Beneficial Bacteria Identified for the Control of Botrytis cinerea in Petunia Greenhouse Production

Botrytis cinerea infects most major greenhouse crops worldwide. With its increasing resistance to conventional fungicides and the movement of the greenhouse industry toward more sustainable production practices, alternative methods of control are needed. The objective of this study was to evaluate a collection of 60 bacterial strains through both a dual-culture assay and greenhouse trials to identify strains with biocontrol activity against B. cinerea. For the dual-culture assay, each bacterial strain was streaked on potato dextrose agar medium with B. cinerea. The B. cinerea growth reduction and the zone of inhibition were measured. Thirty-five strains reduced the growth of B. cinerea. All strains were also tested in an initial greenhouse trial in which Petunia × hybrida ‘Carpet Red Bright’ was sprayed and drenched with the bacteria biweekly for 6 weeks. All open flowers were tagged, and plants were inoculated with B. cinerea (1 × 104 conidia per 1 ml). Disease severity indices calculated from the daily flower gray mold severity ratings of all tagged flowers were used to identify the seven top-performing strains. These seven strains were then evaluated in a greenhouse validation trial. The methods were similar to those of the initial greenhouse trials except that replicate numbers were increased. Three strains (Pseudomonas protegens AP54, Pseudomonas chlororaphis 14B11, and Pseudomonas fluorescens 89F1) were selected for the ability to reduce B. cinerea infection in a greenhouse production setting. These strains can be used in future studies to develop additional biocontrol products for the management of B. cinerea in floriculture crops.

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First Report of Gray Mold Caused by Botrytis cinerea on Greenhouse-Grown Zucchini in Korea

Plant Disease ◽

10.1094/pdis-01-13-0005-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1116-1116 ◽

Cited By ~ 1

Author(s):

W. Cheon ◽

Y. H. Jeon

Keyword(s):

New York ◽

Botrytis Cinerea ◽

Severe Disease ◽

Morphological Characteristics ◽

Its Region ◽

Gray Mold ◽

Greenhouse Production ◽

First Report ◽

Plastic Bags ◽

Diseased Fruit

In the winter of 2011, greenhouse-grown zucchini (Cucurbita pepo) in Andong City, Korea, showed severe disease symptoms on fruits and dying leaves of zucchini plants that resembled gray mold disease with about 20% yield loss. Symptoms included extensive growth of mycelia and gray conidia on stem and fruit lesions. Lesions expanded rapidly under cool, humid conditions. As the disease progressed, leaves, stems, and fruits became necrotic and were covered by an abundant, soft, gray, sporulating mycelium. Diseased fruit tissue was excised and surface sterilized by immersion in 2% NaOCl for 1 min, placed on PDA (potato dextrose agar), and incubated at 22°C. Fungal colonies were initially white and became gray to brown after 72 h. Analysis of light micrographs showed the presence of elliptical conidia on PDA that was 7.5 to 16.0 μm long and 5 to 10.5 μm wide. In culture, a few, black, small and large irregular sclerotia were produced. Microsclerotia were round, spherical or irregular in shape, and ranged from 1.0 to 3.3 and 1.2 to 3.4 mm (width and length). Conidiophores were slender and branched with enlarged apical cells bearing smooth, ash-colored conidia. These morphological characteristics identified the fungus as Botrytis cinerea (1). The internal transcribed spacer (ITS) region of rDNA was amplified using the ITS1 (forward) and ITS4 (reverse) primer set (ITS1: 5′-TCCGTAGGTGAACCTGCGG-3′, ITS4: 5′-TCCTCCGCTTATTGATATGC-3′) and sequenced (2). BLAST analysis of the PCR product showed that the sequence had 100% identity with the nucleotide sequences for B. cinerea. Pathogenicity tests were performed by placing mycelium fragments (1 cm2) of PDA cultures on zucchini fruits. Controls were treated with PDA alone. Five replicates for the inoculated and control plants were used. All fruits were covered with plastic bags and incubated in a growth chamber to maintain 90 to 100% relative humidity at 22°C. Typical symptoms appeared 2 to 6 days after inoculation. The inoculated plants developed typical gray mold symptoms with gray sporulating lesions, while controls remained healthy with no lesions. B. cinerea reisolated from the inoculated tissues was morphologically identical to the original isolates. In a cold outside (below 0°C), wet greenhouse, plants are likely to be exposed to resident Botrytis populations and if the gray mold disease occurs, it can spread on zucchini plants very fast, in 2 days to a week inside a 100 m2 greenhouse. Therefore, gray mold disease could have a significant impact on greenhouse production of zucchini. To our knowledge, this is the first report of B. cinerea causing gray mold of greenhouse-grown zucchini in Korea. References: (1) H. L. Barnett and B. B. Hunter. Illustrated Genera of Imperfect Fungi. Burgess Publishing Company, Minneapolis, MN, 1972. (2) T. J. White et al. PCR Protocols. Academic Press, Inc., New York, 1990.

