Inhibition of mycelial growth, conidial germination, and Botrytis cinerea Pers: Fr colonization in begonia with biocompatible products

This study evaluated the effects of potassium and sodium carbonate and bicarbonate, Bacillus subtilis (Cohn, 1872) QST-713, Bacillus pumilus (Meyer & Gottheil, 1901) QST-2808, and crude and roasted coffee oils on the inhibition of mycelial growth and conidial germination in Botrytis cinerea Pers.: Fr and the colonization of begonia (Begonia elatior Hort. ex Steud) leaf discs by B. cinerea inoculated before, simultaneously and after with these alternative products. The assays were carried out using the Baladin begonia cultivar. The inhibition of B. cinerea mycelial growth and conidial germination was proportional to increases in the concentration of all the products. The inhibition of conidial germination was directly proportional to the concentrations of B. pumilus QST-2808 and B. subtilis QST-713. Coffee oils were less efficient in inhibiting germination than the other products. The crude and roasted coffee oils, potassium and sodium carbonates and bicarbonates, and B. pumilus and B. subtilis sprayed 24 h before, simultaneously, or 24 h after pathogen inoculation inhibited the colonization of begonia leaf discs by B. cinerea. The positive results for the suppression of B. cinerea by the alternative products tested herein merit scrutiny. There is a pressing need to evaluate these products in the management of gray mold, as the severity of this disease is usually high under favorable conditions in greenhouses.

Prohexadione-calcium on downy mildew and botrytis bunch rot epidemiology, and yield and technological and phenolic parameters of the Merlot vine cultivar in the highland region of southern Brazil / Efeitos da prohexadiona de cálcio na epidemiologia do mildio e podridão de botrytis e no desempenho vitícola da cultivar de videira Merlot em região de altitude do sul do Brasil

Prohexadione-calcium (ProCa) is a gibberellin biosynthesis inhibitor and the objective of this study was to evaluate the effect of ProCa on the downy mildew (DM) and Botrytis bunch rot (BBR) epidemiology, and the yield and technological and phenolic parameters of grape Merlot cultivar. Experiments were carried out in a commercial vineyard in São Joaquim Municipality/SC, Southern Brazil, during the 2017-2018 and 2018-2019 growing seasons. ProCa was applied at four doses and three phenological stages: A) 0 g ha1 (control); B) 1000 g ha1 (inflorescence fully developed); C) 500 + 500 g ha1 (inflorescence fully developeted and full flowering); D) 500 + 500 + 500 g ha1 (inflorescence fully developed, full flowering, and berries pea-sized). DM and BBR incidence and severity were quantified weekly from the first symptom appearance until harvest, and their epidemiology was compared according to: a) the beginning of symptom appearance; b) the time to reach the maximum disease incidence and severity; c) the maximum value of disease incidence and severity; d) the area under the disease progress curve. In general, there were significant differences in the ProCa doses for all epidemiologic parameters of DM and BBR compared with the control plot; however, there was no significant difference among the ProCa doses. The principal epidemiological variables that differentiated the effect of ProCa on the DM and BBR control were the Smax and AUSDPC. Some yield and technological and phenolic parameters were negatively affected by different doses of ProCa, but it was still a good option for DM and BBR control in highland region of southern Brazil during the 2017-2018 and 2018-2019 growing seasons.(AU)

A prohexadiona-cálcio (ProCa) é um inibidor da biossíntese de giberelina e o objetivo desse estudo foi avaliar o efeito do ProCa na epidemiologia do míldio da videira (MV) e da podridão de Botrytis (PB) e no desempenho vitícola da cultivar de videira Merlot. Experimentos foram realizados em um vinhedo comercial no município de São Joaquim/SC, Sul do Brasil, durante as safras 2017-2018 e 2018-2019. A ProCa foi aplicada em quatro doses e três estágios fenológicos: A) 0 g ha-1 (controle); B) 1000 g ha-1 (inflorescência totalmente desenvolvida); C) 500 + 500 g ha-1 (inflorescência totalmente desenvolvida e floração completa); D) 500 + 500 + 500 g ha-1 (inflorescência totalmente desenvolvida, floração completa e baga tipo ervilha). A incidência e severidade de MV e PB foram quantificadas semanalmente a partir do aparecimento dos primeiros sintomas até a colheita e a epidemiologia das doenças foram comparadas de acordo com: a) o início do aparecimento dos sintomas; b) tempo para atingir a máxima incidência e severidade da doença; c) valor máximo da incidência e severidade da doença; d) área abaixo da curva de progresso da doença. Em geral, houve diferenças significativas na dose de ProCa para todos os parâmetros epidemiológicos de MV e PB em comparação com o controle; no entanto, não houve diferença significativa entre as doses de ProCa. As principais variáveis epidemiológicas que diferenciaram o efeito da ProCa no controle do MV e PB foram o Smax e o AUSDPC. Algumas variáveis vitícolas da cultivar Merlot foram afetados negativamente por diferentes doses de ProCa, mas ainda foi uma boa opção o uso da ProCa para o controle do MV e PB em região de altitude do sul do Brasil durante as safras 2017-2018 e 2018-2019.(AU)

Arabidopsis adc-silenced line exhibits differential defense responses to Botrytis cinerea and Pseudomonas syringae infection.

