Ornamental plant Pachystachys lutea as a source of promising endophytes for plant growth and phytoprotective activity

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskaya’s agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).

Ornamental plant Pachystachys lutea as a source of promising endophytes for plant growth and phytoprotective activity

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskayas agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).(AU)

Endophytic fungi from Brachiaria grasses in Brazil and preliminary screening of Sclerotinia sclerotiorum antagonists

Fungal endophytes of Brachiaria, a nonhost of Sclerotinia sclerotiorum, may harbor species with antagonistic effects against this plant pathogen. The objective of this work was to investigate the diversity of endophytic fungi associated with different Brachiaria species and hybrids and evaluate their potential to inhibit the plant pathogen S. sclerotiorum. Stem samples from 39 Brachiaria spp. plants were collected in pasture fields and experimental areas of three states of Brazil resulting in 74 endophytes isolated. Twenty-eight species were identified by sequences of the Internal Transcribed Spacer (ITS) and 18S rDNA regions. Paraconiothyrium sp. was the most abundant endophyte, accounting for 24 % (14 isolates) of total, and it was isolated from B. ruziziensis, B. decumbens, B. humidicola, and B. brizantha. Phoma sorghina was the second most abundant taxon, followed by Sarocladium strictum, and Plenodomus sp. In vitro analyses showed that Paraconiothyrium sp., Sarocladium kiliense, Acremonium curvulum, Setophoma terrestris, Dissoconium sp., and Cladosporium flabelliforme exhibited antagonistic activity against S. sclerotiorum, with percentages of growth inhibition ranging from 25 to 60 (p < 0.05). Paraconiothyrium sp. BBXE1 (60 %), BBPB4.1 (60 %), BCMT4.1 (54 %), and S. kiliense (54 %) showed the highest values of Antagonism Percentages (AP). Therefore, fungi with inhibitory activity against S. sclerotiorum such as Paraconiothyrium sp. are naturally endophytic in Brachiaria grasses.

Endophytic fungi from Brachiaria grasses in Brazil and preliminary screening of Sclerotinia sclerotiorum antagonists

Fungal endophytes of Brachiaria, a nonhost of Sclerotinia sclerotiorum, may harbor species with antagonistic effects against this plant pathogen. The objective of this work was to investigate the diversity of endophytic fungi associated with different Brachiaria species and hybrids and evaluate their potential to inhibit the plant pathogen S. sclerotiorum. Stem samples from 39 Brachiaria spp. plants were collected in pasture fields and experimental areas of three states of Brazil resulting in 74 endophytes isolated. Twenty-eight species were identified by sequences of the Internal Transcribed Spacer (ITS) and 18S rDNA regions. Paraconiothyrium sp. was the most abundant endophyte, accounting for 24 % (14 isolates) of total, and it was isolated from B. ruziziensis, B. decumbens, B. humidicola, and B. brizantha. Phoma sorghina was the second most abundant taxon, followed by Sarocladium strictum, and Plenodomus sp. In vitro analyses showed that Paraconiothyrium sp., Sarocladium kiliense, Acremonium curvulum, Setophoma terrestris, Dissoconium sp., and Cladosporium flabelliforme exhibited antagonistic activity against S. sclerotiorum, with percentages of growth inhibition ranging from 25 to 60 (p < 0.05). Paraconiothyrium sp. BBXE1 (60 %), BBPB4.1 (60 %), BCMT4.1 (54 %), and S. kiliense (54 %) showed the highest values of Antagonism Percentages (AP). Therefore, fungi with inhibitory activity against S. sclerotiorum such as Paraconiothyrium sp. are naturally endophytic in Brachiaria grasses.(AU)

Action of Trichodermil 1306 in the control of Fusarium oxysporum f. sp. lycopersici in tomato crops / Ação do Trichodermil 1306 no controle de Fusarium oxysporum f. sp. lycopersici na cultura do tomateiro

Fusarium wilt, caused by Fusarium oxysporum f. sp. lycopersici, is one of the most important diseases of tomato crop in Brazil. Several alternatives to fungicides have been evaluated in search for products that control this disease and have low environmental impact and toxicity to living beings. In this context, the use of Trichoderma spp. is an alternative to agrochemicals, since they are bioregulators and antagonists of several phytopathogens. This work aimed to evaluate the antagonistic and mycoparasitory action of the Trichodermil 1306 biofungicide against F. oxysporum, and its action in the control of fusarium wilt in tomato seeds and seedlings. The biofungicide was used at concentrations of 0, 50, 100, and 150% of the dose recommended for application of 20 L with a backpack sprayer. The experiment was conducted in a completely randomized design with five replications. Data were submitted to analysis of variance and mean compared by Tukey's test at 5% probability. The 100% Trichodermil concentration was the most efficient for treating tomato seeds. Although this concentration did not show differences in relation to the lowest dose used (50%) in the tomato seed antagonism, mycoparasitism and F. oxysporum incidence tests were recommended because that the lowest concentration resulted in 18% lower root protrusion than that observed with 100% dosage. The in vitro antagonistic and mycoparasitory action of this biological fungicide indicates its potential in the control of fusarium wilt in tomato crops.(AU)

