Methyl jasmonate does improve postharvest conservation of 'Golden' papaya fruit / Metil jasmonato contribui para a conservação de mamão 'Golden'

Methyl jasmonate (MeJA) is recognized as a plant hormone that induces several biochemical changes related to improving fruit quality, but it is evident that the effect of MeJA during postharvest is very variable upon concentration, plant species, and even cultivars. The objective of this research was to relate the application of this regulator at low concentration (10 µmol L-1 for 24 h) to changes in postharvest physiology, on the incidence of anthracnose and quality of 'Golden' papaya fruit during storage at 24 ± 1°C. From the seventh day of storage, anthracnose incidence was reduced by more than 50% with MeJA treatment. The lesion area after infection was also reduced. Although, MeJA reduced fruit acidity, the ascorbic acid and soluble solids content were not altered during storage with the hormonal treatment. The skin color and pulp firmness showed slight retention. The reduction in ethylene production was accompanied by a reduction of respiration in treated fruits. MeJa increased CAT activity only in the skin whereas SOD activity was not induced by MeJA in both skin and pulp. Although, the increase of CAT from the third day of storage may have contributed to the reduction of lipid peroxidation in the skin, the MDA reduction in the pulp cannot be explained only by CAT activity. In summary, the application of MeJA in 'Golden' papaya reduced the incidence and severity of anthracnose, decreased respiration, ethylene production and lipid peroxidation. It is concluded that the application of MeJA at a low concentration (10 µmol L-1) may contribute to anthracnose control in ´Golden`papaya and slows the ripening of fruits.

O metil jasmonato (MeJA) é reconhecido como um hormônio vegetal que induz várias alterações bioquímicas relacionadas à qualidade dos frutos, mas é evidente que o efeito do MeJA durante a pós-colheita é muito variável quanto à concentração, espécies de plantas e até mesmo cultivares. O objetivo deste trabalho foi relacionar a aplicação deste regulador em baixa concentração (10 µmol L-1 por 24 h) às mudanças na fisiologia pós-colheita, na incidência de antracnose e na qualidade do mamão 'Golden' durante o armazenamento a 24 ± 1 ° C. A partir do sétimo dia de armazenamento, a incidência de antracnose foi reduzida em mais de 50% com o tratamento com MeJA. Incidência da doença também foi reduzida. Embora o MeJA tenha reduzido a acidez dos frutos, os teores de ácido ascórbico e sólidos solúveis não foram alterados durante o armazenamento com o tratamento hormonal. A cor da casca e a firmeza da polpa apresentaram ligeira retenção. A redução na produção de etileno foi acompanhada pela redução da respiração nos frutos tratados. MeJa aumentou a atividade da CAT apenas na casca, enquanto a atividade da SOD não foi induzida por MeJA na casca e na polpa. Embora o aumento da CAT a partir do terceiro dia de armazenamento possa ter contribuído para a redução da peroxidação lipídica na casca, a redução do MDA na polpa não pode ser explicada apenas pela atividade da CAT. Em resumo, a aplicação de MeJA em mamão 'Golden' reduziu a incidência e severidade da antracnose, diminuiu a respiração, a produção de etileno e a peroxidação lipídica. Conclui-se que a aplicação de MeJA em baixa concentração (10 µmol L-1) pode contribuir para o controle da antracnose em mamão ´Golden` e retardar o amadurecimento dos frutos.

Methyl jasmonate does improve postharvest conservation of 'Golden' papaya fruit / Metil jasmonato contribui para a conservação de mamão 'Golden'

ABSTRACT: Methyl jasmonate (MeJA) is recognized as a plant hormone that induces several biochemical changes related to improving fruit quality, but it is evident that the effect of MeJA during postharvest is very variable upon concentration, plant species, and even cultivars. The objective of this research was to relate the application of this regulator at low concentration (10 µmol L-1 for 24 h) to changes in postharvest physiology, on the incidence of anthracnose and quality of 'Golden' papaya fruit during storage at 24 ± 1°C. From the seventh day of storage, anthracnose incidence was reduced by more than 50% with MeJA treatment. The lesion area after infection was also reduced. Although, MeJA reduced fruit acidity, the ascorbic acid and soluble solids content were not altered during storage with the hormonal treatment. The skin color and pulp firmness showed slight retention. The reduction in ethylene production was accompanied by a reduction of respiration in treated fruits. MeJa increased CAT activity only in the skin whereas SOD activity was not induced by MeJA in both skin and pulp. Although, the increase of CAT from the third day of storage may have contributed to the reduction of lipid peroxidation in the skin, the MDA reduction in the pulp cannot be explained only by CAT activity. In summary, the application of MeJA in 'Golden' papaya reduced the incidence and severity of anthracnose, decreased respiration, ethylene production and lipid peroxidation. It is concluded that the application of MeJA at a low concentration (10 µmol L-1) may contribute to anthracnose control in ´Golden`papaya and slows the ripening of fruits.

