Navegando por Autor "Alfenas, Acelino C."
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Item Ceratocystis fimbriata isolates on Mangifera indica have different levels of aggressiveness(European Journal of Plant Pathology, 2016-02-06) Oliveira, Leonardo S. S.; Damacena, Michelle B.; Guimarães, Lúcio M. S.; Siqueira, Dalmo L.; Alfenas, Acelino C.Mango is amongst the most exported fruits worldwide and the mango tree has been cultivated in many countries with tropical and subtropical climates. Currently, Ceratocystis wilt caused by Ceratocystis fimbriata is among the most important diseases on mango tree. Planting of resistant genotypes is the best strategy for disease control. In the selection of resistant plants a highly aggressive isolate should be used in inoculations under controlled conditions. Thus, the objective of this study was to evaluate whether there are differences in aggressiveness among isolates of C. fimbriata obtained from diseased mango trees in Brazil. Before inoculation, five randomly selected isolates were tested for sporulation and mycelial growth in different culture media in order to select the best conditions for inoculum production. Twenty-four isolates from the States of São Paulo, Rio de Janeiro, Minas Gerais and Mato Grosso do Sul were inoculated in the susceptible mango cultivar Espada. Subsequently, the four most aggressive isolates were inoculated in other five different mango cultivars. The culture media MEA provided the best levels of sporulation and mycelial growth for four of the five tested isolates. Isolates SEMG1-1 from Minas Gerais and SESP8-3 from São Paulo were the most aggressive. The response of cultivars to the most aggressive isolates tested in the present work varied from highly resistant to highly susceptible, ‘Ubá’ being the most resistant regardless of the isolate tested. These results demonstrate that there are significant differences in aggressiveness among C. fimbriata isolates from mango in Brazil, as well as cultivar × isolate interaction.Item Effects of sublater rates of dazomet and metam- sodium applied under low- permeability films on Calonectria microsclerotia survival(Plant Disease, 2018-04) Aiello, Dalia; Vitale, Alessandro; Alfenas, Rafael F.; Alfenas, Acelino C.; Cirvilleri, Gabriella; Polizzi, GiancarloInfested soil is the primary inoculum source for Calonectria spp. for initiating disease in ornamental and forestry crops. The effects of dazomet and metam-sodium on survival of microsclerotia of 28 isolates belonging to 19 Calonectria spp. were evaluated in this study under nursery conditions. Two experiments with exotic Calonectria spp. in plastic containers in a greenhouse and three trials with endemic species in field plots were performed during different seasons. The containers and plots were artificially infested with Calonectria microsclerotia differentiated on carnation leaf tissues. Basamid (dazomet) was applied at 100, 160, 200, 400, and 500 kg/ha, while Divapan (metam-sodium) was applied at 250, 350, 400, 700, and 1,000 liters/ha in both the containers and plots. The fumigants were applied under virtually and totally impermeable films. Fungal survival was evaluated after 21 days using leaf tissues collected from treated soil and plated on potato dextrose agar, and the ability of microsclerotia to cause infection was tested on red clover. The survival of Calonectria inocula and microsclerotia decreased with increasing fumigant rates. In the greenhouse trials, where Basamid was applied at 200, 400, and 500 kg/ha and Divapan at 400, 700, and 1,000 liters/ha, no viable microsclerotia were recovered for 14 exotic Calonectria spp., whereas viable inocula of Calonectria hongkongensis, C. naviculata, and C. sulawesiensis were retrieved from the fumigated plots. Low rates of Basamid (100 and 160 kg/ha) and Divapan (250 and 350 liters/ha) were less effective at reducing Calonectria viability and, for these treatments, the rate of microsclerotia survival was highly variable among the different isolates and species. Furthermore, totally impermeable film significantly enhanced fumigant performance. Relative to endemic Calonectria spp., all of the treatments killed microsclerotia of C. polizzii and C. pauciramosa independent from fumigant, rate, and film. This research demonstrated the possibility of reducing the application rates by up to 160 kg/ha for Basamid and 400 liters/ha for Divapan under low-permeability films (virtually impermeable film or totally impermeable film) for eradicating or reducing the primary inoculum of Calonectria spp. in soil.Infested soil is the primary inoculum source for Calonectria spp. for