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Biological Control of Tomato Gray Mold Caused by Botrytis Cinerea with the Entomopathogenic Fungus Metarhizium Anisopliae

Pathogens ◽

10.3390/pathogens9030213 ◽

2020 ◽

Vol 9 (3) ◽

pp. 213 ◽

Cited By ~ 1

Author(s):

Most.Sinthia Sarven ◽

Qiuyan Hao ◽

Junbo Deng ◽

Fang Yang ◽

Gaofeng Wang ◽

...

Keyword(s):

Botrytis Cinerea ◽

Culture Filtrate ◽

Metarhizium Anisopliae ◽

Entomopathogenic Fungus ◽

Radial Growth ◽

Tomato Fruit ◽

Inhibitory Effect ◽

Gray Mold ◽

Dual Culture ◽

Cell Organelles

Gray mold disease caused by Botrytis cinerea is a devastating disease that leads to serious financial loss. In this study, the entomopathogenic fungus Metarhizium anisopliae that acts against the gray mold pathogen B. cinerea was evaluated. M. anisopliae produced a significant inhibition zone in front of the B. cinerea colony in the dual culture test. In addition, volatile organic compounds generated by M. anisopliae were shown to have an inhibitory effect on B. cinerea mycelia growth and reduced 41% of gray mold severity of postharvest tomatoes. The 10% concentration of the culture filtrate of M. anisopliae inhibited 88.62% of colony radial growth as well as 63.85% of sclerotia germination and all conidia germination of B. cinerea. Furthermore, the culture filtrate of M. anisopliae retained its inhibitory effect against the radial growth of B. cinerea even after heating for 15 min at 100 °C. Feasible mechanisms of M. anisopliae involved in the control of B. cinerea were explored, and it was demonstrated that the plasma membrane of B. cinerea conidia was damaged by the product of metabolism of M. anisopliae. In addition, after treating with culture filtrate of M. anisopliae, the B. cinerea phenotype was shown to be abnormal, and cell organelles of B. cinerea mycelia were damaged significantly. A significant control efficacy of M. anisopliae against tomato gray mold was detected on both the detached leaf assay (84.24%) as well as the whole plant (72.38%). In addition, a 78% reduction in tomato fruit mold was detected at a 10% treated concentration of M. anisopliae. These findings suggest that M. anisopliae possesses potential as a biocontrol agent against tomato gray mold in the greenhouse and during the postharvest stage.

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Post-Harvest Non-Conventional and Traditional Methods to Control Cadophora luteo-olivacea: Skin Pitting Agent of Actinidia chinensis var. deliciosa (A. Chev.)

Horticulturae ◽

10.3390/horticulturae7070169 ◽

2021 ◽

Vol 7 (7) ◽

pp. 169

Author(s):

Alessandra Di Francesco ◽

Michele Di Foggia ◽

Alessio Vittoria ◽

Elena Baraldi

Keyword(s):

