Cinerea Isolates Research Articles

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).

Characterization of Botrytis cinerea populations associated with treated and untreated cv. Moscato vineyards.

Three Botrytis cinerea populations, isolated from three vineyards, one untreated and two treated twice a year, respectively, with fenhexamid or cyprodinil+fludioxonil, were investigated to evaluate the effect of repeated fungicide treatments on the presence and distribution of the transposons Boty and Flipper , and on the phenotypic traits of each pathogen community. The vacuma individuals lacking the two transposons represented the majority of the 390 B. cinerea isolates followed by transposa strains containing Boty and Flipper, while the remaining 67 isolates harboured respectively only Boty (60) or Flipper (7). This research has demonstrated that fungicide application did not influence the transposon distribution patterns, the sensitivity towards various botryticides, or the growth rate of the isolates belonging to the three different populations, but did induced overall reduction of the population size and selected isolates characterized by an enhanced pathogenicity, especially on Vitis vinifera leaves.

Analyses of genetic and pathogenic variability among Botrytis cinerea isolates

Seventy nine isolates of Botrytis cinerea were collected from different host plants and different locations of India and Nepal. All the isolates were identified as B. cinerea based on morphological features and were confirmed using B. cinerea specific primers. Differentiation among the isolates was assessed using morphological, genetic and biochemical approaches. To analyze morphological variability, differences in conidial size, presence or absence of sclerotia and their arrangement were observed. Genetic variability was characterized using RAPD analysis, presence or absence of transposons and mating type genes. Cluster analysis based on RAPD markers was used for defining groups on the basis of geographical region and host. The biochemical approach included determining differences in concentration of oxalic acid and activity of lytic enzymes. All the isolates were categorized into different pathogenic groups on the basis their variable reaction towards chickpea plants. Isolates with higher concentration of oxalic acid and greater activity of lytic enzymes were generally more pathogenic. Pathogenicity was also correlated to transposons. Isolates containing transposa group showed some degree of correlation with pathogenic behavior. However, isolates could not be grouped on the basis of a single approach which provides evidence of their wide diversity and high evolution potential. Sensitivity of sampled isolates was also tested against five botryticides. Most of the isolates from same region were inhibited by a particular fungicide. This feature provided interesting cues and would assist in devising novel and more effective measures for managing the disease.

High‐resolution melting analysis for rapid detection and characterization of Botrytis cinerea phenotypes resistant to fenhexamid and boscalid

A novel, high‐resolution melting (HRM) analysis was developed to detect single nucleotide polymorphisms (SNPs) associated with resistance to fenhexamid (hydroxyanilides) and boscalid (succinate dehydrogenase inhibitors) in Botrytis cinerea isolates. Thirty‐six single‐spore isolates arising from 13 phenotypes were selected and tested for fungicide sensitivity. Germ tube elongation assays showed two distinct sensitivity levels for each fungicide. Sequencing revealed that resistance to fenhexamid was due to a nucleotide change in the erg27 gene, resulting in an amino acid replacement of phenylalanine (F) with serine (S) or valine (V) at position 412 of the protein, whereas in isolates resistant to boscalid, a nucleotide change in the sdhB gene resulted in the replacement of histidine (H) with arginine (R) or tyrosine (Y) at position 272 of the respective protein. In each case, melting curve analysis generated three distinct profiles corresponding to the presence of each nucleotide in the targeted areas. HRM analysis successfully detected and differentiated the substitutions associated with resistance to both fungicides. In vitro bioassays, direct sequencing and high‐resolution melting analysis showed a 100% correlation with detection of resistance. The results demonstrate the utility of HRM analysis as a potential molecular tool for routine detection of fungicide resistance using known polymorphic genes of B. cinerea populations.

Selection, Fitness, and Control of Grape Isolates of Botrytis cinerea Variably Sensitive to Fenhexamid.