A murcha-de-fusário, causada por Fusarium oxysporum f. sp. lycopersici, é uma das mais importantes doenças da cultura do tomate no Brasil. Várias alternativas aos fungicidas têm sido avaliadas na busca de produtos que controlem a doença e tenham baixo impacto ambiental e toxicidade aos seres vivos. Nesse contexto, o uso de Trichoderma spp. é uma alternativa a agrotóxicos, pois são biorreguladores e antagonistas de vários fitopatógenos. O objetivo do trabalho foi avaliar a ação antagonista e micoparasitória do biofungicida Trichodermil 1306 em F. oxysporum, e no controle da murcha-de-fusário em sementes e mudas de tomate. O biofungicida foi utilizado nas concentrações de 0, 50, 100 e 150% da dose recomendada para aplicação com pulverizador costal de 20 L. O experimento foi conduzido em delineamento inteiramente casualizado, com cinco repetições. Os dados foram submetidos à análise de variância e as médias foram comparadas pelo teste de Tukey a 5% de probabilidade. A concentração de Trichodermil 100% foi a mais eficaz para o tratamento de sementes de tomateiro. Embora essa concentração não tenha se diferenciado da menor dosagem utilizada (50%) para os testes de antagonismo, micoparasitismo e incidência de F. oxysporum em mudas de tomate, sua recomendação se justifica pelo fato da menor concentração ter propiciado protrusão radicular 18% menor do que a observada na concentração de 100%. A ação antagonista e micoparasitória in vitro do fungicida biológico testado no presente estudo indica potencialidade de uso para o controle de murcha-de-fusário em tomateiro.(AU)

Action of Trichodermil 1306 in the control of Fusarium oxysporum f. sp. lycopersici in tomato crops / Ação do Trichodermil 1306 no controle de Fusarium oxysporum f. sp. lycopersici na cultura do tomateiro

Fusarium wilt, caused by Fusarium oxysporum f. sp. lycopersici, is one of the most important diseases of tomato crop in Brazil. Several alternatives to fungicides have been evaluated in search for products that control this disease and have low environmental impact and toxicity to living beings. In this context, the use of Trichoderma spp. is an alternative to agrochemicals, since they are bioregulators and antagonists of several phytopathogens. This work aimed to evaluate the antagonistic and mycoparasitory action of the Trichodermil 1306 biofungicide against F. oxysporum, and its action in the control of fusarium wilt in tomato seeds and seedlings. The biofungicide was used at concentrations of 0, 50, 100, and 150% of the dose recommended for application of 20 L with a backpack sprayer. The experiment was conducted in a completely randomized design with five replications. Data were submitted to analysis of variance and mean compared by Tukey's test at 5% probability. The 100% Trichodermil concentration was the most efficient for treating tomato seeds. Although this concentration did not show differences in relation to the lowest dose used (50%) in the tomato seed antagonism, mycoparasitism and F. oxysporum incidence tests were recommended because that the lowest concentration resulted in 18% lower root protrusion than that observed with 100% dosage. The in vitro antagonistic and mycoparasitory action of this biological fungicide indicates its potential in the control of fusarium wilt in tomato crops.(AU)

A murcha-de-fusário, causada por Fusarium oxysporum f. sp. lycopersici, é uma das mais importantes doenças da cultura do tomate no Brasil. Várias alternativas aos fungicidas têm sido avaliadas na busca de produtos que controlem a doença e tenham baixo impacto ambiental e toxicidade aos seres vivos. Nesse contexto, o uso de Trichoderma spp. é uma alternativa a agrotóxicos, pois são biorreguladores e antagonistas de vários fitopatógenos. O objetivo do trabalho foi avaliar a ação antagonista e micoparasitória do biofungicida Trichodermil 1306 em F. oxysporum, e no controle da murcha-de-fusário em sementes e mudas de tomate. O biofungicida foi utilizado nas concentrações de 0, 50, 100 e 150% da dose recomendada para aplicação com pulverizador costal de 20 L. O experimento foi conduzido em delineamento inteiramente casualizado, com cinco repetições. Os dados foram submetidos à análise de variância e as médias foram comparadas pelo teste de Tukey a 5% de probabilidade. A concentração de Trichodermil 100% foi a mais eficaz para o tratamento de sementes de tomateiro. Embora essa concentração não tenha se diferenciado da menor dosagem utilizada (50%) para os testes de antagonismo, micoparasitismo e incidência de F. oxysporum em mudas de tomate, sua recomendação se justifica pelo fato da menor concentração ter propiciado protrusão radicular 18% menor do que a observada na concentração de 100%. A ação antagonista e micoparasitória in vitro do fungicida biológico testado no presente estudo indica potencialidade de uso para o controle de murcha-de-fusário em tomateiro.(AU)

In vitro control of Botrytis cinerea and Penicillium italicum by antagonistic yeasts / Controle in vitro de Botrytis cinerea e Penicillium italicum por leveduras antagonistas

