Causal Agent Of Gray Mold Research Articles

Effect of the sowing date and cultivar reaction on the gray mold progress on castor bean cultivars

Amphobotrys ricini, the causal agent of gray mold, is the most important plant pathogen of castor bean. As there are no effective control measures for gray mold, the objective of this study was to evaluate the progress of the disease on representative castor bean cultivars (BRS Energia, IAC Guarani, and IAC 2028) sown in October, November, and December in Paraná state, Brazil. Field experiments were conducted under minimum tillage in the Instituto Agronômico do Paraná Research Station in the 2010/11 and 2011/12 seasons. Evaluations of gray mold incidence and severity were initiated at the onset of first disease symptoms on castor bean bunches. Bunch weight was estimated. The weather conditions were favorable to gray mold epidemics in both planting seasons. A significant interaction between sowing dates and cultivar genotypes was found for the area under the disease progress curve of gray mold by incidence (AUGMI) and severity (AUGMS). Cultivar IAC Guarani sown in December 2010 and October 2011 had the lowest values of AUGMI and AUGMS. Highest values of AUGMI and AUGMS were estimated for cv. BRS Energia in both seasons. Although there were no significant interactions between sowing dates and cultivar genotypes, highest bunch weights were found with cvs. IAC Guarani and IAC 2028 in the 2010/11 season. However, there were no differences for bunch weights among cultivars in the 2011/12 season. Therefore, cultivar IAC Guarani is a suitable option for castor bean production in Paraná state, combining lowest susceptibility and highest yields.

Exposure to volatiles of essential oils alone or under hypobaric treatment to control postharvest gray mold of table grapes

After harvest, table grapes can easily undergo fungal spoilage, which is mainly caused by Botrytis cinerea, the causal agent of gray mold. To reduce such losses, table grapes are usually treated with conventional fungicides during the season, and cold stored in the presence of sulfur dioxide. However, these applications are not permitted in organic agriculture, and at the same time, there is a growing demand from consumers for fresh fruit free from pesticide residues. The application of essential oils and hypobaric treatments are promising alternatives to sulfur dioxide with minimal environmental impacts and limited concerns about human health risks. The aim of this study was to determine the effectiveness for control of postharvest gray mold of table grapes of 24-h exposure to volatiles of essential oils of Rosmarinus officinalis (rosemary), Mentha piperita (peppermint), and Thymus vulgaris (thyme) individually and in combinations with hypobaric treatment at 50kPa (0.5atm). Exposure to volatiles of rosemary essential oils under atmospheric pressure and hypobaric conditions reduced by around 65% the incidence and McKinney’s Index of gray mold for table grapes that were then stored at room temperature for 9 d and 5 d, respectively, or that were stored at 4°C for 7 d and followed by 3 d shelf life at 20°C. Peppermint essential oils similarly controlled gray mold for grapes stored at room temperature and under hypobaric conditions for 24h. Panel tasting revealed perception of the essential oils soon after the treatments and 24h later for grape berries exposed to vapors of rosemary, peppermint, and Lavandula×ibrida (lavender). Then 48h after treatment, the rosemary and peppermint essential oils were no longer perceived on grapes stored at 4°C and at 20°C. Exposure to volatiles of the rosemary and peppermint essential oils alone or in combination with hypobaric treatment might represent an innovative method to control postharvest gray mold of table grapes, although at least 48h were needed between exposure to volatiles of essential oils and presentation to consumers.

First Report of Resistance to Multiple Chemical Classes of Fungicides in Botrytis cinerea, the Causal Agent of Gray Mold From Greenhouse-Grown Petunia in Florida

First Report of Resistance to Multiple Chemical Classes of Fungicides in <i>Botrytis cinerea</i>, the Causal Agent of Gray Mold From Greenhouse-Grown Petunia in Florida

Fungal adaptation to contemporary fungicide applications: the case of Botrytis cinerea populations from Champagne vineyards (France).