During plant-microbe interactions, polyamines participate in the plant defense response. Previously, we reported that silencing of ADC genes in Arabidopsis thaliana causes a drastic reduction of polyamine levels as well as increments in reactive oxygen species content. In this study, we examined the response of the adc-silenced line to Botrytis cinerea and Pseudomonas syringae infection. The adc-silenced line was more susceptible to Botrytis cinerea, showing larger lesion length and a higher incidence of fungal infection. Pre-treatments with putrescine reestablished the response of the adc-silenced line to Botrytis cinerea, resulting in a similar phenotype to the parental plant. Expression levels of defense-related genes were analyzed during fungal infection showing that the salicylic acid-induced gene PR1 was up-regulated, while the jasmonic acid-related genes LOX3 and PDF1.2, as well as, the camalexin biosynthetic gene PAD3 were down-regulated in the adc-silenced line. Furthermore, methyl jasmonate pre-treatments reduced Botrytis cinerea infection in the adc-silenced line. On the other hand, the adc-silenced line showed an increased resistance to Pseudomonas syringae infection. SA-related genes such as PR1, ZAT1.2, WRKY54 and WRKY70 were highly expressed in the adc-silenced line upon bacterial interaction. Our data show that the adc-silenced line has altered the defense-response against Botrytis cinerea and Pseudomonas syringae, that is consistent with deregulation of SA- and JA-mediated response pathways.

Defects in the Ferroxidase That Participates in the Reductive Iron Assimilation System Results in Hypervirulence in Botrytis Cinerea.

The plant pathogen Botrytis cinerea is responsible for gray-mold disease, which infects a wide variety of species. The outcome of this host-pathogen interaction, a result of the interplay between plant defense and fungal virulence pathways, can be modulated by various environmental factors. Among these, iron availability and acquisition play a crucial role in diverse biological functions. How B. cinerea obtains iron, an essential micronutrient, during infection is unknown. We set out to determine the role of the reductive iron assimilation (RIA) system during B. cinerea infection. This system comprises the BcFET1 ferroxidase, which belongs to the multicopper oxidase (MCO) family of proteins, and the BcFTR1 membrane-bound iron permease. Gene knockout and complementation studies revealed that, compared to the wild type, the bcfet1 mutant displays delayed conidiation, iron-dependent sclerotium production, and significantly reduced whole-cell iron content. Remarkably, this mutant exhibited a hypervirulence phenotype, whereas the bcftr1 mutant presents normal virulence and unaffected whole-cell iron levels and developmental programs. Interestingly, while in iron-starved plants wild-type B. cinerea produced slightly reduced necrotic lesions, the hypervirulence phenotype of the bcfet1 mutant is no longer observed in iron-deprived plants. This suggests that B. cinerea bcfet1 knockout mutants require plant-derived iron to achieve larger necrotic lesions, whereas in planta analyses of reactive oxygen species (ROS) revealed increased ROS levels only for infections caused by the bcfet1 mutant. These results suggest that increased ROS production, under an iron sufficiency environment, at least partly underlie the observed infection phenotype in this mutant.IMPORTANCE The plant-pathogenic fungus B. cinerea causes enormous economic losses, estimated at anywhere between $10 billion and $100 billion worldwide, under both pre- and postharvest conditions. Here, we present the characterization of a loss-of-function mutant in a component involved in iron acquisition that displays hypervirulence. While in different microbial systems iron uptake mechanisms appear to be critical to achieve full pathogenic potential, we found that the absence of the ferroxidase that is part of the reductive iron assimilation system leads to hypervirulence in this fungus. This is an unusual and rather underrepresented phenotype, which can be modulated by iron levels in the plant and provides an unexpected link between iron acquisition, reactive oxygen species (ROS) production, and pathogenesis in the Botrytis-plant interaction.

The fungal subtilase AsES elicits a PTI-like defence response in Arabidopsis thaliana plants independently of its enzymatic activity.

Acremonium strictum elicitor subtilisin (AsES) is a 34-kDa serine-protease secreted by the strawberry fungal pathogen A. strictum. On AsES perception, a set of defence reactions is induced, both locally and systemically, in a wide variety of plant species and against pathogens of alternative lifestyles. However, it is not clear whether AsES proteolytic activity is required for triggering a defence response or if the protein itself acts as an elicitor. To investigate the necessity of the protease activity to activate the defence response, AsES coding sequences of the wild-type gene and a mutant on the active site (S226A) were cloned and expressed in Escherichia coli. Our data show that pretreatment of Arabidopsis plants with inactive proteins, i.e. inhibited with phenylmethylsulphonyl fluoride (PMSF) and mutant, resulted in an increased systemic resistance to Botrytis cinerea and expression of defence-related genes in a temporal manner that mimics the effect already reported for the native AsES protein. The data presented in this study indicate that the defence-eliciting property exhibited by AsES is not associated with its proteolytic activity. Moreover, the enhanced expression of some immune marker genes, seedling growth inhibition and the involvement of the co-receptor BAK1 observed in plants treated with AsES suggests that AsES is being recognized as a pathogen-associated molecular pattern by a leucine-rich repeat receptor. The understanding of the mechanism of action of AsES will contribute to the development of new breeding strategies to confer durable resistance in plants.

