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 Effects of Origanum vulgare essential oil and its two main components, carvacrol and thymol, on the plant pathogen Botrytis cinerea

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9626
Author(s):  
Huiyu Hou ◽  
Xueying Zhang ◽  
Te Zhao ◽  
Lin Zhou
Keyword(s):  
Antifungal Activity ◽  
Essential Oil ◽  
Botrytis Cinerea ◽  
Spore Germination ◽  
Gray Mold ◽  
Origanum Vulgare ◽  
Mycelium Growth ◽  
Main Components

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.

scholarly journals Survival of Spray-Dried Rhodotorula mucilaginosa Isolated from Natural Microbiota of Murta Berries and Antagonistic Effect on Botrytis cinerea

2019 ◽  
Vol 57 (2) ◽  
pp. 222-229
Author(s):  
Alexandra González-Esparza ◽  
Kong S. Ah-Hen ◽  
Osvaldo Montenegro ◽  
Erika Briceño ◽  
Joaquín Stevenson ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Spray Drying ◽  
Grey Mould ◽  
Antagonistic Activity ◽  
Antagonistic Effect ◽  
Inlet Temperature ◽  
Rhodotorula Mucilaginosa ◽  
Natural Microbiota

The aim of this study is to evaluate the survival rate and effective antagonistic activity against Botrytis cinerea, responsible for grey mould on harvested fruits and vegetables, of yeast Rhodotorula mucilaginosa, isolated and identified from the natural microbiota of murta (Chilean guava) berries, after spray drying at different inlet air temperatures, mass per volume ratio of encapsulating agent (maltodextrin) and feed flow rates. The 100 % survival of the yeast was obtained after spray drying with 18 % maltodextrin at 130 °C inlet temperature and a feed flow rate of 9.25 mL/min. The dried yeast obtained under such conditions had the highest antagonistic activity in vitro and in vivo on apples, which showed that spray drying is a valid method to produce active dried cells of R. mucilaginosa that can be used for biocontrol of grey mould spoilage. It was also found that the encapsulating agent maltodextrin improved the in vitro antagonistic activity of R. mucilaginosa.

scholarly journals Identification of epiphytic yeasts and bacteria with potential for biocontrol of grey mold disease on table grapes caused by Botrytis cinerea

2018 ◽  
Vol 16 (1) ◽  
pp. e1002 ◽  
Author(s):  
Kazem Kasfi ◽  
Parissa Taheri ◽  
Behrooz Jafarpour ◽  
Saeed Tarighi
Keyword(s):  
Botrytis Cinerea ◽  
Disease Incidence ◽  
Gray Mold ◽  
Antagonistic Activity ◽  
Antagonistic Effect ◽  
Bacterial Isolates ◽  
Thompson Seedless ◽  
Grape Berries ◽  
Table Grapes

The objective of this study was to identify grapevine epiphytic yeasts and bacteria for biocontrol of Botrytis cinerea on grapes. Antagonistic yeasts and bacteria were isolated from the epiphytic flora associated with grape berries and leaves cv. ‘Thompson seedless’ from vineyards in Iran and identified by sequencing the conserved genomic regions. A total of 130 yeast and bacterial isolates from the surface of grapevine were screened in vitro for determining their antagonistic effect against B. cinerea and used to control postharvest gray mold. Among the 130 isolates, five yeasts and four bacterial isolates showed the greatest antagonistic activity in vitro against B. cinerea. Two yeasts species including Meyerozyma guilliermondii and Candida membranifaciens had high antagonistic capability against the pathogen. Also, 4 bacterial isolates belonging to Bacillus sp. and Ralstonia sp. showed significant biocontrol effect against B. cinerea. The isolates were capable of producing volatile and non-volatile substances, which suppressed the pathogen growth. The antagonistic activity of selected yeasts and bacteria against the pathogen was investigated on wounded berries of ‘Thompson seedless’. On small clusters with intact berries, all of the antagonistic isolates considerably reduced the decay on grape berries and inhibition of gray mold incidence on fruits treated by these isolates was less than 50%, except for the isolate N1, which had higher capability in inhibiting the disease incidence. These results suggest that antagonist yeasts and bacteria with potential to control B. cinerea on grape can be found in the microflora of grape berries and leaves.

scholarly journals Genomic and Metabolomic Insights into Secondary Metabolites of the Novel Bacillus halotolerans Hil4, an Endophyte with Promising Antagonistic Activity against Gray Mold and Plant Growth Promoting Potential