Gray mold in grapes (caused by Botrytis cinerea) and blue mold in citrus (caused by Penicillium italicum) are post-harvest diseases that cause major losses in these crops. The control of these diseases is based on the use of synthetic chemical fungicides. The increase in regulatory policies and demand to reduce the application of pesticides, due to harmful effects on the environment and humans, have led to the search for more ecofriendly alternatives, such as biological control agents. Thus, the present work aims to verify the antagonistic potential of four yeast strains, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762), and Lachancea thermotolerans (CCMA0763), against of B cinerea and P. italicum. To assess the antagonism of volatile compounds, Petri plates with two divisions containing potato-dextrose-agar (PDA) were used by placing a fungal mycelial disc and yeast suspension (3.0 × 106 cells mL-¹) on opposite sides of the plate. The colony diameter and mycelial growth rate index of the fungi were evaluated via comparisons with the control plate without yeast. For the evaluation of the antagonism of diffusible substances in the medium, yeasts were striated 3 cm from the center of the plates containing PDA. After 48 h, a mycelial disc of each phytopathogen was placed in the center of the plates. The colony growth, inhibition halo, and mycelial growth rate index were evaluated via comparisons with the control plate. All yeast strains showed an antagonistic effect on the mycelial growth of B. cinerea in both tests. In the volatile compounds test, H. opuntiae, L. thermotolerans, P. caribbica, and P. manshurica inhibited mycelial growth by approximately 82%, 75%, 72%, and 50%, respectively. In the antagonism test of the diffusible substances in the medium, [...].

O mofo cinzento na uva (causada por Botrytis cinerea) e o bolor azul nos citros (causada por Penicillium italicum) são doenças de pós-colheita que causam perdas importantes nestas culturas. O controle destas doenças é baseado na utilização de fungicidas químicos sintéticos. O aumento das políticas regulatórias e a demanda para reduzir a aplicação de agrotóxicos, devido a efeitos prejudiciais ao meio ambiente e aos seres humanos, levam à procura de alternativas mais ecológicas como o uso de agentes de controle biológico. Desta forma, o presente trabalho teve por finalidade analisar o potencial antagônico de quatro cepas de leveduras, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762) e Lachancea thermotolerans (CCMA 0763) no controle de B. cinerea e P. italicum. Para avaliar o antagonismo por compostos voláteis, foram utilizadas placas bipartidas contendo meio batata-dextrose-ágar (BDA) onde foram colocados um disco micelial dos fungos e uma suspensão de 3,0x106 cél. mL-¹ das leveduras em lados opostos da placa. Avaliou-se o diâmetro da colônia e índice de velocidade de crescimento micelial dos fungos, comparando-a com a placa controles em a levedura. Para a avaliação do antagonismo de substâncias difusíveis no meio, as leveduras foram estriadas a 3 cm do centro de placas contendo meio BDA e após 48 h colocou-se um disco micelial de cada fitopatógeno no centro das mesmas. Avaliou-se o crescimento da colônia, a formação do halo de inibição e o índice de velocidade de crescimento micelial comparando-os com a placa controle. Todos os isolados apresentaram efeito antagônico no crescimento micelial de B. cinerea nos dois testes realizados. H. opuntiae, L. thermotolerans, P. caribbica e P. manshurica inibiram o crescimento micelial em aproximadamente 82%, 75%, 72% e 50%, respectivamente, no teste de compostos voláteis. P. caribbica e P. manshurica inibiram o crescimento micelial em 58% e 33%, [...].

In vitro control of Botrytis cinerea and Penicillium italicum by antagonistic yeasts / Controle in vitro de Botrytis cinerea e Penicillium italicum por leveduras antagonistas

Gray mold in grapes (caused by Botrytis cinerea) and blue mold in citrus (caused by Penicillium italicum) are post-harvest diseases that cause major losses in these crops. The control of these diseases is based on the use of synthetic chemical fungicides. The increase in regulatory policies and demand to reduce the application of pesticides, due to harmful effects on the environment and humans, have led to the search for more ecofriendly alternatives, such as biological control agents. Thus, the present work aims to verify the antagonistic potential of four yeast strains, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762), and Lachancea thermotolerans (CCMA0763), against of B cinerea and P. italicum. To assess the antagonism of volatile compounds, Petri plates with two divisions containing potato-dextrose-agar (PDA) were used by placing a fungal mycelial disc and yeast suspension (3.0 × 106 cells mL-¹) on opposite sides of the plate. The colony diameter and mycelial growth rate index of the fungi were evaluated via comparisons with the control plate without yeast. For the evaluation of the antagonism of diffusible substances in the medium, yeasts were striated 3 cm from the center of the plates containing PDA. After 48 h, a mycelial disc of each phytopathogen was placed in the center of the plates. The colony growth, inhibition halo, and mycelial growth rate index were evaluated via comparisons with the control plate. All yeast strains showed an antagonistic effect on the mycelial growth of B. cinerea in both tests. In the volatile compounds test, H. opuntiae, L. thermotolerans, P. caribbica, and P. manshurica inhibited mycelial growth by approximately 82%, 75%, 72%, and 50%, respectively. In the antagonism test of the diffusible substances in the medium, [...].(AU)

