Control of infection of tomato fruits by <Emphasis Type="Italic">Alternaria</Emphasis> and mycotoxin production using plant extracts

Tomato fruits are susceptible to infection by Alternaria species. In addition, Alternaria species may contaminate the fruits with mycotoxins. There is thus interest in control systems to minimise pathogenicity and control toxin production. The objectives of this study were to examine the effect of plant extracts of Eucalyptus globulus and Calendula officinalis on the growth of strains of Alternaria alternata and A. arborescens, on pathogenicity of tomato fruits and mycotoxin production. The growth bioassays showed that the ethanolic and chloroformic fractions of E. globulus were the most effective in reducing growth of A. alternata (66–74 %) and A. arborescens (86–88 %), respectively at 2500 μg/g. The effects of plant extracts on mycotoxin biosynthesis were variable and strain dependent. The most effective fractions in decreasing mycotoxin accumulation were the ethanolic and chloroformic extracts of E. globulus, which reduced tenuazonic acid by 89 %, alternariol by 75–94 % and almost complete inhibition of alternariol monomethyl ether. All the tested fractions reduced percentage of infected tomato fruits when compared to the controls. The ethanolic and chloroformic fractions of E. globulus completely inhibited growth of A. alternata and A. arborescens on unwounded fruits and reduced the aggressiveness on wounded fruits of strains of both species significantly.     

Identification and characterization of <Emphasis Type="Italic">Alternaria</Emphasis> species causing leaf spot on cabbage,cauliflower, wild and cultivated rocket by using molecular and morphological features and mycotoxin production

Alternaria species are common pathogens of fruit and vegetables able to produce secondary metabolites potentially affecting human health. Twenty-nine isolates obtained from cabbage, cauliflower, wild and cultivated rocket were characterized and identified based on sporulation pattern and virulence; the phylogenetic analysis was based on the β-tubulin gene. Isolates were identified as A. alternata, A. tenuissima, A. arborescens, A. brassicicola and A. japonica. Pathogenicity was evaluated on plants under greenhouse conditions. Two isolates showed low level of virulence on cultivated rocket while the other isolates showed medium or high level of virulence. Isolates were also characterized for their mycotoxin production on a modified Czapek-Dox medium. Production of the five Alternaria toxins, tenuazonic acid, alternariol, alternariol monomethyl ether, altenuene and tentoxin were evaluated. Under these conditions, about 80% of the isolates showed the ability to produce at least one mycotoxin.     

Mycotoxins in Alternaria alternata - infected olive fruits and their possible transfer into oil1

Alternaria species, mostly A. alternata, have occasionally been found in some years on olive samples collected in Puglia (southern Italy). A survey was consequently made on the occurrence of the major Alternaria mycotoxins, i.e. alternariol (AOH), alternariol methyl ether (AME), altenuene (ALT), altertoxin-I (ATX-I), and tenuazonic acid (TA) in olives and products of their processing (oil and husks). The toxigenicity of Alternaria strains isolated from olives, and the possible transfer of mycotoxins into the oil, were also investigated. Four out of 13 olive samples were contaminated by 2–4 Alternaria mycotoxins. The highest contamination was found in a badly damaged sample containing 2.9, 2.3, 1.4 and 0.3 mg kg^?1 of AME, AOH, ALT and ATX-I, respectively. No mycotoxins were detected in olive oil destined for human consumption (6 samples) or olive husks (3 samples) collected from oil-mills after the first pressing of olives. An oil sample produced in our laboratory by processing the most contaminated olive sample, contained AOH (0.79 mg kg^?1) and AME (0.29 mg kg^?1). The estimated amount of mycotoxin transferred into the oil was 4%, for AME, 1.8% for AOH, and zero for ALT and TA (considering oil yield as 15%). The A. alternata strains isolated from olive produced much more mycotoxins when cultured on rice (up to 3 orders of magnitude for TA) than on olive.     