2021 ◽  
Vol 9 (12) ◽  
pp. 2508
Author(s):  
Eirini-Evangelia Thomloudi ◽  
Polina C. Tsalgatidou ◽  
Eirini Baira ◽  
Konstantinos Papadimitriou ◽  
Anastasia Venieraki ◽  
...  
Keyword(s):  
Plant Growth ◽  
Growth Promotion ◽  
Gene Clusters ◽  
Gray Mold ◽  
Antagonistic Activity ◽  
Future Research ◽  
Single Culture ◽  
Plant Growth Promoting ◽  
Growth Promoting

The endophytic bacterial strain Hil4 was isolated from leaves of the medicinal plant Hypericum hircinum. It exhibited antifungal activity against Botrytis cinerea and a plethora of plant growth promoting traits in vitro. Whole genome sequencing revealed that it belongs to Bacillus halotolerans and possesses numerous secondary metabolite biosynthetic gene clusters and genes involved in plant growth promotion, colonization, and plant defense elicitation. The Mojavensin cluster was present in the genome, making this strain novel among plant-associated B. halotolerans strains. Extracts of secreted agar-diffusible compounds from single culture secretome extracts and dual cultures with B. cinerea were bioactive and had the same antifungal pattern on TLC plates after bioautography. UHPLC-HRMS analysis of the single culture secretome extract putatively annotated the consecutively produced antimicrobial substances and ISR elicitors. The isolate also proved efficient in minimizing the severity of gray mold post-harvest disease on table grape berries, as well as cherry tomatoes. Finally, it positively influenced the growth of Arabidopsis thaliana Col-0 and Solanum lycopersicum var. Chondrokatsari Messinias after seed biopriming in vitro. Overall, these results indicate that the B. halotolerans strain Hil4 is a promising novel plant growth promoting and biocontrol agent, and can be used in future research for the development of biostimulants and/or biological control agents.

scholarly journals A Polygalacturonase-Inhibiting Protein from Grapevine Reduces the Symptoms of the Endopolygalacturonase BcPG2 from Botrytis cinerea in Nicotiana benthamiana Leaves Without Any Evidence for In Vitro Interaction

2007 ◽  
Vol 20 (4) ◽  
pp. 392-402 ◽  
Author(s):  
Dirk A. Joubert ◽  
Ilona Kars ◽  
Lia Wagemakers ◽  
Carl Bergmann ◽  
Gabré Kemp ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Nicotiana Benthamiana ◽  
Substantial Reduction ◽  
In Vitro Assays ◽  
Physical Interaction ◽  
In Planta ◽  
Polygalacturonase Inhibiting Protein ◽  
Inhibiting Protein

Six endopolygalacturonases from Botrytis cinerea (BcPG1 to BcPG6) as well as mutated forms of BcPG1 and BcPG2 were expressed transiently in leaves of Nicotiana benthamiana using agroinfiltration. Expression of BcPG1, BcPG2, BcPG4, BcPG5, and mutant BcPG1-D203A caused symptoms, whereas BcPG3, BcPG6, and mutant BcPG2-D192A caused no symptoms. Expression of BcPG2 caused the most severe symptoms, including wilting and necrosis. BcPG2 previously has been shown to be essential for B. cinerea virulence. The in vivo effect of this enzyme and the inhibition by a polygalacturonase-inhibiting protein (PGIP) was examined by coexpressing Bcpg2 and the Vvpgip1 gene from Vitis vinifera in N. benthamiana. Coinfiltration resulted in a substantial reduction of the symptoms inflicted by the activity of BcPG2 in planta, as evidenced by quantifying the variable chlorophyll fluorescence yield. In vitro, however, no interaction between pure VvPGIP1 and pure BcPG2 was detected. Specifically, VvPGIP1 neither inhibited BcPG2 activity nor altered the degradation profile of polygalacturonic acid by BcPG2. Furthermore, using surface plasmon resonance technology, no physical interaction between VvPGIP1 and BcPG2 was detected in vitro. The data suggest that the in planta environment provided a context to support the interaction between BcPG2 and VvPGIP1, leading to a reduction in symptom development, whereas neither of the in vitro assays detected any interaction between these proteins.

scholarly journals The Endophytic Fungus Albifimbria verrucaria from Wild Grape as an Antagonist of Botrytis cinerea and Other Grape Pathogens