initiating disease in ornamental and forestry crops. The effects of dazomet and metam-sodium on survival of microsclerotia of 28 isolates belonging to 19 Calonectria spp. were evaluated in this study under nursery conditions. Two experiments with exotic Calonectria spp. in plastic containers in a greenhouse and three trials with endemic species in field plots were performed during different seasons. The containers and plots were artificially infested with Calonectria microsclerotia differentiated on carnation leaf tissues. Basamid (dazomet) was applied at 100, 160, 200, 400, and 500 kg/ha, while Divapan (metam-sodium) was applied at 250, 350, 400, 700, and 1,000 liters/ha in both the containers and plots. The fumigants were applied under virtually and totally impermeable films. Fungal survival was evaluated after 21 days using leaf tissues collected from treated soil and plated on potato dextrose agar, and the ability of microsclerotia to cause infection was tested on red clover. The survival of Calonectria inocula and microsclerotia decreased with increasing fumigant rates. In the greenhouse trials, where Basamid was applied at 200, 400, and 500 kg/ha and Divapan at 400, 700, and 1,000 liters/ha, no viable microsclerotia were recovered for 14 exotic Calonectria spp., whereas viable inocula of Calonectria hongkongensis, C. naviculata, and C. sulawesiensis were retrieved from the fumigated plots. Low rates of Basamid (100 and 160 kg/ha) and Divapan (250 and 350 liters/ha) were less effective at reducing Calonectria viability and, for these treatments, the rate of microsclerotia survival was highly variable among the different isolates and species. Furthermore, totally impermeable film significantly enhanced fumigant performance. Relative to endemic Calonectria spp., all of the treatments killed microsclerotia of C. polizzii and C. pauciramosa independent from fumigant, rate, and film. This research demonstrated the possibility of reducing the application rates by up to 160 kg/ha for Basamid and 400 liters/ha for Divapan under low-permeability films (virtually impermeable film or totally impermeable film) for eradicating or reducing the primary inoculum of Calonectria spp. in soil.Item Etiologia do oídio do eucalipto(Fitopatologia Brasileira, 2001-06) Silva, Maria D. D.; Alfenas, Acelino C.; Maffia, Luiz A.; Zauza, Edival A. V.Compararam-se isolados de Oidium sp. obtidos de mudas de eucalipto (Eucalyptus spp.) com isolados de Erysiphe cichoracearum, originário de dália, e Sphaerotheca pannosa de roseira (Rosa sp.), por meio de características morfológicas. Com base nas características morfológicas (fase anamórfica) do tubo germinativo, do micélio, do apressório, de conidióforos e conídios e na presença de corpos de fibrosina, concluiu-se que o isolado de Oidium, obtido de eucalipto é similar ao de roseira, descrito como S. pannosa. Inoculações dos isolados do fungo originários de todos os hospedeiros estudados, indicaram que S. pannosa, obtido de roseira, e E. cichoracearum, de dália (Dahlia sp.), foram patogênicos a mudas de Eucalyptus pellita, uma das espécies mais suscetíveis a esta doença, em condições de casa de vegetação.Item Etiology of bacterial leaf blight of eucalyptus in Brazil(Tropical Plant Pathology, 2008-05) Oliveira, José R.; Maffia, Luiz A.; Alfenas, Acelino C.; Gonçalves, Rivadalve C.; Lau, Douglas; Cascardo, Júlio C. M.Bacterial leaf blight of eucalyptus is initially characterized by water soaked, angular, amphigenous and interveinal lesions, concentrated along the main vein, at the edges or scattered on the leaf blade. As the disease progresses, the lesions become brown to pale, and when young leaves are infected leaf cut areas at the edges or perforations at the center of the lesions may appear due to abortion of the necrotic area. Eventually, necrosis may be found on petiole and twigs. Leaf fall commonly occurs on highly susceptible genotypes due to the early senescence of diseased leaves. Precise diagnosis is accomplished by bacterial exudation from leaf sections placed in a water drop under light microscope (200 x). Twenty-five bacterial isolates from Amapá (2), Bahia (4), Minas Gerais (2), São Paulo (9), Pará (3), Mato Grosso do Sul (1), and Rio Grande do Sul (4) States, which induced hypersensitive reaction (HR) in non-host plants and were pathogenic to eucalyptus, when inoculated by inoculum injection, were identified by biochemical assays, using carbon sources (MicroLogTM BIOLOG) and sequence analysis (16S rDNA). Ten isolates were identified as Xanthomonas axonopodis, four as X. campestris, four as Pseudomonas syringae, two as P. putida, two as P. cichorii, one