Ethylene production by <Emphasis Type="Italic">Alternaria alternata</Emphasis> and its association with virulence on inoculated grape berries

Ethylene has been shown to promote spore germination and hyphal growth in the phytopathogenic fungus Alternaria alternata. However, little is known about the ethylene biosynthetic pathway in this fungus. In the present study, the ethylene biosynthetic pathway in A. alternata was investigated to explore ethylene-associated virulence of this fungus. The strain A0 of A. alternata did not produce ethylene on basal medium with different possible precursors or intermediates for ethylene biosynthesis (glutamate, aspartate, 2-oxoglutarate and 1-aminocyclopropano ?1-carboxylic acid). However, ethylene production was observed when methionine was added as a precursor to the medium and was further promoted by continuous light illumination. Furthermore, addition of 2-keto-4-methylthiobutyric acid (KMBA) promoted ethylene production in the absence of methionine, indicating that the KMBA pathway was mainly responsible for ethylene biosynthesis in this fungus. The strain A0 was inoculated into grape berries to examine the effect of ethylene production on its virulence (as assessed by lesion formation at the inoculation site). The results indicated that higher ethylene production caused larger lesion formation. Similar results were also obtained when isolates of A. alternata, obtained from infected grapes, were inoculated. Thus, the present study thus demonstrated that A. alternata produces ethylene via the KMBA pathway and utilizes it for enhanced virulence expression during infection.     

<Emphasis Type="Italic">Alternaria alternata</Emphasis>: A new seed-transmitted disease of coriander in South Africa

This is the first report of Alternaria leaf spot disease on coriander (Coriandrum sativum L.) in South Africa. Using the agar plate method, Alternaria alternata was isolated from coriander seed lots together with four other fungal genera, which included Aspergillus, Fusarium, Penicillium and Rhizopus. Standard seed germination tests of coriander seed lots infected with seed-borne mycoflora showed a positive correlation with the number of diseased seedlings (r?=?0.239, p?<?0.01). Pathogenicity tests demonstrated that this seed-borne A. alternata was pathogenic on coriander and symptoms on leaves first appeared as small, dark brown to black, circular lesions (<5 mm diam.) that enlarged and coalesced to form dark brown blotches as time progressed. Leaf spot disease was most severe (64%) on wounded leaves inoculated with A. alternata. Re-isolation of A. alternata from diseased coriander plants satisfied the Koch’s postulates, thus confirming it as the causal agent of Alternaria leaf spot disease. Parsimony analysis based on rpb2 (GenBank Accession No. KT895947), gapdh (KT895949) and tef-1α (KT895945) sequences confirmed identity of the Alternaria isolate, which grouped within the A. alternata clade. Alternaria alternata was shown to be transmitted from infected coriander seed to the developing plants.     

Sensitivity of Red Delicious Apple Fruit at Various Phenologic Stages to Infection by Alternaria alternata and Moldy-core Control

The fungus Alternaria alternata, is considered to be the predominant fungus involved in moldy-core of Red Delicious strains of apple. In this paper, we report on the sensitivity of various phenologic stages to infection by A. alternata, and on the efficacy of various fungicides in controlling moldy-core disease in apple orchards. Artificial inoculations conducted in the orchard during 1999 and 2000 seasons revealed that the beginning of bloom (10–30%) and full bloom were the most susceptible developmental stages for infection. Natural infection with A. alternata in fruits was relatively high, reaching 44% and 46% of the fruits on control non-treated trees in 1999 and 2000. Four foliar applications of polyoxin B, difenoconazole and azoxystrobin, starting from the beginning of bloom until fruit set, reduced the number of infected fruits by 54–70%, 61–70% and 50–65%, respectively, compared with non-treated trees. Four or eight foliar applications of each fungicide provided similar levels of control. There were no significant differences between two, four or six foliar applications of difenoconazole, neither between two or four applications of polyoxin B. Adding CaCl₂, as a tank mixture with difenoconazole at full rate, did not improve efficacy. Alternaria was recovered from the inner part of the core region of 71–88% of the fruits of the non-treated control, but was recovered less frequently from the outside part of the core region. Fruits of difenoconazole and polyoxin B treated trees were less colonized with A. alternata at both the inner and outside parts of the core region, as compared with controls. Results indicate that a control programme based on spray applications of difenoconazole or polyoxin B, during bloom period, can effectively reduce Alternaria on Red Delicious.     