RESUMO: O metil jasmonato (MeJA) é reconhecido como um hormônio vegetal que induz várias alterações bioquímicas relacionadas à qualidade dos frutos, mas é evidente que o efeito do MeJA durante a pós-colheita é muito variável quanto à concentração, espécies de plantas e até mesmo cultivares. O objetivo deste trabalho foi relacionar a aplicação deste regulador em baixa concentração (10 µmol L-1 por 24 h) às mudanças na fisiologia pós-colheita, na incidência de antracnose e na qualidade do mamão 'Golden' durante o armazenamento a 24 ± 1 ° C. A partir do sétimo dia de armazenamento, a incidência de antracnose foi reduzida em mais de 50% com o tratamento com MeJA. Incidência da doença também foi reduzida. Embora o MeJA tenha reduzido a acidez dos frutos, os teores de ácido ascórbico e sólidos solúveis não foram alterados durante o armazenamento com o tratamento hormonal. A cor da casca e a firmeza da polpa apresentaram ligeira retenção. A redução na produção de etileno foi acompanhada pela redução da respiração nos frutos tratados. MeJa aumentou a atividade da CAT apenas na casca, enquanto a atividade da SOD não foi induzida por MeJA na casca e na polpa. Embora o aumento da CAT a partir do terceiro dia de armazenamento possa ter contribuído para a redução da peroxidação lipídica na casca, a redução do MDA na polpa não pode ser explicada apenas pela atividade da CAT. Em resumo, a aplicação de MeJA em mamão 'Golden' reduziu a incidência e severidade da antracnose, diminuiu a respiração, a produção de etileno e a peroxidação lipídica. Conclui-se que a aplicação de MeJA em baixa concentração (10 µmol L-1) pode contribuir para o controle da antracnose em mamão ´Golden` e retardar o amadurecimento dos frutos.

Eco-friendly fungicide based on chitosan and pecan nut oil: development and evaluation in anthracnose control

Industrial processing of pecan nuts results in edible oil ­ with fractions of fatty acids, polyphenols and phytosterols - that may be used for managing phytopathogens. Besides, chitosan may interfere with pathogen development directly and/or activate mechanisms of defense in plant tissues. This study aimed at developing a novel, natural and eco-friendly fungicide based on pecan nut oil immobilized in chitosan and at evaluating its activity against Colletotrichum gloeosporioides, an agent of anthracnose in guava trees and other fruit trees. Changes in the immobilized material exhibited at bands were identified by infrared spectroscopy. Additionally, micrographs by scanning electron microscopy (SEM) showed efficient changes in the immobilized material, by comparison with the polymer chitosan. Decreases in Colletotrichum gloeosporioides sporulation in vitro were 37% in the case of pecan nut oil immobilized in chitosan (IO) and 39% in the case of non-immobilized pecan nut oil (NIO). These results are positive because this fungus produces a large number of spores which may disseminate and survive on surfaces of plant tissues infected by anthracnose. Immobilization of pecan oil in chitosan showed benefits in the synthesis of a novel, stable and eco-friendly material which may be applied to guava trees to control anthracnose.(AU)

Morphological characterization and molecular identification of Colletotrichum species associated to sweet persimmon anthracnose in Southern Brazil / Caracterização morfológica e identificação molecular de espécies de Colletotrichum associadas à antracnose de caqui doce no Sul do Brasil

The highlands of Southern Brazil contribute with 40% of Brazilian persimmon production. Although expanding, persimmon production faces major problems caused by anthracnose disease (black spot), including fruit rot and necrosis of leaves. Several Colletotrichum species (C. horii, C. gloeosporioides, among others) are implicated in persimmon anthracnose around the world. To identify Colletotrichum species associated with persimmon anthracnose in the highlands of Southern Brazil, 34 isolates were analyzed by ITS-rDNA partial region, GAPDH, and TUB2 partial gene sequences, morphological characteristics, and virulence on persimmon fruits and leaves. Data showed a high prevalence of C. horii (85.3%), that associated with its high virulence on fruits and leaves, confirm a considerable degree of host preference. Moreover, other species C. aenigma, C. asianum, C. fructicola, and C. nymphaeae, were identified, but the last three ones exhibited low virulence on fruits and were not able to produce symptoms on leaves. As far as we know this is the first reference on C. asianum in persimmon. The present data may contribute to a better understanding of the etiology of anthracnose in sweet persimmon in Southern Brazil, and it will be useful for epidemiological studies and the development of disease management measures.

As terras altas do sul do Brasil contribuem com 40% da produção brasileira de caqui. Embora em expansão, a produção de caqui enfrenta grandes problemas causados ​​pela antracnose (mancha preta), incluindo o apodrecimento dos frutos e necrose das folhas. Várias espécies de Colletotrichum (C. horii, C. gloeosporioides, entre outras) estão envolvidas com a antracnose do caqui em todo mundo. Para identificar espécies de Colletotrichum associadas à antracnose de caqui nas terras altas do sul do Brasil, 34 isolados foram analisados ​​através da região parcial de ITS-rDNA e sequências parciais dos genes GAPDH e TUB2, características morfológicas e virulência em frutos e folhas de caqui. Os dados mostraram uma alta prevalência de C. horii (85,3%), que associada à sua alta virulência em frutos e folhas, confirma um grau considerável de preferência pelo hospedeiro. Além disso, foram identificadas outras espécies C. aenigma, C. asianum, C. fructicola, e C. nymphaeae, mas as três últimas exibiram baixa virulência nos frutos e não foram capazes de produzir sintomas nas folhas. Até onde sabemos, esta é a primeira referência sobre C. asianum em caqui. Os presentes dados podem contribuir para uma melhor compreensão da etiologia da antracnose em caqui doce no sul do Brasil e isso pode ser útil para estudos epidemiológicos e para o desenvolvimento de medidas de controle da doença.