as Erwinia sp., and two were similar to bacterial genera of Rhizobiaceae. When spray inoculated on intact plants of eucalyptus, only X. axonopodis, P. cichorii and isolates of the Rhizobiaceae family induced typical symptoms of the disease and were considered pathogenic. In Brazil, X. axonopodis seems to be the most widespread species causing the bacterial leaf blight of Eucalyptus spp.Item Etiology of bacterial leaf blight of eucalyptus in Brazil(Tropical Plant Pathology, 2008-05) Oliveira, José R.; Maffia, Luiz A.; Cascardo, Júlio C. M.; Alfenas, Acelino C.; Gonçalves, Rivadalve C.; Lau, DouglasBacterial leaf blight of eucalyptus is initially characterized by water soaked, angular, amphigenous and interveinal lesions, concentrated along the main vein, at the edges or scattered on the leaf blade. As the disease progresses, the lesions become brown to pale, and when young leaves are infected leaf cut areas at the edges or perforations at the center of the lesions may appear due to abortion of the necrotic area. Eventually, necrosis may be found on petiole and twigs. Leaf fall commonly occurs on highly susceptible genotypes due to the early senescence of diseased leaves. Precise diagnosis is accomplished by bacterial exudation from leaf sections placed in a water drop under light microscope (200 x). Twenty-five bacterial isolates from Amapá (2), Bahia (4), Minas Gerais (2), São Paulo (9), Pará (3), Mato Grosso do Sul (1), and Rio Grande do Sul (4) States, which induced hypersensitive reaction (HR) in non-host plants and were pathogenic to eucalyptus, when inoculated by inoculum injection, were identified by biochemical assays, using carbon sources (MicroLogTM BIOLOG) and sequence analysis (16S rDNA). Ten isolates were identified as Xanthomonas axonopodis, four as X. campestris, four as Pseudomonas syringae, two as P. putida, two as P. cichorii, one as Erwinia sp., and two were similar to bacterial genera of Rhizobiaceae. When spray inoculated on intact plants of eucalyptus, only X. axonopodis, P. cichorii and isolates of the Rhizobiaceae family induced typical symptoms of the disease and were considered pathogenic. In Brazil, X. axonopodis seems to be the most widespread species causing the bacterial leaf blight of Eucalyptus spp.Item Evidências de indução de resistência sistêmica à ferrugem do eucalipto mediada por rizobactérias promotoras do crescimento de plantas(Fitopatologia Brasileira, 2005-07) Teixeira, Débora A.; Alfenas, Acelino C.; Mafia, Reginaldo G.; Maffia, Luiz A.; Ferreira, Eraclides M.A indução de resistência sistêmica mediada por rizobactérias promotoras do crescimento de plantas foi avaliada para a ferrugem do eucalipto (Eucalyptus spp.) causada por Puccinia psidii. Para isso, mudas com cerca de 80 dias de idade, previamente enraizadas em substrato tratado com diferentes isolados de rizobactérias foram inoculadas com uma suspensão de inóculo de P. psidii ajustada para 2 x 104 urediniósporos/ml. As plantas inoculadas foram mantidas em câmara de nevoeiro com nebulização intermitente a 25 ºC, no escuro por 24 h, e posteriormente transferidas para câmara de crescimento a 22 ºC, com fotoperíodo de 12 h e intensidade luminosa de 40 mmoles.s-1.m-2. Utilizou-se um delineamento inteiramente casualizado com quatro repetições, cada uma delas com quatro plantas. Após 13 dias da inoculação, avaliaram-se número médio de pústulas/folha, número de uredínias/amostra e número médio de esporos produzidos/uredínia. Os isolados FL2 e MF4 foram eficientes na redução da severidade da ferrugem. Além disso, o tratamento das mudas com rizobactérias, apenas uma semana antes da inoculação, com P. psidii foi menos eficiente em reduzir a severidade da doença do que o tratamento em que foram utilizadas mudas produzidas em substrato previamente rizobacterizadas, ou seja, com 80 dias. Estes resultados indicam que estes isolados de rizobactérias podem induzir maior resistência a doenças foliares diminuindo a necessidade de aplicação de fungicidas e otimizando a produção de mudas clonais de eucalipto.Item Genetic variation, morphology and pathogenicity of Ceratocystis fimbriata on Hevea brasiliensis in Brazil(Tropical Plant Pathology, 2015-06) Valdetaro, Denise C. O. F.; Oliveira, Leonardo S. S.; Guimarães, Lúcio M. S.; Freitas, Rodrigo G.; Alfenas, Acelino C.; Ferreira, Maria A.; Harrington, Thomas C.Ceratocystis fimbriata causes diseases on a wide variety of plants in Brazil, including rubber tree (Hevea brasiliensis), on which it causes gray mold or moldy rot on tapping panels affecting latex yield. However, C. fimbriata isolated from rubber tree have not been critically studied. Phylogenetic