O mofo cinzento na uva (causada por Botrytis cinerea) e o bolor azul nos citros (causada por Penicillium italicum) são doenças de pós-colheita que causam perdas importantes nestas culturas. O controle destas doenças é baseado na utilização de fungicidas químicos sintéticos. O aumento das políticas regulatórias e a demanda para reduzir a aplicação de agrotóxicos, devido a efeitos prejudiciais ao meio ambiente e aos seres humanos, levam à procura de alternativas mais ecológicas como o uso de agentes de controle biológico. Desta forma, o presente trabalho teve por finalidade analisar o potencial antagônico de quatro cepas de leveduras, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762) e Lachancea thermotolerans (CCMA 0763) no controle de B. cinerea e P. italicum. Para avaliar o antagonismo por compostos voláteis, foram utilizadas placas bipartidas contendo meio batata-dextrose-ágar (BDA) onde foram colocados um disco micelial dos fungos e uma suspensão de 3,0x106 cél. mL-¹ das leveduras em lados opostos da placa. Avaliou-se o diâmetro da colônia e índice de velocidade de crescimento micelial dos fungos, comparando-a com a placa controles em a levedura. Para a avaliação do antagonismo de substâncias difusíveis no meio, as leveduras foram estriadas a 3 cm do centro de placas contendo meio BDA e após 48 h colocou-se um disco micelial de cada fitopatógeno no centro das mesmas. Avaliou-se o crescimento da colônia, a formação do halo de inibição e o índice de velocidade de crescimento micelial comparando-os com a placa controle. Todos os isolados apresentaram efeito antagônico no crescimento micelial de B. cinerea nos dois testes realizados. H. opuntiae, L. thermotolerans, P. caribbica e P. manshurica inibiram o crescimento micelial em aproximadamente 82%, 75%, 72% e 50%, respectivamente, no teste de compostos voláteis. P. caribbica e P. manshurica inibiram o crescimento micelial em 58% e 33%, [...].(AU)

Bioprospection and molecular phylogeny of culturable endophytic fungi associated with yellow passion fruit

Endophytic microorganisms live inside the plants without causing any damage to their hosts. In the agricultural field, these endophytes might be a strategy of biological control for phytopathogens. We aimed to isolate endophytic fungifrom yellowpassion fruit (Passiflora edulis) leaves, evaluating its biocontrol capacity by in vitroantagonism against phytopathogen Colletotrichum sp. CNPU378. We also carried out greenhouse experiments in bean seedlings. A high colonization frequency was obtained (89%), and the molecular identification based on DNA sequencing attested Colletotrichumas the most frequent genus and minor occurrence of Curvulariaendophytes. The endophytes tested showed different types of competitive interactions in in vitro antagonism inhibition rate ranging from 28.8 to 48.8%. There were 10 promising antagonists tested for their antagonist activity of crude extracts of secondary metabolites, in which strain PE-36 (20.8%) stood out among the other strains evaluated. In the greenhouse assay, plants inoculated only with endophyte Colletotrichumsp. PE-36 was symptomless and suggest that the endophyte strengthened the growth promotion in common bean plants, especially in the root length and number of leaves when compared to control plantsand other treatments. Despite many fungiof Colletotrichumgenus being described as causative agents of anthracnose, in this study, the plant sampled was colonized predominantly by Colletotrichumendophytes living in asymptomatic relationship. By the way,we come across a Colletotrichumsp. endophyte able to antagonize a Colletotrichumsp. pathogen

Identification and susceptibility of clinical isolates of Candida spp. to killer toxins / Identificação e susceptibilidade de isolados clínicos de Candida spp. para toxinas assassinas

Although invasive infections and mortality caused by Candida species are increasing among compromised patients, resistance to common antifungal agents is also an increasing problem. We analyzed 60 yeasts isolated from patients with invasive candidiasis using a PCR/RFLP strategy based on the internal transcribed spacer (ITS2) region to identify different Candida pathogenic species. PCR analysis was performed from genomic DNA with a primer pair of the ITS2-5.8S rDNA region. PCR-positive samples were characterized by RFLP. Restriction resulted in 23 isolates identified as C. albicans using AlwI, 24 isolates as C. parapsilosis using RsaI, and 13 as C. tropicalis using XmaI. Then, a group of all isolates were evaluated for their susceptibility to a panel of previously described killer yeasts, resulting in 75% being susceptible to at least one killer yeast while the remaining were not inhibited by any strain. C. albicans was the most susceptible group while C. tropicalis had the fewest inhibitions. No species-specific pattern of inhibition was obtained with this panel of killer yeasts. Metschnikowia pulcherrima, Pichia kluyveri and Wickerhamomyces anomalus were the strains that inhibited the most isolates of Candida spp.(AU)

Embora as infecções invasivas e a mortalidade causada por espécies de Candida estejam aumentando entre pacientes comprometidos, a resistência a agentes antifúngicos comuns também é um problema crescente. Analisamos 60 leveduras isoladas de pacientes com candidíase invasiva utilizando como estratégia PCR/RFLP baseada na região espaçadora transcrita interna (ITS2) para identificar diferentes espécies patogênicas de Candida. A análise por PCR foi realizada a partir de ADN genómico com um par de iniciadores da região ITS2-5.8S rDNA. As amostras PCR-positivas foram caracterizadas por RFLP. A restrição resultou em 23 isolados identificados como C. albicans usando AlwI, 24 isolados como C. parapsilosis usando RsaI e 13 como C. tropicalis usando XmaI. Em seguida, avaliou-se o grupo de todos os isolados quanto à sua susceptibilidade a um painel de leveduras killer previamente descritas, resultando em 75% sendo suscetíveis a pelo menos uma levedura killer, enquanto que as restantes não foram inibidas por qualquer cepa. C. albicans foi o grupo mais suscetível enquanto C. tropicalis teve o menor número de inibições. Não se obteve um padrão de inibição específico da espécie com este painel de leveduras killer. Metschnikowia pulcherrima, Pichia kluyveri e Wickerhamomyces anomalus foram as cepas que inibiram a maioria dos isolados de Candida spp.(AU)