Morphological characterization and molecular identification of Colletotrichum species associated to sweet persimmon anthracnose in Southern Brazil / Caracterização morfológica e identificação molecular de espécies de Colletotrichum associadas à antracnose de caqui doce no Sul do Brasil

The highlands of Southern Brazil contribute with 40% of Brazilian persimmon production. Although expanding, persimmon production faces major problems caused by anthracnose disease (black spot), including fruit rot and necrosis of leaves. Several Colletotrichum species (C. horii, C. gloeosporioides, among others) are implicated in persimmon anthracnose around the world. To identify Colletotrichum species associated with persimmon anthracnose in the highlands of Southern Brazil, 34 isolates were analyzed by ITS-rDNA partial region, GAPDH, and TUB2 partial gene sequences, morphological characteristics, and virulence on persimmon fruits and leaves. Data showed a high prevalence of C. horii (85.3%), that associated with its high virulence on fruits and leaves, confirm a considerable degree of host preference. Moreover, other species C. aenigma, C. asianum, C. fructicola, and C. nymphaeae, were identified, but the last three ones exhibited low virulence on fruits and were not able to produce symptoms on leaves. As far as we know this is the first reference on C. asianum in persimmon. The present data may contribute to a better understanding of the etiology of anthracnose in sweet persimmon in Southern Brazil, and it will be useful for epidemiological studies and the development of disease management measures.(AU)

As terras altas do sul do Brasil contribuem com 40% da produção brasileira de caqui. Embora em expansão, a produção de caqui enfrenta grandes problemas causados ​​pela antracnose (mancha preta), incluindo o apodrecimento dos frutos e necrose das folhas. Várias espécies de Colletotrichum (C. horii, C. gloeosporioides, entre outras) estão envolvidas com a antracnose do caqui em todo mundo. Para identificar espécies de Colletotrichum associadas à antracnose de caqui nas terras altas do sul do Brasil, 34 isolados foram analisados ​​através da região parcial de ITS-rDNA e sequências parciais dos genes GAPDH e TUB2, características morfológicas e virulência em frutos e folhas de caqui. Os dados mostraram uma alta prevalência de C. horii (85,3%), que associada à sua alta virulência em frutos e folhas, confirma um grau considerável de preferência pelo hospedeiro. Além disso, foram identificadas outras espécies C. aenigma, C. asianum, C. fructicola, e C. nymphaeae, mas as três últimas exibiram baixa virulência nos frutos e não foram capazes de produzir sintomas nas folhas. Até onde sabemos, esta é a primeira referência sobre C. asianum em caqui. Os presentes dados podem contribuir para uma melhor compreensão da etiologia da antracnose em caqui doce no sul do Brasil e isso pode ser útil para estudos epidemiológicos e para o desenvolvimento de medidas de controle da doença.(AU)

Evaluation of the production of antifungal metabolites against Colletotrichum gloeosporioides in Streptomyces 5.1 by random mutagenesis

Streptomyces 5.1 is a bacterium isolated from rice soils in the south of the Tolima department (Colombia). This microorganism is characterized by its antagonistic activity against rubber tree phytopathogens like Colletotrichum gloeosporioides, the causal agent of leaf anthracnose. The antifungal activity of this Streptomyces isolate has been associated with secondary metabolites production. However, the identity of those metabolites is unknown because its purification and identification have not been possible through classic chemical studies. Therefore, aiming to contribute in the study of the secondary metabolites produced by 5.1 from a molecular approach, this research seeks to identify -preliminarily- the genomic fingerprint changes associated with the production of antifungal secondary metabolites produced by Streptomyces 5.1 through the evaluation of a mutant library of 5.1 obtained by random mutagenesis using controlled ultraviolet light exposure. The antifungal activity of obtained mutants was evaluated using Colletotrichum gloeosporioides (C1) fungus as a biosensor, isolated by the Biotechnology Institute of Universidad Nacional de Colombia. In this way, the library of mutants of 5.1, initially formed by 300 isolations, was classified into two phenotypic groups of interest: enhanced mutants (1 isolate) and null mutants (11 isolates) of secondary metabolites. The genomic changes in both groups were analyzed by obtaining the genomic profile of the isolates using Repetitive Extragenic Palindromic (Rep-PCR). The obtained profiles evidenced the presence of one additional band in the enhanced mutant, and the absence of a specific band in the non-producing mutants, both in comparison with the original strain. These bands are proposed for a future sequencing study which will define their role in the production process of metabolites with antifungal activity in Streptomyces 5.1.