Control of postharvest gray mold of 'BRS Nubia' table grape under cold storage / Controle de mofo cinzento na pós colheita da uva de mesa 'BRS Nubia' sob armazenamento refrigerado

The demand for high-quality nutritional products has increased fruit consumption, as grapes, for this reason postharvest techniques are required to prevent losses, to preserve quality, to extend shelf life, and to attend to consumer needs. In this way, the objective of this study was to evaluate strategies to control gray mold caused by Botrytis cinerea in 'BRS Nubia' grapes during cold storage and shelf life periods. Grape bunches were harvested from a commercial vineyard in Marialva, Parana, Brazil. Grapes were subjected to the following treatments: cold storage at 2 ºC (control), cold storage at 2 ºC with SO2-generating pads, cold storage at 2 ºC and inoculated with B. cinere a suspension, and cold storage at 2 ºC with SO2-generating pads and inoculated with B. cinerea suspension. The experiment was conducted in a complete randomized design with five replications per treatment using four bunches per experiment al unit. A factorial arrangement (absence/presence of SO2 pads × absence/presence of Botrytis inoculation) was applied. At the end of 30 days of cold storage and 7 days of shelf life (22 ºC), gray mold incidence, shattered berries, and physicochemical parameters were evaluated. The gray mold incidence on 'BRS Nubia' grapes decreased when SO2-generating pads were used during cold storage. Berry weight loss was greater in the treatments without SO2-generating pads after 30 days of cold storage followed by 7 days of shelf life. Berry firmness, soluble solids content (SS), total acidity (TA), SS/TA ratio, and anthocyanins concentration were not negatively affected by SO2-generating pad treatments. However, a slight increase in the shattered berries percentage was recorded for the SO2-generating pad treatments. No significant quality loss of 'BRS Nubia' grape was evident after 30 days of cold storage [...].(AU)

A demanda por produtos de alta qualidade nutricional tem aumentado o consumo de frutas, como as uvas, por esse motivo, são necessárias a adoção de técnicas pós-colheita para evitar perdas, preservara qualidade e prolongar a vida útil, além de atender às necessidades do consumidor. Dessa forma, o objetivo deste estudo foi avaliar estratégias para o controle do mofo cinzento causado por Botrytis cinerea em uvas 'BRS Nubia' durante o armazenamento refrigerado e a vida de prateleira. Cachos da uva 'BRS Nubia' foram colhidos em um vinhedo comercial localizado em Marialva, Paraná, Brasil. As uvas foram submetidas aos seguintes tratamentos: armazenamento refrigerado à 2 ºC (controle);armazenamento refrigerado à 2 ºC com folha geradora de SO2; armazenamento refrigerado à 2 ºC, inoculado com suspensão de B. cinerea; e armazenamento refrigerado à 2 ºC com folha geradora deSO2, inoculado com suspensão de B. cinerea; empregando o delineamento inteiramente casualizado, arranjado em esquema fatorial (ausência / presença de SO2 x ausência / presença de Botrytis), com cinco repetições e quatro cachos por repetição. Após 30 dias de armazenamento refrigerado e sete dias sob temperatura ambiente (22 ºC), avaliou-se a incidência de mofo cinzento, degrana e características físico-química das bagas. A incidência de mofo cinzento em uvas ‘BRS Nubia diminuiu com o uso de folhas geradoras de SO2 durante o armazenamento refrigerado. A perda de massa das bagas foi maior nos tratamentos sem folhas geradoras de SO2 após 30 dias de armazenamento refrigerado e 7 dias de prateleira. A firmeza das bagas, a concentração de sólidos solúveis (SS), acidez total (TA), SS/TA e antocianinas totais não foram afetadas pelos tratamentos com folhas geradoras de SO2. No entanto, houve aumento na porcentagem de degrana das bagas nesses tratamentos. Não foi observada perda significativa de qualidade [...].(AU)

Control of postharvest gray mold of 'BRS Nubia' table grape under cold storage / Controle de mofo cinzento na pós colheita da uva de mesa 'BRS Nubia' sob armazenamento refrigerado

The demand for high-quality nutritional products has increased fruit consumption, as grapes, for this reason postharvest techniques are required to prevent losses, to preserve quality, to extend shelf life, and to attend to consumer needs. In this way, the objective of this study was to evaluate strategies to control gray mold caused by Botrytis cinerea in 'BRS Nubia' grapes during cold storage and shelf life periods. Grape bunches were harvested from a commercial vineyard in Marialva, Parana, Brazil. Grapes were subjected to the following treatments: cold storage at 2 ºC (control), cold storage at 2 ºC with SO2-generating pads, cold storage at 2 ºC and inoculated with B. cinere a suspension, and cold storage at 2 ºC with SO2-generating pads and inoculated with B. cinerea suspension. The experiment was conducted in a complete randomized design with five replications per treatment using four bunches per experiment al unit. A factorial arrangement (absence/presence of SO2 pads × absence/presence of Botrytis inoculation) was applied. At the end of 30 days of cold storage and 7 days of shelf life (22 ºC), gray mold incidence, shattered berries, and physicochemical parameters were evaluated. The gray mold incidence on 'BRS Nubia' grapes decreased when SO2-generating pads were used during cold storage. Berry weight loss was greater in the treatments without SO2-generating pads after 30 days of cold storage followed by 7 days of shelf life. Berry firmness, soluble solids content (SS), total acidity (TA), SS/TA ratio, and anthocyanins concentration were not negatively affected by SO2-generating pad treatments. However, a slight increase in the shattered berries percentage was recorded for the SO2-generating pad treatments. No significant quality loss of 'BRS Nubia' grape was evident after 30 days of cold storage [...].

