanthes capitata and S. macrocephala, components of cv. Estilosantes Campo Grande, were examined. The extractions were made by centrifugal flotation in solution of sucrose with kaolin. Aphelenchoides and Ditylenchus genera were detected in forage grasses. The highest number of Aphelenchoides spp. specimens was observed in seeds of cv. Marandu, followed by seeds of cvs. Xaraes, Basilisk, and Tanzania. Regarding Ditylenchus spp. population density, higher values were detected in seeds of cvs. Marandu, Xaraes, and Basilisk. The results suggest that two or more species of Aphelenchoides and Ditylenchus are associated to forage grass seeds destined for Brazilian market. Phytopathogenic nematodes are not detected in Estilosantes Campo Grande seeds, neither in seeds of cvs. Marandu, Mombaca and Tanzania destined for exportation.

Reação de acessos e cultivares de Brachiaria spp. e Panicum maximum à Pratylenchus brachyurus

The use of forage grass in integrated crop-livestock systems has been increasingly frequent in Brazil. However, some forage genotypes may host plant parasitic nematodes, contributing to the maintenance of large populations of these organisms in the soil. This study was conducted with the aim of evaluating the reaction of accessions and cultivars of Brachiaria spp. (B. ruziziensis, B. brizantha cvs. BRS Piata and BRS Paiaguas, B. humidicola cv. BRS Tupi, and the genotypes B4 and HBGC 331) and Panicum maximum (cvs. Tanzânia-1 and Massai, and the genotypes PM32, PM36, PM45 and PM46) to Pratylenchus brachyurus. Corn BRS-2020 (susceptible) and millet ADR-300 (resistant) were used as controls. The experiments were conducted in a greenhouse at Embrapa Beef Cattle, Campo Grande-MS, in randomized blocks with seven replicates, from May to September 2011 and from March to May 2012. Five plants per pot were inoculated with 1,000 specimens of P. brachyurus. After 90 days, nematode populations in the roots and the soil were evaluated to obtain the final nematode population and the reproduction factor (RF). The reaction of genotypes in relation to the percentage of reduction in the RF was also assessed. Except for B. humidicola cv. BRS Tupi, which showed RF of 0.98 and 0.44 in the first and in the second experiments, respectively, all evaluated materials allowed the nematode multiplication. Regarding the percentage of reduction in the RF, only B. humidicola cv. BRS Tupi and millet ADR-300 were classified as moderately resistant, showing reductions of up to 90.58% and 94.73% in the first and in the second experiment, respectively. Most of the studied forage genotypes were susceptible to P. brachyurus, Even though the susceptibility degree varied among them. Thus, in areas with history of the presence of P. brachyurus, B. humidicola cv. BRS Tupi can be indicated for integrated and/or crop rotation system as a management strategy to reduce P. brachyurus population.

Fungos e nematoides associados a sementes de forrageiras tropicais

This study aimed to assess the incidence of fungi and nematodes in Brachiaria sp. and Panicum maximum seeds produced in the Brazilian states of Mato Grosso do Sul (MS), Mato Grosso (MT), Goias (GO), Minas Gerais (MG) and Sao Paulo (SP). The main fungi found in the seeds were Bipolaris sp., Curvularia sp. and Phoma sp.. The lowest incidence of these fungi was found for seeds of Brachiaria brizantha cultivars BRS Piata and Xaraes, and Brachiaria decumbens cv. Basilisk, from the states of GO, MG and MS, respectively. The cultivars Marandu and BRS Piata, from several regions, exhibited high occurrence of Aphelenchoides sp. and Ditylenchus sp.. Seeds of B. humidicola cultivar Humidicola, produced in MS and SP, did not show association with nematodes. The seeds of Panicum maximum cv. Massai and cv. Mombaca showed higher incidence of Bipolaris sp., Cladosporium sp., Curvularia sp., Fusarium sp. and Phoma sp., as well as Aphelenchoides sp. and Ditylenchus sp., especially for seeds produced in MT. Some of the detected pathogens are causative agents of diseases of major importance in forage plants, such as Bipolaris sp., causing leaf spot in Panicum, of high severity in Tanzânia, which provides serious compromising of the pasture sustainability.

Densidade populacional de Pratylenchus spp. em pastagens de Brachiaria spp. e sua influência na disponibilidade e na qualidade da forragem

Population density of Pratylenchus spp. in pastures of Brachiaria spp. and its influence on the forage availability and quality The aim of this work was to evaluate the population density of Pratylenchus brachyurus and Pratylenchus zeae associated with Brachiaria brizantha, B. decumbens and B. humidicola and their influence on forage availability and quality. The experiment was conducted in the Hisaeda Farm, Terenos, MS, Brazil. Soil, roots and plant aerial part were harvest, with ten replications each, in one square meter randomized sets encompassing three treatments: Good, Intermediary and Bad, visually characterized, considering the percentage of green material. P. brachyurus and P. zeae density were evaluated in soil and plant roots. Dry matter of green, dead and re-growth materials, plant nutritional status and forage quality were assessed in the aerial plant part. Soil fertility was determined in all harvested samples. Both nematode species were identified from all samples, with a lar ger number in the roots (between 87-311 P. brachyurus and 1-61 P. zeae.10 g -1 ) than in the soil (0-8 P. brachyurus and 1-39 P. zeae.200 cm - ≥), however, no significant differences were found in the number of specimens between treatments. Considering that these forage species are perennial and host Pratylenchus spp, there is a tendency to increase the population of these pathogens over time, becoming a serious phytosanitary problem.