Evaluation of the production of antifungal metabolites against Colletotrichum gloeosporioides in Streptomyces 5.1 by random mutagenesis

Streptomyces 5.1 is a bacterium isolated from rice soils in the south of the Tolima department (Colombia). This microorganism is characterized by its antagonistic activity against rubber tree phytopathogens like Colletotrichum gloeosporioides, the causal agent of leaf anthracnose. The antifungal activity of this Streptomyces isolate has been associated with secondary metabolites production. However, the identity of those metabolites is unknown because its purification and identification have not been possible through classic chemical studies. Therefore, aiming to contribute in the study of the secondary metabolites produced by 5.1 from a molecular approach, this research seeks to identify -preliminarily- the genomic fingerprint changes associated with the production of antifungal secondary metabolites produced by Streptomyces 5.1 through the evaluation of a mutant library of 5.1 obtained by random mutagenesis using controlled ultraviolet light exposure. The antifungal activity of obtained mutants was evaluated using Colletotrichum gloeosporioides (C1) fungus as a biosensor, isolated by the Biotechnology Institute of Universidad Nacional de Colombia. In this way, the library of mutants of 5.1, initially formed by 300 isolations, was classified into two phenotypic groups of interest: enhanced mutants (1 isolate) and null mutants (11 isolates) of secondary metabolites. The genomic changes in both groups were analyzed by obtaining the genomic profile of the isolates using Repetitive Extragenic Palindromic (Rep-PCR). The obtained profiles evidenced the presence of one additional band in the enhanced mutant, and the absence of a specific band in the non-producing mutants, both in comparison with the original strain. These bands are proposed for a future sequencing study which will define their role in the production process of metabolites with antifungal activity in Streptomyces 5.1.(AU)

Atividades antioxidante e antifúngica dos óleos essenciais de Cochlospermum regium frente à Sclerotinia sclerotiorum e Colletotrichum gloeosporioides / Antioxidant and antifungal activities of Cochlospermum regium essential oils against Sclerotinia sclerotiorum and Colletotrichum gloeosporioides

O objetivo do trabalho foi avaliar os óleos essenciais de Cochlospermum regium e suas atividades antioxidante e antifúngica frente à Sclerotinia sclerotiorum e Colletotrichum gloeosporioides. Foram coletados raízes e folhas em indivíduos de C. regiumem uma área de Cerrado, o material foi processado, pesado e a extração dos óleos se deram pelo método de hidrodestilação em Clevenger. Os óleos tiveram seu rendimento avaliado, bem como as atividades antioxidante e antifúngica frente à S. sclerotiorum e C. gloeosporioides. Os rendimentos foram de 0,168 e 0,301% para folhas e raízes; a atividade antioxidante apresentou 100% de redução do radical DPPH foi de 100% nas concentrações entre 50 a 30 μL mL-1e a inibição micelial de 14,2 e 60,1% na concentração de 100 μL mL-1para S. sclerotiorum e de 84,4 e 81,3% para C. gloeosporioides para os óleos das raízes e folhas, respectivamente.

The objective of this work was to evaluate the essential oils of Cochlospermum regium and its antioxidant and antifungal activities against Sclerotinia sclerotiorum and Colletotrichum gloeosporioides. Roots and leaves were collected in individuals of C. regiumin a Cerrado area, the material was processed, weighed and extraction of the oils were by the hydroventilation method in Clevenger. The oils had their evaluated yield, as well as the antioxidant and antifungal activities against S. sclerotiorum and C. gloeosporioides. Yields were 0.168 and 0.301% for leaves and roots; the antioxidant activity showed 100% reduction of the DPPH radical was 100% in the concentrations between 50 and 30 μL mL-1and the mycelial inhibition of 14.2 and 60.1% in the concentration of 100 μL mL-1for S. sclerotiorum and 84.4 and 81.3% for C. gloeosporioides for root and leaf oils, respectively.

Atividades antioxidante e antifúngica dos óleos essenciais de Cochlospermum regium frente à Sclerotinia sclerotiorum e Colletotrichum gloeosporioides / Antioxidant and antifungal activities of Cochlospermum regium essential oils against Sclerotinia sclerotiorum and Colletotrichum gloeosporioides

O objetivo do trabalho foi avaliar os óleos essenciais de Cochlospermum regium e suas atividades antioxidante e antifúngica frente à Sclerotinia sclerotiorum e Colletotrichum gloeosporioides. Foram coletados raízes e folhas em indivíduos de C. regiumem uma área de Cerrado, o material foi processado, pesado e a extração dos óleos se deram pelo método de hidrodestilação em Clevenger. Os óleos tiveram seu rendimento avaliado, bem como as atividades antioxidante e antifúngica frente à S. sclerotiorum e C. gloeosporioides. Os rendimentos foram de 0,168 e 0,301% para folhas e raízes; a atividade antioxidante apresentou 100% de redução do radical DPPH foi de 100% nas concentrações entre 50 a 30 μL mL-1e a inibição micelial de 14,2 e 60,1% na concentração de 100 μL mL-1para S. sclerotiorum e de 84,4 e 81,3% para C. gloeosporioides para os óleos das raízes e folhas, respectivamente.(AU)

The objective of this work was to evaluate the essential oils of Cochlospermum regium and its antioxidant and antifungal activities against Sclerotinia sclerotiorum and Colletotrichum gloeosporioides. Roots and leaves were collected in individuals of C. regiumin a Cerrado area, the material was processed, weighed and extraction of the oils were by the hydroventilation method in Clevenger. The oils had their evaluated yield, as well as the antioxidant and antifungal activities against S. sclerotiorum and C. gloeosporioides. Yields were 0.168 and 0.301% for leaves and roots; the antioxidant activity showed 100% reduction of the DPPH radical was 100% in the concentrations between 50 and 30 μL mL-1and the mycelial inhibition of 14.2 and 60.1% in the concentration of 100 μL mL-1for S. sclerotiorum and 84.4 and 81.3% for C. gloeosporioides for root and leaf oils, respectively.(AU)