2020 ◽  
Vol 110 (4) ◽  
pp. 843-850 ◽  
Author(s):  
Zhi Li ◽  
Pingping Chang ◽  
Linlin Gao ◽  
Xiping Wang
Keyword(s):  
Botrytis Cinerea ◽  
Chitinase Activity ◽  
Gray Mold ◽  
Vitis Amurensis ◽  
Grape Quality ◽  
Wide Range ◽  
Biocontrol Activity ◽  
Elsinoe Ampelina ◽  
Conidium Germination

Gray mold, caused by Botrytis cinerea, is one of the most prevalent fungal diseases in table and wine grapes, affecting grape quality and yields. In this study, we isolated several endophytic fungi, including Alternaria alternata, Bipolaris cynodontis, Phoma sp., and Albifimbria verrucaria, from leaves of Amur grape (Vitis amurensis) cultivar Shuangyou and investigated their biocontrol activity against B. cinerea. In vitro dual assay showed that A. verrucaria isolate SYE-1 inhibited growth of B. cinerea. The isolate also had a wide range of biocontrol activity against Lasiodiplodia theobromae and Elsinoë ampelina. Mycelial growth and conidium germination of B. cinerea were significantly inhibited by metabolites of A. verrucaria in agar plates and culture extracts of A. verrucaria from liquid culture. The isolate produced a total chitinase activity of 0.4 U/ml after incubation for 10 days in Czapek’s liquid medium. In addition, application of culture extracts of A. verrucaria prior to B. cinerea inoculation significantly reduced disease severity on grape leaves of the susceptible cultivar Red Globe. Taken together, our results indicate that A. verrucaria has potential as a biocontrol agent to control grape gray mold.

scholarly journals First Report of Fenpyrazamine Resistance in Botrytis cinerea from Strawberry Fields in Spain

2018 ◽  
Vol 19 (1) ◽  
pp. 45-45
Author(s):  
Dolores Fernández-Ortuño ◽  
Alejandra Vielba-Fernández ◽  
Alejandro Pérez-García ◽  
Juan A. Torés ◽  
Antonio de Vicente
Keyword(s):  
Botrytis Cinerea ◽  
Gray Mold ◽  
In Vivo Studies ◽  
Cross Resistance ◽  
In Vitro Tests ◽  
Class Iii ◽  
Single Spore ◽  
First Report

Botrytis cinerea Pers. is an important fungal pathogen responsible for gray mold, one of the most economically important diseases of strawberry (Fragaria × ananassa) worldwide. The primary disease management strategy involves the application of different classes of fungicides, including the sterol biosynthesis inhibitor class III fungicide fenpyrazamine. In 2014 and 2015, strawberries affected with gray mold symptoms were collected from eight locations in Huelva, where fenhexamid had been used extensively. Twenty-five B. cinerea single-spore isolates were examined to determine EC50 values and to determine a discriminatory dose to monitor fenpyrazamine resistance in the field in future studies. The in vitro tests divided the isolates into two groups: 15 sensitive (EC50 from 0.02 to 1.3 μg/ml) and 10 resistant (EC50 from 50.1 to 172.6 μg/ml), which showed cross-resistance with fenhexamid. Performance of fenpyrazamine in in vivo studies was also carried out. Only the fenpyrazamine-resistant isolates developed gray mold on the fungicide-treated fruit. This is the first report of fenpyrazamine resistance in B. cinerea from strawberry fields in Spain and cross-resistance with fenhexamid.

scholarly journals Fitness and Competitive Ability of Botrytis cinerea Field Isolates with Dual Resistance to SDHI and QoI Fungicides, Associated with Several sdhB and the cytb G143A Mutations

2014 ◽  
Vol 104 (4) ◽  
pp. 347-356 ◽  
Author(s):  
T. Veloukas ◽  
P. Kalogeropoulou ◽  
A. N. Markoglou ◽  
G. S. Karaoglanidis
Keyword(s):  
Botrytis Cinerea ◽  
Selection Pressure ◽  
Competitive Ability ◽  
Storage Conditions ◽  
Gray Mold ◽  
Apple Fruit ◽  
Competition Experiment ◽  
Individual Fitness