Characterization of iprodione-resistant Alternaria isolates from pistachio in California

Alternaria late blight caused by Alternaria spp. in the alternata, tenuissima, and arborescens species-groups is one of the most common fungal diseases of pistachio in California. In this study, a field iprodione-resistant (FIR) isolate of the arborescens species-group and a laboratory-induced iprodione-resistant (LIIR) isolate of the alternata species-group were characterized by fungicide and osmotic sensitivity, virulence on detached pistachio leaflets, and sequence of the coiled-coil region (six repeats of approximately 90-amino-acid domain) of the two-component histidine kinase (HK) gene. Both FIR and LIIR isolates were sensitive to azoxystrobin and tebuconazole, and azoxystrobin-resistant isolates were sensitive to iprodione and tebuconazole. The LIIR isolate showed more sensitivity to osmotic stress than its wild-type parent. However, the FIR isolate did not show higher osmotic sensitivity compared to field iprodione-sensitive (FIS) isolates. Laboratory inoculation tests showed that both FIR and LIIR isolates remained highly virulent on pistachio. Analysis of DNA sequences of the HK coiled-coil region showed that there were no differences in deduced amino acid sequence of this region from the LIIR, FIR, and FIS Alternaria isolates from pistachio in California.     

Assessing the Belgian potato <Emphasis Type="Italic">Alternaria</Emphasis> population for sensitivity to fungicides with diverse modes of action

Early blight and brown spot, caused by respectively Alternaria solani and Alternaria alternata, can lead to severe yield losses in potato-growing areas. To date, fungicide application is the most effective measure to control the disease. However, in recent years, a reduced sensitivity towards several active ingredients has been reported. To shed light on this issue, Alternaria isolates were collected from different potato fields in Belgium during two growing seasons. Subsequently, the sensitivity of these isolates was assessed using four widely used fungicides with different modes of action. Demethylation inhibitors, quinone outside inhibitors, a dithiocarbamate and a carboxylic acid amide were included in this study. Although all fungicides reduced spore germination and vegetative growth of Alternaria species to some extent, the interspecies sensitivity was very variable. In general, A. solani was more suppressed by the fungicides compared to A. alternata. The effectiveness of the dithiocarbamate mancozeb was high, whereas the quinone outside inhibitor azoxystrobin showed a limited activity, especially towards A. alternata. Therefore, a subset of the A. alternata and A. solani isolates was tested for the presence of, respectively, the G143A substitution and the F129L substitution in the cytochrome b. The frequency of A. alternata isolates bearing the resistant G143A allele (approximately 65%) was comparable in both sampling years, although sensitivity of isolates decreased during the growing season. This finding points to a shift of the population towards resistant isolates. Both the European genotype I and American genotype II were present in the A. solani population, with genotype I being the most prevalent. None of the genotype I isolates carried the F129L substitution, whereas in 83% of the genotype II isolates this substitution was present. Our results demonstrate for the first time that the Belgian Alternaria population on potato comprises a considerable broad spectrum of isolates with different sensitivity to fungicides.     