Colletotrichum nymphaeae var. entomophilum var. nov. a natural enemy of the citrus scale insect, Praelongorthezia praelonga (Hemiptera: Ortheziidae)

The citrus scale insect Praelongorthezia praelonga (Douglas), a major pest of citrus and other economically important crops, has only two commonly documented natural enemies: an entomopathogenic strain of the fungus Colletotrichum nymphaeae (Pass.) Aa and several parasitoids. The entomopathogenic strain of C. nymphaeae, formerly recognized under the synonym C. gloeosporioides f. sp. ortheziidae, is under development for commercial application as a biological control agent in citrus in Brazil-the top exporter of citrus globally. The synonomy of C. gloeosporioides f. sp. ortheziidae with C. nymphaeae remains based on limited DNA sequence data and without morphological study. To qualify for future approval as a biological control agent by federal agencies in Brazil and the European Union, the circumscription of a microorganism must be explicit and without ambiguities. Herein, through morphological study and phylogenetic analysis of five DNA regions we clarify the circumscription and affinity of entomopathogenic C. nymphaeae and describe it as a new variety.

Colletotrichum nymphaeae var. entomophilum var. nov. a natural enemy of the citrus scale insect, Praelongorthezia praelonga (Hemiptera: Ortheziidae)

The citrus scale insect Praelongorthezia praelonga (Douglas), a major pest of citrus and other economically important crops, has only two commonly documented natural enemies: an entomopathogenic strain of the fungus Colletotrichum nymphaeae (Pass.) Aa and several parasitoids. The entomopathogenic strain of C. nymphaeae, formerly recognized under the synonym C. gloeosporioides f. sp. ortheziidae, is under development for commercial application as a biological control agent in citrus in Brazil-the top exporter of citrus globally. The synonomy of C. gloeosporioides f. sp. ortheziidae with C. nymphaeae remains based on limited DNA sequence data and without morphological study. To qualify for future approval as a biological control agent by federal agencies in Brazil and the European Union, the circumscription of a microorganism must be explicit and without ambiguities. Herein, through morphological study and phylogenetic analysis of five DNA regions we clarify the circumscription and affinity of entomopathogenic C. nymphaeae and describe it as a new variety.(AU)

Secondary metabolites produced by endophytic fungi: novel antifungal activity of fumiquinone B

Fungi are present in the most diverse environments including the interior of plant tissues, living as endophytes without causing apparent damage. These endophytes are producers of secondary metabolites, also known as natural products, such as fungicides. Here, we evaluated the ethyl acetate fractions obtained from endophytic fungiisolated from plants in the genus Begonia. The fractions were submitted to inhibitorytest against the plant pathogens Diaporthe phaseolorum and Colletotrichum gloeosporioides. From the 88 ethyl acetate fractions evaluated, 14.7 % inhibited C. gloeosporioidesand 11.3 %inhibited D. phaseolorum. One fungal isolate displaying an active fraction was selected for chemical investigation. The fungus identified as Neopestalotiopsissp., produced a compound that was active against D. phaseolorum, with a MIC of 312 μg mL-1(1,695.3 μM). The compound was identified by mass spectrometry and 1H NMR as the known compound fumiquinone B. The results highlight that the endophytes are capable of producing compounds that may be used to control plant pathogens. The compound fumiquinone B is reported for the first time as an antifungal agent against D. phaseolorum, a relevant plant pathogen worldwide. This is also the first report of the production of fumiquinone B by the genus Neopestalotiopsis.

The potential of compounds isolated from Xylaria spp. as antifungal agents against anthracnose

Anthracnose is a crop disease usually caused by fungi in the genus Colletotrichum or Gloeosporium. These are considered one of the main pathogens, causing significant economic losses, such as in peppers and guarana. The current forms of control include the use of resistant cultivars, sanitary pruning and fungicides. However, even with the use of some methods of controlling these cultures, the crops are not free of anthracnose. Additionally, excessive application of fungicides increases the resistance of pathogens to agrochemicals and cause harm to human health and the environment. In order to find natural antifungal agents against guarana anthracnose, endophytic fungi were isolated from Amazon guarana. The compounds piliformic acid and cytochalasin D were isolated by chromatographic techniques from two Xylaria spp., guided by assays with Colletotrichum gloeosporioides. The isolated compounds were identified by spectrometric techniques, as NMR and mass spectrometry. This is the first report that piliformic acid and cytochalasin D have antifungal activity against C. gloeosporioides with MIC 2.92 and 2.46 µmol mL-1 respectively. Captan and difenoconazole were included as positive controls (MIC 16.63 and 0.02 µmol mL-1, respectively). Thus, Xylaria species presented a biotechnological potential and production of different active compounds which might be promising against anthracnose disease.(AU)

In vitro antimicrobial activity and alternative control of anthracnose in papaya / Atividade antimicrobiana in vitro e controle alternativo da antracnose em mamão

