scholarly journals SELEÇÃO IN VITRO DE FUNGOS ENDOFÍTICOS PARA O CONTROLE BIOLÓGICO DE Botrytis cinerea EM Eucalyptus benthamii

FLORESTA ◽  
2013 ◽  
Vol 43 (1) ◽  
pp. 145 ◽  
Author(s):  
José Antonio Sbravatti Junior ◽  
Celso Garcia Auer ◽  
Ida Chapaval Pimentel ◽  
Álvaro Figueredo dos Santos ◽  
Bruno Schultz

   O Eucalyptus benthamii é uma das principais espécies de eucalipto plantadas na região Sul do Brasil, por sua resistência a geadas e por seu uso na produção florestal de madeira para fins energéticos. Na produção de mudas, uma das principais doenças ocorrentes em viveiros é o mofo-cinzento, causado pelo fungo Botrytis cinerea. Uma das alternativas para o controle dessa doença é o controle biológico com fungos endofíticos, os quais podem competir com os patógenos foliares de mudas de eucalipto. O objetivo deste trabalho foi isolar os fungos endofíticos provenientes de mudas de E. benthamii, identificá-los e selecioná-los para o controle de B. cinerea. Eles foram isolados do interior de tecidos vegetais desinfectados, identificados de acordo com critérios macro e micromorfológicos e classificados a partir de testes de controle biológico in vitro. Os resultados evidenciaram o potencial antagonista dos fungos Aspergillus sp., Penicillium sp. e Trichoderma sp. Nenhum desses fungos causou lesões em mudas de E. benthamii.Palavras-chave: Mofo-cinzento; eucalipto; viveiro.AbstractIn vitro selection of endophytes for biological control of Botrytis cinerea in Eucalyptus benthamii. Eucalyptus benthamii is one of the main eucalypt species planted in Southern Brazil, due to its resistance to frost and its use in the production of forest wood for energy purposes. During the production of seedlings, the main disease occurring in forest nurseries is gray-mold caused by the fungus Botrytis cinerea. One alternative for control this disease is biological control with fungal endophytes, which can compete with the foliar pathogens of eucalypt seedlings. The objective of this study was to isolate endophytic fungi from seedlings of Eucalyptus benthamii, identify and select them for B. cinerea control. These were isolated from the interior of disinfected plant tissues, identified according to macro and micromorphological criteria, and based on tests of biological control in vitro. The results revealed the potential antagonist of Aspergillus sp., Penicillium sp. and Trichoderma sp. No fungi caused lesions in E. benthamii seedlings.Keywords: Gray-mold; eucalypt; nursery.    

2014 ◽  
Vol 23 (6) ◽  
pp. 3-9
Author(s):  
René Dionisio Cupull-Santana ◽  
Remigio Cortés-Rodríguez ◽  
Ervelio Eliseo Olazábal-Manso ◽  
Carlos Alberto Hernández Medina

La investigación se realizó con el objetivo de evaluar las características fisicoquímicas y la actividad antifúngica de propóleos de Apis mellifera, provenientes de las provincias de Villa Clara, Cienfuegos y Sancti Spíritus, Cuba. Se utilizó el método de recolección por raspado y se estableció el contenido de cera, ceniza, material insoluble y resina de los propóleos crudos. Además, se evaluó la actividad antimicrobiana in vitro frente a los hongos Aspergillus sp., Penicillium sp., Fusarium sp., Trichoderma sp., Colletotrichum sp. y Monilia sp.; los extractos presentaron alta actividad antifúngica en un amplio rango de concentraciones; además, los parámetros fisicoquímicos presentaron valores semejantes y dentro de los establecidos por regulaciones internacionales.


Plant Disease ◽  
2020 ◽  
Vol 104 (5) ◽  
pp. 1298-1304 ◽  
Author(s):  
Ting-ting Li ◽  
Jing-di Zhang ◽  
Jia-quan Tang ◽  
Zhi-cheng Liu ◽  
Ya-qian Li ◽  
...  