Pathogenicity and toxigenicity of <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> isolates collected from maize roots,stems and ears in South Africa

Sunflower (Helianthus annuus L.) is an important oilseed crop in South Africa, and is grown in rotation with maize in some parts of North West, Limpopo, Free State, Mpumalanga and Gauteng provinces. Alternaria leaf blight is currently one of the major potential disease threats of sunflower and is capable of causing yield losses in all production regions. Alternaria helianthi was reported as the main cause of Alternaria leaf blight of sunflower in South Africa; however small-spored Alternaria species have been consistently isolated from leaf blight symptoms during recent surveys. The aim of this study was to use morphological and molecular techniques to identify the causal agent(s) of Alternaria blight isolated from South African sunflower production areas. Alternaria helianthi was not recovered from any of the sunflower lesions or seeds, with only Alternaria alternata retrieved from the symptomatic tissue. Molecular identification based on a combined phylogenetic dataset using the partial internal transcribed spacer regions, RNA polymerase second largest subunit, glyceraldehyde-3-phosphate dehydrogenase, translation elongation factor and Alternaria allergen gene regions was done to support the morphological identification based on the three-dimensional sporulation patterns of Alternaria. Furthermore, this study aimed at evaluating the pathogenicity of the recovered Alternaria isolates and their potential as causal agents of Alternaria leaf blight of sunflower. Pathogenicity tests showed that all the Alternaria alternata isolates tested were capable of causing Alternaria leaf blight of sunflower as seen in the field. This is the first report of A. alternata causing leaf blight of sunflower in South Africa.     

Characterization of Alternaria species associated with leaf blight of sunflower in China

Nine Alternaria species have been reported to be associated with sunflower leaf blight worldwide, and A. helianthi has been recognized as the most prevalent and damaging species. However, the population structure of Alternaria species causing leaf blight of sunflower in China had not been examined thoroughly prior to this study. During 2010 to 2013, a total of 272 Alternaria isolates were obtained from infected sunflower leaves in 11 provinces, autonomous regions, and municipalities of China. Based on morphological traits and sequence analyses of the internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) and the partial coding sequences of the histone 3 gene, 227 (83.5 %) isolates were identified as belonging to Alternaria tenuissima, the remainder 45 isolates were grouped to A. alternata (16.5 %). Compared with the ITS regions of rDNA, sequence analyses of the partial coding sequences of histone 3 gene displayed a critical role in discrimination of the small-spored Alternaria species. Phylogenetic analysis of the partial coding sequence of histone 3 gene clearly divided the representative Alternaria isolates into two main clades, A. tenuissima and A. alternata. The pathogenicity of A. tenuissima and A. alternata on detached leaves of sunflower cv. Gankui No.2 did not significantly differ between the two species or among isolates from different geographical origins. Our results indicate that the population structure of Alternaria species associated with sunflower leaf blight differed from that reported previously in China since A. helianthi was not found in this study. In addition, this is the first report about A. tenuissima causing leaf blight on sunflower in China.     

Cytological and biochemical changes induced by chitosan in the pathosystem Alternaria alternata-tomato

The cytological and biochemical response of the fungus Alternaria alternata to chitosan application in tomato fruits was evaluated. The research was developed in the following stages: microscopically to observe the degree of damage that chitosan causes over the conidia and hyphae of the fungus at the structural level and during the infection process in tomato tissue. Biochemically we tried to identify the elicitation of the phytoalexin rhisitin and other compounds involved in resistance. At the microscopic level, mycelium and conidia of chitosan-treated of A. alternata showed cell wall disintegration, plasma membrane retraction, cellular distortion, release of the apical portion of the conidia and lysis of fungal cells. Hyphae and conidia were susceptible to chitosan application. Infection always took place in chitosan treated and inoculated tomatoes and it was difficult to observe ultrastructural alterations due to chitosan application. The phytoalexin rhisitin was not isolated from any of the treatments but other compounds such as alkenes, fatty acids and vitamin E whose antimicrobial effects have been reported were detected. The elicitation of precursor compounds in the pathosystem A. alternata-tomato was more associated with the infection process than with the chitosan application. Further studies are necessary to confirm these findings.     