Biological control of bacterial spot of tomato by saprobe fungi from semi-arid areas of northeastern Brazil / Controle biológico da mancha bacteriana do tomateiro usando fungos sapróbios do semiárido nordestino do Brasil

Tomato bacterial spot caused by Xanthomonas spp., is a common disease in tomato fields that causes significant economic losses. Due to the difficulty with control of bacterial spot by conventional methods, new techniques such as biological control and induction of resistance are gaining prominence. This study aimed to select saprobe fungi from semi-arid regions of the Brazilian Northeast for the biological control of bacterial spot of tomato. To select the best isolates to control bacterial spot, a greenhouse experiment was initially conducted. Tomato plants (‘Santa Cruz Kada) were treated with filtrates of 25 saprobe fungi and inoculated three days later with Xanthomonas euvesicatoria. Filtrates of Memnoniella levispora, Periconia hispidula, Zygosporium echinosporum, and Chloridium virescens var. virescens were selected as the most effective. Filtrates and volatile compounds from these four isolates were tested for their antibacterial activity in cultures of X. euvesicatoria and in tomato plants (‘Santa Cruz Kada) inoculated with X. euvesicatoria. In vitro, the addition of nonvolatile fungal metabolites into the culture medium at 5% and 50% (v/v) inhibited bacterial growth by 28.9% and 53.8%, respectively. The volatile compounds produced by C. virescens var. virescens reduced the number of colony-forming units of X. euvesicatoria by 25.9%. In vivo, all treatments reduced from 62.4 to 71.3% the area under bacterial spot progress curve, showing the same control efficacy as the commercial resistance inducer used as a positive control (acibenzolar-S-methyl). Systemicity of the fungal filtrates was confirmed in a separate experiment, where application of the treatments exclusively to the third leaf decreased the severity of the disease on the fourth leaf (except for C. virescens var. virescens)...(AU)

A mancha bacteriana do tomateiro causada por Xanthomonas spp. é uma doença frequente nos campos de cultivo, provocando importantes perdas econômicas. Devido à dificuldade de controle com os métodos tradicionais, novas ferramentas de manejo para esta doença, como o controle biológico e a indução de resistência vêm ganhando destaque. O objetivo desse trabalho foi a seleção de fungos sapróbios do semiárido nordestino brasileiro para o controle biológico da mancha bacteriana do tomateiro. Inicialmente, com o objetivo de selecionar os isolados que apresentassem melhor controle da mancha bacteriana, foi montado um ensaio em casa de vegetação com tomateiros (‘Santa Cruz Kada) tratados com os filtrados de 25 fungos sapróbios e inoculados com a bactéria Xanthomonas euvesicatoria três dias depois. Foram selecionados os isolados de Memnoniella levispora, Periconia hispidula, Zygosporium echinosporum e Chloridium virescens var. virescens. Filtrados e voláteis fúngicos desses quatro isolados foram testados in vitro, para se determinar a sensibilidade da bactéria fitopatogênica, e in vivo em tomateiros (‘Santa Cruz Kada) tratados e inoculados com X. euvesicatoria. A incorporação de metabólitos não voláteis dos fungos no meio de cultura a 5% e 50% (v/v) controlou o crescimento da bactéria em até 28,9% e 53,8%, respectivamente. Os compostos voláteis produzidos por C. virescens var. virescens inibiram até 25,9% das unidades formadoras de colônias. Com relação ao experimento in vivo, todos os tratamentos testados reduziram de 62,4 a 71,3% a área abaixo da curva de progresso da mancha bacteriana, mostrando nível de controle igual ao indutor de resistência comercial utilizado como controle positivo (acibenzolar-S-metílico). A sistemicidade deste controle foi confirmada em outro experimento, onde a aplicação dos tratamentos somente na terceira folha determinou menor severidade da doença na quarta folha (exceto para C. virescens var. virescens)...(AU)

Biological control of bacterial spot of tomato by saprobe fungi from semi-arid areas of northeastern Brazil / Controle biológico da mancha bacteriana do tomateiro usando fungos sapróbios do semiárido nordestino do Brasil

