scholarly journals Deletion of the Bcnrps1 Gene Increases the Pathogenicity of Botrytis cinerea and Reduces Its Tolerance to the Exogenous Toxic Substances Spermidine and Pyrimethanil

2021 ◽  
Vol 7 (9) ◽  
pp. 721
Author(s):  
Ana Fernández-Morales ◽  
María Carbú ◽  
Victoria González-Rodríguez ◽  
Sokratis Papaspyrou ◽  
Carlos Garrido ◽  
...  
Keyword(s):  
Osmotic Stress ◽  
Vitis Vinifera ◽  
Botrytis Cinerea ◽  
Mycelial Growth ◽  
Culture Medium ◽  
Gene Deletion ◽  
Toxic Substances ◽  
Toxic Compounds ◽  
Sublethal Concentrations

During the infection of grapevine (Vitis vinifera) by the fungus Botrytis cinerea, the concentration of polyamines, which are toxic substances for the phytopathogen, increases in the grape. Nine NRPS genes have been identified in the genome of B. cinerea, yet the function of five of them remains unknown. For this reason, we have studied the expression of the 9 NRPS genes by RT-qPCR in a medium supplemented with sublethal concentrations of three polyamines (1,3-diaminopropane (1,3-DAP), spermidine (SPD), and spermine (SPM)). Our results show that the presence of polyamines in the culture medium triggered the overexpression of the Bcnrps1 gene in the pathogen. Deleting Bcnrps1 did not affect mycelial growth or adaptation to osmotic stress, and we show that its expression is not essential for the cycle of infection of the B. cinerea. However, mutating the Bcnrps1 gene resulted in overexpression of the Bcnrps6 gene, which encodes for the excretion of siderophores of the coprogen family. Moreover, gene deletion has reduced the tolerance of B. cinerea B05.10 to toxic substances such as the polyamine SPD and the fungicide pyrimethanil, and its virulence has increased. Our findings provide new insights into the function of the Bcnrps1 gene and its involvement in the tolerance of B. cinerea against exogenous toxic compounds.

An Attempt at Biological Control of Blossom Blight of Rose Caused by Botrytis cinerea Using some Local Trichoderma spp. Strains

2020 ◽  
Vol 55 (1) ◽  
pp. 27-34
Author(s):  
G. Zadehdabagh ◽  
K. Karimi ◽  
M. Rezabaigi ◽  
F. Ajamgard
Keyword(s):  
Botrytis Cinerea ◽  
Mycelial Growth ◽  
Limiting Factor ◽  
Dual Culture ◽  
Trichoderma Spp ◽  
Khuzestan Province ◽  
Inhibitory Potential ◽  
Cut Rose

The northern of Khuzestan province in Iran is mainly considered as one of the major areas of miniature rose production. Blossom blight caused by Botrytis cinerea has recently become a serious limiting factor in rose production in pre and post-harvest. In current study, an attempt was made to evaluate the inhibitory potential of some local Trichoderma spp. strains against B. cinerea under in vitro and in vivo conditions. The in vitro results showed that all Trichoderma spp. strains were significantly able to reduce the mycelial growth of the pathogen in dual culture, volatile and non-volatile compounds tests compared with control, with superiority of T. atroviride Tsafi than others. Under in vivo condition, the selected strain of T. atroviride Tsafi had much better performance than T. harzianum IRAN 523C in reduction of disease severity compared with the untreated control. Overall, the findings of this study showed that the application of Trichoderma-based biocontrol agents such as T. atroviride Tsafi can be effective to protect cut rose flowers against blossom blight.

scholarly journals Combination of Suspension Array and Mycelial Growth Assay for Detecting Multiple-Fungicide Resistance in Botrytis cinerea in Hebei Province in China