We evaluated the efficacy of natural products in the control of papaya anthracnose, in vitro and in vivo. The in vitro experiments for evaluation of mycelial growth used a completely randomized 10 × 4 factorial design (treatments × evaluation periods) with eight replicates, with sporulation evaluated at the end of the experiment. The treatments involved the use of aqueous extract at concentrations of 5 and 15% for Syzygium aromaticum (L.) Merr. & Perry (clove), Cinnamomum zeylanicum Breym (cinnamon), and Zingiber officinalis Rox. (ginger); 1 and 3% chitosan; the fungicide Prochloraz at 100 µg.mL-1; and a control (no treatment). For evaluating conidia germination, we used six treatments with five replicates. The treatments included 7.5% of each extract (clove, cinnamon, and ginger), 1.5% chitosan, and 50 µg.mL-1 of Prochloraz. For the in vivo experiment, "Formosa" papaya "Tainung 1" was used in a completely randomized design, with six treatments and four replicates to evaluate the severity of anthracnose caused by Colletotrichum gloeosporioides. The fruits were treated by immersion for 5 min with 15% clove, cinnamon, and ginger extracts, 8% chitosan, and control with distilled water, and immersion for 2 min in Prochloraz (33.75 g a.i./100 L). The treatments with 15% clove extract and 8% chitosan were effective in all evaluations, resulting in a viable alternative to the fungicide Prochloraz. The treatments with ginger extract were less effective and those with cinnamon offered intermediate control.(AU)

Este trabalho teve por objetivo avaliar, in vitro e in vivo, a eficácia de produtos naturais no controle da antracnose do mamão. Os experimentos in vitro foram instalados em delineamento inteiramente casualizado, sendo para a avaliação de crescimento micelial considerado o esquema fatorial 10 × 4 (tratamentos x períodos de avaliação) com oito repetições, com a esporulação avaliada ao final do experimento. Os tratamentos utilizados foram: extrato aquoso nas concentrações de 5 e 15% de Syzygium aromaticum (L.) Merr. & Perry (cravo-da-índia), Cinnamomum zeylanicum Breym (canela) e Zingiber officinalis Rox (gengibre), quitosana 1 e 3%, fungicida Procloraz a 100 µg.mL-1 e testemunha (sem tratamento). Para a avaliação da germinação de conídios, foram considerados seis tratamentos com cinco repetições. Foram utilizados os mesmos tratamentos, porém, nas concentrações de 7,5% de cada extrato (cravo-da-índia, canela e gengibre), 1,5% de quitosana, e 50 µg.mL-1 de Procloraz. Para o experimento in vivo, foram utilizados mamões "Formosa" "Tainung 1", em delineamento inteiramente casualizado, sendo seis tratamentos com quatro repetições para avaliação de severidade de antracnose causada por Colletotrichum gloeosporioides. Os frutos foram tratados por imersão durante 5 minutos com extratos de cravo-da-índia, canela e gengibre a 15%, quitosana a 8% e testemunha com água destilada, e imersão por 2 minutos em fungicida Procloraz (33,75 g i.a./100 L). Os tratamentos com extrato de cravo-da-índia a 15% e quitosana a 8% foram eficazes em todas as avaliações, sendo uma alternativa viável ao fungicida Procloraz. Os tratamentos com extrato de gengibre foram menos eficientes, com controle intermediário para o extrato de canela.(AU)

In vitro antimicrobial activity and alternative control of anthracnose in papaya / Atividade antimicrobiana in vitro e controle alternativo da antracnose em mamão

We evaluated the efficacy of natural products in the control of papaya anthracnose, in vitro and in vivo. The in vitro experiments for evaluation of mycelial growth used a completely randomized 10 × 4 factorial design (treatments × evaluation periods) with eight replicates, with sporulation evaluated at the end of the experiment. The treatments involved the use of aqueous extract at concentrations of 5 and 15% for Syzygium aromaticum (L.) Merr. & Perry (clove), Cinnamomum zeylanicum Breym (cinnamon), and Zingiber officinalis Rox. (ginger); 1 and 3% chitosan; the fungicide Prochloraz at 100 µg.mL-1; and a control (no treatment). For evaluating conidia germination, we used six treatments with five replicates. The treatments included 7.5% of each extract (clove, cinnamon, and ginger), 1.5% chitosan, and 50 µg.mL-1 of Prochloraz. For the in vivo experiment, "Formosa" papaya "Tainung 1" was used in a completely randomized design, with six treatments and four replicates to evaluate the severity of anthracnose caused by Colletotrichum gloeosporioides. The fruits were treated by immersion for 5 min with 15% clove, cinnamon, and ginger extracts, 8% chitosan, and control with distilled water, and immersion for 2 min in Prochloraz (33.75 g a.i./100 L). The treatments with 15% clove extract and 8% chitosan were effective in all evaluations, resulting in a viable alternative to the fungicide Prochloraz. The treatments with ginger extract were less effective and those with cinnamon offered intermediate control.(AU)