Tomato bacterial spot caused by Xanthomonas spp., is a common disease in tomato fields that causes significant economic losses. Due to the difficulty with control of bacterial spot by conventional methods, new techniques such as biological control and induction of resistance are gaining prominence. This study aimed to select saprobe fungi from semi-arid regions of the Brazilian Northeast for the biological control of bacterial spot of tomato. To select the best isolates to control bacterial spot, a greenhouse experiment was initially conducted. Tomato plants (‘Santa Cruz Kada’) were treated with filtrates of 25 saprobe fungi and inoculated three days later with Xanthomonas euvesicatoria. Filtrates of Memnoniella levispora, Periconia hispidula, Zygosporium echinosporum, and Chloridium virescens var. virescens were selected as the most effective. Filtrates and volatile compounds from these four isolates were tested for their antibacterial activity in cultures of X. euvesicatoria and in tomato plants (‘Santa Cruz Kada’) inoculated with X. euvesicatoria. In vitro, the addition of nonvolatile fungal metabolites into the culture medium at 5% and 50% (v/v) inhibited bacterial growth by 28.9% and 53.8%, respectively. The volatile compounds produced by C. virescens var. virescens reduced the number of colony-forming units of X. euvesicatoria by 25.9%. In vivo, all treatments reduced from 62.4 to 71.3% the area under bacterial spot progress curve, showing the same control efficacy as the commercial resistance inducer used as a positive control (acibenzolar-S-methyl). Systemicity of the fungal filtrates was confirmed in a separate experiment, where application of the treatments exclusively to the third leaf decreased the severity of the disease on the fourth leaf (except for C. virescens var. virescens)...

A mancha bacteriana do tomateiro causada por Xanthomonas spp. é uma doença frequente nos campos de cultivo, provocando importantes perdas econômicas. Devido à dificuldade de controle com os métodos tradicionais, novas ferramentas de manejo para esta doença, como o controle biológico e a indução de resistência vêm ganhando destaque. O objetivo desse trabalho foi a seleção de fungos sapróbios do semiárido nordestino brasileiro para o controle biológico da mancha bacteriana do tomateiro. Inicialmente, com o objetivo de selecionar os isolados que apresentassem melhor controle da mancha bacteriana, foi montado um ensaio em casa de vegetação com tomateiros (‘Santa Cruz Kada’) tratados com os filtrados de 25 fungos sapróbios e inoculados com a bactéria Xanthomonas euvesicatoria três dias depois. Foram selecionados os isolados de Memnoniella levispora, Periconia hispidula, Zygosporium echinosporum e Chloridium virescens var. virescens. Filtrados e voláteis fúngicos desses quatro isolados foram testados in vitro, para se determinar a sensibilidade da bactéria fitopatogênica, e in vivo em tomateiros (‘Santa Cruz Kada’) tratados e inoculados com X. euvesicatoria. A incorporação de metabólitos não voláteis dos fungos no meio de cultura a 5% e 50% (v/v) controlou o crescimento da bactéria em até 28,9% e 53,8%, respectivamente. Os compostos voláteis produzidos por C. virescens var. virescens inibiram até 25,9% das unidades formadoras de colônias. Com relação ao experimento in vivo, todos os tratamentos testados reduziram de 62,4 a 71,3% a área abaixo da curva de progresso da mancha bacteriana, mostrando nível de controle igual ao indutor de resistência comercial utilizado como controle positivo (acibenzolar-S-metílico). A sistemicidade deste controle foi confirmada em outro experimento, onde a aplicação dos tratamentos somente na terceira folha determinou menor severidade da doença na quarta folha (exceto para C. virescens var. virescens)...

Exploring plant growth-promotion actinomycetes from vermicompost and rhizosphere soil for yield enhancement in chickpea

The main objective of the present study was to isolate and characterize actinomycetes for their plant growth-promotion in chickpea. A total of 89 actinomycetes were screened for their antagonism against fungal pathogens of chickpea by dual culture and metabolite production assays. Four most promising actinomycetes were evaluated for their physiological and plant growth-promotion properties under in vitro and in vivo conditions. All the isolates exhibited good growth at temperatures from 20 °C to 40 °C, pH range of 711 and NaCl concentrations up to 8%. These were also found highly tolerant to Bavistin, slightly tolerant to Thiram and Captan (except VAI-7 and VAI-40) but susceptible to Benlate and Ridomil at field application levels and were found to produce siderophore, cellulase, lipase, protease, chitinase (except VAI-40), hydrocyanic acid (except VAI-7 and VAI-40), indole acetic acid and -1,3-glucanase. When the four actinomycetes were evaluated for their plant growth-promotion properties under field conditions on chickpea, all exhibited increase in nodule number, shoot weight and yield. The actinomycetes treated plots enhanced total N, available P and organic C over the un-inoculated control. The scanning electron microscope studies exhibited extensive colonization by actinomycetes on the root surface of chickpea. The expression profiles for indole acetic acid, siderophore and -1,3-glucanase genes exhibited up-regulation for all three traits and in all four isolates. The actinomycetes were identified as Streptomyces but different species in the 16S rDNA analysis. It was concluded that the selected actinomycetes have good plant growth-promotion and biocontrol potentials on chickpea. (AU)

Antifungal and proteolytic activities of endophytic fungi isolated from Piper hispidum Sw