Tomato gray mold caused by Botrytis cinerea is one of the main diseases of tomato and significantly impacts the yield and quality of tomato fruit. The overuse of chemical fungicides has resulted in the development of fungicide-resistant strains. Biological control is becoming an alternative method for the control of plant diseases to replace or decrease the application of traditional synthetic chemical fungicides and genus Trichoderma is widely used as a biological agent for controlling tomato gray mold. Brassinolide (BR) is a plant-growth-promoting steroid. To enhance the efficiency and stability of Trichoderma activity against B. cinerea, an optimal combination of Trichoderma atroviride CCTCCSBW0199 and BR that controls B. cinerea infection in tomato was identified. Strain CCTCCSBW0199 was found to have antagonistic activity against B. cinerea both in vitro and in vivo. In addition, a fermented culture of chlamydospores and metabolites, or metabolites only of strain CCTCCSBW0199 also reduced growth of B. cinerea. BR reduced growth of B. cinerea and had no effect on the sporulation and growth of Trichoderma spp. An application of metabolites of a Trichoderma sp. + BR reduced gray mold on tomato leaves by approximately 70.0%. Furthermore, the activities of induced defense response-related enzyme, such as peroxidase, superoxide dismutase, catalase, and phenylalanine ammonia-lyase were increased in tomato plants treated with a Trichoderma sp. + BR. Our data suggested that applying a mix of metabolites of T. atroviride CCTCCSBW0199 + BR was effective at reducing gray mold of tomato and may lay a theoretical foundation for the development of novel biofungicides.


2017 ◽  
Vol 107 (3) ◽  
pp. 362-368 ◽  
Author(s):  
Wayne M. Jurick ◽  
Otilia Macarisin ◽  
Verneta L. Gaskins ◽  
Eunhee Park ◽  
Jiujiang Yu ◽  
...  

Botrytis cinerea causes gray mold and is an economically important postharvest pathogen of fruit, vegetables, and ornamentals. Fludioxonil-sensitive B. cinerea isolates were collected in 2011 and 2013 from commercial storage in Pennsylvania. Eight isolates had values for effective concentrations for inhibiting 50% of mycelial growth of 0.0004 to 0.0038 μg/ml for fludioxonil and were dual resistant to pyrimethanil and thiabendazole. Resistance was generated in vitro, following exposure to a sublethal dose of fludioxonil, in seven of eight dual-resistant B. cinerea isolates. Three vigorously growing B. cinerea isolates with multiresistance to postharvest fungicides were further characterized and found to be osmosensitive and retained resistance in the absence of selection pressure. A representative multiresistant B. cinerea strain caused decay on apple fruit treated with postharvest fungicides, which confirmed the in vitro results. The R632I mutation in the Mrr1 gene, associated with fludioxonil resistance in B. cinerea, was not detected in multipostharvest fungicide-resistant B. cinerea isolates, suggesting that the fungus may be using additional mechanisms to mediate resistance. Results from this study show for the first time that B. cinerea with dual resistance to pyrimethanil and thiabendazole can also rapidly develop resistance to fludioxonil, which may pose control challenges in the packinghouse environment and during long-term storage.


2003 ◽  
Vol 28 (1) ◽  
pp. 67-75 ◽  
Author(s):  
Carolina Leoni ◽  
Raquel Ghini

Uma alternativa de manejo das doenças causadas por Phytophthora spp. é o uso de matéria orgânica. No presente trabalho foi avaliada a potencialidade do lodo de esgoto na indução de supressividade in vitro a P. nicotianae. O efeito do lodo de esgoto incorporado ao solo na sobrevivência de P. nicotianae foi avaliado mediante um experimento fatorial com dois fatores: doses de lodo de esgoto (0, 10, 20 e 40% p/p) e concentrações de inóculo [0, 10 ou 20 g de grãos de trigo (Triticum aestivum) colonizados kg-1]. Aos 21 dias, quando aumentaram as doses de lodo de esgoto, a sobrevivência de P. nicotianae e os pHs das misturas diminuíram, e as condutividades elétricas (CE) aumentaram. As correlações entre a CE e a sobrevivência do patógeno foram negativas e significativas (P>0,05). Para estudar o efeito dos compostos químicos envolvidos na supressividade, foram obtidos extratos em água, H2SO4 2N e KOH 0,4N de misturas de areia – lodo de esgoto (20% p/p), e foram acrescentados ao meio de cultura e seu efeito avaliado no crescimento das colônias de P. nicotianae. O extrato ácido (H2SO4 2N) do tratamento com 20% de lodo de esgoto inibiu significativamente (P>0,05) o crescimento da colônia do patógeno. O efeito biológico foi estudado mediante isolamento de microrganismos em meio de cultura e seleção por antagonismo. No bioensaio com plântulas de alfafa (Medicago sativa) destacaram-se os isolados F9.1 (Aspergillus sp.) e A12.1 (actinomiceto, não identificado); e no teste de culturas pareadas destacou-se um Trichoderma sp. e dois actinomicetos por antibiose, e um Trichoderma sp. e três Aspergillus sp. por hiperparasitismo.