Alternative Methods ◽

In Vitro Assays ◽

Dual Culture ◽

Critical Problem ◽

Post Harvest ◽

Dual Culture Assay ◽

Ft Ir ◽

Alternative Solutions

Cadophora luteo-olivacea represents a critical problem for kiwifruit in the post-harvest phase, mainly for its little note epidemiology. The study presented some results about the possibility of preserving kiwifruit from skin pitting symptoms using alternative methods to fungicides. By in vitro assays, antagonist mechanisms of action against pathogen isolates were tested. Trichoderma harzianum (Th1) showed the highest inhibitory activity against C. luteo-olivacea isolates by volatile, non-volatile, and by dual culture assay, displaying an inhibition respectively by 90%, 70.6%, and 78.8%, and with respect to Aureobasidium pullulans (L1 and L8) by 23.3% and 25.8%, 50% and 34.7%, and 22.5% and 23.6%, respectively. Further, the sensitivity on CFU and mycelial growth of C. luteo-olivacea isolates to fludioxonil, and CaCl2 was tested, displaying interesting EC50 values (0.36 and 0.92 g L−1, 22.5 g L−1, respectively). The effect of Brassica nigra defatted meal was tested as biofumigation assays and through FT-IR (Fourier-Transform Infrared) spectroscopy. The above-mentioned treatments were applied in vivo to evaluate their efficacy on kiwifruits. Our data demonstrated that alternative solutions could be considered to control postharvest pathogens such as C. luteo-olivacea.

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Potencial de extratos etanólicos de propólis e extratos aquosos de plantas espontâneas no controle de doenças pós-colheita do morango

Revista Verde de Agroecologia e Desenvolvimento Sustentável ◽

10.18378/rvads.v11i5.4175 ◽

2016 ◽

Vol 11 (1) ◽

pp. 57

Author(s):

Gabriela Silva Moura ◽

Jonas Marcelo Jaski ◽

Gilmar Franzener

Keyword(s):

Antifungal Activity ◽

Botrytis Cinerea ◽

Gray Mold ◽

Alternative Methods ◽

Wild Plants ◽

Rhizopus Nigricans ◽

Post Harvest ◽

Control Water

A cultura do morangueiro é severamente acometida por várias doenças, dentre elas o mofo cinzento, causado por Botrytis cinerea é considerada a doença mais severa na pós-colheita. Visando reduzir o uso de fungicidas sintéticos, vem sendo realizadas pesquisas propondo a utilização de métodos alternativos de controle de patógenos pós-colheita envolvendo a utilização de extratos vegetais, uso de biofungicidas e óleos essenciais. Assim, o presente trabalho teve como objetivo avaliar o potencial de diferentes extratos de própolis e plantas espontâneas no controle de podridão pós-colheita causada pelo fungo B. cinerea em morangos. Para avaliar a atividade antifúngica direta dos extratos etanólico de própolis e extratos aquosos de plantas espontâneas sobre B. cinerea, foi realizado o experimento in vitro, utilizando-se os tratamentos própolis verde 0,5%; própolis verde 2,5%; própolis marrom 0,5%; própolis marrom 2,5%; língua-de-vaca 10%; assa-peixe 10%; rubim 10%; tansagem 10%; testemunha (água). As medições do diâmetro das colônias foram iniciadas 48, 72 e 96 horas após a instalação do experimento. No experimento in vivo os frutos foram imersos nos tratamentos descritos acima. Após cinco dias avaliou-se a incidência e severidade da doença mofo cinzento e das doenças pós-colheita como antracnose e podridão de Rhizopus que apareceram no experimento. Utilizou-se o delineamento experimental inteiramente casualizado (DIC) com quatro e cinco repetições para o ensaio in vitro e in vivo, respectivamente. Os resultados mostram que os extratos etanólicos de própolis verde e marrom a 2,5% apresentaram in vitro e in vivo atividade antifúngica a B. cinerea e Rhizopus nigricans, respectivamente.Potential of propolis extracts and extracts etanol spontaneous plants aqueous in control of diseases of strawberry post-harvestAbstract: The strawberry crop is severely affected by various diseases, including gray mold, caused by Botrytis cinerea is considered the most severe disease in post-harvest fruit. To reduce the use of synthetic fungicides, has been carried out research proposing the use of alternative methods of control postharvest pathogens involving the use of plant extracts, use of biofungicides, essential oils among others. Thus, this study aimed to evaluate the potential of different propolis extracts and wild plants in the control of post-harvest rot caused by the fungus Botrytis cinerea in strawberries. To evaluate the direct antifungal activity of ethanolic extracts of propolis and aqueous extracts of wild plants of B. cinerea, the in vitro experiment was performed, using treatments propolis 0.5%; propolis 2.5%; brown propolis 0.5%; brown propolis 2.5%; control (water + alcohol 2%); cow tongue 10%; assa-fish 10%; rubim 10%; tansagem 10%; control (water). Measurements of the diameter of the colonies were started 48, 72 and 96 hours after installation of the experiment. Conducted the in vivo experiment in which the fruits of strawberry plants were immersed in the treatments described above. After five days we evaluated the incidence and severity of gray mold disease and post-harvest diseases such as anthracnose and Rhizopus rot appearing in the experiment. We used a completely randomized design (CRD) with four and five replicates for the in vitro assay and in vivo, respectively. The results show that ethanol extracts of green and brown propolis 2.5% presented in vitro and in vivo antifungal activity to B. cinerea and Rhizopus nigricans respectively.