Plant Disease ◽  
2019 ◽  
Vol 103 (6) ◽  
pp. 1213-1219
Author(s):  
Zehua Su ◽  
Xin Zhang ◽  
Jianjiang Zhao ◽  
Wenqiao Wang ◽  
Lei Shang ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
High Throughput ◽  
Fungicide Resistance ◽  
Mycelial Growth ◽  
Accurate Method ◽  
Hebei Province ◽  
Combination Method ◽  
Major Type ◽  
Suspension Array ◽  
Growth Assay

To provide a high-throughput, efficient, and accurate method to monitor multiple-fungicide resistance of Botrytis cinerea in the field, we used the suspension array, sequencing, and mycelial growth assay in our research. Discriminating-dose bioassays for detecting carbendazim, diethofencarb, boscalid, and iprodione resistance (CarR, DieR, BosR, and IprR, respectively) were used to analyze 257 isolates collected from Hebei Province in China during 2016 and 2017. High resistance frequencies to carbendazim (100%), diethofencarb (92.08%), and iprodione (86.59%) were detected. BosR isolates accounted for 11.67% of the total. In addition, 103 isolates were randomly selected for phenotype and genotype detection. The high-throughput suspension array was utilized to detect eight genotypes simultaneously, including BenA-E198, BenA-198A, SdhB-H272, SdhB-272Y, BcOS1-I365, BcOS1-365S, erg27-F412, and erg27-412S, which were associated with resistance toward carbendazim or diethofencarb, boscalid, iprodione, and fenhexamid (FenR), respectively. Most of the benzimidazole-resistant isolates (81.55%) possessed the E198V mutation in the BenA gene. Ninety-three isolates with dual resistance to carbendazim and diethofencarb showed the E198V/K mutation. All BosR isolates carried the H272R mutation in the SdhB gene. The I365S and Q369P+N373S (66.99%) mutations in the BcOS1 gene were more frequently observed. No mutation was detected in the erg27 gene in Hebei isolates. There were 13 resistance profile phenotypes. Phenotypes with triple resistance were the most common (83.50%), and CarRDieRBosSIprRFenS was the major type. CarR isolates that carried E198V/K/A were all highly resistant (HR) and only one F200Y mutant was moderately resistant (MR) to carbendazim. Isolates that possessed E198V/K were MR or HR to diethofencarb. BosR isolates that possessed H272R mutation were lowly resistant (LR). IprR isolates were all LR or MR. The distribution of half maximal effective concentrations of CarR isolates with E198V/K mutations and IprR isolates with Q369P+N373S mutations significantly increased from 2016 to 2017. Combined with our observations, a combination method of the high-throughput suspension array and the mycelial growth assay was suggested to accurately monitor multiple resistance of B. cinerea in the field.

Native and sulfated oligoglucuronans as elicitors of defence-related responses inducing protection against Botrytis cinerea of Vitis vinifera

2012 ◽  
Vol 87 (2) ◽  
pp. 1728-1736 ◽  
Author(s):  
Sébastien Caillot ◽  
Stéphanie Rat ◽  
Marie-Laure Tavernier ◽  
Philippe Michaud ◽  
José Kovensky ◽  
...  
Keyword(s):  
Vitis Vinifera ◽  
Botrytis Cinerea

scholarly journals Maintenance culture medium and inoculum based on peach palm leaves for Pleurotus spp. production

2016 ◽  
Vol 83 (0) ◽  
Author(s):  
Elisabeth Wisbeck ◽  
Endi Pricila Alves ◽  
Styfanie Gonçalves de Lima ◽  
Regina Maria Miranda Gern ◽  
Marcia Luciane Lange Silveira ◽  
...  
Keyword(s):  
Radial Velocity ◽  
Mycelial Growth ◽  
Culture Medium ◽  
Agricultural Waste ◽  
Production Costs ◽  
Fruiting Bodies ◽  
Wheat Grains ◽  
Peach Palm ◽  
Maintenance Medium ◽  
Production Parameters