analyses of sequences of ITS rDNA and a mating type gene placed rubber tree isolates from Acre and Bahia among Brazilian isolates of C. fimbriata from other hosts. In the analyses of 14 microsatellite loci, the rubber tree isolates from Bahia were identical to each other and had alleles similar to those of Brazilian isolates from mango and eucalyptus. The microsatellite alleles of the Acre rubber tree isolates were identical to each other but distinct from other Brazilian isolates. The rubber tree isolates were morphologically indistinguishable from each other and very similar to the isolates of C. fimbriata on Ipomoea batatas, on which the species was originally described. Based on inoculation experiments results, the Bahia and Acre rubber tree isolates do not appear to be host specialized, which is typical for Brazilian isolates of C. fimbriata sensu stricto.Item A high-density transcript linkage map with 1,845 expressed genes positioned by microarray-based Single Feature Polymorphisms (SFP) in Eucalyptus(BMC Genomics, 2011) Neves, Leandro G.; Mamani, Eva MC.; Alfenas, Acelino C.; Kirst, Matias; Grattapaglia, DarioTechnological advances are progressively increasing the application of genomics to a wider array of economically and ecologically important species. High-density maps enriched for transcribed genes facilitate the discovery of connections between genes and phenotypes. We report the construction of a high-density linkage map of expressed genes for the heterozygous genome of Eucalyptus using Single Feature Polymorphism (SFP) markers. SFP discovery and mapping was achieved using pseudo-testcross screening and selective mapping to simultaneously optimize linkage mapping and microarray costs. SFP genotyping was carried out by hybridizing complementary RNA prepared from 4.5 year-old trees xylem to an SFP array containing 103,000 25-mer oligonucleotide probes representing 20,726 unigenes derived from a modest size expressed sequence tags collection. An SFP-mapping microarray with 43,777 selected candidate SFP probes representing 15,698 genes was subsequently designed and used to genotype SFPs in a larger subset of the segregating population drawn by selective mapping. A total of 1,845 genes were mapped, with 884 of them ordered with high likelihood support on a framework map anchored to 180 microsatellites with average density of 1.2 cM. Using more probes per unigene increased by two-fold the likelihood of detecting segregating SFPs eventually resulting in more genes mapped. In silico validation showed that 87% of the SFPs map to the expected location on the 4.5X draft sequence of the Eucalyptus grandis genome. The Eucalyptus 1,845 gene map is the most highly enriched map for transcriptional information for any forest tree species to date. It represents a major improvement on the number of genes previously positioned on Eucalyptus maps and provides an initial glimpse at the gene space for this global tree genome. A general protocol is proposed to build high-density transcript linkage maps in less characterized plant species by SFP genotyping with a concurrent objective of reducing microarray costs. HIgh-density gene-rich maps represent a powerful resource to assist gene discovery endeavors when used in combination with QTL and association mapping and should be especially valuable to assist the assembly of reference genome sequences soon to come for several plant and animal species.Item A host specialized form of Ceratocystis fimbriata causes seed and seedling blight on native Carapa guianensis (andiroba) in Amazonian rainforests(Fungal Biology, 2019-02) Valdetaro, Denise C. O. F.; Oliveira, Leonardo S. S.; McNew, Douglas L.; Pimenta, Lucas V. A.; Gonçalves, Rivadalve C.; Schurt, Daniel A.; Alfenas, Acelino C.; Harrington, Thomas C.; Guimarães, Lúcio M. S.Ceratocystis fimbriata Ellis & Halsted recently was recorded causing seed and seedling blight on Carapa guianensis Aubl. (andiroba), a tree species native to the Amazon Rainforest and prized for its valuable timber and medicinal seed oil. C. fimbriata more commonly causes wilt type diseases in woody hosts, especially on non-native host trees. However, on andiroba the disease occurs on seedlings and seeds, affecting the species regeneration. We studied 73 isolates of C. fimbriata on andiroba from three regions of the Amazon Basin to see if they represented natural or introduced populations. Analysis of ITS rDNA sequences and phylogenetic analysis of mating type genes revealed new haplotypes of C. fimbriata from the Latin American Clade that were closely related to other Brazilian populations of the fungus. In mating experiments, andiroba