Molecular characterization and pathogenicity of <Emphasis Type="Italic">Alternaria</Emphasis> species on wheat and date palms in Oman

This study was conducted to investigate the Alternaria species associated with leaf spot of date palm and wheat in Oman. Out of 98 date palm leaf samples and 146 wheat leaf samples, Alternaria was isolated from 27 and 23% of the samples developing leaf spot symptoms, respectively. Identification of Alternaria isolates using sequences of the internal transcribed spacer region of the ribosomal RNA (ITS rRNA), glyceraldehyde-3-phosphate dehydrogenase (GPDH), translation elongation factor (TEF) and RNA polymerase II subunit (RPB2) genes, showed that the isolates belong to seven Alternaria species or species complexes. A. burnsii - A. tomato and A. arborescens species complexes (58 and 4%, respectively) and A. alternata (38%) were the species recovered from the symptomatic date palm leaves. A. alternata (67%), A. burnsii - A. tomato species complex (15%), A. jacinthicola (3%), A. ventricosa (3%), A. slovaca (6%) and Alternaria caespitosa (6%) were isolated from wheat. Pathogenicity test showed that tested isolates of A. alternata (DPM19, WDK12), A. burnsii - A. tomato species complex (DPM31), A. jacinthicola (WBR4) and A. slovaca (WDK9, WDK7) were pathogenic on date palm, while A. alternata (DPM19, WDK12), A. burnsii - A. tomato species complex (DPM31, WDK11) and A. slovaca (WDK9, WDK7) were pathogenic on wheat. This is the first report of date palm and wheat as new hosts for A. burnsii - A. tomato species complex and the first reports of A. burnsii - A. tomato species complex, A. caespitosa A. slovaca, and A. ventricosa in Oman. The study shows that several species of Alternaria are associated with leaf spot in date palm and wheat in Oman, with some isolates having the ability to cause infection in both hosts.     

Chlorocholine chloride mediated resistance mechanism and protection against leaf spot disease of Stevia rebaudiana Bertoni

Investigations have been carried out to determine the influence of chlorocholine chloride on induction of the resistance mechanisms of Stevia rebaudiana against leaf spot disease, caused by Alternaria alternata. The paper also focuses an impact of chlorocholine chloride induced resistance on reduction of leaf spot disease. Chlorocholine chloride is attributed to its significant role in defence responses through augmentation of phenol and salicylic acid content as well as stimulation of phenylalanine ammonia lyase and peroxidase activity in S. rebaudiana following inoculation with A. alternata. Histochemical studies revealed that fungal invasion as well as infection process was appreciably reduced in chlorocholine chloride treated plants through peroxidase-H₂O₂ mediated strengthening of cell wall. The overall study highlights the significant role of chlorocholine chloride in induction of resistance in S. rebaudiana against A. alternata.     

Infection and colonization of peanut pods by Aspergillus parasiticus and the expression of the aflatoxin biosynthetic gene,nor-1, in infection hyphae

The aflatoxigenic fungi, Aspergillus flavus and A. parasiticus infect a wide variety of crops, all of which produce oil-rich seed. A histological study of the host–pathogen interaction between peanut,Arachis hyphogea , and A. parasiticus was performed in a system where peanuts remained attached to the plant and were inoculated without wounding. For infection studies, a genetically-tagged strain of A. parasiticus, G5, was engineered to harbor the β-glucuronidase (GUS) reporter gene under control of the nor-1 promoter from the aflatoxin biosynthetic pathway. There was a similar temporal pattern of aflatoxin B1 production and appearance of GUS activity in cultures ofA. parasiticus G5. This strain was used to follow infection and aflatoxin production during colonization of undamaged, drought-stressed peanuts. The fungus colonized all tissues of the peanut pod and appeared to gain ingress through the corky layer of the pericarp. Both intra- and inter-cellular colonization were observed. Fungal colonization of the cotyledons resulted in visible depletion of storage bodies within cells. Two morphologically distinct types of hyphae, wider hyphae and narrower hyphae, were seen throughout the pod tissues. Statistical analysis revealed that the narrower hyphae were significantly more likely to produce GUS activity than wider ones. GUS activity was found in hyphae infecting the pericarp, embryo and cotyledons indicating expression of aflatoxin biosynthetic genes in these tissues. Interestingly, GUS activity was not observed in the hyphae colonizing the testa.     