Of 683 Botrytis cinerea isolates collected from a fungicide-trial vineyard, 31 were classified as putatively resistant to fenhexamid (50% effective concentration [EC50] ≥ 0.1 μg/ml). For the resistant isolates that survived and sporulated in culture, colony expansion and conidial germination frequency was significantly reduced relative to the mean of 30 representative baseline isolates (EC50 = 0.03 μg/ml). Grape berries were inoculated with four isolates representing a range of fenhexamid sensitivities and treated preventively or curatively with fenhexamid concentrations (150 to 600 mg/liter) representing 25 to 100% of the recommended rate. All treatments significantly delayed disease onset and progress caused by isolates with EC50 values of 0.03 and 0.15 μg/ml but provided little to no control of isolates with EC50 values of 0.32 and 62.5 μg/ml. The latter isolate exhibited a previously unreported F427V mutation of ERG27, an enzyme of ergosterol biosynthesis. In a duplex quantitative polymerase chain reaction test, the ratio of pathogen/host DNA increased significantly for 14 days after inoculation of untreated berries with a baseline isolate but declined slightly in berries treated with fenhexamid at 600 mg/liter 1 day post inoculation. In the vineyard, disease control was affected by the number and rate of fenhexamid applications but B. cinerea isolates with EC50 ≥ 0.1 μg/ml were not preferentially selected.

Presence of Transposons and Mycoviruses in Botrytis cinerea Isolates Collected from a German Grapevine Growing Region

AbstractTransposons and infection of fungal strains with mycoviruses can have significant effects on distinctive phenotypic traits of phytopathogenic fungi such as mycelial growth and sporulation, pathogenicity or fungicide resistance. Two transposable elements (TE), Boty and Flipper, are known to be associated with the ubiquitous fungus Botrytis cinerea. In addition, the presence of two types of ssRNAsRNA viruses, BVX and BVF, has been reported in B. cinerea. In this study, we assessed the genetic diversity of B. cinerea isolates, all sampled within a small‐sized German viticultural area (‘Rheingau’) by examining and classifying them according to the presence of TEs and mycoviruses. A subset of the isolates was further analysed with microsatellite markers to determine the origin of particular isolates with or without one or both mycoviruses. Virtually all isolates (98%) sampled in two different years (2008 and 2010) were screened positive for the presence of a transposon. Presence of one or both B. cinerea mycoviruses was confirmed for 37% of the analysed isolates sampled in 2010, representing the first record of B. cinerea mycoviruses in German isolates. Assignment on individual B. cinerea isolates to different genetic groups was independent of the presence or absence of a mycovirus or a transposable element, respectively. Furthermore, we found no correlation between the presence of either a mycovirus or a transposable element and different viticultural management practices, soil properties or levels of nitrogen fertilization applied to the respective vineyards. However, mycelial growth of B. cinerea strains containing mycovirus BVF was significantly reduced at lower temperatures.

Assessing the effects of light on differentiation and virulence of the plant pathogen Botrytis cinerea: characterization of the White Collar Complex.

Organisms are exposed to a tough environment, where acute daily challenges, like light, can strongly affect several aspects of an individual's physiology, including pathogenesis. While several fungal models have been widely employed to understand the physiological and molecular events associated with light perception, various other agricultural-relevant fungi still remain, in terms of their responsiveness to light, in the dark. The fungus Botrytis cinerea is an aggressive pathogen able to cause disease on a wide range of plant species. Natural B. cinerea isolates exhibit a high degree of diversity in their predominant mode of reproduction. Thus, the majority of naturally occurring strains are known to reproduce asexually via conidia and sclerotia, and sexually via apothecia. Studies from the 1970′s reported on specific developmental responses to treatments with near-UV, blue, red and far-red light. To unravel the signaling machinery triggering development – and possibly also connected with virulence – we initiated the functional characterization of the transcription factor/photoreceptor BcWCL1 and its partner BcWCL2, that form the White Collar Complex (WCC) in B. cinerea. Using mutants either abolished in or exhibiting enhanced WCC signaling (overexpression of both bcwcl1 and bcwcl2), we demonstrate that the WCC is an integral part of the mentioned machinery by mediating transcriptional responses to white light and the inhibition of conidiation in response to this stimulus. Furthermore, the WCC is required for coping with excessive light, oxidative stress and also to achieve full virulence. Although several transcriptional responses are abolished in the absence of bcwcl1, the expression of some genes is still light induced and a distinct conidiation pattern in response to daily light oscillations is enhanced, revealing a complex underlying photobiology. Though overlaps with well-studied fungal systems exist, the light-associated machinery of B. cinerea appears more complex than those of Neurospora crassa and Aspergillus nidulans.