A demanda por produtos de alta qualidade nutricional tem aumentado o consumo de frutas, como as uvas, por esse motivo, são necessárias a adoção de técnicas pós-colheita para evitar perdas, preservara qualidade e prolongar a vida útil, além de atender às necessidades do consumidor. Dessa forma, o objetivo deste estudo foi avaliar estratégias para o controle do mofo cinzento causado por Botrytis cinerea em uvas 'BRS Nubia' durante o armazenamento refrigerado e a vida de prateleira. Cachos da uva 'BRS Nubia' foram colhidos em um vinhedo comercial localizado em Marialva, Paraná, Brasil. As uvas foram submetidas aos seguintes tratamentos: armazenamento refrigerado à 2 ºC (controle);armazenamento refrigerado à 2 ºC com folha geradora de SO2; armazenamento refrigerado à 2 ºC, inoculado com suspensão de B. cinerea; e armazenamento refrigerado à 2 ºC com folha geradora deSO2, inoculado com suspensão de B. cinerea; empregando o delineamento inteiramente casualizado, arranjado em esquema fatorial (ausência / presença de SO2 x ausência / presença de Botrytis), com cinco repetições e quatro cachos por repetição. Após 30 dias de armazenamento refrigerado e sete dias sob temperatura ambiente (22 ºC), avaliou-se a incidência de mofo cinzento, degrana e características físico-química das bagas. A incidência de mofo cinzento em uvas ‘BRS Nubia’ diminuiu com o uso de folhas geradoras de SO2 durante o armazenamento refrigerado. A perda de massa das bagas foi maior nos tratamentos sem folhas geradoras de SO2 após 30 dias de armazenamento refrigerado e 7 dias de prateleira. A firmeza das bagas, a concentração de sólidos solúveis (SS), acidez total (TA), SS/TA e antocianinas totais não foram afetadas pelos tratamentos com folhas geradoras de SO2. No entanto, houve aumento na porcentagem de degrana das bagas nesses tratamentos. Não foi observada perda significativa de qualidade [...].

Diagrammatic scale for measuring severity of gray mould in thornless Castilla blackberry (Rubus glaucus Benth) / Escala diagramática para medir a severidade do mofo cinzento em amora de Castilla (Rubus glaucus Benth) sem espinhos

Gray mould, caused by the fungus Botrytis cinerea Pers., is a disease that largely affects the crops of Rubus glaucus Benth. (Castilla blackberry) in Colombia. In spite of the economic losses that it causes in the production of Castilla blackberry at national level, a standardized method to quantify the disease severity caused by this fungus in the fruits has not yet been reported. In the present work, a diagrammatic scale was prepared to assess gray mould severity in thornless fruits of R. glaucus. The proposed scale showed the levels of 5, 10, 20, 40, 60, 80, and 100%. Photographic images were obtained for each of these levels, which were processed using the ImageJ software. The scale was validated by 13 evaluators who assessed affected fruits with and without the scale. The precision and accuracy of each evaluator was determined by simple linear regression between actual and estimated severity. The evaluators showed better precision, accuracy, and reproducibility in the assessments performed with the scale. The proposed scale is appropriate to estimate the severity of gray mould in R. glaucus fruits.(AU)

O mofo cinzento, causado pelo fungo Botrytis cinerea Pers., é uma doença que afeta amplamente as culturas de Rubus glaucus Benth. (Amora preta) na Colômbia. Apesar das perdas econômicas causadas na produção de amora preta em nível nacional, ainda não foi relatado um método padronizado para quantificar a severidade da doença causada por esse fungo nos frutos. No presente trabalho, uma escala diagramática foi preparada para avaliar a severidade do mofo cinza em frutos sem espinho de R. glaucus. A escala proposta apresentou os níveis de 5, 10, 20, 40, 60, 80 e 100%. Imagens fotográficas foram obtidas para cada um desses níveis, processadas com o software ImageJ. A escala foi validada por 13 avaliadores que analisaram os frutos afetados com e sem a escala. A precisão e exatidão de cada avaliador foram determinadas por regressão linear simples entre a severidade real e a estimada. Os avaliadores mostraram melhor precisão, exatidão e reprodutibilidade nas avaliações realizadas com a escala. A escala proposta é apropriada para estimar a severidade do mofo cinza em frutos de R. glaucus.(AU)

Enhanced Tolerance against a Fungal Pathogen and Insect Resistance in Transgenic Tobacco Plants Overexpressing an Endochitinase Gene from Serratia marcescens.

In this study we cloned a chitinase gene (SmchiC), from Serratia marcescens isolated from the corpse of a Diatraea magnifactella lepidopteran, which is an important sugarcane pest. The chitinase gene SmchiC amplified from the S. marcescens genome was cloned into the transformation vector p2X35SChiC and used to transform tobacco (Nicotiana tabacum L. cv Petit Havana SR1). The resistance of these transgenic plants to the necrotrophic fungus Botrytis cinerea and to the pest Spodoptera frugiperda was evaluated: both the activity of chitinase as well as the resistance against B. cinerea and S. frugiperda was significantly higher in transgenic plants compared to the wild-type.

Mechanisms of plant protection against two oxalate-producing fungal pathogens by oxalotrophic strains of Stenotrophomonas spp.