Este trabalho teve por objetivo avaliar, in vitro e in vivo, a eficácia de produtos naturais no controle da antracnose do mamão. Os experimentos in vitro foram instalados em delineamento inteiramente casualizado, sendo para a avaliação de crescimento micelial considerado o esquema fatorial 10 × 4 (tratamentos x períodos de avaliação) com oito repetições, com a esporulação avaliada ao final do experimento. Os tratamentos utilizados foram: extrato aquoso nas concentrações de 5 e 15% de Syzygium aromaticum (L.) Merr. & Perry (cravo-da-índia), Cinnamomum zeylanicum Breym (canela) e Zingiber officinalis Rox (gengibre), quitosana 1 e 3%, fungicida Procloraz a 100 µg.mL-1 e testemunha (sem tratamento). Para a avaliação da germinação de conídios, foram considerados seis tratamentos com cinco repetições. Foram utilizados os mesmos tratamentos, porém, nas concentrações de 7,5% de cada extrato (cravo-da-índia, canela e gengibre), 1,5% de quitosana, e 50 µg.mL-1 de Procloraz. Para o experimento in vivo, foram utilizados mamões "Formosa" "Tainung 1", em delineamento inteiramente casualizado, sendo seis tratamentos com quatro repetições para avaliação de severidade de antracnose causada por Colletotrichum gloeosporioides. Os frutos foram tratados por imersão durante 5 minutos com extratos de cravo-da-índia, canela e gengibre a 15%, quitosana a 8% e testemunha com água destilada, e imersão por 2 minutos em fungicida Procloraz (33,75 g i.a./100 L). Os tratamentos com extrato de cravo-da-índia a 15% e quitosana a 8% foram eficazes em todas as avaliações, sendo uma alternativa viável ao fungicida Procloraz. Os tratamentos com extrato de gengibre foram menos eficientes, com controle intermediário para o extrato de canela.(AU)

Phytosterols isolated from endophytic fungus Colletotrichum gloeosporioides (Melanconiaceae) / Fotoesteróis isolados do fungo endofítico Colletotrichum gloeosporioides (Melanconiaceae)

Endophytic fungi are fungi that colonize internal tissues of plants. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, the aim this study was the isolation and structural identification of sitosterol (1), stigmasterol (2), sitostenone (3), squalene (4), ergosterol (5) and ergosterol peroxide (6) from fungus Colletotrichum gloeosporioidesisolated as endophytic from Virola michelli, a typical Amazon plant, used in folk medicine against skin infection. Compounds were isolated by chromatography column on silica and identified by 1H and 13C NMR and MS. The presence of phytosterols in fungi is rare and this is the first report of the isolation of the phytosterols sitosterol, stigmasterol and sitostenone from the genus Colletotrichum.

Fungos endofíticos são fungos que colonizam os tecidos internos das plantas. Existem poucos estudos de compostos isolados de fungos endofíticos de plantas da Amazônia. Assim, o objetivo deste estudo foi o isolamento e identificação estrutural de sitosterol (1), estigmasterol (2), sitostenona (3), esqualeno (4), ergosterol (5) e peroxido de ergosterol (6) do fungo Colletotrichum gloeosporioidesisolado como endofítico de Virola michelli, uma planta típica da Amazônia, usada na medicina popular no combate a infecções de pele. Os compostos foram isolados por cromatografia em coluna de sílica e identificados por RMN 1H e 13C e EM. A presença de fitoesteróis em fungos é rara e este é o primeiro relato do isolamento dos fitoesteróides sitosterol, estigmasterol e sitostenona do gêreno Colletotrichum.

Phytosterols isolated from endophytic fungus Colletotrichum gloeosporioides (Melanconiaceae) / Fotoesteróis isolados do fungo endofítico Colletotrichum gloeosporioides (Melanconiaceae)

Endophytic fungi are fungi that colonize internal tissues of plants. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, the aim this study was the isolation and structural identification of sitosterol (1), stigmasterol (2), sitostenone (3), squalene (4), ergosterol (5) and ergosterol peroxide (6) from fungus Colletotrichum gloeosporioidesisolated as endophytic from Virola michelli, a typical Amazon plant, used in folk medicine against skin infection. Compounds were isolated by chromatography column on silica and identified by 1H and 13C NMR and MS. The presence of phytosterols in fungi is rare and this is the first report of the isolation of the phytosterols sitosterol, stigmasterol and sitostenone from the genus Colletotrichum.(AU)

Fungos endofíticos são fungos que colonizam os tecidos internos das plantas. Existem poucos estudos de compostos isolados de fungos endofíticos de plantas da Amazônia. Assim, o objetivo deste estudo foi o isolamento e identificação estrutural de sitosterol (1), estigmasterol (2), sitostenona (3), esqualeno (4), ergosterol (5) e peroxido de ergosterol (6) do fungo Colletotrichum gloeosporioidesisolado como endofítico de Virola michelli, uma planta típica da Amazônia, usada na medicina popular no combate a infecções de pele. Os compostos foram isolados por cromatografia em coluna de sílica e identificados por RMN 1H e 13C e EM. A presença de fitoesteróis em fungos é rara e este é o primeiro relato do isolamento dos fitoesteróides sitosterol, estigmasterol e sitostenona do gêreno Colletotrichum.(AU)

Cgl-SLT2 is required for appressorium formation, sporulation and pathogenicity in Colletotrichum gloeosporioide