Site‐directed mutagenesis of the P225, N230 and H272 residues of succinate dehydrogenase subunit B from Botrytis cinerea highlights different roles in enzyme activity and inhibitor binding

Carboxamide fungicides target succinate dehydrogenase (SDH). Recent field monitoring studies have identified Botrytis cinerea isolates resistant to one or several SDH inhibitors (SDHIs) with amino acid substitutions in the SDH B subunit. We confirmed, by site-directed mutagenesis of the sdhB gene, that each of the mutations identified in field strains conferred resistance to boscalid in B.cinerea, and in some cases cross-resistance to other SDHIs (fluopyram, carboxin). Enzyme inhibition studies showed that the studied modifications (SdhB_P225T/L/F, N230I, H272Y/R/L) affected the inhibition of SDH activity by SDHIs, directly contributing to resistance. Our results confirm the importance of H272, P225 and N230 for carboxamide binding. Modifications of P225 and N230 conferred resistance to the four carboxamides tested (boscalid, fluopyram, carboxin, bixafen). Modifications of H272 had differential effects on the susceptibility of SDH to SDHIs. SdhB(H272L) , affected susceptibility to all SDHIs, SdhB(H272R) conferred resistance to all SDHIs tested except fluopyram, and SdhB(H272Y) conferred fluopyram hypersensitivity. Affinity-binding studies with radiolabelled fluopyram revealed strong correlations among the affinity of SDHIs for SDH, SDH inhibition and in vivo growth inhibition in the wild type. The sdhB(H272Y) mutation did not affect SDH and respiration activities, whereas all the other mutations affected respiration by decreasing SDH activity.

Mycoviruses ofBotrytis cinereaisolates from different hosts

Botrytis cinerea (teleomorph Botryotinia fuckeliana) is a necrotrophic plant pathogenic fungus that causes grey mould and enormous economic losses worldwide in different crops. Control of B. cinerea is difficult due to the appearance of fungicide-resistant isolates, and the diversity in virulence due to genetic variability and, perhaps, the infection with mycoviruses or fungal viruses. The discovery of mycoviruses and their possible application as biocontrol agents, as well as their use as tools to study the plant–pathogen interaction, has encouraged their study in B. cinerea. Herein, we have analysed the occurrence of mycoviruses in Spanish B. cinerea isolates to approach a better understanding of the interactions among viruses, fungi and plants in this pathosystem. Fifty-five percent of the B. cinerea isolates analysed contained double-stranded RNA (dsRNA) elements, and the number of dsRNA elements, their relative concentration and size were variable among isolates. Some of these dsRNAs were related to the presence of virus like rod or isometric particles, and to cellular degeneration and malformed mitochondria. We have also demonstrated that a 3 kb dsRNA present in 55% of the isolates having dsRNA elements was a mycovirus genome. Partial sequence of that mycovirus presented high identity in nucleotide and amino acid sequence with Botrytis cinerea mitovirus 1 (BcMV1). Analysis of the genetic distance within Spanish BcMV1 sequences showed the existence of different isolates of this mitovirus inside the Spanish B. cinerea population analysed. This is the first report of the variability of dsRNA elements and the partial genome sequence of a mitovirus associated with Spanish B. cinerea isolates and the genetic diversity within Spanish isolates of BcMV1.

The Effects of Glucosinolates and Their Breakdown Products on Necrotrophic Fungi

Glucosinolates are a diverse class of S- and N-containing secondary metabolites that play a variety of roles in plant defense. In this study, we used Arabidopsis thaliana mutants that contain different amounts of glucosinolates and glucosinolate-breakdown products to study the effects of these phytochemicals on phytopathogenic fungi. We compared the fungus Botrytis cinerea, which infects a variety of hosts, with the Brassicaceae-specific fungus Alternaria brassicicola. B. cinerea isolates showed variable composition-dependent sensitivity to glucosinolates and their hydrolysis products, while A. brassicicola was more strongly affected by aliphatic glucosinolates and isothiocyanates as decomposition products. We also found that B. cinerea stimulates the accumulation of glucosinolates to a greater extent than A. brassicicola. In our work with A. brassicicola, we found that the type of glucosinolate-breakdown product is more important than the type of glucosinolate from which that product was derived, as demonstrated by the sensitivity of the Ler background and the sensitivity gained in Col-0 plants expressing epithiospecifier protein both of which accumulate simple nitrile and epithionitriles, but not isothiocyanates. Furthermore, in vivo, hydrolysis products of indole glucosinolates were found to be involved in defense against B. cinerea, but not in the host response to A. brassicicola. We suggest that the Brassicaceae-specialist A. brassicicola has adapted to the presence of indolic glucosinolates and can cope with their hydrolysis products. In contrast, some isolates of the generalist B. cinerea are more sensitive to these phytochemicals.