KEY MESSAGE: Oxalotrophic Stenotrophomonas isolated from tomato rhizosphere are able to protect plants against oxalate-producing pathogens by a combination of actions including induction of plant defence signalling callose deposition and the strengthening of plant cell walls and probably the degradation of oxalic acid. Oxalic acid plays a pivotal role in the virulence of the necrotrophic fungi Botrytis cinerea and Sclerotinia sclerotiorum. In this work, we isolated two oxalotrophic strains (OxA and OxB) belonging to the bacterial genus Stenotrophomonas from the rhizosphere of tomato plants. Both strains were capable to colonise endophytically Arabidopsis plants and protect them from the damage caused by high doses of oxalic acid. Furthermore, OxA and OxB protected Arabidopsis from S. sclerotiorum and B. cinerea infections. Bacterial inoculation induced the production of phenolic compounds and the expression of PR-1. Besides, both isolates exerted a protective effect against fungal pathogens in Arabidopsis mutants affected in the synthesis pathway of salicylic acid (sid2-2) and jasmonate perception (coi1). Callose deposition induced by OxA and OxB was required for protection against phytopathogens. Moreover, B. cinerea and S. sclerotiorum mycelial growth was reduced in culture media containing cell wall polysaccharides from leaves inoculated with each bacterial strain. These findings suggest that cell walls from Arabidopsis leaves colonised by these bacteria would be less susceptible to pathogen attack. Our results indicate that these oxalotrophic bacteria can protect plants against oxalate-producing pathogens by a combination of actions and show their potential for use as biological control agents against fungal diseases.

Expression of disease resistance in genetically modified grapevines correlates with the contents of viral sequences in the T-DNA and global genome methylation.

Increased tolerance to pathogens is an important goal in conventional and biotechnology-assisted grapevine breeding programs worldwide. Fungal and viral pathogens cause direct losses in berry production, but also affect the quality of the final products. Precision breeding strategies allow the introduction of resistance characters in elite cultivars, although the factors determining the plant's overall performance are not fully characterized. Grapevine plants expressing defense proteins, from fungal or plant origins, or of the coat protein gene of grapevine leafroll-associated virus 3 (GLRaV-3) were generated by Agrobacterium-mediated transformation of somatic embryos and shoot apical meristems. The responses of the transformed lines to pathogen challenges were investigated by biochemical, phytopathological and molecular methods. The expression of a Metarhizium anisopliae chitinase gene delayed pathogenesis and disease progression against the necrotrophic pathogen Botrytis cinerea. Modified lines expressing a Solanum nigrum osmotin-like protein also exhibited slower disease progression, but to a smaller extent. Grapevine lines carrying two hairpin-inducing constructs had lower GLRaV-3 titers when challenged by grafting, although disease symptoms and viral multiplication were detected. The levels of global genome methylation were determined for the genetically engineered lines, and correlation analyses demonstrated the association between higher levels of methylated DNA and larger portions of virus-derived sequences. Resistance expression was also negatively correlated with the contents of introduced viral sequences and genome methylation, indicating that the effectiveness of resistance strategies employing sequences of viral origin is subject to epigenetic regulation in grapevine.

Pathogenicity and transmission of fungi detected on Passiflora alata seeds / Patogenicidade e transmissão de fungos detectados em sementes de Passiflora alata

Passion fruit is usually propagated by seeds because of the ease and lower cost in seedling production. However, the seed is the most efficient agent for the spread of pathogens. The damages from seed-borne diseases occur mainly during the germination stages or at the formation of seedlings in nurseries. Considering the need for knowledge on the pathology of sweet passion fruit seeds, the objective was to evaluate the transmission and pathogenicity of the fungi Alternaria sp., Botrytis fabae, Cladosporium cladosporioides, Fusarium spp. and Lasiodiplodia theobromae, known as potentially pathogenic to this crop, and isolated from sweet passion fruit seeds. Therefore, tests on seed health, germination and seedling emergence in a sterilized commercial substrate were conducted using seeds from this species, inoculated with those fungal isolates. Leaves, stems and fruit from this plant were also inoculated with the same fungi. Alternaria sp., Fusarium spp. and L. theobromae were identified in seedlings obtained from inoculated seeds, confirming the transmission of these fungi by seeds. L. theobromae was also considered the most harmful fungus to passion fruit crop, as it causes seed rot and other disease symptoms on the leaves, stem and fruit. These findings inferred that healthy seeds of sweet passion fruit are essential for producing seedlings and to prevent the spread of the diseases caused by these fungi to exempt areas.(AU)

O maracujazeiro geralmente é propagado por meio de sementes em virtude da facilidade e do menor custo na produção de mudas. No entanto, a semente é o agente mais eficiente de disseminação de patógenos, sendo que os danos decorrentes das doenças transmitidas por elas ocorrem principalmente durante os estágios de germinação ou na formação de mudas nos viveiros. Considerando a necessidade de informações acerca da patologia de sementes de maracujá-doce nesse contexto, objetivou-se obter informações sobre a transmissão e a patogenicidade dos fungos Alternaria sp., Botrytis fabae, Cladosporium cladosporioides, Fusarium spp. e Lasiodiplodia theobromae, isolados de sementes de maracujá-doce e potencialmente patogênicos à cultura. Para tanto, testes de sanidade, germinação e emergência de plântulas em substrato comercial esterilizado foram conduzidos com sementes dessa espécie, inoculadas com esses isolados. Folhas, colo e frutos dessa planta também foram inoculados com os mesmos fungos. Alternaria sp., Fusarium spp. e L. theobromae foram identificados em plântulas obtidas de sementes inoculadas, confirmando a transmissão por sementes. L. theobromae foi considerado o mais agressivo à cultura do maracujá, por ter causado podridão nas sementes, além de maiores lesões nas folhas, no colo da planta e nos frutos. Dessa forma, infere-se que a obtenção de sementes de maracujá-doce sadias é imprescindível para a produção de mudas, evitando-se assim a disseminação desses patógenos em áreas isentas.(AU)