isolates were inter-fertile with tester strains of C. fimbriata from Brazil and elsewhere, confirming that they belong to a single biological species. Using microsatellite markers, 14 genotypes and populations with intermediate levels of genetic variability were found, suggesting that the fungus is indigenous to the Amazon Basin. Inoculation tests indicated that the andiroba isolates are host-specialized on andiroba, supporting the proposition of the special form C. fimbriata f. sp. carapa.Item Mapping global potential risk of mango sudden decline disease caused by Ceratocystis fimbriata(Public Library of Science One, 2016-07-14) Galdino, Tarcísio Visintin da Silva; Kumar, Sunil; Oliveira, Leonardo S. S.; Alfenas, Acelino C.; Neven, Lisa G.; Al-Sad, Abdullah M.; Picanço, Marcelo C.The Mango Sudden Decline (MSD), also referred to as Mango Wilt, is an important disease of mango in Brazil, Oman and Pakistan. This fungus is mainly disseminated by the mango bark beetle, Hypocryphalus mangiferae (Stebbing), by infected plant material, and the infested soils where it is able to survive for long periods. The best way to avoid losses due to MSD is to prevent its establishment in mango production areas. Our objectives in this study were to: (1) predict the global potential distribution of MSD, (2) identify the mango growing areas that are under potential risk of MSD establishment, and (3) identify climatic factors associated with MSD distribution. Occurrence records were collected from Brazil, Oman and Pakistan where the disease is currently known to occur in mango. We used the correlative maximum entropy based model (MaxEnt) algorithm to assess the global potential distribution of MSD. The MaxEnt model predicted suitable areas in countries where the disease does not already occur in mango, but where mango is grown. Among these areas are the largest mango producers in the world including India, China, Thailand, Indonesia, and Mexico. The mean annual temperature, precipitation of coldest quarter, precipitation seasonality, and precipitation of driest month variables contributed most to the potential distribution of MSD disease. The mango bark beetle vector is known to occur beyond the locations where MSD currently exists and where the model predicted suitable areas, thus showing a high likelihood for disease establishment in areas predicted by our model. Our study is the first to map the potential risk of MSD establishment on a global scale. This information can be used in designing strategies to prevent introduction and establishment of MSD disease, and in preparation of efficient pest risk assessments and monitoring programs.Item Microsatellite analysis indicates that Puccinia psidii in Australia is mutating but not recombining(Australasian Plant Pathology, 2015-07) Machado, Patrícia da S.; Alfenas, Acelino C.; Alfenas, Rafael F.; Mohammed, Caroline L.; Glen, MoragPuccinia psidii is considered a biosecurity threat in Australia because of its broad host range that includes many species of Myrtaceae which dominate Australian ecosystems. Since it was first reported in this country, in April 2010, there has been little information about the population structure of the pathogen. In this study, six microsatellite loci were analysed to determine the genetic relationship among rust specimens from different hosts and locations in Australia, New Caledonia, Hawaii and China. The Chinese and New Caledonian specimens share a multi-locus genotype with the majority of the Australian specimens. The results also indicated a close relationship between Australian and Hawaiian samples. At present, the P. psidii population in Australia is genetically uniform with no evidence of sexual recombination. Five of the 104 collections varied by one allele at single loci, indicating that mutations are common but persistence of the mutants in the population may be less common.Item Movement of genotypes of Ceratocystis fimbriata within and among Eucalyptus plantations in Brazil(Phytopathology, 2011-04-05) Ferreira, Maria A.; Harrington, Thomas C.; Alfenas, Acelino C.; Mizubuti, Eduardo S. G.Ceratocystis wilt on eucalyptus, caused by Ceratocystis fimbriata, was first recognized in 1997 in the state of Bahia, Brazil, but is now known in five other states and in four other countries. C. fimbriata is a native, soilborne pathogen in some parts of Brazil but we hypothesized that genotypes of the pathogen have been moved among plantations in rooted cuttings collected from diseased trees and within plantations on cutting tools. We used six microsatellite markers to identify 78 genotypes of C. fimbriata among 177 isolates from