FLORESTA ◽  
2013 ◽  
Vol 43 (2) ◽  
pp. 225
Author(s):  
Miriam Machado Cunico ◽  
Celso Garcia Auer ◽  
Marlon Wesley Machado Cunico ◽  
Obdulio Gomes Miguel ◽  
Patricio Peralta Zamora ◽  
...  

 Extratos etanólicos de anestesia, Ottonia martiana Miq., foram reavaliados quanto à inibição do crescimento micelial dos fungos Cylindrocladium spathulatum (pinta-preta da erva-mate) e Botrytis cinerea (mofo-cinzento do eucalipto), por meio do planejamento fatorial. A ocorrência de decomposição de bioativos no processo de autoclavagem também foi investigada, por meio de teste de eficiência de extratos filtrados (filtro Millipore) e esterilizados (autoclave) no controle dos fitopatógenos, nas concentrações de 1, 10, 100 e 1000 ppm. Os extratos etanólicos filtrado e esterilizado inibiram o crescimento micelial dos fungos e foram mais ativos frente a B. cinerea.O extrato filtrado exibiu maior potencial antifúngico que o extrato esterilizado. O processo de esterilização por autoclavagem causou pequena decomposição dos bioativos presentes no extrato de anestesia.Palavras-chave: Anestesia; mofo-cinzento; pinta-preta. Abstract Fungitoxic potential of ethanolic extracts of anestesia in the control of phytopathogenic diseases. The antifungal potential of anestesia, Ottonia martiana Miq. was reassessed by factorial design, in vitro testing of fungal mycelial growth compared to the pathogenic isolates Cylindrocladium spathulatum, causal agent of black spot onyerba mate, and Botrytis cinerea causal agent of gray-mold on eucalypts. Occurrence of decomposition of bioactive of the autoclaving process was investigated using foliar detached test compared to the pathogens (1000 ppm). Ethanolic extracts - EBEtOH (filtered and autoclaved) inhibited the mycelial growth of C. spathulatum and B. cinerea (1000 ppm) and were more pronounced against B. cinerea (43.6 % and 68.9 %). EBEtOH filtered (0.22 µm) presented higher activity than EBEtOH autoclaved (C. spathulatum: 52.8 % and 43.6 %, B. cinerea: 68.9 % and 43.6 %), suggesting little decomposition ofbioactive after autoclaving. EBEtOH filtrate presented potential inhibition of 28 % in eucalypt leaves against B. cinerea.  Keywords: Ottonia martiana; black spot; gray-mold.


2011 ◽  
Vol 101 (12) ◽  
pp. 1433-1445 ◽  
Author(s):  
Anne-Sophie Walker ◽  
Angélique Gautier ◽  
Johann Confais ◽  
Daniel Martinho ◽  
Muriel Viaud ◽  
...  

Botrytis cinerea is a major crop pathogen infesting >220 hosts worldwide. A cryptic species has been identified in some French populations but the new species, B. pseudocinerea, has not been fully delimited and established. The aim of this study was to distinguish between the two species, using phylogenetic, biological, morphological, and ecological criteria. Multiple gene genealogies confirmed that the two species belonged to different, well-supported phylogenetic clades. None of the morphological criteria tested (spore size, germination rate, or mycelial growth) was able to discriminate between these two species. Sexual crosses between individuals from the same species and different species were carried out. Only crosses between individuals from the same species were successful. Moreover, population genetics analysis revealed a high level of diversity within each species and a lack of gene flow between them. Finally, a population survey over time showed that B. cinerea was the predominant species but that B. pseudocinerea was more abundant in spring, on floral debris. This observation could not be explained by temperature adaptation in tests carried out in vitro or by aggressiveness on tomato or bean leaves. This study clearly establishes that B. cinerea and B. pseudocinerea constitute a complex of two cryptic species living in sympatry on several hosts, including grapevine and blackberry. We propose several biological or molecular tools for unambiguous differentiation between the two species. B. pseudocinerea probably makes a negligible contribution to gray mold epidemics on grapevine. This new species has been deposited in the MycoBank international database.


2019 ◽  
Vol 10 (4) ◽  
pp. 779-788
Author(s):  
Aidé González Ruiz ◽  
Abiel Sánchez Arizpe ◽  
Yisa Maria Ochoa Fuentes ◽  
Ma. Elizabeth Galindo Cepeda ◽  
Raúl Rodríguez Guerra ◽  
...  