In addition to being one of the most acute problems impeding chemical control of fungal diseases, the evolution of fungicide resistance is an emblematic case of local adaptation to spatially heterogeneous and temporally variable selection pressures. Here we dissected the adaptation of Botrytis cinerea (the causal agent of grey mould) populations on grapes to several fungicides. We carried out a 2-year survey (four collection dates) on three treated/untreated pairs of plots from vineyards in Champagne (France) and monitored the frequency of four resistant phenotypes that are unambiguously associated with four distinct genotypes. For two loci under selection by currently used fungicides (MDR1 and MDR2), the frequencies of resistant mutations at vintage were greater in treated plots compared to untreated plots, showing that the effect of selection is detectable even at the plot scale. This effect was not detectable for two other loci under selection by previously used fungicides (BenR1 and ImiR1). We also found that treatment with currently used fungicides reduced B.cinerea effective population size, leading to a significant decrease in genic diversity and allelic richness in treated vs. untreated plots. We further highlight that even under ample drift and migration, fungal populations can present an efficient response to selection. Finally, for the four studied loci, the costs of fungicide resistance were estimated by modelling the decrease in the frequency of resistant mutations in the absence of treatment. We discuss the importance of these estimates for defining strategies for limiting or counteracting the local adaptation of pests to fungicides.

Combining biocontrol agents with different mechanisms of action in a strategy to control Botrytis cinerea on grapevine

The use of several microbial biocontrol agents to combat Botrytis cinerea, the causal agent of grey mould, has been studied. However, only a few microorganisms have been developed as biofungicides, which are currently used in some countries, mostly in organic farming. The main reason for the limited market uptake of microbial biofungicides is their debated variable efficacy. To cope with poor survival in the canopy, due to unfavourable environmental conditions or their intrinsic lower level of disease control compared to synthetic chemical fungicides, use of a mixture of two or more microorganisms with different environmental requirements and mechanisms of action has been proposed with contrasting results. However, their use in strategies involving calculated timing of the microbial biocontrol agents, taking into consideration their mechanism of action in relation to the epidemiology and pathogenesis of the disease, has never been attempted in relation to combating grey mould on grapes. The results of four years of trials in three locations in Northern and Central Italy show that Trichoderma atroviride, Aureobasidium pullulans and Bacillus subtilis, applied at bunch-closure, veraison and pre-harvest, respectively, controlled B. cinerea on bunches very satisfactorily, and the results did not differ from those obtained with a strategy combining the three biofungicides, applied at the aforementioned stages. Colonisation of berries by each of the different microbial biocontrol agents at harvest time did not differ for individual treatments or when applied in the combined strategy, suggesting that the microorganisms did not negatively interfere with each other and that they may possibly occupy different ecological niches. The high level of efficacy of the tested biocontrol agents against grey mould can be explained with the relatively low-medium level of the disease, their integration with agronomic practices or the optimal timing of the treatment.

First Report of Fludioxonil Resistance in Botrytis cinerea, the Causal Agent of Gray Mold, From Strawberry Fields in Spain

First Report of Fludioxonil Resistance in <i>Botrytis cinerea</i>, the Causal Agent of Gray Mold, From Strawberry Fields in Spain

Filamentous fungi isolated from grape marc as antagonists of Botrytis cinerea

In this paper we report on the isolation and identification of three filamentous fungi from grape marc, and antifungal effect of their cell-free culture filtrates on the growth of Botrytis cinerea, causal agent of gray mold. Grape marc is a waste material that has been used as soil amendment in sustainable agriculture. Isolates originating from grape marc were identified on the basis of morphological features and internal transcribed spacer rDNA or ?-tubulin gene sequencing. The presence of three different species, Penicillium paneum, Penicillium chrysogenum and Aspergillus fumigatus has been detected expressing different effect on the growth of B. cinerea. The effect of crude culture filtrates of selected fungi on B. cinerea growth was tested. Heat sensitivity of the established inhibition effect was examined by autoclaving the crude culture filtrate prior to testing. Additional aim was to determine whether antifungal effect was influenced by previous exposure to B. cinerea in dual liquid cultures. Crude culture filtrate of A. fumigatus K16/2 showed the lowest suppression of B. cinerea growth. A maximal percentage inhibition achieved within the study was 38.2%, 39.8% and 23.8 for crude filtrates of P. paneum K7/1, P. chrysogenum K11/1 and A. fumigatus K16/2, respectively. Presence of B. cinerea in dual liquid culture induced significant increase in antifungal capacity of the culture filtrates in comparison to pure culture filtrates of the chosen isolates. The antifungal activity of all of the isolates? culture filtrates retained after heat treatment suggesting the presence of some thermostable antifungal metabolites. The results indicate the complexity and specificity of the interaction between filamentous fungi and B. cinerea. Grape marc is a good source for isolation od B. cinerea fungal antagonists and their antifungal metabolites. Specificity of fungal-fungal interactions suggests that further research on the antagonistic mechanisms and factors affecting them should be studied separately for each pair of antagonists.