individual trees in 20 eucalyptus plantations. The highest gene and genotypic diversity values were found in plantations on formerly wild Cerrado forest in Minas Gerais, suggesting that the fungus was in the soil prior to planting eucalyptus. In contrast, one or only a few genotypes were found in plantations on previous pastureland (with no woody hosts) in Bahia and São Paulo, and most of these genotypes were found in a Bahian nursery or in one of two Bahian plantations that were sources for rooted cuttings. Sources of cuttings tended to be dominated by one or a few genotypes that may have been spread within the plantation on cutting tools.Item Mycoparasitic species of Sphaerellopsis, and allied lichenicolous and other genera(IMA Fungus The Global Mycological Journal, 2014-11-27) Trakunyingcharoen, Thippawan; Lombard, Lorenzo; Groenewald, Johannes Z.; Cheewangkoon, Ratchadawan; Alfenas, Acelino C.Species of Sphaerellopsis (sexual morph Eudarluca) are well-known cosmopolitan mycoparasites occurring on a wide range of rusts. Although their potential role as biocontrol agents has received some attention, the molecular phylogeny of the genus has never been resolved. Based on morphology and DNA sequence data of the large subunit nuclear ribosomal RNA gene (LSU, 28S) and the internal transcribed spacers (ITS) and 5.8S rRNA gene of the nrDNA operon, the genus Sphaerellopsis is shown to belong to Leptosphaeriaceae in Dothideomycetes. Sphaerellopsis is circumscribed, and the sexually typified generic name Eudarluca treated as a synonym on the basis that Sphaerellopsis is more commonly used in literature, is the older generic name, and is the morph commonly encountered by plant pathologists in the field. A neotype is designated for Sphaerellopsis filum, and two new species are introduced, S. macroconidialis and S. paraphysata spp. nov. Species previously incorrectly placed in Sphaerellopsis are allocated to Neosphaerellopsis gen. nov. as N. thailandica, and to the genus Acrocalymma, as A. fici. The genus Rhizopycnis is nestled among species of Acrocalymma, and reduced to synonymy based on its morphology and DNA phylogeny, while Acrocalymmaceae is introduced as novel family to accommodate members of this genus in the Dothideomycetes. Furthermore, Sphaerellopsis proved to be phylogenetically closely allied to a lichenicolous complex of phoma-like taxa, for which the new genera Diederichomyces and Xenophoma are established. Several new combinations are introduced, namely D. xanthomendozae, D. ficuzzae, D. caloplacae, D. cladoniicola, D. foliaceiphila, and X. puncteliae combs. nov, while Paraphaeosphaeria parmeliae sp. nov. is newly described.Item A new species of Calonectria causing leaf blight and cutting rot of three forest tree species in Brazil(Tropical Plant Pathology, 2013-08-13) Alfenas, Rafael F.; Pereira, Olinto L.; Jorge, Vanessa L.; Crous, Pedro W.; Alfenas, Acelino C.Several species of Calonectria cause diseases on a wide range of forest tree species that are propagated either via seedlings or rooted cuttings. In nurseries these fungi cause damping-off, cutting and root rots, stem lesions, and leaf blights. Recently a Calonectria sp. was isolated from rooted cuttings of Anadenanthera peregrina (Fabaceae), Piptadenia gonoacantha (Fabaceae), and Azadirachta indica (Meliaceae) exhibiting leaf blight and cutting rot in a forest nursery at the Universidade Federal de Viçosa, Brazil. Morphological comparisons and DNA sequences of three loci containing partial gene sequences of β-tubulin (TUB2), calmodulin (CAL), and elongation factor (TEF-1α) indicated that these isolates represent an unnamed species of Calonectria, described here as C. hodgesii sp. nov. Sprayinoculated plants of all three hosts with a suspension at 1x104 conidia mL-1 induced leaf lesions, cutting rot, and intense defoliation as observed under natural conditions. Calonectria hodgesii was re-isolated from infected tissue, which fulfilled Koch's postulates and confirmed its status as a pathogen with a wide host range.Item Positioning of the major locus for Puccinia psidii rust resistance (Ppr1) on the Eucalyptus reference map and its validation across unrelated pedigrees(Tree Genetics & Genomes, 2010-06-12) Guimarães, Lucio M. S.; Mamani, Eva M. C.; Bueno, Nathalia W.; Faria, Danielle A.; Lau, Douglas; Alfenas, Acelino C.; Grattapaglia, DarioIn this report the major locus for Puccinia psidii rust resistance, Ppr1, was positioned on the reference genetic map for Eucalyptus. Additionally, its position was validated by association genetics in a related and two unrelated pedigrees involving different Eucalyptus grandis resistant