Functional characterization of the recombinant antimicrobial peptide Trx-<Emphasis Type="Italic">Ace</Emphasis>-AMP1 and its application on the control of tomato early blight disease

Ace-AMP1 is a potent antifungal peptide found in onion (Allium cepa) seeds with sequence similarity to plant lipid transfer proteins. Transgenic plants over-expressing Ace-AMP1 gene have enhanced disease resistance to some fungal pathogens. However, mass production in heterologous systems and in vitro application of this peptide have not been reported. In this study, Ace-AMP1 was highly expressed in a prokaryotic Escherichia coli system as a fusion protein. The purified protein inhibited the growth of many plant fungal pathogens, especially Alternaria solani, Fusarium oxysporum f. sp. vasinfectum, and Verticillium dahliae. The inhibitory effect was accompanied by hyphal hyperbranching for V. dahliae but not for F. oxysporum f. sp. vasinfectum and A. solani, suggesting that the mode of action of Ace-AMP1 on different fungi might be different. Application of Ace-AMP1 on tomato leaves showed that the recombinant protein conferred strong resistance to the tomato pathogen A. solani and could be used as an effective fungicide.     

Mycotoxin-producing potential of fungi associated with qat (Catha edulis) leaves in yemen

Forty-four fungal species belonging to 20 genera were isolated from 30 samples of qat leaves. The most frequent genera wereAspergillus, Alternaria, Penicillium, andCladosporium followed byFusarium, Drechslera, Chœtomium, andMucor. The most prevalent species in above genera wereAspergillus niger, A. flavus, A fumigatus, Alternaria alternata, Penicillium chrysogenum, P. citrinum, Cladosporium cladosporioides, andFusarium verticillioides. From these fungi, 17 species (39%) related to 7 genera (35%) proved to be true endophytes. Eleven out of 75 isolates were mycotoxigenic.A. alternata produced alternariol and alternariol monomethyl ether whereasA. flavus produced aflatoxins B₁ and B₂. Ochratoxin A, sterigmatocystin, citrinin and T-2 toxin were produced byA. ochraceus, A. versicolor, P. citrinum andF. oxysporum, respectively. The presence of such toxigenic fungi associated with qat leaves is considered to be a threat to public health.     

Effect of flavonoids on the development of Fusarium oxysporum f. sp. lycopersici

Abstract

In the present study the effect of flavonoid compounds on the germination and fungal growth of the soil-borne tomato pathogen Fusarium oxysporum f. sp. lycopersici was studied. Out of 12 flavonoid compounds only myricetin and luteolin exhibited a low stimulating activity on microconidia germination of Fusarium oxysporum f. sp. lycopersici, whereas the other flavonoids tested were inactive when applied at five different concentrations. In our study the tested flavonoids affect fungal growth differently to microconidia germination. Individual flavonoid concentrations resulted in a small increase of fungal growth, but the lowest flavonoid concentrations showed an inhibiting effect on fungal growth for all flavonoids tested. There is evidence to suggest, that low flavonoid concentrations exhibit slight antimicrobial properties against Fusarium oxysporum f. sp. lycopersici.     

Phytopathogenic Fusarium oxysporum Schlecht, as a Necrotrophic Mycoparasite

Fusarium oxysporum f. sp. dianthi, f. sp. lycopersici, f. sp. cepae, f. sp. niveum and one unidentified F. oxysporum isolate proved to be active necrotrophic mycoparasites. In dual cultures hyphae of Trichoderma hamatum, T. longibrachiatum, T. pseudokoningii, T. harzianum, Botrytis cinerea and Rhizoctonia solani were parasitized and destroyed by F. oxysporum. One isolate of Phytophthora sp. was not affected. Mutual parasitism between F. oxysporum and T. pseudokoningii and T. longibrachiatum has been observed, too. Details of parasitic hyphal interactions: hyphal coiling, penetration sites, resistance sheat formation, hyphal invasion and internal growing are described. The mycoparasitic feature as well as antimicrobial metabolic production of F. oxysporum is probably a common phenomenon to ensure this important plant pathogenic species to compete successfully against other soil-borne fungal pathogens and saprophytes.     