Genetic characterization of Botrytis cinerea isolates collected from pine and eucalyptus nurseries in Bio‐Bio Region, Chile

SummaryA total of 55 Botrytis cinerea isolates collected from Pinus radiata and Eucalyptus globulus plants cultivated in nurseries located in the Bio‐Bio Region, Chile, as well as isolates collected from native plants such as Rubus spp and Aristotelia chilensis located near the nurseries were genetically characterized. All isolates carried the Bc‐hch2 allele, thus belonging to genetic Group II, which is now referred to as B. cinerea. Genotyping based on the presence of transposons Boty and Flipper showed differences between isolates related to the plant host. Thus, transposa isolates (containing both transposons) were detected in P. radiata and E. globulus, while vacuma isolates (containing neither transposon) were detected in all plants except E. globulus. Notably, boty isolates (containing just the Boty transposon) were detected at high frequencies in all plant hosts. Analyses to detect mutations involved in resistance to fungicides such as benzimidazoles (BZ), dicarboximides and QoIs also showed differences in the studied isolates. Isolates collected from E. globulus were shown to carry mutations for all tree fungicides, while those collected from P. radiata presented mutations involved in resistance to BZ only. Isolates collected from native plant hosts did not carry any of the mutations analysed.

Resistance to polyoxin AL and other fungicides in Botrytis cinerea collected from sweet basil crops in Israel

A total of 568 B. cinerea isolates were collected from diseased sweet basil plants and the air in 10 sweet basil greenhouses. Mycelial growth tests were used to evaluate the sensitivity of these isolates to benomyl, fenhexamid, iprodione, polyoxin AL and pyrimethanil. EC50 values for polyoxin, the main botryticide on sweet basil in Israel, ranged from 0.4 to 6.5 μg ml−1 and had a bimodal distribution; the EC50 values for sensitive isolates ranged from 0.4 to 1.5 μg ml−1 and the EC50 values for low-level resistant isolates ranged from 4 to 6.5 μg ml−1. Among populations that had not been exposed to polyoxin treatments, 20 to 35 % of the collected isolates were low-level resistant for polyoxin. Polyoxin treatments in an experimental greenhouse shifted the equilibrium in favour of low-level resistant isolates, and the change occurred rapidly: from a frequency of 20 % low-level resistant isolates in the population that had never been treated with polyoxin to a frequency of 72 % after a few treatments over two seasons. Prolonged use of polyoxin in Israeli basil crops (in some sites for more than 10 years) does not appear to have led to the development of high-level resistance, but low-level resistant isolates were found in commercial greenhouses with the frequency of up to 73 %. High-level resistance to benzimidazoles was common (60 to 80 % of isolates) in greenhouses with a history of benzimidazole treatments; whereas 15–25 % of the isolates from greenhouses in which fungicides were not used were resistant. Low-level resistance to dicarboximides was fairly widespread (frequency of 30 to 80 % depending on the greenhouse) and a few cases of moderate resistance to dicarboximides were also noted (frequency of 0 to 9 %). Neither high- nor low-level resistance to anilinopyrimidines was common in sweet basil commercial greenhouses (0 to 7 %). However, 34 % of the isolates were strongly resistant in the experimental greenhouse, following a few treatments with anilinopyrimidine fungicides during the previous season. Before those treatments, the proportion of anilinopyrimidines resistant isolates had been 1 %. About 3 % of the isolates exhibited low-level resistance to fenhexamid and no isolates were found to be strongly resistant to fenhexamid. Low-level resistance to one fungicide was often associated with low-level resistances to other fungicides. Thirty-two phenotypes exhibiting resistance to one or more of the tested fungicides were noted among B. cinerea isolates. Resistant isolates showed similar or reduced fitness parameters in comparison to wild-type isolates.