ABSTRACT: This study evaluated the use of immersion water from peach palm leaves (PPLDA) as a component of the culture medium for the maintenance of Pleurotus spp. and the use of agricultural waste, peach palm leaves, as inoculum support for the fungi. The performance of the inoculum based on peach palm leaves (PPL) for the production of Pleurotus spp. fruiting bodies was compared with that using wheat grains (WG) as inoculum support. PPLDA culture medium (immersion water of peach palm leaves, dextrose, and agar) showed a lower radial velocity of mycelial growth for both fungi than that obtained with the culture medium WDA (wheat extract, dextrose and agar), commonly used as maintenance medium for Pleurotus spp. However, the type of inoculum support does not significantly influence the linear velocity of P. ostreatus mycelial growth, reaching 6.71 mm/day on wheat grains and 6.18 mm/day on peach palm leaves. Thus, when the inoculum based on peach palm leaves is utilized, the immersion water used for preparing this support can be used for preparing the PPLDA maintenance culture medium, diminishing the production costs of Pleurotus mushrooms. Data also showed that when Pleurotus sajor-caju was cultivated on peach palm leaves, using PPL as inoculum support, the fruiting bodies production parameters (Y = 47%, BE = 3% and Pr = 0.2 g/day) did not differ from that obtained using WG.

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 First Report of Botrytis cinerea Causing Gray Mold Disease on Peach from Pakistan

2018 ◽  
Vol 7 (3) ◽  
pp. 131-131
Author(s):  
Raees Ahmed ◽  
Amjad S. Gondal ◽  
Muhammad Tariq Khan ◽  
Shazia Shahzaman ◽  
Sajjad Hyder
Keyword(s):  
Botrytis Cinerea ◽  
Mycelial Growth ◽  
Agar Medium ◽  
Management Strategies ◽  
Gray Mold ◽  
First Report ◽  
Average Production ◽  
Conducive Environment ◽  
Hyphal Tip ◽  
Important Disease

Gray mold caused by Botrytis cinerea is an important disease that attacks fruits, leaves and twigs of peach. Peach is grown on an area of 18,008 ha with an average production of 72,085 tons per year in Pakistan (FAO, 2017). During May 2017, brown spots on 33% of the peach fruits examined were observed in Swat district of KPK province of Pakistan. Infected fruits were incubated at 25±2 °C in a humid chamber resulted in greyish mycelial growth with light brown lesions. Hyphal growths on infected fruits were cultured on PDA media and purified by hyphal tip method. Morphologically whitish grey growth was observed on PDA and later on dark sclerotia were observed after 6-7 days of incubation. Hyphae were found septate with branched hyaline conidiophores having a bunch of ovoid conidia at their tips. Further confirmations were done by amplifying internal transcribed spacer regions (Andrew et al., 2009) and glyceraldehyde-3-phosphate dehydrogenase (G3PDH) region of the isolates (Li et al., 2012). Amplicons sequenced from Macrogen Korea were blasted and submitted in NCBI showed that ITS sequences (Accessions MH049690 and MH049691) were 99% identical with already reported (MG878388 and MG654661) sequences and the G3PDH gene sequences (Accessions MH560352 and MH560353) were 99 % identical with already reported (Accessions MG204876) sequences of B. cinerea. Pathogenicity was confirmed on healthy peach fruits disinfected with 50% ethanol, inoculated by placing a plug of about 1cm2 taken from the edge of actively growing B. cinerea isolate (BTS-16). Fruits were incubated at 25±2 °C in a humid chamber (Abata et al., 2016). A set of healthy fruits mock-inoculated with a plug of agar medium were used as control. Three days after inoculation, inoculated fruits showed sunken lesions with cottony greyish mycelial growth on their surface. Fungus isolated from these infections was re-confirmed as B. cinerea. Conducive environment for the disease progression in nearby areas can result into a huge loss in peach produce so there is a need to devise management strategies to cope with the pathogen. This is the first report of gray mold disease of peach caused by B. cinerea from Pakistan. 