Results of two consecutive years on mould prevention in viticulture by means of UV C application of vines (Vitis viniferaL.)

In the last decade, impacts of climate change on cool climate viticulture were most obvious concerning cluster infections caused by Botrytis cinerea , the causal agent of grey mould. The control of primary and secondary fungal pathogens is economically most important since bunch rot may severely affect the annual harvest with regard to quality and quantity. Trials with UV C applications, a new approach to control bunch rot, were conducted in 2013 and 2014, representing years with intensive bunch rot epidemics. UV C irradiation was applied by a prototype at an experimental site of Hochschule Geisenheim University. At both canopy sides, leaves and cluster zones were irradiated repeatedly at a dose of ≤160 mWs/cm2 alone or in combination with chemical control. The results of our experiments indicated that irradiation with UV C, in addition to fungicide treatments at reduced number of applications, was able to significantly decrease infection with grey mould to a degree of efficiency of 82% compared to standard chemical treatment that achieved 51% efficiency in 2014. Accordingly, the new technology is suitable for reducing cluster infection with mould fungi allowing a prolonged grape ripening time, which leads to a better harvest quality and a decreased yield loss.

First Report of Fludioxonil Resistance in Botrytis cinerea, the Causal Agent of Gray Mold, from Strawberry Fields in Maryland and South Carolina.

Botrytis cinerea Pers. is the causal agent of gray mold and one of the most economically important plant-pathogenic fungi affecting strawberry (Fragaria × ananassa). Control of gray mold mainly depends on the use of site-specific fungicides, including the phenylpyrrole fludioxonil. This fungicide is currently registered in combination with cyprodinil in form of Switch 62.5WG (Syngenta Crop Protection, Greensboro, NC) for gray mold control of small fruits in the United States. In June 2013, strawberries affected with symptoms resembling gray mold were observed despite the application of Switch in one field located in Federalsburg, MD, and one located near Chesnee, SC. Ten single-spore isolates, each from a different fruit, were obtained from each location and confirmed to be B. cinerea using cultural and molecular tools as described previously (3). In vitro sensitivity to fludioxonil (Scholar SC, 20.4% [v/v] active ingredient, Syngenta Crop Protection, Greensboro, NC) was determined using a conidial germination assay as previously described (4). Eight of the 20 isolates (six from Maryland and two from South Carolina) were moderately resistant to fludioxonil, i.e., they grew on medium amended with 0.1 μg/ml fludixonil and showed residual growth at 10 μg/ml (4). The in vitro assay was repeated obtaining the same results. To assess in vivo sensitivity on fungicide-treated fruit, commercially grown strawberries were rinsed with water, dried, and sprayed 4 h prior to inoculation with either water or 2.5 ml/liter of Scholar SC to runoff using a hand mister. Fruit was stab-wounded with a sterile syringe and inoculated with a 30-μl droplet of conidia suspension (106 spores/ml) of either two sensitive or four resistant isolates (two isolates from Maryland and two isolates from South Carolina). Each isolate/treatment combination consisted of 24 mature but still firm strawberry fruit with three 8-fruit replicates. The fruit were kept at 22°C and lesion diameters were measured after 4 days of inoculation. The sensitive isolates developed gray mold symptoms on nontreated (2.5 cm lesion diameter) but not on Scholar SC-treated fruit. The resistant isolates developed gray mold on both, the water-treated control (2.3 cm lesion diameter), and the fungicide-treated fruit (1.8 cm lesion diameter). The experiment was performed twice. To our knowledge this is the first report of fludioxonil resistance in B. cinerea from strawberry fields in Maryland and South Carolina. Resistance to fludioxonil is still rare in the United States and has only been reported in B. cinerea isolates from a Virginia strawberry field (1). The increase in occurrence of resistance to fludioxonil may be a result of increased use of Switch following reports of resistance to other chemical classes in this pathogen in southern strawberry fields (2).