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

Acta Phytopathologica et Entomologica Hungarica ◽

10.1556/038.55.2020.010 ◽

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.

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Cyclophilin BcCyp2 Regulates Infection-Related Development to Facilitate Virulence of the Gray Mold Fungus Botrytis cinerea

International Journal of Molecular Sciences ◽

10.3390/ijms22041694 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1694

Author(s):

Jiao Sun ◽

Chen-Hao Sun ◽

Hao-Wu Chang ◽

Song Yang ◽

Yue Liu ◽

...

Keyword(s):

Botrytis Cinerea ◽

Cyclic Adenosine Monophosphate ◽

Adenosine Monophosphate ◽

Gray Mold ◽

Gene Encoding ◽

Histone 3 ◽

Mold Fungus ◽

Related Development ◽

Hyphal Morphology ◽

Cellular Components

Cyclophilin (Cyp) and Ca2+/calcineurin proteins are cellular components related to fungal morphogenesis and virulence; however, their roles in mediating the pathogenesis of Botrytis cinerea, the causative agent of gray mold on over 1000 plant species, remain largely unexplored. Here, we show that disruption of cyclophilin gene BcCYP2 did not impair the pathogen mycelial growth, osmotic and oxidative stress adaptation as well as cell wall integrity, but delayed conidial germination and germling development, altered conidial and sclerotial morphology, reduced infection cushion (IC) formation, sclerotial production and virulence. Exogenous cyclic adenosine monophosphate (cAMP) rescued the deficiency of IC formation of the ∆Bccyp2 mutants, and exogenous cyclosporine A (CsA), an inhibitor targeting cyclophilins, altered hyphal morphology and prevented host-cell penetration in the BcCYP2 harboring strains. Moreover, calcineurin-dependent (CND) genes are differentially expressed in strains losing BcCYP2 in the presence of CsA, suggesting that BcCyp2 functions in the upstream of cAMP- and Ca2+/calcineurin-dependent signaling pathways. Interestingly, during IC formation, expression of BcCYP2 is downregulated in a mutant losing BcJAR1, a gene encoding histone 3 lysine 4 (H3K4) demethylase that regulates fungal development and pathogenesis, in B. cinerea, implying that BcCyp2 functions under the control of BcJar1. Collectively, our findings provide new insights into cyclophilins mediating the pathogenesis of B. cinerea and potential targets for drug intervention for fungal diseases.

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

Phytopathology ◽

10.1094/phyto-07-16-0250-r ◽

2017 ◽

Vol 107 (3) ◽

pp. 362-368 ◽

Cited By ~ 16

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.

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Promotion of Tomato Growth by the Volatiles Produced by the Hypovirulent Strain QT5-19 of the Plant Gray Mold Fungus Botrytis cinerea

Microbiological Research ◽

10.1016/j.micres.2021.126731 ◽

2021 ◽

pp. 126731

Author(s):

Md Kamaruzzaman ◽

Ze Wang ◽

Mingde Wu ◽

Long Yang ◽

Yongchao Han ◽

...

