scholarly journals Functional Analysis of the Cytochrome P450 Monooxygenase Gene bcbot1 of Botrytis cinerea Indicates That Botrydial Is a Strain-Specific Virulence Factor

2005 ◽  
Vol 18 (6) ◽  
pp. 602-612 ◽  
Author(s):  
Verena Siewers ◽  
Muriel Viaud ◽  
Daniel Jimenez-Teja ◽  
Isidro G. Collado ◽  
Christian Schulze Gronover ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Functional Characterization ◽  
Gray Mold ◽  
Infection Process ◽  
In Planta ◽  
P450 Monooxygenase ◽  
Α Subunit ◽  
Monooxygenase Gene

The micrographic phytopathogen Botrytis cinerea causes gray mold diseases in a large number of dicotyledonous crop plants and ornamentals. Colonization of host tissue is accompanied by rapid killing of plant cells ahead of the growing hyphen, probably caused by secretion of nonspecific phytotoxins, e.g., the sesquiterpene botrydial. Although all pathogenic strains tested so far had been shown to secrete botrydial and although the toxin causes comparable necrotic lesions as infection by the fungus, the role of botrydial in the infection process has not been elucidated so far. Here, we describe the functional characterization of bcbot1, encoding a P450 monooxygenase and provide evidence that it is involved in the botrydial pathway, i.e., it represents the first botrydial biosynthetic gene identified. We show that bcbot1 is expressed in planta and that expression in vitro and in planta is controlled by an α-subunit of a heterotrimeric GTP-binding protein, BCG1. Deletion of bcbot1 in three standard strains of B. cinerea shows that the effect on virulence (on several host plants) is strain-dependent; only deletion in one of the strains (T4) led to reduced virulence.

scholarly journals Actinomycete as biocontrol agents against tomato gray mold disease caused by Botrytis cinerea

2021 ◽  
Vol 48 (3) ◽  
Author(s):  
Hind Lahmyed ◽  
◽  
Rachid Bouharroud ◽  
Redouan Qessaoui ◽  
Abdelhadi Ajerrar ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Extracellular Enzymes ◽  
Chitinase Activity ◽  
Gray Mold ◽  
Antagonistic Activity ◽  
Future Research ◽  
In Planta ◽  
Tomato Fruits

The present work aims to isolate actinomycete bacteria with antagonistic abilities towards Botrytis cinerea, the causal agent of gray mold, from a soil sample collected from the rhizosphere of a healthy tomato grove. In vitro confrontation led to the isolation of 104 actinomycete isolates; fifteen isolates have shown the most significant mortality rate of the mycelial growth of B. cinerea (>50%). Based on the results of this screening, representative strains were selected to verify their in vivo antagonistic activity on tomato fruits; the reduction of B. cinerea has a percentage ranging from 52.38% to 96.19%. Furthermore, the actinomycete isolates were evaluated for their plant growth-promoting (PGP) properties and their ability to produce biocontrol-related extracellular enzymes viz., amylase, protease, cellulase, chitinase, esterases, and lecithinase. Indeed, Ac70 showed high β-1,3-glucanase activity and siderophore production (17U/ml and 43% respectively), and the highest chitinase activity (39μmol/ml) was observed for Ac24. These results indicated that these actinomycetes might potentially control gray mold caused by B. cinerea on tomato fruits. Investigations on enhancing the efficacy and survival of the biocontrol agent in planta and finding out the best formulation are recommended for future research.

scholarly journals The Role of G Protein Alpha Subunits in the Infection Process of the Gray Mold Fungus Botrytis cinerea

2001 ◽  
Vol 14 (11) ◽  
pp. 1293-1302 ◽  
Author(s):  
Christian Schulze Gronover ◽  
Daniela Kasulke ◽  
Paul Tudzynski ◽  
Bettina Tudzynski
Keyword(s):  
Botrytis Cinerea ◽  
Magnaporthe Grisea ◽  
Gray Mold ◽  
Infection Process ◽  
Colony Morphology ◽  
Gene Copy ◽  
Wild Type ◽  
Mold Fungus ◽  
Protease Secretion