Perception of soft mechanical stress in Arabidopsis leaves activates disease resistance

BackgroundIn a previous study we have shown that wounding of Arabidopsis thaliana leaves induces a strong and transient immunity to Botrytis cinerea, the causal agent of grey mould. Reactive oxygen species (ROS) are formed within minutes after wounding and are required for wound–induced resistance to B. cinerea.ResultsIn this study, we have further explored ROS and resistance to B. cinerea in leaves of A. thaliana exposed to a soft form of mechanical stimulation without overt tissue damage. After gentle mechanical sweeping of leaf surfaces, a strong resistance to B. cinerea was observed. This was preceded by a rapid change in calcium concentration and a release of ROS, accompanied by changes in cuticle permeability, induction of the expression of genes typically associated with mechanical stress and release of biologically active diffusates from the surface. This reaction to soft mechanical stress (SMS) was fully independent of jasmonate (JA signaling). In addition, leaves exposed soft mechanical stress released a biologically active product capable of inducing resistance to B. cinerea in wild type control leaves.ConclusionArabidopsis can detect and convert gentle forms of mechanical stimulation into a strong activation of defense against the virulent fungus B. cinerea.

First Report of Gray Mold of Blackberry Caused by Botrytis cinerea in South Carolina.

Botrytis cinerea Pers.: Fr. is a causal agent of gray mold of blackberry but may also affect grapevine, tomato, bulb flowers, and ornamental crops (2). In August 2010, blackberries (Rubus fruticosus and other species) showing gray mold symptoms were found in Longcreek, Six Mile, and Cheddar, SC. Symptomatic blackberry fruit exhibited patterns of brown-to-gray mycelia and conidiophores. Upon isolation, the mycelium grew at a rate of 12.3 mm per day at 22°C on potato dextrose agar, forming pale white-to-gray colonies with concentric rings and conidiophores (less than 12 h of fluorescent light per day). Some isolates formed dark brown sclerotia in the dark after 18 days. The lemon-shaped spores averaged 12 × 9 μm and were consistent with descriptions of B. cinerea. (1) The ribosomal internal transcribed spacer (ITS) ITS1-5.8S-ITS2 region was amplified via PCR from genomic DNA obtained from mycelia using primers ITS1 and ITS4. A BLAST search in GenBank revealed highest similarity (99 to 100%) to sequences from various Botrytis spp. collected in China, Canada, and Spain (GenBank Accession Nos. FJ169666.1, GU934505.1, and EF207414.1). The ITS sequence amplified from the blackberry isolate was submitted to GenBank (Accession No. JN164269). The pathogen was further identified to the species level as B. cinerea using glyceraldehyde-3-phosphate dehydrogenase, heat-shock protein 60 (HSP60), and DNA-dependent RNA polymerase subunit II (RPB2) gene sequences (2) (GenBank Accession Nos. JN164270, JN164271, JN164272). Pathogenicity was confirmed by inoculating three surface-sterilized (soaked in 5% bleach for 15 min), mature blackberry fruit (R. fruticosus) with a conidial suspension (105 spores/ml) of the blackberry isolate. A 20-μl droplet was placed on the fruit; control fruit received sterile water without conidia. After 5 days of incubation at room temperature in an air-tight Magenta box, the inoculated fruit developed typical signs and symptoms of gray mold. The developing spores on inoculated fruit were confirmed to be B. cinerea. All control fruit remained healthy. To our knowledge, this is the first report of B. cinerea on blackberry in South Carolina. The disease must be managed with fungicides to obtain high quality fruit with market-requested shelf life.