Pathogenicity and transmission of fungi detected on Passiflora alata seeds / Patogenicidade e transmissão de fungos detectados em sementes de Passiflora alata

Passion fruit is usually propagated by seeds because of the ease and lower cost in seedling production. However, the seed is the most efficient agent for the spread of pathogens. The damages from seed-borne diseases occur mainly during the germination stages or at the formation of seedlings in nurseries. Considering the need for knowledge on the pathology of sweet passion fruit seeds, the objective was to evaluate the transmission and pathogenicity of the fungi Alternaria sp., Botrytis fabae, Cladosporium cladosporioides, Fusarium spp. and Lasiodiplodia theobromae, known as potentially pathogenic to this crop, and isolated from sweet passion fruit seeds. Therefore, tests on seed health, germination and seedling emergence in a sterilized commercial substrate were conducted using seeds from this species, inoculated with those fungal isolates. Leaves, stems and fruit from this plant were also inoculated with the same fungi. Alternaria sp., Fusarium spp. and L. theobromae were identified in seedlings obtained from inoculated seeds, confirming the transmission of these fungi by seeds. L. theobromae was also considered the most harmful fungus to passion fruit crop, as it causes seed rot and other disease symptoms on the leaves, stem and fruit. These findings inferred that healthy seeds of sweet passion fruit are essential for producing seedlings and to prevent the spread of the diseases caused by these fungi to exempt areas.(AU)

O maracujazeiro geralmente é propagado por meio de sementes em virtude da facilidade e do menor custo na produção de mudas. No entanto, a semente é o agente mais eficiente de disseminação de patógenos, sendo que os danos decorrentes das doenças transmitidas por elas ocorrem principalmente durante os estágios de germinação ou na formação de mudas nos viveiros. Considerando a necessidade de informações acerca da patologia de sementes de maracujá-doce nesse contexto, objetivou-se obter informações sobre a transmissão e a patogenicidade dos fungos Alternaria sp., Botrytis fabae, Cladosporium cladosporioides, Fusarium spp. e Lasiodiplodia theobromae, isolados de sementes de maracujá-doce e potencialmente patogênicos à cultura. Para tanto, testes de sanidade, germinação e emergência de plântulas em substrato comercial esterilizado foram conduzidos com sementes dessa espécie, inoculadas com esses isolados. Folhas, colo e frutos dessa planta também foram inoculados com os mesmos fungos. Alternaria sp., Fusarium spp. e L. theobromae foram identificados em plântulas obtidas de sementes inoculadas, confirmando a transmissão por sementes. L. theobromae foi considerado o mais agressivo à cultura do maracujá, por ter causado podridão nas sementes, além de maiores lesões nas folhas, no colo da planta e nos frutos. Dessa forma, infere-se que a obtenção de sementes de maracujá-doce sadias é imprescindível para a produção de mudas, evitando-se assim a disseminação desses patógenos em áreas isentas.(AU)

Differential control and function of Arabidopsis ProDH1 and ProDH2 genes on infection with biotrophic and necrotrophic pathogens.

Arabidopsis contains two proline dehydrogenase (ProDH) genes, ProDH1 and ProDH2, encoding for homologous and functional isoenzymes. Although ProDH1 has been studied extensively, especially under abiotic stress, ProDH2 has only started to be analysed in recent years. These genes display distinctive expression patterns and show weak transcriptional co-regulation, but are both activated in pathogen-infected tissues. We have demonstrated previously that Arabidopsis plants with silenced ProDH1/2 expression fail to trigger defences against the hemibiotrophic bacterial pathogen Pseudomonas syringae pv. tomato AvrRpm1 (Pst-AvrRpm1), and that ProDH1 and ProDH2 are differentially regulated by salicylic acid (SA). In the current work, we used prodh1 and prodh2 single-mutant plants to assess the particular contribution of each gene to resistance against Pst-AvrRpm1 and the necrotrophic fungal pathogen Botrytis cinerea. In addition, we studied the sensitivity of ProDH1 and ProDH2 to the jasmonic acid (JA) defence pathway. We found that ProDH1 and ProDH2 are both necessary to achieve maximum resistance against Pst-AvrRpm1 and B. cinerea. However, ProDH2 has a major effect on early restriction of B. cinerea growth. Interestingly, ProDH1 is up-regulated by SA and JA, whereas ProDH2 is only activated by JA, and both genes display transcriptional inter-regulation at basal and infection conditions. These studies provide the first evidence of the contribution of ProDH2 to disease resistance, and describe the differential regulation and non-redundant but complementary function of both enzyme isoforms in infected tissues, providing support for a fundamental role of ProDH in the control of biotrophic and necrotrophic pathogens.

