Ethylene production of Botrytis cinerea in vitro and during in planta infection of tomato fruits

Author(s):  
S. M. Cristescu ◽  
F. J. M. Harren ◽  
Ernst Woltering
2021 ◽  
Vol 48 (3) ◽  
Author(s):  
Hind Lahmyed ◽  
◽  
Rachid Bouharroud ◽  
Redouan Qessaoui ◽  
Abdelhadi Ajerrar ◽  
...  

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.


2008 ◽  
Vol 98 (12) ◽  
pp. 1334-1339 ◽  
Author(s):  
R. M. Patel ◽  
J. A. L. van Kan ◽  
A. M. Bailey ◽  
G. D. Foster

Gene silencing is a powerful tool utilized for identification of gene function and analysis in plants, animals, and fungi. Here, we report the silencing of superoxide dismutase (bcsod1) in Botrytis cinerea through sense and antisense-mediated silencing mechanisms. Because superoxide dismutase (SOD) is a virulence factor, transformants were tested for phenotypic silencing in vitro and reduction in pathogenicity in planta. Plate-based assays with and without paraquat were performed to screen initial silencing efficiency, and a subset of transformants was used for in planta studies of virulence. Transformants exhibiting strongly decreased transcripts levels were recovered with both constructs but none of those exhibited a reduction in virulence in planta. Our investigations may help optimize a high-throughput gene silencing system useful for identifying potential gene targets for future fungal control.


2007 ◽  
Vol 20 (4) ◽  
pp. 392-402 ◽  
Author(s):  
Dirk A. Joubert ◽  
Ilona Kars ◽  
Lia Wagemakers ◽  
Carl Bergmann ◽  
Gabré Kemp ◽  
...  

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.


2002 ◽  
Vol 68 (11) ◽  
pp. 5342-5350 ◽  
Author(s):  
Simona M. Cristescu ◽  
Domenico De Martinis ◽  
Sacco te Lintel Hekkert ◽  
David H. Parker ◽  
Frans J. M. Harren

ABSTRACT A laser-based ethylene detector was used for on-line monitoring of ethylene released by the phytopathogenic fungus Botrytis cinerea in vitro and in tomato fruit. Ethylene data were combined with the results of a cytological analysis of germination of B. cinerea conidia and hyphal growth. We found that aminoethoxyvinylglycine and aminooxyacetic acid, which are competitive inhibitors of the 1-aminocyclopropane-1-carboxylic acid pathway, did not inhibit the ethylene emission by B. cinerea and that the fungus most likely produces ethylene via the 2-keto-4-methylthiobutyric acid pathway. B. cinerea is able to produce ethylene in vitro, and the emission of ethylene follows the pattern that is associated with hyphal growth rather than the germination of conidia. Ethylene production in vitro depended on the l-methionine concentration added to the plating medium. Higher values and higher emission rates were observed when the concentration of conidia was increased. Compared with the ethylene released by the fungus, the infection-related ethylene produced by two tomato cultivars (cultivars Money Maker and Daniela) followed a similar pattern, but the levels of emission were 100-fold higher. The time evolution of enhanced ethylene production by the infected tomatoes and the cytological observations indicate that ethylene emission by the tomato-fungus system is not triggered by the ethylene produced by B. cinerea, although it is strongly synchronized with the growth rate of the fungus inside the tomato.


Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 970
Author(s):  
Danai Gkizi ◽  
Eirini G. Poulaki ◽  
Sotirios E. Tjamos

Grapevine bunch rot, caused by Botrytis cinerea and Aspergillus carbonarius, causes important economic losses every year in grape production. In the present study, we examined the plant protective activity of the biological control agents, Paenibacillus alvei K165, Blastobotrys sp. FP12 and Arthrobacter sp. FP15 against B. cinerea and A. carbonarius on grapes. The in vitro experiments showed that strain K165 significantly reduced the growth of both fungi, while FP15 restricted the growth of A. carbonarius and FP12 was ineffective. Following the in vitro experiments, we conducted in planta experiments on grape berries. It was shown that K165, FP12 and FP15 reduced A. carbonarius rot severity by 81%, 57% and 37%, respectively, compared to the control, whereas, in the case of B. cinerea, the only protective treatment was that with K165, which reduced rot by 75%. The transcriptomic analysis of the genes encoding the pathogenesis-related proteins PR2, PR3, PR4 and PR5 indicates the activation of multiple defense responses involved in the biocontrol activity of the examined biocontrol agents.