Biocontrol of Botrytis cinerea in table grapes by non-pathogenic indigenous Saccharomyces cerevisiae yeasts isolated from viticultural environments in Argentina

Botrytis cinerea, the causal agent of gray mold, is an important disease of grapes. Yeasts are members of the epiphytic microbial community on surfaces of fruits and vegetables and because some yeasts inhibit fungi they are used as biocontrol agents. The major objective of the present work was to isolate yeasts from grapes, vineyard soil, and grape must and select them for their ability to prevent gray mold onset after harvest. Yeasts that were found effective against the fungus were also assayed for their possible pathogenicity in humans. Two antagonism experiments were performed to study the effect of yeasts on B. cinerea, an in vitro study with Czapeck Yeast Extract Agar and an in vivo study with grape berries at 2 °C and 25 °C; both experiments were conducted at different yeast concentrations (10 5, 10 6 and 10 7 cfu/mL). Antagonists were subsequently assayed for their ability to colonize and grow in fruit wounds. The biocontrol yeasts were also examined for their possible pathogenicity in humans: phospholipase and proteolytic activity, growth at 37 °C and 42 °C, pseudohyphal formation and invasive growth. A total of 225 yeasts belonging to 41 species were isolated from must and grape berries and 65 of them, representing 15 species, exhibited in vitro inhibition of B. cinerea at 25 °C. These 65 biocontrol yeasts were subsequently assayed in vivo and 16 of them (15 Saccharomyces cerevisiae and 1 Schizosaccharomyces pombe) showed antagonistic properties against B. cinerea at 25 °C. Only one isolate ( S. cerevisiae BSc68) was able to inhibit mycelial growth of B. cinerea on grape berries at both 2 °C and 25 °C. The biomass of this strain in grape wounds increased 221.5-fold at 25 °C after 3 d and 325.5-fold at 2 °C after 10 d of incubation. An increase in the concentration of certain yeasts significantly enhanced their antagonistic activity. All yeast isolates determined as biocontrol agents under in vivo conditions were isolated from fermenting musts. Twelve biocontrol agents ( S. cerevisiae) revealed one or more phenotypical characteristics associated with pathogenicity in humans but none of them showed all characteristics together. The fact that there exist few reports on S. cerevisiae and none on Sch. pombe as biocontrol agents against B. cinerea makes our results even more relevant.

A Permeable Cuticle Is Associated with the Release of Reactive Oxygen Species and Induction of Innate Immunity

Wounded leaves of Arabidopsis thaliana show transient immunity to Botrytis cinerea, the causal agent of grey mould. Using a fluorescent probe, histological staining and a luminol assay, we now show that reactive oxygen species (ROS), including H2O2 and O2 −, are produced within minutes after wounding. ROS are formed in the absence of the enzymes Atrboh D and F and can be prevented by diphenylene iodonium (DPI) or catalase. H2O2 was shown to protect plants upon exogenous application. ROS accumulation and resistance to B. cinerea were abolished when wounded leaves were incubated under dry conditions, an effect that was found to depend on abscisic acid (ABA). Accordingly, ABA biosynthesis mutants (aba2 and aba3) were still fully resistant under dry conditions even without wounding. Under dry conditions, wounded plants contained higher ABA levels and displayed enhanced expression of ABA-dependent and ABA-reporter genes. Mutants impaired in cutin synthesis such as bdg and lacs2.3 are already known to display a high level of resistance to B. cinerea and were found to produce ROS even when leaves were not wounded. An increased permeability of the cuticle and enhanced ROS production were detected in aba2 and aba3 mutants as described for bdg and lacs2.3. Moreover, leaf surfaces treated with cutinase produced ROS and became more protected to B. cinerea. Thus, increased permeability of the cuticle is strongly linked with ROS formation and resistance to B. cinerea. The amount of oxalic acid, an inhibitor of ROS secreted by B. cinerea could be reduced using plants over expressing a fungal oxalate decarboxylase of Trametes versicolor. Infection of such plants resulted in a faster ROS accumulation and resistance to B. cinerea than that observed in untransformed controls, demonstrating the importance of fungal suppression of ROS formation by oxalic acid. Thus, changes in the diffusive properties of the cuticle are linked with the induction ROS and attending innate defenses.