Pathogenic variation of Alternaria species associated with leaf blotch and fruit spot of apple in Australia

Four Alternaria species groups (A. longipes, A. arborescens, A. alternata/A. tenuissima and A. tenuissima/A. mali) are associated with leaf blotch and fruit spot of apple in Australia. There is no information on the variability of pathogenicity among the species and isolates within each species causing leaf blotch or fruit spot. We used a detached leaf assay and an in planta fruit inoculation assay to determine the pathogenicity and virulence of the four Alternaria species. Our results showed that isolates within the same species were not specific to either leaf or fruit tissue and showed great variability in pathogenicity and virulence, indicating cross-pathogenicity, which may be isolate dependent rather than species dependent. Generally, virulence of A. tenuissima and A. alternata isolates on leaf and fruit was higher than other species. Isolates of all species groups were pathogenic on leaves of different cultivars, but pathogenicity on fruit of different cultivars varied among isolates and species. Implications of our findings on prevalence of the diseases in different apple-producing regions in Australia and the development of targeted disease management of the diseases are discussed.     

Sweet potato stem blight caused by Alternaria sp.: A new disease in Ethiopia

A new sweet potato disease was discovered in Ethiopia. The disease mainly affects the stems and petioles of sweet potato and the name sweet potato stem blight is proposed. The pathogen is a species ofAlternaria, for which no definite name has been found so far. Sweet potatoes were most susceptible, tomatoes were slightly susceptible, and muskmelons and chilli peppers were resistant. Thorn apples and onions showed slight to moderate symptoms, butAlternaria was not reisolated from these two species. Although all sweet potato varieties tested were susceptible, there were significant differences in sensitivity.     

Detoxification of α-tomatine by tomato pathogens Alternaria alternata tomato pathotype and Corynespora cassiicola and its role in infection

In tomato plants, α-tomatine, a steroidal glycoalkaloid saponin, inhibits fungal growth. Tomato pathogens that produce host-specific toxins, Alternaria alternata tomato pathotype causing Alternaria stem canker and Corynespora cassiicola causing Corynespora target spot, were investigated for sensitivity to α-tomatine. Although spore germination of A. alternata pathogenic and nonpathogenic to tomato and of C. cassiicola pathogenic to tomato was not affected by 0.1 mM α-tomatine, spore germination of C. cassiicola nonpathogenic to tomato was significantly inhibited. This result showed that A. alternata, regardless of its pathogenicity, and only the C. cassiicola pathogenic to tomato are resistant to α-tomatine. Germinating spores of A. alternata and C. cassiicola resistant to α-tomatine detoxified α-tomatine by degrading it to a less polar product. After inoculation of tomato leaves, spores of A. alternata and C. cassiicola nonpathogenic to tomato germinated and formed appressoria, but did not form infection hyphae in host tissues. When a host-specific toxin (CCT-toxin) produced by C. cassiicola pathogenic to tomato was added to nonpathogenic spores, colonization within leaves was observed in A. alternata, but not in C. cassiicola. On the other hand, when spores of C. cassiicola nonpathogenic to tomato were suspended in spore germination fluid of nonpathogenic A. alternata with α-tomatine detoxification activity, the fungus could be induced to colonize leaves in the presence of CCT-toxin. These results indicate that A. alternata tomato pathotype and C. cassiicola pathogenic to tomato detoxify α-tomatine during infection and that this detoxification is essential for host colonization by pathogens that produce host-specific toxins.     