trees crossed to individuals of two other species, Eucalyptus tereticornis and Eucalyptus camaldulensis. These results are consistent with the hypothesis that Ppr1 controls a large proportion of the variation for rust resistance, strengthening its role as a major locus in Eucalyptus and providing its unequivocal genomic position on linkage group 3. A localized map with 19 microsatellite loci was built around Ppr1. Multiallelic profiles were observed at several mapped microsatellites suggesting recent tandem duplications in the genomic landscape surrounding Ppr1. Markers EMBRA125 and EMBRA1071 flank Ppr1 at 9.5% and 7% recombination, respectively, and were found to be in linkage equilibrium in a E. grandis breeding population, consistent with the expectations in outcrossed Eucalyptus. Their potential use for MAS will specifically be directed to identifying resistant offspring of P. psidii resistant parent trees that are heterozygous at Ppr1. In these circumstances, a significant amount of LD is expected to occur between specific alleles at flanking microsatellites and the resistance allele at Ppr1. Moreover, the positional information of Ppr1 paves the way for prospective undertakings in this genomic region with the upcoming availability of a draft genome for E. grandis.Item Purification and characterization of xylanases from the fungus Chrysoporthe cubensis for production of xylooligosaccharides and fermentable sugars(Applied Biochemistry and Biotechnology, 2016-12-24) Sousa Gomes, Kamila de; Maitan-Alfenas, Gabriela P.; Andrade, Lorena G. A. de; Falkoski, Daniel Luciano; Guimarães, Valéria Monteze; Alfenas, Acelino C.; Rezende, Sebastião Tavares deXylanases from the pathogen fungus Chrysoporthe cubensis were produced under solid state fermentation (SSF) using wheat bran as carbon source. The enzymatic extracts were submitted to ion exchange (Q Sepharose) and gel filtration chromatography methods (Sephadex S-200) for purification. The xylanases were divided into three groups: P1 showed better performance at 60 °C and pH 4.0, P2 at 55 °C and pH 3.0, and P3 at 80 °C and pH 3.0. Oat spelt xylan was the best substrate hydrolyzed by P1 and P3, while beechwood xylan was better degraded by P2. Carboxymethyl cellulose (CMC) and p-nitrophenyl-β-d-xylopyranoside (p-NPβXyl) were not hydrolyzed by any of the xylanases. The K M ’ or K M values, using oat spelt xylan as substrate, were 2.65 mg/mL for P1, 1.81 mg/mL for P2, and 1.18 mg/mL for P3. Xylobiose and xylotriose were the main xylooligosaccharides of oat spelt xylan degradation, indicating that the xylanases act as endo-β-1,4-xylanases. Xylanases also proved to be efficient for hydrolysis of sugarcane bagasse when used as supplement of a commercial cocktail due to the increase of the reducing sugar release.Item Rust disease of eucalyptus, caused by Puccinia psidii, did not originate via host jump from guava in Brazil(Molecular Ecology, 2013-12) Graça, Rodrigo N.; Ross-Davis, Amy L.; Klopfenstein, Ned B.; Kim, Mee-Sook; Peever, Tobin L.; Cannon, Phil G.; Aun, Cristina P.; Mizubuti, Eduardo S. G.; Alfenas, Acelino C.The rust fungus, Puccinia psidii, is a devastating pathogen of introduced eucalypts (Eucalyptus spp.) in Brazil where it was first observed in 1912. This pathogen is hypothesized to be endemic to South and Central America and to have first infected eucalypts via a host jump from native guava (Psidium guajava). Ten microsatellite markers were used to genotype 148 P. psidii samples from eucalypts and guava plus five additional myrtaceous hosts across a wide geographic range of south-eastern Brazil and Uruguay. Principal coordinates analysis, a Bayesian clustering analysis and a minimum-spanning network revealed two major genetic clusters among the sampled isolates, one associated with guava and another associated with eucalypts and three additional hosts. Multilocus genotypes infecting guava differed by multiple mutational steps at eight loci compared with those infecting eucalypts. Approximate Bayesian computation revealed that evolutionary scenarios involving a coalescence event between guava- and eucalypt-associated pathogen populations within the past 1000 years are highly unlikely. None of the analyses supported the hypothesis that eucalypt-infecting P. psidii in Brazil originated via host jump from guava following the introduction of eucalypts to Brazil approximately 185 years ago. The existence of host-associated biotypes of P. psidii in Brazil indicates that this diversity must be considered when assessing the invasive threat posed by this pathogen to myrtaceous hosts worldwide.Item Species or genotypes? reassessment of four