Botrytis cinerea control and the problem of fungicide resistance

Botrytis cinerea, the causal agent of grey mould, greatly affects fruit, grapevine, vegetable and ornamental crops production. It is a common causal agent of diseases in plants grown in protected areas, as well as fruit decay during storage and transport. The fungus invades almost all parts of the plant in all developmental stages, and the symptoms are usually described as grey mould, grey mildew, brown rot and seedling blight. The paper reviews the current knowledge on control possibilities of this necrotrophic pathogen. The attention is particularly paid to the mode of action of novel fungicides and to the problem of resistance. It is pointed out that by limiting the number of treatments in the growing season, avoiding the use of only one fungicide with a high risk for resistance development, appropriate application rate and timing, using mixtures of pesticides with different modes of action, as well as by alternative use of pesticides from different resistance groups, a longterm preservation of pesticide efficacy is provided.

Analysis of Polygalacturonase Enzyme Production in Isolates of Botrytis cinerea Sensitive to Different Fungicides Used in Vineyards

AbstractReduction of mycelial growth and spore germination of Botrytis cinerea, the causal agent of grey mould, are common criteria to determine the sensitivity of this fungus to fungicides. In this article, the influence of nine different fungicides on polygalacturonase (PG) activity and mycelial dry weight (MDW) was analysed on culture filtrates from B. cinerea, obtained from grapes. All fungicides except triadimenol and tebuconazole inhibited MDW of isolates &lt;50%. Cyprodinil + fludioxonil, myclobutanil and imazalil inhibited PG activity more than 50%. Fenhexamid had a lower inhibitory effect (&lt;50%) on PG activity. Procymidone and pyrimethanil induced both PG activity and isoenzyme banding profile of isolates sensitive to these fungicides. This study provides a new additional tool for determining sensitivity to fungicides and monitoring the effect of fungicide resistance management policies.

Exploring Mechanisms of Resistance to Respiratory Inhibitors in Field Strains of Botrytis cinerea , the Causal Agent of Gray Mold

Respiratory inhibitors are among the fungicides most widely used for disease control on crops. Most are strobilurins and carboxamides, inhibiting the cytochrome b of mitochondrial complex III and the succinate dehydrogenase of mitochondrial complex II, respectively. A few years after the approval of these inhibitors for use on grapevines, field isolates of Botrytis cinerea, the causal agent of gray mold, resistant to one or both of these classes of fungicide were recovered in France and Germany. However, little was known about the mechanisms underlying this resistance in field populations of this fungus. Such knowledge could facilitate resistance risk assessment. The aim of this study was to investigate the mechanisms of resistance occurring in B. cinerea populations. Highly specific resistance to strobilurins was correlated with a single mutation of the cytb target gene. Changes in its intronic structure may also have occurred due to an evolutionary process controlling selection for resistance. Specific resistance to carboxamides was identified for six phenotypes, with various patterns of resistance levels and cross-resistance. Several mutations specific to B. cinerea were identified within the sdhB and sdhD genes encoding the iron-sulfur protein and an anchor protein of the succinate dehydrogenase complex. Another as-yet-uncharacterized mechanism of resistance was also recorded. In addition to target site resistance mechanisms, multidrug resistance, linked to the overexpression of membrane transporters, was identified in strains with low to moderate resistance to several respiratory inhibitors. This diversity of resistance mechanisms makes resistance management difficult and must be taken into account when developing strategies for Botrytis control.

Improved leaf pruning reduces development of stem cankers caused by grey mould in greenhouse tomatoes

Botrytis cinerea, the causal agent of grey mould, is considered as a major burden on greenhouse tomato production. In heated greenhouses, where leaf pruning is a routine practice and microclimatic conditions are favourable for grey mould development, infections of the pruning wounds result in stem cankers that can rapidly kill the plants. Chemical control is not always sufficiently effective and growers face increasing demands for reducing their dependency on pesticides. Several techniques can be used for the removal of leaves. They differ mainly on the presence or absence and the size of petiole stubs remaining on the stems after pruning. In this study, leaf pruning techniques were compared in an experimental greenhouse and the results validated in four commercial greenhouses, in a variety of agronomic conditions and in various situations of inoculum pressure. All experiments clearly showed that a careful removal of the leaves without leaving any petiole stubs on the stems can significantly decrease the incidence of stem cankers even under high disease pressure. As up to 10-fold differences were observed, this suggests the possibility of improving pruning practices in tomato greenhouses by removing leaves systematically close to the stem. This environmentally-friendly cultural practice appears compatible for integration with other preventative methods for the objective of sustainable crop protection in greenhouse tomatoes.