Characterization of Alternaria species associated with potato foliar diseases in China

The population structure of Alternaria species associated with potato foliar diseases in China has not been previously examined thoroughly. Between 2010 and 2013, a total of 511 Alternaria isolates were obtained from diseased potato leaves sampled in 16 provinces, autonomous regions or municipalities of China. Based on morphological traits and molecular characteristics, all the isolates were identified as Alternaria tenuissima, A. alternata or A. solani. Of the three species, A. tenuissima was the most prevalent (75·5%), followed by A. alternata (18·6%) and A. solani (5·9%). Phylogenetic analysis based on sequences of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) of representative Alternaria isolates showed that A. solani was distinct from the two small‐spored Alternaria species. Phylogenetic analysis of the partial coding sequence of the histone 3 gene divided the same collection of isolates into three main clades representing A. tenuissima, A. alternata and A. solani, respectively. The pathogenicity of the isolates on detached leaves of potato cv. Favorite did not differ significantly between the three species or between isolates from different geographical origins. The results indicate that the population structure of Alternaria species associated with potato foliar diseases differs from that reported previously in China. This is the first report of A. tenuissima causing potato foliar diseases in China.     

Sulphur supply impairs spread of Verticillium dahliae in tomato

Vascular wilt caused by the soil-borne fungus Verticillium dahliae is a major yield and quality-limiting disease across a broad spectrum of crop plants worldwide. Sulphur-enhanced plant defence mechanisms provide an opportunity to effectively and environmentally safely constrain the wilt disease levels in planta. To evaluate the influence of sulphur nutrition on the protective potential of these mechanisms, two near-isogenic tomato genotypes differing in fungal susceptibility, were treated with low or supra-optimal sulphur supply. Microscopic analysis revealed a significant sulphur-induced decrease in the amount of infected vascular cells in both genotypes. However, plant shoot and severely pathogen-affected root growth did not display this distinct alleviating influence of sulphur nutrition. Rates of leaf photosynthesis were impeded by Verticillium dahliae infection in both genotypes especially under low sulphur nutrition. However, assimilate transport rates in the phloem sap were enhanced by fungal infection more in the resistant genotype and under high sulphur nutrition suggesting a stronger sink for assimilates in infected plant tissues possibly involved in sugar-induced defence. A SYBR Green-based absolute quantitative Real-Time assay using a species-specific primer was developed which sensitively reflected sulphur nutrition-dependent changes in fungal colonization patterns. High sulphur nutrition significantly reduced fungal spread in the stem in both tomato genotypes. Concentrations of selected sulphur-containing metabolites revealed an increase of the major anti-oxidative redox buffer glutathione under high sulphur nutrition in response to fungal colonization. Our study demonstrates the existence of sulphur nutrition-enhanced resistance of tomato against Verticillium dahliae mediated by sulphur-containing defence compounds.     

Modes of action of the protective strain Fo47 in controlling verticillium wilt of pepper

The protective fungus Fusarium oxysporum Fo47 reduces the severity of wilt caused by the soilborne pathogen Verticillium dahliae in pepper. Modes of action responsible for the biocontrol activity were studied. Microscopic observations of fluorescent protein‐transformed strains colonizing the root surface show that the colonization patterns of Fo47 and V. dahliae were similar. Pixel counting of the images obtained by confocal microscopy showed that Fo47 reduces colonization of the root surface by V. dahliae, suggesting a possible role of competition for nutrients at the root surface. Besides these effects on surface colonization, the hormonal pathways activated during priming of plant defence responses were identified by measuring the amount of some phytohormones and their derivatives in roots and stems of pepper. Results showed an early, slight increase of jasmonyl isoleucine, followed by a transient increase of salicylic acid during the pre‐challenged phase of priming and an increase of 12‐oxo‐phytodienoic acid during the challenge phase of priming. The caffeic, ferulic and chlorogenic acids, known to play a role in plant defence reactions, showed a strong antimicrobial activity against V. dahliae in vitro. In pepper roots, Fo47 stimulated the biosynthesis of caffeic acid and primed that of chlorogenic acid. These results demonstrated that the effective control of V. dahliae provided by Fo47 is based on different but complementary mechanisms.