2016 ◽  
Vol 29 (4) ◽  
pp. 287-298 ◽  
Author(s):  
Jason A. Corwin ◽  
Anushriya Subedy ◽  
Robert Eshbaugh ◽  
Daniel J. Kliebenstein

The modern evolutionary synthesis suggests that both environmental variation and genetic diversity are critical determinants of pathogen success. However, the relative contribution of these two sources of variation is not routinely measured. To estimate the relative contribution of plasticity and genetic diversity for virulence-associated phenotypes in a generalist plant pathogen, we grew a population of 15 isolates of Botrytis cinerea from throughout the world, under a variety of in vitro and in planta conditions. Under in planta conditions, phenotypic differences between the isolates were determined by the combination of genotypic variation within the pathogen and environmental variation. In contrast, phenotypic differences between the isolates under in vitro conditions were predominantly determined by genetic variation in the pathogen. Using a correlation network approach, we link the phenotypic variation under in vitro experimental conditions to phenotypic variation during plant infection. This study indicates that there is a high level of phenotypic variation within B. cinerea that is controlled by a mixture of genetic variation, environment, and genotype × environment. This argues that future experiments into the pathogenicity of B. cinerea must account for the genetic and environmental variation within the pathogen to better sample the potential phenotypic space of the pathogen.


2018 ◽  
Vol 108 (6) ◽  
pp. 691-701 ◽  
Author(s):  
Xuefei Wang ◽  
Dean A. Glawe ◽  
Elizabeth Kramer ◽  
David Weller ◽  
Patricia A. Okubara

Native yeasts are of increasing interest to researchers, grape growers, and vintners because of their potential for biocontrol activity and their contributions to the aroma, flavor, and mouthfeel qualities of wines. To assess biocontrol activity, we tested 11 yeasts from Washington vineyards, representing isolates of Candida saitoana, Curvibasidium pallidicorallinum, Metschnikowia chrysoperlae, M. pulcherrima, Meyerozyma guilliermondii, Saccharomyces cerevisiae, and Wickerhamomyces anomalus, for ability to colonize Thompson Seedless grape berries, inhibit the growth of Botrytis cinerea in vitro, and suppress disease symptoms on isolated berries. The yeast-like fungus Aureobasidium pullulans was also included based on its known biocontrol activity against B. cinerea in studies on apple and grape. All yeast strains multiplied rapidly in grape berries and reached densities of over log 6 cells per wound as early as 2 days after inoculation with 200 cells. One of the Botrytis isolates used in this study was much less virulent than the others and was provisionally identified as B. prunorum based on multilocus sequence analysis. Suppression of the growth of B. cinerea isolates 111bb, 207a, 207cb, and 407cb occurred on berries treated with A. pullulans P01A006, Metschnikowia chrysoperlae P34A004 and P40A002, M. pulcherrima P01A016 and P01C004, Meyerozyma guilliermondii P34D003, and S. cerevisiae HNN11516. Inhibition of Botrytis isolates by the yeast strains was more common on berries than in vitro, suggesting the possibility that niche competition was a more likely biocontrol mechanism than antibiosis in planta. Metabolic profiling of yeast strains and B. cinerea isolates using Biolog YT plates revealed seven distinct metabolic groups. Furthermore, the yeast strains showed partial to complete tolerance to the commonly used fungicides fluopyram, triflumizole, metrafenone, pyraclostrobin, and boscalid. Implications of these findings for field deployment of native Washington yeasts as biocontrol agents against B. cinerea are discussed.


2009 ◽  
Vol 22 (8) ◽  
pp. 987-998 ◽  
Author(s):  
Nora Temme ◽  
Paul Tudzynski

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.


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