scholarly journals Selection of filamentous fungi that are resistant to the herbicides atrazine, glyphosate and pendimethalin

2021 ◽  
Vol 43 ◽  
pp. e51656
Author(s):  
Nara Priscila Barbosa Bravim ◽  
Anatércia Ferreira Alves ◽  
José Fábio França Orlanda ◽  
Patricia Barbosa Rodrigues Silva
Keyword(s):  
Filamentous Fungi ◽  
Mycelial Growth ◽  
Culture Medium ◽  
Agricultural Soils ◽  
Culture Media ◽  
Fusarium Verticillioides ◽  
Fungal Growth ◽  
Relative Growth ◽  
Maximum Inhibition ◽  
Selection Of

The objective of the present study was to isolate fungi from agricultural soils and evaluate fungal growth in culture medium contaminated with atrazine, glyphosate and pendimethalin. Filamentous fungi were isolated from agricultural soils and cultured in a modified culture medium containing 0, 10, 20, 50, and 100 μg mL-1 atrazine, glyphosate and pendimethalin for 14 days at 28°C. The fungi that presented optimal and satisfactory growth were plated in Sabouraud culture medium with 4% dextrose and containing the herbicides at concentrations of 0, 10, 20, 50, and 100 μg mL-1 for seven days at 28°C. The mean mycelial growth values were submitted to analysis of variance and the Tukey test (p < 0.05%) for comparison and relative growth determination, and maximum inhibition rates were calculated. The isolated fungi Aspergillus fumigatus, Fusarium verticillioides and Penicillium citrinum were shown to be resistant to atrazine, glyphosate and pendimethalin. F. verticillioides showed higher mean mycelial growth in the culture media contaminated with atrazine and glyphosate than the other two fungi. In the culture medium contaminated with pendimethalin, F. verticillioides, and A. fumigatus presented the highest mean mycelial growth values.

scholarly journals Saprophytic microorganisms with potential for biological control of Botrytis cinerea on Geraldton waxflower flowers

2001 ◽  
Vol 41 (5) ◽  
pp. 697 ◽  
Author(s):  
D. R. Beasley ◽  
D. C. Joyce ◽  
L. M. Coates ◽  
A. H. Wearing
Keyword(s):  
Botrytis Cinerea ◽  
Mycelial Growth ◽  
Germ Tube ◽  
Conidial Germination ◽  
Pseudomonas Sp ◽  
Trichoderma Spp ◽  
Fusarium Sp ◽  
Flower Abscission ◽  
Germ Tube Elongation

Saprophytic bacteria, yeasts and filamentous fungi were isolated from Geraldton waxflower flowers and screened to identify potential antagonism towards Botrytis cinerea. Isolates from other sources (e.g. avocado) were also tested. Isolates were initially screened in vitro for inhibition of B. cinerea conidial germination, germ tube elongation and mycelial growth. The most antagonistic bacteria, yeasts and fungi were selected for further testing on detached waxflower flowers. Conidia of the pathogen were mixed with conidia or cells of the selected antagonists, co-inoculated onto waxflower flowers, and the flowers were sealed in glass jars and incubated at 20˚C. The number of days required for the pathogen to cause flower abscission was determined. The most antagonistic bacterial isolate, Pseudomonas sp. 677, significantly reduced conidial germination and retarded germ tube elongation of B. cinerea. None of the yeast or fungal isolates tested was found to significantly reduce conidial germination or retard germ tube elongation, but several significantly inhibited growth of B. cinerea. Fusarium sp., Epicoccum sp. and Trichoderma spp. were the most antagonistic of these isolates. Of the isolates tested on waxflower, Pseudomonas sp. 677 was highly antagonistic towards B. cinerea and delayed waxflower abscission by about 3 days. Trichoderma harzianum also significantly delayed flower abscission. However, as with most of the fungal antagonists used, inoculation of waxflower flowers with this isolate resulted in unsightly mycelial growth.

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