To identify signal transduction pathways of the gray mold fungus Botrytis cinerea involved in host infection, we used heterologous hybridization and a polymerase chain reaction (PCR)-based approach to isolate two genes (bcg1 and bcg2) encoding α subunits of heterotrimeric GTP-binding proteins. Both genes have homologues in other fungi: bcg1 is a member of the Gαi class, whereas bcg2 has similarities to the magC gene of Magnaporthe grisea and the gna-2 gene of Neurospora crassa. Reverse-transcription (RT)-PCR experiments showed clearly that both genes are expressed at very early stages in infected bean leaves. Gene replacement experiments were performed for both genes. bcg1 null mutants differ in colony morphology from the wild-type strain, do not secrete extracellular proteases, and show clearly reduced pathogenicity on bean and tomato. Conidia germination and penetration of plant tissue is not disturbed in bcg1 mutants, but the infection process stops after formation of primary lesions. In contrast, bcg2 mutants show wild-type colony morphology in axenic culture and are only slightly reduced in pathogenicity. Complementation of bcg1 mutants with the wild-type gene copy led to the full recovery of colony morphology, protease secretion, and pathogenicity on both host plants. Application of exogenous cyclic AMP restored the wild-type growth pattern of bcg1 mutants, but not the protease secretion, implicating an essential role of BCG1 in different signaling pathways.

Characterization of Botrytis cinerea and B. prunorum from Healthy Floral Structures and Decayed ‘Hayward’ Kiwifruit During Post-Harvest Storage

Plant Disease ◽  
2020 ◽  
Author(s):  
Danae Riquelme ◽  
Zdenka Aravena ◽  
Hector Valdes ◽  
Bernardo Antonio Latorre ◽  
Gonzalo A Díaz ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Central Valley ◽  
Gray Mold ◽  
Heat Shock Protein 60 ◽  
Post Harvest ◽  
Diseased Fruit ◽  
Apparently Healthy

Gray mold is the primary post-harvest disease of ‘Hayward’ kiwifruit (Actinidia deliciosa) in Chile, with a prevalence of 33.1% in 2016 and 7.1% in 2017. Gray mold develops during postharvest storage, which is characterized by a soft, light to brown watery decay that is caused by Botrytis cinerea and B. prunorum. However, there is no information related to the role of B. prunorum during the development and storage of kiwifruit in Chile. For this purpose, asymptomatic flowers and receptacles were collected throughout fruit development and harvest from five orchards over two seasons in the Central Valley of Chile. Additionally, diseased kiwifruits were selected after storage for 100 days at 0°C plus 2 days at 20° C. High (HCP) and low conidial production (LCP) colonies of Botrytis sp. were consistently obtained from apparently healthy petals, sepals, receptacles, styles, and diseased kiwifruit. Morphological and phylogenetic analysis using three partial gene sequences encoding glyceraldehyde-3-phosphate dehydrogenase (G3PDH), heat shock protein 60 (HSP60), and DNA-dependent RNA polymerase subunit II (RPB2) were able to identify and separate B. cinerea and B. prunorum species. Consistently, B. cinerea was predominantly isolated from all floral parts and fruit in apparently healthy tissue and diseased kiwifruit. During full bloom, the highest colonization by B. cinerea and B. prunorum was obtained from petals followed by sepals. In storage, both Botrytis species were isolated from the diseased fruit (n=644), of which 6.8% (n=44) were identified as B. prunorum. All Botrytis isolates grew from 0°C to 30°C in vitro and were pathogenic on kiwifruit leaves and fruit. Notably, B. cinerea isolates were always more virulent than B. prunorum isolates. This study confirms the presence of B. cinerea and B. prunorum colonizing apparently healthy flowers and floral parts in fruit and causing gray mold during kiwifruit storage in Chile. Therefore, B. prunorum plays a secondary role in the epidemiology of gray mold developing in kiwifruit during cold storage.

scholarly journals Does Botrytis cinerea Ignore H2O2-Induced Oxidative Stress During Infection? Characterization of Botrytis Activator Protein 1

2009 ◽  
Vol 22 (8) ◽  
pp. 987-998 ◽  
Author(s):  
Nora Temme ◽  
Paul Tudzynski
Keyword(s):  
Oxidative Stress ◽  
Stress Response ◽  
Botrytis Cinerea ◽  
Oxidative Burst ◽  
Target Genes ◽  
Oxidative Stress Response ◽  
Infection Process ◽  
Defense Reaction ◽  
In Planta

Botrytis cinerea is a phytopathogen infecting a broad range of plants including strawberries and grapevine. During infection, the necrotrophic fungus is exposed to reactive oxygen species (ROS) released by the oxidative burst, an early plant defense reaction. B. cinerea even produces ROS itself in planta. This raises questions about how the pathogen senses and responds to the host defense reaction and which role the pathogen's oxidative stress response systems play. Functional analysis of the AP-1 transcription factor Bap1 confirmed its role as a pivotal regulator of ROS detoxification in vitro. Macroarray analysis revealed 99 H2O2-induced Bap1 target genes, of which several genes encoded ROS-degrading enzymes as well as other central components of the cellular redox status. However, Bap1 is not essential for pathogenesis. In planta analyses revealed that the Bap1 target genes were not expressed 2 days postinoculation although H2O2 was detectable, proving that the normal virulence of the Δbap1 mutant is not due to alternative regulation of the major oxidative stress response system in planta. The fungus obviously does not suffer H2O2-induced oxidative stress in planta, questioning classical ideas about the role of the oxidative burst in the infection process.