Endophytes are being considered for use in biological control, and the enzymes they secrete might facilitate their initial colonization of internal plant tissues and direct interactions with microbial pathogens. Microbial proteases are also biotechnologically important products employed in bioremediation processes, cosmetics, and the pharmaceutical, photographic and food industries. In the present study, we evaluated antagonism and competitive interactions between 98 fungal endophytes and Alternaria alternata, Colletotrichum sp., Phyllosticta citricarpa and Moniliophthora perniciosa. We also examined the proteolytic activities of endophytes grown in liquid medium and conducted cup plate assays. The results showed that certain strains in the assemblage of P. hispidum endophytes are important sources of antifungal properties, primarily Lasiodiplodia theobromae JF766989, which reduced phytopathogen growth by approximately 54 to 65%. We detected 28 endophytes producing enzymatic halos of up to 16.40 mm in diameter. The results obtained in the present study highlight the proteolytic activity of the endophytes Phoma herbarum JF766995 and Schizophyllum commune JF766994, which presented the highest enzymatic halo diameters under at least one culture condition tested. The increased activities of certain isolates in the presence of rice or soy flour as a substrate (with halos up to 17.67 mm in diameter) suggests that these endophytes have the potential to produce enzymes using agricultural wastes.(AU)

Synergistic effects of tacrolimus and azole antifungal compounds in fluconazole-susceptible and fluconazole-resistant Candida glabrata isolates

In vitro interaction between tacrolimus (FK506) and four azoles (fluconazole, ketoconazole, itraconazole and voriconazole) against thirty clinical isolates of both fluconazole susceptible and -resistant Candida glabrata were evaluated by the checkerboard microdilution method. Synergistic, indifferent or antagonism interactions were found for combinations of the antifungal agents and FK506. A larger synergistic effect was observed for the combinations of FK506 with itraconazole and voriconazole (43%), followed by that of the combination with ketoconazole (37%), against fluconazole-susceptible isolates. For fluconazole-resistant C. glabrata, a higher synergistic effect was obtained from FK506 combined with ketoconazole (77%), itraconazole (73%), voriconazole (63%) and fluconazole (60%). The synergisms that we observed in vitro, notably against fluconazole-resistant C. glabrata isolates, are promising and warrant further analysis of their applications in experimental in vivo studies.(AU)

Production of lytic enzymes by Trichoderma isolates during in vitro antagonism with Aspergillus niger, the causal agent of collar rot of peanut

Twelve isolates of Trichoderma (six of T. harzianum, five of T. viride, one of T. virens), which reduced variably the incidence of collar rot disease caused in peanut by Aspergillus niger Van Tieghem, were evaluated for their potential to produce lytic enzymes during in vitro antagonism. T. viride 60 inhibited highest (86.2%) growth of test fungus followed by T. harzianum 2J (80.4%) at 6 days after inoculation (DAI) on PDA media. The specific activities of chitinase, -1,3-glucanase and protease were 11, 3.46 and 9 folds higher in T6 antagonist (T. viride 60 and A. niger interactions) followed by 8.72, 2.85 and 9 folds in T8 antagonist (T. harzianum 2J and A. niger interactions), respectively, compared to the activity produced by control petri plate T13 (A. niger alone) at 6 DAI. Activity of these lytic enzymes induced in antagonists' plates comprises the growth of Trichoderma isolates. However, cellulase and poly galacturonase were found least amount in these antagonists treatment. A significant positive correlation (p=0.01) between percentage growth inhibition of test fungus and lytic enzymes - (chitinase, -1,3-glucanase and protease) in the culture medium of antagonist treatment established a relationship to inhibit growth of fungal pathogen by increasing the levels of these enzymes. Among the Trichoderma isolates, T. viride 60 was found best strain to be used in biological control of plant pathogen A. niger.

Production of lytic enzymes by Trichoderma isolates during in vitro antagonism with Aspergillus niger, the causal agent of collar rot of peanut

Twelve isolates of Trichoderma (six of T. harzianum, five of T. viride, one of T. virens), which reduced variably the incidence of collar rot disease caused in peanut by Aspergillus niger Van Tieghem, were evaluated for their potential to produce lytic enzymes during in vitro antagonism. T. viride 60 inhibited highest (86.2%) growth of test fungus followed by T. harzianum 2J (80.4%) at 6 days after inoculation (DAI) on PDA media. The specific activities of chitinase, -1,3-glucanase and protease were 11, 3.46 and 9 folds higher in T6 antagonist (T. viride 60 and A. niger interactions) followed by 8.72, 2.85 and 9 folds in T8 antagonist (T. harzianum 2J and A. niger interactions), respectively, compared to the activity produced by control petri plate T13 (A. niger alone) at 6 DAI. Activity of these lytic enzymes induced in antagonists' plates comprises the growth of Trichoderma isolates. However, cellulase and poly galacturonase were found least amount in these antagonists treatment. A significant positive correlation (p=0.01) between percentage growth inhibition of test fungus and lytic enzymes - (chitinase, -1,3-glucanase and protease) in the culture medium of antagonist treatment established a relationship to inhibit growth of fungal pathogen by increasing the levels of these enzymes. Among the Trichoderma isolates, T. viride 60 was found best strain to be used in biological control of plant pathogen A. niger.