Gray mold in immature fig fruit: pathogenicity and growth temperature / Mofo cinzento em frutos imaturos de figo: patogenicidade e temperatura para crescimento

Several diseases can be associated with figs but recently a fruit rot was observed in green fruit. The purpose of this study was to determine the pathogenicity of Botrytis sp., to quantify incubation period (IP) and latent period (LP), to verify the optimum temperature for mycelial growth, and to identify the different species of Botrytis sp. isolated from immature figs. Botrytis sp. isolated from figs proved to be pathogenic to immature fruit with and without wounding the fruit surface and ostiole. The IP period was 3 days on fruit with wounds and 5 days on fruit inoculated within the ostiole (without wound). The LP was 6 days in all treatments. The optimum temperature for mycelial growth was 18°C. Inferred from sequences of a segment comprising the ITS region of ribosomal DNA concluded that the isolates are Botrytis cinerea.(AU)

Várias doenças podem estar associadas com figos, mas recentemente uma podridão dos frutos foi observada em frutos verdes. O objetivo deste estudo foi determinar a patogenicidade de Botrytis sp., quantificar o período de incubação (PI) e o período de latência (PL), verificar a temperatura ótima para o crescimento micelial e identificar as diferentes espécies de Botrytis sp. isoladas a partir de frutos imaturos de figo. Botrytis sp. isolado a partir de figos provou ser patogênico em frutos imaturos com e sem ferimento na superfície dos frutos e no ostíolo. O PI foi de 3 dias em frutos com ferimento e 5 dias em frutos inoculados no ostíolo e sem ferimento. O PL foi de 6 dias em todos os tratamentos. A temperatura ótima de crescimento micelial foi18°C. A partir de sequências de um segmento que compreende a região de ITS do DNA ribossomal, concluiu-se que os isolados são Botrytis cinerea.(AU)

Relationship between production, nematodes and "redness" in strawberries / Relação entre produção, nematoide e o sintoma de "vermelhão" em morangueiros

In recent years "redness" has increasingly appeared in strawberry plants with leaves taking on a reddish color. No causal agent has been associated with plants. Since strawberries presented problems due to the incidence of nematodes, the purpose of this study was to look at the relationship between production, resistance to the Meloidogyne hapla nematode and the "redness" symptom in strawberry cultivars. Two experiments were performed, both with the 'Camino Real', 'Festival', 'Oso Grande', 'Albion' and 'Camarosa' cultivars. The first experiment was performed in the field, where the following were evaluated: strawberry production, fruit quality, macro and micronutrient contents in fruit and leaves, percentage of plant survival, incidence of nematodes, quantity of eggs in the roots and juveniles in the soil, and the incidence of Botrytis cinerea . In the second experiment, the strawberries were transplanted into pots and filled with pinus bark-based commercial substrate. Half the pots were inocculated with Meloidogyne hapla . Cultivars presented differences in fruit production and also in the incidence of "redness". Lowest performance in production was related to the high incidence of the nematode Meloidogyne hapla. 'Oso Grande' and 'Albion' presented nematode-resistant behavior. It was possible find a relationship between the incidence of the Meloidogyne hapla nematode, and the incidence of "redness" only 'Camino Real' cultivar.(AU)

Nos últimos anos, vem aumentando o aparecimento do "vermelhão" nos morangueiros, que ficam com as folhas avermelhadas. Nenhum agente causal foi associado às plantas. Devido aos plantios de morangueiros apresentarem problemas pela incidência de nematoide, o objetivo do presente trabalho foi verificar a relação entre produção, resistência ao nematoide Meloidogyne hapla e o sintoma de "vermelhão" em cultivares de morangueiro. Foram realizados dois experimentos, ambos com as cultivares 'Camino Real', 'Festival', 'Oso Grande', 'Albion' e 'Camarosa'. O primeiro experimento foi realizado a campo, onde se avaliaram a produção de morangos, qualidade dos frutos, teores de macro e micronutrientes em frutos e folhas, porcentagem de sobrevivência das plantas, incidência de nematoides, quantidade de ovos nas raízes e juvenis no solo e a incidência de Botrytis cinerea. No segundo experimento, os morangueiros foram transplantados em vasos e preenchidos com substrato comercial a base de casca de pinus. Metade dos vasos foi inoculada com Meloidogyne hapla. As cultivares apresentaram diferenças na produção de frutos e também na incidência de "vermelhão". O menor desempenho produtivo relaciona-se com a alta incidência de nematoide Meloidogyne hapla. 'Oso Grande' e 'Albion' se comportaram como resistentes ao nematoide. Foi possível relacionar a incidência de nematoide Meloidogyne hapla com a de "vermelhão" apenas na cultivar 'Camino Real'.(AU)

Characterization of a novel Kazal-type serine proteinase inhibitor of Arabidopsis thaliana.

Many different types of serine proteinase inhibitors have been involved in several kinds of plant physiological processes, including defense mechanisms against phytopathogens. Kazal-type serine proteinase inhibitors, which are included in the serine proteinase inhibitor family, are present in several organisms. These proteins play a regulatory role in processes that involve serine proteinases like trypsin, chymotrypsin, thrombin, elastase and/or subtilisin. In the present work, we characterized two putative Kazal-type serine proteinase inhibitors from Arabidopsis thaliana, which have a single putative Kazal-type domain. The expression of these inhibitors is transiently induced in response to leaf infection by Botrytis cinerea, suggesting that they play some role in defense against pathogens. We also evaluated the inhibitory specificity of one of the Kazal-type serine proteinase inhibitors, which resulted to be induced during the local response to B. cinerea infection. The recombinant Kazal-type serine proteinase inhibitor displayed high specificity for elastase and subtilisin, but low specificity for trypsin, suggesting differences in its selectivity. In addition, this inhibitor exhibited a strong antifungal activity inhibiting the germination rate of B. cinerea conidia in vitro. Due to the important role of proteinase inhibitors in plant protection against pathogens and pests, the information about Kazal-type proteinase inhibitors described in the present work could contribute to improving current methods for plant protection against pathogens.