Relative Contribution of Various Sources of Botrytis cinerea Inoculum in Strawberry Fields in Norway.

To identify the most important sources of inoculum of Botrytis cinerea (causal agent of gray mold) in commercial strawberry (Fragaria × ananassa) fields in Norway, soil and overwintered plant material were collected from planting beds and alleys at five locations in 2000 to 2002 (13 samples altogether). Plant material was sorted by category (e.g., leaves, stems, mulch, and weeds). After subsamples of each material were incubated for 5 days at 20°C at high humidity, conidiophores of B. cinerea growing from mycelia and sclerotia were counted. Overwintered plant debris within planting beds yielded more than 96% of total conidiophores counted, the remainder originating from plant debris collected from alleys or soil. Overwintered strawberry plant debris produced 98% of the conidiophores within planting beds and 80% of the conidiophores in the alleys, while the remaining was produced by weeds. Senescing and dead leaf laminae produced 45% of the conidiophores while stem residues (i.e., petioles, stolons, inflorescences, and unidentifiable stem parts) produced 50% and mummified fruit produced 5% within planting beds. The contribution of sclerotia, compared with mycelia, to conidiophore production varied greatly between fields and years. Overall, 47% of the total number of conidiophores produced in plant material within planting beds originated from sclerotia. More than 90% of the conidiophores from sclerotia were found in dead stem residues.

Mapping of loci from Solanum lycopersicoides conferring resistance or susceptibility to Botrytis cinerea in tomato

Cultivated tomato (Solanum lycopersicum, syn. Lycopersicon esculentum) is susceptible to the necrotrophic ascomycete and causal agent of gray mold, Botrytis cinerea. Resistance to this fungal pathogen is elevated in wild relatives of tomato, including Solanum lycopersicoides. An introgression line population (IL) containing chromosomal segments of S. lycopersicoides within the background of tomato cv. VF36 was used to screen the genome for foliar resistance and susceptibility to B. cinerea. Based on this screen, putative quantitative trait loci (QTL) were identified, five for resistance and two for susceptibility. Four resistance QTL decreased infection frequency while the fifth reduced lesion diameter. One susceptibility QTL increased infection frequency whereas the other increased lesion diameter. Overlapping chromosomal segments provided strong evidence for partial resistance on chromosomes 1 and 9 and for elevated susceptibility on chromosome 11. Segregation analysis confirmed the major resistance QTL on the long arm of chromosome 1 and susceptibility on chromosome 11. Linkage of partial resistance to chromosome 9 could not be confirmed. The usefulness of these data for resistance breeding and for map-based cloning of foliar resistance to B. cinerea is discussed.

Wounding of Arabidopsis leaves causes a powerful but transient protection against Botrytis infection

Physical injury inflicted on living tissue makes it vulnerable to invasion by pathogens. Wounding of Arabidopsis thaliana leaves, however, does not conform to this concept and leads to immunity to Botrytis cinerea, the causal agent of grey mould. In wounded leaves, hyphal growth was strongly inhibited compared to unwounded controls. Wound-induced resistance was not associated with salicylic acid-, jasmonic acid- or ethylene-dependent defence responses. The phytoalexin camalexin was found to be involved in this defence response as camalexin-deficient mutants were not protected after wounding and the B. cinerea strains used here were sensitive to this compound. Wounding alone did not lead to camalexin production but primed its accumulation after inoculation with B. cinerea, further supporting the role of camalexin in wound-induced resistance. In parallel with increased camalexin production, genes involved in the biosynthesis of camalexin were induced faster in wounded and infected plants in comparison with unwounded and infected plants. Glutathione was also found to be required for resistance, as mutants deficient in gamma-glutamylcysteine synthetase showed susceptibility to B. cinerea after wounding, indicating that wild-type basal levels of glutathione are required for the wound-induced resistance. Furthermore, expression of the gene encoding glutathione-S-transferase 1 was primed by wounding in leaves inoculated with B. cinerea. In addition, the priming of MAP kinase activity was observed after inoculation of wounded leaves with B. cinerea compared to unwounded inoculated controls. Our results demonstrate how abiotic stress can induce immunity to virulent strains of B. cinerea, a process that involves camalexin and glutathione.