Characterization of Bacillus isolates of potato rhizosphere from andean soils of Peru and their potential PGPR characteristics

Bacillus spp. are well known rhizosphere residents of many crops and usually show plant growth promoting (PGP) activities that include biocontrol capacity against some phytopatogenic fungi. Potato crops in the Andean Highlands of Peru face many nutritional and phytophatogenic problems that have a significant impact on production. In this context is important to investigate the natural presence of these microorganisms in the potato rhizosphere and propose a selective screening to find promising PGP strains. In this study, sixty three Bacillus strains isolated from the rhizosphere of native potato varieties growing in the Andean highlands of Peru were screened for in vitro antagonism against Rhizoctonia solani and Fusarium solani. A high prevalence (68%) of antagonists against R. solani was found. Ninety one percent of those strains also inhibited the growth of F. solani. The antagonistic strains were also tested for other plant growth promotion activities. Eighty one percent produced some level of the auxin indole-3-acetic acid, and 58% solubilized tricalcium phosphate. Phylogenetic analysis revealed that the majority of the strains belonged to the B. amyloliquefaciens species, while strains Bac17M11, Bac20M1 and Bac20M2 may correspond to a putative new Bacillus species. The results suggested that the rhizosphere of native potatoes growing in their natural habitat in the Andes is a rich source of Bacillus fungal antagonists, which have a potential to be used in the future as PGP inoculants to improve potato crop.

Bacillus subtilis induces morphological changes in Fonsecaea pedrosoi in vitro resulting in more resistant fungal forms in vivo

Interactions among microorganisms may be the cause of morphological modifications, particularly in fungal cells. The aim of this work was to examine the changes that occur in cells of the fungus Fonsecaea pedrosoi after in vitro co-culturing with Bacillus subtilis and to explore the results of this interaction in vivo in an experimental murine infection. B. subtilis strain was inoculated into a 15-day pure culture of F. pedrosoi. In vitro, after 48 hours of co-culturing, the fungal cells were roundish. The secretion of fungal dark pigments and production of terminal chlamydoconidia were observed in hyphae after one week. In the in vivo study, two animal groups of 30 BALB/c mice each were employed. One group was inoculated intraperitoneally with hyphal fragments from the co-culture of bacteria and fungi; the other group was infected only with F. pedrosoi hyphae. After seven days of infection, both animal groups developed neutrophilic abscesses. Phagocytosis of bacilli by macrophages occurred at three days. At later periods, generally after 25 days, only roundish cells similar to sclerotic bodies remained in the tissues while hyphae were eliminated by 15 to 20 days. These fungal forms originated mainly from terminal chlamydoconidia. The co-culturing between bacteria and fungi may constitute a mechanism to rapidly obtain resistant fungal forms for host defenses, especially for chromoblastomycosis (CBM) experimental infections.(AU)

Antagonistic in vitro activity of Trichoderma spp. isolates to the fungi Phytophthora citrophthora / Atividade antagônica in vitro de isolados de Trichoderma spp. ao fungo Phytophthora citrophthora

Gummosis  is  among  the main  fungal  diseases  of  the  citrus.  It  is  caused  by Phytophthora sp.  and usually shows up in the lap of the plant, provoking rottenness and gum exudation, and expands causing  the plant death for constrictions in the cambium or phloem which interrupts the descending fow of sap. The objective of this work was to evaluate the antagonistic in vitro activity of Trichoderma spp. to the fungi Phytophthora citrophthora. Phytophthora citrophthora was exposed to fve environments of antagonism (without antagonist and with four strains of Trichoderma viride, T. virens, T. harzianu and T. stromaticum). The  in vitro essay was accomplished through the method of paired cultures. A completely randomized desing was used with fve treatments and three replications, and each plot was represented by three petri dishes. The isolates of Trichoderma demonstrated signifcant effect in the inhibition of the mycelial growth of the fungi Phytophthora citrophthora, and the fungi Trichoderma stromaticum presented larger antagonism to the fungi P. citrophthora while the T. harzianum presented antagonism smaller.

A citricultura  nacional  apresenta  inúmeras  pragas  e  doenças  que  limitam  sua  produção  dentre  as  principais doenças fúngicas da cultura dos citros pode se destacar a gomose. A gomose causada por  Phytophthora  sp. geralmente  se manifesta no colo da planta, provocando podridão e exsudação de  goma, podendo expandir-se ocasionando a morte da planta por estrangulamento devido ao ataque do  cambio ou foema, o que  interrompe o fuxo descendente de seiva. Este  trabalho  foi  realizado com  o objetivo de  se avaliar a atividade antagônica  in vitro, de  isolados de Trichoderma  spp.  ao  fungo  Phytophthora citrophthora. O fator em estudo foi um patógeno (Phytophthora citrophthora) em cinco  ambientes de antagonismo  (sem antagonista e com os  isolados de Trichoderma viride, T. virens, T.  harzianu e T. stromaticum). O ensaio in vitro foi realizado por meio do método de cultura pareada. O delineamento experimental utilizado foi inteiramente casualizado, com 5 tratamentos e 3 repetições, e  cada parcela foi representada por três placas de Petri. Os isolados de Trichoderma demonstraram efeito  signifcativo na  inibição do crescimento micelial do  fungo Phytophthora citrophthora, sendo que o  fungo Trichoderma stromaticum apresentou maior antagonismo ao fungo P. citrophthora enquanto que o T. harzianum apresentou menor antagonismo.