The mitogen-activated protein (MAP) kinase pathways has been implicated in the pathogenicity of various pathogenic fungi and plays important roles in regulating pathogenicity-related morphogenesis. This work describes the isolation and characterization of MAP kinase gene, Cgl-SLT2, from Colletotrichum gloeosporioides. A DNA sequence, including 1,633 bp of Cgl-SLT2 open-reading frame and its promoter and terminator regions, was isolated via DNA walking and cloned. To analyze gene function, a gene disruption cassette containing hygromycin-resistant gene was constructed, and Cgl-SLT2 was inactivated via gene deletion. Analysis on Cgl-slt2 mutant revealed a defect in vegetative growth and sporulation as compared to the wild-type strain. When grown under nutrient-limiting conditions, hyperbranched hyphal morphology was observed in the mutant. Conidia induction for germination on rubber wax-coated hard surfaces revealed no differences in the percentage of conidial germination between the wild-type and Cgl-slt2 mutant. However, the percentage of appressorium formation in the mutant was greatly reduced. Bipolar germination in the mutant was higher than in the wild-type at 8-h post-induction. A pathogenicity assay revealed that the mutant was unable to infect either wounded or unwounded mangoes. These results suggest that the Cgl-SLT2 MAP kinase is required for C. gloeosporioides conidiation, polarized growth, appressorium formation and pathogenicity.

Genetic transformation with the gfp gene of Colletotrichum gloeosporioides isolates from coffee with blister spot

Blister spot (Colletotrichum gloeosporioides) is now widespread in most coffee producing states of Brazil, becoming a limiting factor for production. The lack of data relating to the reproduction of typical symptoms (light green, oily patches) leaves a gap within the pathosystem, forcing the search for new methodologies for monitoring the disease. Monitoring of genetically modified organisms has proven to be an effective tool in understanding the host x pathogen interactions. Thus, the present study was carried out to evaluate the effectiveness of two systems of genetic transformation in obtaining mutants using the gfp reporter gene. Using the two transformation systems (PEG and electroporation) revealed the efficiency of both, confirmed by fluorescence microscopy and resistance to the antibiotic hygromycin-B, when incorporated into the culture medium. The fungus maintained its cultural and morphological characteristics when compared to wild strains. When inoculated on coffee seedlings, it was found that the pathogenicity of the processed isolates had not changed.

Controle da antracnose em maracujá-amarelo por derivados de capim-limão (Cymbopogon citratus) / Control of anthracnose in yellow passion fruit by lemon grass (Cymbopogon citratus) derivatives

O maracujá-amarelo é uma das principais frutíferas cultivadas no Brasil. A conservação póscolheita de frutos de maracujá tem merecido grande atenção visto que o fruto é perecível e está sujeito à rápida deterioração e incidência de micro-organismos patogênicos. Este trabalho teve por objetivo avaliar a atividade antifúngica in vitro de derivados de capim-limão (Cymbopogon citratus) sobre o crescimento micelial, esporulação, germinação de esporos e tamanho de tubos germinativos de Colletotrichum gloeosporioides e in vivo no controle da antracnose sobre frutos de maracujá-amarelo em pós-colheita. Os tratamentos para o ensaio in vitro foram óleo essencial (OE) (0,5, 1, 5, 10, 15, 25, 50 e 60 μL mL-1), extrato bruto aquoso autoclavado (EAA), extrato bruto aquoso filtrado (EAF), hidrolato e citral (1, 5, 10, 20 e 25%) e no ensaio in vivo, os frutos foram tratados com: OE (0,025; 0,05 e 0,1%), hidrolato (5 e 10%), EAA (10%) e citral (0,1%). Água destilada foi utilizada como testemunha. Foram avaliadas a incidência e a severidade da antracnose a cada três dias. Os quatro derivados de capim-limão inibiram o crescimento micelial, a esporulação e a germinação de esporos, sendo que maiores valores de inibição foram obtidos nas maiores concentrações testadas e, no crescimento micelial, atingiram 55,8; 54,5; 30,0 e 19,9% para OE, EAF, EAA e hidrolato, respectivamente. Na avaliação in vivo, OE 0,1% e EAA 10% promoveram significativa redução na severidade da antracnose.

CONTROL OF ANTHRACNOSE IN YELLOW PASSION FRUIT BY LEMON GRASS (CYMBOPOGON CITRATUS) DERIVATIVES. The yellow passion fruit is a major fruit crop in Brazil. The post-harvest preservation of yellow passion fruits has deserved great attention since the fruit is perishable and subject to a fast deterioration and incidence of pathogenic microorganisms. The aim of this work was to evaluate the in vitro antifungal activity of lemon grass derivatives on the mycelial growth, sporulation, spore germination and length of germ tubes of Colletotrichum gloeosporioides and the effect of essential oil, extract, hydrolate and citral lemon grass (Cymbopogon citratus) for control of anthracnose on the yellow passion fruits. The treatments for the in vitro assay were essential oil (OE) (0.5, 1, 5, 10, 15, 25, 50 and 60 μL mL-1), autoclaved crude aqueous extract (EAA), filtered crude aqueous extract (EAF), hydrolate and citral (1, 5, 10, 20 and 25%). For the in vivo assay the fruits were treated with: OE (0.025, 0.05 and 0.1%), hydrolate (5 and 10%), EAA (10%) and citral (0.1%). Distilled water was used as a control. The anthracnose incidence and severity were evaluated every three days. The in vitro test verified that the 4 evaluated derivatives of lemon grass inhibited the mycelial growth, sporulation and spore germination. The higher the concentration of the derivatives, the higher the inhibition, which for the mycelia growth reached 55.8; 54.5; 30.0 and 19.9% for OE, EAF, EAA and hydrolate, respectively. In vivo tests showed that the OE 0.1% and EAA 10% promoted a reduction in the anthracnose severity.