Progresso e controle da mela-das-sementes (Claviceps maximensis) de Brachiaria brizantha

The efficiency of one or two applications of Pyraclostrobin + Epoxyconazole, Mancozeb, Triadimenol, Azoxystrobin + Cyproconazole, Trifloxystrobin + Cyproconazole or Tebuconazole in the control of honeydew in seeds of Brachiaria brizantha cvs. Marandu and Xaraes was verified during crop 2004-05. The inductors of resistance Acibenzolar-S-Methyl and Potassium Silicate (applied in aerial part or in soil) were evaluated. Triadimenol, with one or two applications, Pyraclostrobin + Epoxyconazole, Azoxystrobin + Cyproconazole, Trifloxystrobin + Cyproconazole or Tebuconazole, with two applications, were the fungicides most promising in the control of disease in marandu grass. About cv. Xaraes, better control was reached with Pyraclostrobin + Epoxyconazole, independent of number of applications, Triadimenol, Trifloxystrobin + Cyproconazole or Tebuconazole, with two applications. Correlation between production of pure seeds and of disease intensity was not detected. With regard to the progress of honeydew in seeds of B. brizantha cvs. Marandu and Xaraes, observed the disease occurred in cold periods with high humidity. In cv. Marandu the pathogen occurred in final phase of culture, while in cv. Xaraes the honeydew was detected in phase of intense bloom. Constant increases of disease intensity were observed in two genotypes. The honeydew occurred in 64% of panicles of cv. Marandu and in 81% of panicles of cv. Xaraes; about 20% and 18% of flowers/seeds, respectively, was affected. The results verified high positive correlation between the values of incidence and severity of disease.

Atividade pectinolítica de Colletotrichum gloeosporioides e a relação com a agressividade ao Stylosanthes spp

Pectinolytic activity of 40 isolates of Colletotrichum gloeosporioides obtained from Stylosanthes spp.lesions, in distinct producing regions, was estimated through the enzymatic diffusion in pectin-medium free of glucose, and measured by pectin degradation. All isolates showed pectinolytic activity, and were divided in five groups. Moderate production of pectic enzymes was evidenced in the majority of the isolates. In general, isolates from Camapua-MS presented low genetic diversity for the production of pectinases. Taking into account the activity of pectinases, isolates of the pathogen were selected and inoculated in S. capitata (GC1081, GC1084, GC1086 and GC1094), S. scabra (GC1496) and S. macrocephala (GC1511). Thus, the enzyme influence on aggressiveness to the host was evaluated. Initially, aggressiveness test was installed with the isolates CG656, CG657, CG687, CG706 and CG707. Differences in aggressiveness were observed only when the isolates were inoculated in S. scabra. The isolates CG657, CG706 and CG707 were the most aggressive, and CG656 was the least aggressive. Another aggressiveness test was established to evaluate the isolates CG768, CG769, CG772, CG775, CG779 and CG781. These isolates showed differences in their aggressiveness when inoculated in S. capitata (GC1081 and GC1094) and S. macrocephala (GC1511). In general, CG768 exhibited the lowest while, CG772, CG775 and CG781 the highest aggressiveness. In both aggressiveness tests, there were no evidences for a direct correlation of anthracnose increase in severity with the of the pectinolytic activity. For the conditions evaluated, the pectic enzymes did not influence the aggressiveness of C. gloeosporioides to the stylo.

Quimio e termoterapia em sementes e aplicação de fungicidas em Brachiaria brizantha como estratégias no manejo do carvão

This work investigated the potentials of the thermal and chemical treatments in seeds of Brachiaria brizantha cv. Piata, which have been described as efficient strategies to overcome the physical dormancy of seeds, in the reduction of smut (Ustilago operta). Samples of seeds were immersed in sulfuric acid for 0 to 10 min, or in hot water (27 to 60 oC) for 5 or 10 min. The effects of the treatments in the number of remaining teliospores of U. operta and in the germination of treated seed have been evaluated. A decrease of inoculum was observed as the time of chemical scarification of seeds was increased. The reduction on the number of teliospores was superior with 10 min of chemotherapy. In general, the exposition to H2SO4 did not result in a decrease of the germinative power of the seeds. The results of the thermotherapy were not as evident as those observed for the chemotherapy. Reductions on the number of teliospores or on the overcoming of the physical dormancy were less apparent, maybe, because the seeds were not immersed in hot water during enough time. In parallel, the efficiency of the application of fungicides on the aerial part of the plants for the control of the smut incidence was verified. The fungicides tested were as follow: 1 - methylic tiofanato + chlorothalonil; 2 - mancozeb; 3 - tebuconazole; 4 - triadimenol; 5 - azoxystrobin + cyproconazole; 6 - pyraclostrobin + epoxyconazole; 7 - trifloxystrobin + cyproconazole; 8 - carboxin + thiram; 9 - methylic tiofanato + flutriafol; 10 - carbendazin, and 11 - difenoconazole. Although the smut incidence observed in the experimental area was low (maximum of 15%), a differentiation among the fungicides was possible. The tebuconazole, pyraclostrobin + epoxyconazole and trifloxystrobin + cyproconazole were the most promising fungicides.