Keyword(s):

Botrytis Cinerea ◽

Gray Mold ◽

Mold Fungus ◽

Hypovirulent Strain ◽

Tomato Growth

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Computational Fluid Dynamics Modelling of the Microclimate within the Boundary Layer of Leaves Leading to Improved Pest Control Management and Low-Input Greenhouse

Sustainability ◽

10.3390/su13158310 ◽

2021 ◽

Vol 13 (15) ◽

pp. 8310

Author(s):

Hicham Fatnassi ◽

Thierry Boulard ◽

Christine Poncet ◽

Nikolaos Katsoulas ◽

Thomas Bartzanas ◽

...

Keyword(s):

Boundary Layer ◽

Pest Control ◽

Fluid Dynamic ◽

Natural Habitat ◽

Greenhouse Production ◽

Greenhouse Crops ◽

Leaf Surfaces ◽

Measurement Tools ◽

Use Efficiency ◽

Control Management

This work aims at using the Computational Fluid Dynamic (CFD) approach to study the distributed microclimate in the leaf boundary layer of greenhouse crops. Understanding the interactions in this microclimate of this natural habitat of plant pests (i.e., boundary layer of leaves), is a prerequisite for their control through targeted climate management for sustainable greenhouse production. The temperature and humidity simulations, inside the greenhouse, were performed using CFD code which has been adapted to simulate the plant activity within each mesh in the crop canopy. The air temperature and air humidity profiles within the boundary layer of leaves were deduced from the local surrounding climate parameters, based on an analytical approach, encapsulated in a Used Defined Function (UDF), and dynamically linked to the CFD solver, a work that forms an innovative and original task. Thus, this model represents a new approach to investigate the microclimate in the boundary layer of leaves under greenhouses, which resolves the issue of the inaccessibility of this area by the conventionnel measurement tools. The findings clearly showed that (i) contrarily to what might be expected, the microclimate parameters within the boundary layer of leaves are different from the surrounding climate in the greenhouse. This is particularly visible during photoperiods when the plant’s transpiration activity is at its maximum and that (ii) the climatic parameters in the leaf boundary layer are more coupled with leaf surfaces than with those of greenhouse air. These results can help developing localized intervention strategies on the microclimate within boundary layer of plant leaves, leading to improved and sustainable pest control management. The developed climatic strategies will make it possible to optimize resources use efficiency.

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The potential biocontrol agent Paenibacillus polymyxa TP3 produces fusaricidin-type compounds involved in the antagonism against gray mold pathogen Botrytis cinerea

Phytopathology ◽

10.1094/phyto-04-21-0178-r ◽

2021 ◽

Author(s):

Shuen-Huang Tsai ◽

Yu-Ting Chen ◽

Yu-Liang Yang ◽

Bo-Yi Lee ◽

Chien-Jui Huang ◽

...

Keyword(s):

Mass Spectrometry ◽

Botrytis Cinerea ◽

Biological Control Agent ◽

Imaging Mass Spectrometry ◽

Paenibacillus Polymyxa ◽

Field Trials ◽

Gray Mold ◽

Control Agent ◽

Antagonistic Activity ◽

New Variant

Paenibacillus polymyxa is a beneficial bacterium for plant health. Paenibacillus polymyxa TP3 exhibits antagonistic activity toward Botrytis cinerea and alleviates gray mold symptoms on the leaves of strawberry plants. Moreover, suppression of gray mold on the flowers and fruits of strawberry plants in field trials, including vegetative cells and endospores, was demonstrated, indicating the potential of strain TP3 as a biological control agent. To examine the anti-B. cinerea compounds produced by P. polymyxa TP3, matrix‐assisted laser‐desorption/ionization time‐of‐flight mass spectrometry was performed and fusaricidin-corresponding mass spectra were detected. Moreover, fusaricidin-related signals appeared in imaging mass spectrometry of TP3 when confronted with B. cinerea. By using liquid chromatography-mass spectrometry-based molecular networking approach, several fusaricidins were identified including a new variant of m/z 917.5455 with serine in the first position of the hexapeptide. Via advanced mass spectrometry and network analysis, fusaricidin-type compounds produced by P. polymyxa TP3 were efficiently disclosed and were presumed to play roles in the antagonism against gray mold pathogen B. cinerea.

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