scholarly journals Characterization of Postharvest Fungicide-Resistant Botrytis cinerea Isolates From Commercially Stored Apple Fruit

2017 ◽  
Vol 107 (3) ◽  
pp. 362-368 ◽  
Author(s):  
Wayne M. Jurick ◽  
Otilia Macarisin ◽  
Verneta L. Gaskins ◽  
Eunhee Park ◽  
Jiujiang Yu ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Gray Mold ◽  
Apple Fruit ◽  
Sublethal Dose ◽  
Long Term Storage ◽  
Pose Control ◽  
First Time

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.

scholarly journals AVALIAÇÃO DO REAL POTENCIAL INIBIDOR DE EXTRATOS ETANÓLICOS DE Ottonia martiana SOBRE Cylindrocladium spathulatum E Botrytis cinerea

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 ◽  
...  
Keyword(s):  
Factorial Design ◽  
Botrytis Cinerea ◽  
Mycelial Growth ◽  
Black Spot ◽  
Gray Mold ◽  
Causal Agent ◽  
In Vitro Testing ◽  
Ethanolic Extracts ◽  
Antifungal Potential

 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.

scholarly journals Expression of the ToxA and PtrPf2 genes of the phytopathogenic fungus Pyrenophora tritici-repentis at the beginning of the infection process

2019 ◽  
Author(s):  
Nina V. Mironenko ◽  
Alexandra S. Orina ◽  
Nadezhda M. Kovalenko
Keyword(s):  
Transcription Factor ◽  
Wheat Variety ◽  
Infection Process ◽  
Phytopathogenic Fungus ◽  
In Planta ◽  
Effector Gene ◽  
Pyrenophora Tritici Repentis ◽  
Wheat Leaves ◽  
Necrotrophic Effector

This study shows that the necrotrophic effector gene ToxA is differentially expressed in isolates of P. tritici-repentis fungus at different time periods after inoculation of the wheat variety Glenlea which has the gene Tsn1 controlling sensitivity to the necrosis inducing toxin Ptr ToxA. Two P. tritici-repentis isolates with different ability to cause necrosis on the leaves of Glenlea variety (nec + and nec-) and with different expression level of ToxA and gene of factor transcription PtrPf2 in vitro were used for analysis. Isolates of P. tritici-repentis are characterized by the differential expression of ToxA in planta. The expression of the ToxA gene in P. tritici-repentis ToxA+ isolates significantly increased when infected the wheat leaves compared to ToxA expression results obtained in vitro. The levels of ToxA expression in both isolates differed significantly after 24, 48 and 96 hours after inoculation, however, the dynamics of the trait change over time were similar. However, the highest ToxA expression in the virulent (nec+) isolate in contrast with the avirulent (nec-) isolate was observed at a point of 48 hours. Whereas the expression of regulating transcription factor PtrPf2 in planta differed imperceptibly from expression in vitro throughout the observation period. Obviously, the role of the fungal transcription factor in regulating the effector gene expression weakens in planta, and other mechanisms regulating the expression of pathogen genes at the biotrophic stage of the disease develop.

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
Keyword(s):  
Biological Control ◽  
Botrytis Cinerea ◽  
Fungal Endophytes ◽  
Gray Mold ◽  
Penicillium Sp ◽  
Trichoderma Sp ◽  
Forest Nurseries ◽  
Aspergillus Sp ◽  
Foliar Pathogens

   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.    

scholarly journals Botrytis pseudocinerea, a New Cryptic Species Causing Gray Mold in French Vineyards in Sympatry with Botrytis cinerea

2011 ◽  
Vol 101 (12) ◽  
pp. 1433-1445 ◽  
Author(s):  
Anne-Sophie Walker ◽  
Angélique Gautier ◽  
Johann Confais ◽  
Daniel Martinho ◽  
Muriel Viaud ◽  
...  
Keyword(s):  
New Species ◽  
Cryptic Species ◽  
Botrytis Cinerea ◽  
Germination Rate ◽  
Gray Mold ◽  
Temperature Adaptation ◽  
Morphological Criteria ◽  
Ecological Criteria ◽  
High Level

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.

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