Defense responses in plants of Eucalyptus elicited by Streptomyces and challenged with Botrytis cinerea.

MAIN CONCLUSION: Elicitation of E. grandis plants with Streptomyces PM9 reduced the gray-mold disease, through increasing the levels of enzymes directly related to the induction of plant defense responses, and accumulation of specific phenolic compounds. Members of Eucalyptus are economically important woody species, especially as a raw material in many industrial sectors. Species of this genus are susceptible to pathogens such as Botrytis cinerea (gray mold). Biological control of plant diseases using rhizobacteria is one alternative to reduce the use of pesticides and pathogen attack. This study evaluated the metabolic and phenotypic responses of Eucalyptus grandis and E. globulus plants treated with Streptomyces sp. PM9 and challenged with the pathogenic fungus B. cinerea. Metabolic responses were evaluated by assessing the activities of the enzymes polyphenol oxidase and peroxidase as well as the levels of phenolic compounds and flavonoids. The incidence and progression of the fungal disease in PM9-treated plants and challenged with B. cinerea were evaluated. Treatment with Streptomyces sp. PM9 and challenge with B. cinerea led to changes in the activities of polyphenol oxidase and peroxidase as well as in the levels of phenolic compounds in the plants at different time points. Alterations in enzymes of PM9-treated plants were related to early defense responses in E. grandis. Gallic and chlorogenic acids were on average more abundant, although caffeic acid, benzoic acid and catechin were induced at specific time points during the culture period. Treatment with Streptomyces sp. PM9 significantly delayed the establishment of gray mold in E. grandis plants. These results demonstrate the action of Streptomyces sp. PM9 in inducing plant responses against B. cinerea, making this organism a potential candidate for biological control in Eucalyptus.

Identification and Characterization of Botrytis Blossom Blight of Japanese Plums Caused by Botrytis cinerea and B. prunorum sp. nov. in Chile.

Blossom blight is a destructive disease of plums (Prunus salicina) when humid and temperate weather conditions occur in Chile. Disease incidence ranging from 4 to 53% has been observed. Symptoms include light brown petal necrosis, starting as light brown mottles or V-shaped necrosis at the margins of the petals, progressing to the stamen and pistils. In this study, the etiology of blossom blight of plums was determined. High- and low-sporulating isolates of Botrytis were obtained consistently from blighted blossoms and apparently healthy flowers of plums. Based on colony morphology, conidial production and molecular phylogenetic analysis, these high- and low-sporulating isolates were identified as B. cinerea and B. prunorum sp. nov., respectively. Phylogenetic analysis of the genes glyceraldehyde 3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60), and DNA-dependent RNA polymerase subunit II (RPB2) grouped B. prunorum isolates in a single cluster, distantly from B. cinerea and other Botrytis species. The phylogenetic analysis of necrosis and ethylene-inducing protein (NEP1 and NEP2) genes corroborated these results. Analysis of the internal transcribed spacer region and large-subunit (26S) ribosomal DNA and detection of Boty and Flipper transposable elements, were not useful to differentiate between these Botrytis species. Both species were pathogenic on plum flowers and the fruit of plums, apples, and kiwifruits. However, B. prunorum was less virulent than B. cinerea. These pathogens were re-isolated from inoculated and diseased tissues; thus, Koch's postulates were fulfilled, confirming its role in blossom blight of plums. B. cinerea was predominant, suggesting that B. prunorum may play a secondary role in the epidemiology of blossom blight in plums in Chile. This study clearly demonstrated that the etiology of blossom blight of plums is caused by B. cinerea and B. prunorum, which constitute a species complex living in sympatry on plums and possibly on other stone fruit trees.

A circadian oscillator in the fungus Botrytis cinerea regulates virulence when infecting Arabidopsis thaliana.

The circadian clock of the plant model Arabidopsis thaliana modulates defense mechanisms impacting plant-pathogen interactions. Nevertheless, the effect of clock regulation on pathogenic traits has not been explored in detail. Moreover, molecular description of clocks in pathogenic fungi--or fungi in general other than the model ascomycete Neurospora crassa--has been neglected, leaving this type of question largely unaddressed. We sought to characterize, therefore, the circadian system of the plant pathogen Botrytis cinerea to assess if such oscillatory machinery can modulate its virulence potential. Herein, we show the existence of a functional clock in B. cinerea, which shares similar components and circuitry with the Neurospora circadian system, although we found that its core negative clock element FREQUENCY (BcFRQ1) serves additional roles, suggesting extracircadian functions for this protein. We observe that the lesions produced by this necrotrophic fungus on Arabidopsis leaves are smaller when the interaction between these two organisms occurs at dawn. Remarkably, this effect does not depend solely on the plant clock, but instead largely relies on the pathogen circadian system. Genetic disruption of the B. cinerea oscillator by mutation, overexpression of BcFRQ1, or by suppression of its rhythmicity by constant light, abrogates circadian regulation of fungal virulence. By conducting experiments with out-of-phase light:dark cycles, we confirm that indeed, it is the fungal clock that plays the main role in defining the outcome of the Arabidopsis-Botrytis interaction, providing to our knowledge the first evidence of a microbial clock modulating pathogenic traits at specific times of the day.