  Nodulosporium es un hongo ampliamente distribuido y que se encuentra en zonas tropicales y se ha reportado como fitopatógeno ocasionando muerte descendente, pudriciones y resinosis, también se le considera como un potencial agente de control biológico de otros hongos y oomicetos. El estado teleomorfo se encuentra en estado solitario y en racimos, y se ha reportado como endófito, saprofito o fitopatógeno débil. La variedad criolla de cacao Theobroma cacao, es de gran relevancia ecológica, económica y cultural en el estado de Chiapas; sin embargo, presenta alta susceptibilidad a enfermedades a diferencia de otras variedades importantes, afectando la calidad y producción. En Villa de Comaltitlán Chiapas se observaron frutos de cacao con características típicas de síntomas de pudrición. El objetivo fue identificar los hongos asociados a frutos de cacao enfermos con síntomas de pudrición. Se identificaron morfológicamente los hongos Aspergillus sp., Penicillium sp., Rhizopus sp., Trichoderma sp., y Nodulosporium sp. Se informa por primera ocasión la presencia de Nodulosporium sp., en frutos de cacao con síntomas de pudrición de Villa de Comaltitlán, Chiapas, México. Se describe las características morfológicas y morfométricas de Nodulosporium sp. e Hypoxylon sp., y su identificación molecular. Se confirmó la patogenicidad de Nodulosporium sp., en follaje del cultivo causando clorosis y deshidratación de la hoja, y se recuperó en forma de picnidios y micelio con conidios, correspondiendo a su teleomorfo, identificado como Hypoxylon morfológica y molecularmente con los oligonucleótidos ITS4-ITS5.


PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9626
Author(s):  
Huiyu Hou ◽  
Xueying Zhang ◽  
Te Zhao ◽  
Lin Zhou

Background Botrytis cinerea causes serious gray mold disease in many plants. This pathogen has developed resistance to many fungicides. Thus, it has become necessary to look for new safe yet effective compounds against B. cinerea. Methods Essential oils (EOs) from 17 plant species were assayed against B. cinerea, of which Origanum vulgare essential oil (OVEO) showed strong antifungal activity, and accordingly its main components were detected by GC/MS. Further study was conducted on the effects of OVEO, carvacrol and thymol in vitro on mycelium growth and spore germination, mycelium morphology, leakages of cytoplasmic contents, mitochondrial injury and accumulation of reactive oxygen species (ROS) of B. cinerea. The control efficacies of OVEO, carvacrol and thymol on tomato gray mold were evaluated in vivo. Results Of all the 17 plant EOs tested, Cinnamomum cassia, Litsea cubeba var. formosana and O. vulgare EOs had the best inhibitory effect on B. cinerea, with 0.5 mg/mL completely inhibiting the mycelium growth of B. cinerea. Twenty-one different compounds of OVEO were identified by gas chromatography–mass spectrometry, and the main chemical components were carvacrol (89.98%), β-caryophyllene (3.34%), thymol (2.39%), α-humulene (1.38%) and 1-methyl-2-propan-2-ylbenzene isopropyl benzene (1.36%). In vitro experiment showed EC50 values of OVEO, carvacrol and thymol were 140.04, 9.09 and 21.32 μg/mL, respectively. Carvacrol and thymol completely inhibited the spore germination of B. cinerea at the concentration of 300 μg/mL while the inhibition rate of OVEO was 80.03%. EC50 of carvacrol and thymol have significantly (P < 0.05) reduced the fresh and dry weight of mycelia. The collapse and damage on B. cinerea mycelia treated with 40 μg/mL of carvacrol and thymol was examined by scanning electron microscope (SEM). Through extracellular conductivity test and fluorescence microscope observation, it was found that carvacrol and thymol led to increase the permeability of target cells, the destruction of mitochondrial membrane and ROS accumulation. In vivo conditions, 1000 μg/mL carvacrol had the best protective and therapeutic effects on tomato gray mold (77.98% and 28.04%, respectively), and the protective effect was significantly higher than that of 400 μg/mL pyrimethanil (43.15%). While the therapeutic and protective effects of 1,000 μg/mL OVEO and thymol were comparable to chemical control. Conclusions OVEO showed moderate antifungal activity, whereas its main components carvacrol and thymol have great application potential as natural fungicides or lead compounds for commercial fungicides in preventing and controlling plant diseases caused by B. cinerea.


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