Respiration inhibitors such as the succinate dehydrogenase inhibitors (SDHIs) and the quinone outside inhibitors (QoIs) are fungicide classes with increasing relevance in gray mold control. However, recent studies have shown that dual resistance to both fungicide classes is a common trait in Botrytis cinerea populations from several hosts throughout the world. Resistance of B. cinerea to SDHIs is associated with several mutations in the sdhB, sdhC, and sdhD genes, while resistance to QoIs, in most cases, is associated with the G143A mutation in the cytb gene. The objective of the current study was to investigate the fitness and the competitive ability of B. cinerea field strains possessing one of the H272Y/R/L, N230I, or P225F sdhB substitutions and the G143A mutation of cytb. Fitness parameters measured were (i) mycelial growth and conidia germination in vitro, (ii) aggressiveness and sporulation capacity in vivo, (iii) sclerotia production in vitro and sclerotia viability under different storage conditions, and (iv) sensitivity to oxidative stress imposed by diquat treatments. The competitive ability of the resistant isolates was measured in the absence and presence of the SDHI fungicides boscalid and fluopyram selection pressure. The measurements of individual fitness components showed that the H272R/G143A isolates had the lower differences compared with the sensitive isolates. In contrast, the groups of H272Y/L/G143A, N230I/G143A, and P225F/G143A isolates showed reduced fitness values compared with the sensitive isolates. Isolates possessing only the cytb G143A substitution did not show any fitness cost. The competition experiments showed that, in the absence of fungicide selection pressure, after four disease cycles on apple fruit, the sensitive isolates dominated in the population in all the mixtures tested. In contrast, when the competition experiment was conducted under the selection pressure of boscalid, a gradual decrease in the frequency of sensitive isolates was observed, whereas the frequency of H272L and P225F isolates was increased. When the competition experiment was conducted in the presence of fluopyram, the sensitive isolates were eliminated even after the first disease cycle and the P225F mutants dominated in the population. Such results suggest that the sdhB mutations may have adverse effects on the mutants. The observed dominance of sensitive isolates in the competition experiments conducted in the absence of fungicides suggest that the application of SDHIs in alternation schemes may delay the selection or reduce the frequency of SDHI-resistant mutants.

scholarly journals Antagonistic Activity of Bacillus spp. Against Fire Blight Disease In vitro and In planta

2022 ◽  
Vol 9 (sp) ◽  
pp. 2486-2492
Author(s):  
Haris Butt ◽  
Kubilay Kurtulus Bastas
Keyword(s):  
Fire Blight ◽  
Positive Control ◽  
Antagonistic Activity ◽  
In Vivo Studies ◽  
In Planta ◽  
Bacillus Spp ◽  
Apple Cultivars ◽  
Blight Disease

Fire blight, affecting more than one hundred and thirty species in the Rosaceae, is probably the most destructive disease affecting pear and apple cultivars in many countries. Currently, there are no effective synthetic compounds with systemic properties. Other major problem is the occurrence and spread of strains of Erwinia amylovora with resistance to streptomycin and copper. Taken into consideration the human and environmental health, the use of biocontrol agents either as an alternative or as a supplement within an integrated fire blight management strategy has attracted worldwide attention. In this study, E. amylovora solution of 107 CFU ml-1 was treated with bio-control agents, Bacillus subtilis str. QST 713, B. amyloliquefaciens str. MBI 600 and their mixture (at solution densities of 106, 107 and 108 CFU ml-1 for each one) on Petri dishes, containing King’s B medium and, compared with positive (streptomycin sulphate) and negative (sterile distilled water) controls. In vivo studies were performed on two-year-old apple cv. Gala seedlings grown in 45-cm-diameter pots containing a sterilized mix of soil–sand–peat under controlled greenhouse conditions (85% relative humidity, 25°C temperature and 16h of day light). The plants were irrigated as needed by drip-irrigation and each pot received a mineral solution (NPK: 20–20–20) at 2 g l-1 twice. When plant shoots reached a length of 30-35 cm, bio-control agents, individually and their mixture, were applied to the plants by a hand-sprayer. Obtaining the data, 108 CFU ml-1 of Bacillus spp. suspension mixture showed strongest in vitro antibacterial effect (26mm) among the tested treatments after positive control streptomycin (28.6mm). Parallel to in vitro findings, the mixture was most effective against the pathogen on cv. Gala (66.03%). Findings show that the use of mixture of beneficial microorganisms with individual antagonistic properties against the pathogen can be an effective strategy as a natural alternative to agrochemicals in the scope of good agriculture practices.

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