Botrytis cinerea에 의한 무화과 잿빛곰팡이병

In 2010 and 2011, gray mold was found on common fig (Ficus carica) fruit grown at the research field of Jeollabuk-do Agricultural Research and Extension Services, Korea. Gray mold symptoms on common fig fruit mainly occurred after harvest season until December. The typical symptom included brown water-soaked rot and fruit decay. The diseased fruit was covered by gray to brown colored conidiophore and conidia. The conidiophores were tree shape and measured . Conidia on conidiophore were ellipsoidal or lemon shape, colorless, single cell, and measured . The nucleotide sequences of the rDNA ITS region obtained from the pure culture of the gray mold on common fig were 100% similar to the sequences of the GenBank accession number HQ171052, EU519210, HQ171053, FN812726, HM849615, and EU563120 of B. cinerea isolates. In phylogenetic tree, the representative isolate was placed within same clade of B. cinerea. Based on the morphological characteristics and analysis of rDNA ITS sequence data, the fungus was identified as B. cinerea.

Comparison of Botrytis cinerea populations isolated from two open-field cultivated host plants

The necrotrophic fungus Botrytis cinerea is reported to infect more than 220 host plants worldwide. In phylogenetical–taxonomical terms, the pathogen is considered a complex of two cryptic species, group I and group II. We sampled populations of B. cinerea on sympatric strawberry and raspberry cultivars in the North-East of Hungary for three years during flowering and the harvest period. Four hundred and ninety group II B. cinerea isolates were analyzed for the current study. Three different data sets were generated: (i) PCR-RFLP patterns of the ADP-ATP translocase and nitrate reductase genes, (ii) MSB1 minisatellite sequence data, and (iii) the fragment sizes of five microsatellite loci. The structures of the different populations were similar as indicated by Nei's gene diversity and haplotype diversity. The F statistics (Fst, Gst), and the gene flow indicated ongoing differentiation within sympatric populations. The population genetic parameters were influenced by polymorphisms within the three data sets as assessed using Bayesian algorithms. Data Mining analysis pointed towards the five microsatellite loci as the most defining markers to study differentiation in the 490 isolates. The results suggest the occurrence of host-specific, sympatric divergence of generalist phytoparasites in perennial hosts.

Spectral footprint of <i>Botrytis cinerea</i>, a novel way for fungal characterization

Botrytis cinerea affects plant yield and quality. Many Botrytis species are morphologically similar leading to difficulty in pathogen identification. Spectroscopy can be used to identify pathogenic fungi. This study describes a novel method for fungal characterization. Here, we determined the spectral signatures of different B. cinerea isolates as well as various fungal genera. A unique spectral pattern was investigated at both genus and isolate level. The short wave infrared II (2055 - 2315 nm) provided the best discrimination between the fungal samples observed. Moreover, the spectral analysis was performed on non-transformed data and investigated significant differences among fungal genera as well as B. cinerea isolates, while the results investigated high similarity among replicates of the same isolate of B. cinerea. The results of each spectral test were obtained reproducibly without an expensive cost consumable during sample preparation and measurements. This innovative approach would allow us to identify, discriminate and classify fungi rapidly and inexpensively at the genus, species and isolate level.

Identification of Arabidopsis accession with resistance to Botrytis cinerea by natural variation analysis, and characterization of the resistance response

Botrytis cinerea is a ubiquitous necrotrophic fungal pathogen that infects over 200 different plant species. We have analyzed 17 Arabidopsis ecotypes for natural variations in their susceptibility to B. cinerea, and found compatible and incompatible Arabidopsis–Botrytis interactions. We determined that Arabidopsis ecotype Ler is resistant to 5 B. cinerea isolates used in this study. To further investigate the roles of the salicylic acid (SA)-dependent defense response pathways against B. cinerea, we inoculated various Arabidopsis mutants with the pathogen. Arabidopsis Ler plants expressing the nahG gene inoculated with B. cinerea showed as much resistance as the parental plants (Ler-wild type). The sgt1b-1 and rar1-10 mutants also showed resistance to the pathogen. In this study, we discuss the natural variations in the symptoms observed among various ecotypes upon inoculation with B. cinerea. In addition, SA plays only a minor role in preventing systemic infection with B. cinerea.

Differential Effect of SdhB Gene Mutations on the Sensitivity to SDHI Fungicides in Botrytis cinerea.