recently described species of the Ceratocystis wilt pathogen, Ceratocystis fimbriata, on Mangifera indica(Phytopathology, 2015-09) Oliveira, Leonardo S. S.; Harrington, Thomas C.; Ferreira, Maria A.; Damacena, Michelle B.; Al-Sadi, Abdullah M.; Al-Mahmooli, Issa H. S.; Alfenas, Acelino C.Ceratocystis wilt is among the most important diseases on mango (Mangifera indica) in Brazil, Oman, and Pakistan. The causal agent was originally identified in Brazil as Ceratocystis fimbriata, which is considered by some as a complex of many cryptic species, and four new species on mango trees were distinguished from C. fimbriata based on variation in internal transcribed spacer sequences. In the present study, phylogenetic analyses using DNA sequences of mating type genes, TEF-1α, and β-tubulin failed to identify lineages corresponding to the four new species names. Further, mating experiments found that the mango isolates representing the new species were interfertile with each other and a tester strain from sweet potato (Ipomoea batatas), on which the name C. fimbriata is based, and there was little morphological variation among the mango isolates. Microsatellite markers found substantial differentiation among mango isolates at the regional and population levels, but certain microsatellite genotypes were commonly found in multiple populations, suggesting that these genotypes had been disseminated in infected nursery stock. The most common microsatellite genotypes corresponded to the four recently named species (C. manginecans, C. acaciivora, C. mangicola, and C. mangivora), which are considered synonyms of C. fimbriata. This study points to the potential problems of naming new species based on introduced genotypes of a pathogen, the value of an understanding of natural variation within and among populations, and the importance of phenotype in delimiting species.Item Supressão da atividade saprofítica de Rhizoctonia spp. Em solos de jardim clonal de Eucalyptus(Fitopatologia Brasileira, 2002-09) Sanfuentes, Eugenio A.; Alfenas, Acelino C.; Maffia, Luiz A.; Silveira, Silvaldo F.; Penchel, Ricardo; Sartorio, Robert C.Avaliou-se a eficiência de Trichoderma longibranchiatum (UFV-1), de T. inhamatum (UFV-2 e UFV-3), compostos de casca e folhas de eucalipto contra Rhizoctonia spp., aplicados em solo de jardim clonal de eucalipto (Eucalyptus sp.). Em solos artificialmente infestados com Rhizoctonia spp., sob condições controladas, os antagonistas UFV-2 e UFV-3 apresentaram níveis elevados de supressividade, quando se aumentou a fonte alimentar na formulação, de 5 a 50 g de farelo de trigo por litro. No campo, o antagonista UFV-3 não teve efeito significativo na redução do inóculo de Rhizoctonia spp. Compostos de casca de eucalipto apresentaram diferentes graus de supressão a Rhizoctonia spp., dependendo da origem e do lote do composto. A incorporação de folhas de eucalipto ao solo favoreceu o aumento do inóculo de Rhizoctonia spp.Item Variáveis climáticas associadas à incidência de mofo-cinzento em eucalipto(Fitopatologia Brasileira, 2006-03) Mafia, Reginaldo G.; Alfenas, Acelino C.; Ferreira, Eraclides M.; Leite, Fernando Palha; Souza, Flávio Luiz deCom base em dados de infecção natural avaliou-se a incidência do mofo-cinzento, causado por Botrytis cinerea, em 14 clones de Eucalyptus spp. em relação às condições climáticas predominantes em um viveiro clonal localizado em Belo Oriente, Minas Gerais. A temperatura máxima, mínima e média, precipitação pluviométrica e umidade relativa do ar foram coletadas entre 1991 e 2004. A incidência da doença foi avaliada mensalmente em 2004 em todas as fases de produção de mudas clonais de eucalipto. A presença do patógeno foi avaliada na água coletada do efluente de fertirrigação. A incidência do mofo-cinzento correlacionou-se melhor e negativamente com a temperatura máxima. Os resultados indicaram que a temperatura máxima é a variável a ser monitorada para fins de previsão da doença, a qual apresenta alto risco de incidência quando a temperatura máxima registrada for inferior a 27 ºC. Dentre os 14 clones propagados em 2004, o clone 957 (híbrido de Eucalyptus urophylla) apresentou menor incidência da doença, sob condições de infecção natural. Observou-se que o fungo encontra-se comumente associado a mudas de eucalipto e que o desenvolvimento da epidemia é regulado por temperaturas amenas, uma vez que, condições de alta umidade relativa e presença de água livre no hospedeiro ocorrem constantemente, em virtude das freqüentes irrigações requeridas para produção de mudas. Constatou-se que a água reutilizada, coletada no efluente de irrigação, contém inóculo do patógeno.