Succinate dehydrogenase inhibiting (SDHI) fungicides constitute a relatively novel fungicide group used for gray mold control caused mainly by Botrytis cinerea. Shortly after registration, resistance was observed in fungal populations that correlated with several mutations in the succinate dehydrogenase complex (complex II). In the current study, 30 B. cinerea isolates possessing five different mutations at three different codons of SdhB (P225F, N230I, and H272L/R/Y) were characterized for their sensitivities to eight SDHI fungicides. The results show different sensitivities and cross-resistance patterns between structurally different SDHIs. P225F mutants were resistant in vitro to all SDHIs tested. Similarly, isolates possessing the H272L mutation were highly resistant to boscalid but showed low to moderate levels of resistance to other SDHIs. The N230I mutants were moderately resistant to boscalid, fluopyram, and fluxapyroxad and showed low resistance levels to isopyrazam, bixafen, fenfuram, benodanil, and carboxin. The H272R mutants showed moderate levels of resistance to boscalid and low resistance levels to isopyrazam, fenfuram, and carboxin but remained sensitive to fluopyram, bixafen, fluxapyroxad, and benodanil. Similarly, the H272Y showed moderate levels of resistance to boscalid and very low resistance levels to isopyrazam, bixafen, fenfuram, and carboxin but showed increased sensitivity to benodanil and fluopyram. Boscalid provided moderate to high control of H272R/Y and N230I mutants in detached fruit assays but provided little control against the H272L and P225F mutants. In contrast, fluopyram controlled H272R/Y mutants and provided moderate levels of control toward H272L, N230I, and P225F mutants. Our findings suggest that sensitivity to SDHIs may vary greatly, dependent on the point mutation in the sdhb subunit.

VARIATION AMONG ISOLATES OF Botrytis spp. AND INTERACTION BETWEEN B. fabae ISOLATES AND SOME FABA BEAN VARIETIES

Isolates of Botrytis spp. collected from faba bean growing governorates of Northern and Middle Egypt were compared for their morphological and physiological characters. Botrytis spp. isolates showed differences in growth rate and spore production as well as number and size of sclerotia while, growth of Botrytis cinerea isolates was faster than those of Botrytis fabae onto all tested media and most of the fabae type isolates produced number of spores less than that produced the cinerea type. Also, B. fabae isolates were the highest number of small sclerotia. Eight faba bean varieties varied in their reaction to chocolate leaf sport disease caused by B. fabae Sard. Giza 402 is highly susceptible cv. (74.2%) while, Giza-Blanka was 402 the highly resistant (40.0%) whereas, Giza-3 was moderately resistant. Also, isolate obtained from Nubaria was more virulent than those obtained from other governorates.

Chlamydospore formation by Corynespora cassiicola

The fungus Corynespora cassiicola is an important pathogen that causes necrotic lesions in several plant species. Saprophytism and parasitism habits of plants are common survival strategies for this pathogen. Few studies referred to the formation of chlamydospores by C. cassiicola. The objective of this study was to test the formation of chlamydospores by several C. cassiicola isolates from different agronomic crops and weeds. Fifteen isolates were analysed by in vivo and in vitro tests. Six isolates from four host plants (Cucumis sativus, Lantana camara, Malpighia glabra and Vernonia cinerea) were able to produce chlamydospores, both in vitro and in vivo. In vitro, chlamydospore production was highest for the Malpighia glabra isolate and one Cucumis sativus isolate, intermediate for two other C. sativus isolates, and lowest for the Vernonia cinerea and Lantana camara isolates. However, no difference in the relative number of chlamydospores produced among isolates was observed in vivo.

Population genetic results of Hungarian Botrytis cinerea isolates establishing new technologies with decreased chemical usage against grey mould

Botrytis cinerea causes gray mold on a high number of crop plants. Information about the populations of plant pathogen fungi may help to develop new strategies for the effective and economic crop protection with reduced fungicide usage. Hungarian B. cinerea populations were characterized with using different molecular genetic parameters. B. cinerea group I strains, characterized with high rate of fenhexamid resistance, could be detected only in restricted number. The Hungarian B. cinerea populations were characterized with high genetic diversity, and the regular occurrence of sexual reproduction. These results highlight the importance of rotating different type of fungicide in the plant protection technology against grey mould.