Direct effect of biocontrol agents on wilt and root-rot diseases of sesame

In Egypt, sesame cultivation is subject to attack by wilt and root-rot diseases caused by Fusarium oxysporum f.sp. sesami (Zap) Cast. and Macrophomina phaseolina (Maubl) Ashby causing losses in quality and quantity of sesame seed yield. Bacillus subtilis and Trichoderma viride isolates which were isolated from sesame rhizosphere were the most effective to antagonise fungal pathogens, causing high reduction of hyphal fungal growth. Trichoderma viride was found to be mycoparasitic on Fusarium oxysporum f.sp. sesami and M. phaseolina causing morphological atternation of fungal cells and sclerotial formation. In general, Bacillus subtilis, T. viride, avirulent Fusarium oxysporum isolate and Glomus spp. (Amycorrhizae) significantly reduced wilt and root-rot incidence of sesame plants at artificially infested potted soil by each one or two pathogens. Data obtained indicate that Glomus spp significantly reduced wilt and disease severity development on sesame plants followed by T. viride. Meanwhile, avirulent Fusarium oxysporum isolate followed by Glomus spp. were effective against root-rot disease incidence caused by M. phaseolina. Glomus spp. followed by B. subtilis significantly reduced wilt and root-rot disease of sesame plants. All biotic agents significantly reduced F. oxysporum f.sp. sesami and M. phaseolina counts in sesame rhizosphere at the lowest level. Glomus spp. and the avirulent isolate of F. oxysporum eliminated M. phaseolina in sesame rhizosphere. Meanwhile T. viride was the best agent at reducing F. oxysporum at a lower level than other treatments. Application of VA mycorrhizae (Glomus spp.) in fields naturally infested by pathogens significantly reduced wilt and root-rot incidence and it significantly colonised sesame root systems and rhizospheres compared to untreated sesame transplantings.     

The synthetic strigolactone GR24 influences the growth pattern of phytopathogenic fungi

Strigolactones that are released by plant roots to the rhizosphere are involved in both plant symbiosis with arbuscular mycorrhizal fungi and in plant infection by root parasitic plants. In this paper, we describe the response of various phytopathogenic fungi to the synthetic strigolactone GR24. When GR24 was embedded in the growth medium, it inhibited the growth of the root pathogens Fusarium oxysporum f. sp. melonis, Fusarium solani f. sp. mango, Sclerotinia sclerotiorum and Macrophomina phaseolina, and of the foliar pathogens Alternaria alternata, Colletotrichum acutatum and Botrytis cinerea. In the presence of this synthetic strigolactone, intense branching activity was exhibited by S. sclerotiorum, C. acutatum and F. oxysporum f. sp. melonis. Slightly increased hyphal branching was observed for A. alternata, F. solani f. sp. mango and B. cinerea, whereas suppression of hyphal branching by GR24 was observed in M. phaseolina. These results suggest that strigolactones not only affect mycorrhizal fungi and parasitic plants, but they also have a more general effect on phytopathogenic fungi.     

Modulation des Réactions de Défense du Palmier a Huile Contre le Fusarium oxysporum f. sp. elaeidis (Schlecht) Toovey: Applications: Prémunition et Stimulation Chimique

Defence reactions of palm trees to Fusarium oxysporum f. sp. elaeidis (Schlecht) Toovey Cross protection and stimulation of inhibitory substances In palm tree genetic characters of tolerance to Fusarium oxysporum f. sp. elaeidis are correlated with synthesis of fungal inhibitors in infected tissues. Individual variation of synthesis level is also observed among plants of a same line. Defence reactions are triggered by pre-inoculation of an avirulent strain of Fusarium oxysporum. Similar results are obtained by application of analogs of fungal elicitors like arachidonic acid. Quite the contrary, treatment of plants with a competitive inhibitor of PAL downs natural barriers and in a same way the effects of cross protection and arachidonic treatment. Seven phenolic compounds, mainly benzoic acid derivatives, inhibit in vitro the growth of pathogen like spores germination. The variability of the host reaction and its stimulation by elicitors could be used to improve the resistance.     

Interaction of Fusarium Oxysporum f.sp. Cubense with Pseudomonas Fluorescens Precolonized to Banana Roots

Effect of precolonization of banana cv Neeypovan roots with Pseudomonas fluorescens on infection with Fusarium oxysporum f.sp. cubense was studied. Under in vitro conditions Pseudomonas fluorescens clearly inhibited Fusarium oxysporum f.sp. cubense. Fluorescein isothiocyanate-tagged antibodies raised in a rabbit system for Pseudomonas fluorescens and Fusarium oxysporum f.sp. cubense separately were used to study the spread of both organisms in banana root. It was observed that precolonization with Pseudomonas fluorescens could reduce Fusarium oxysporum f.sp. cubense colonization by 72%, and also correlated with a number of structural changes in the cortical cells, mainly with densely stained amorphous material and polymorphic wall thickenings as revealed by light and electron microscopic studies. Massive depositions of unusual structures at sites of fungal entry was also noticed, which clearly indicated that bacterized root cells were signalled to mobilize a number of defence structures for preventing the spread of pathogen in the tissue. This revised version was published online in July 2006 with corrections to the Cover Date.     

Biological Control ofMonilinia laxaandFusarium oxysporumf. sp.lycopersiciby a Lytic Enzyme-ProducingPenicillium purpurogenum

Biological control experiments were conducted with the lytic enzyme-producing fungusPenicillium purpurogenumagainst the plant pathogensMonilinia laxaandFusarium oxysporumf. sp.lycopersici.Applications ofP. purpurogenumto peach shoots previously inoculated withM. laxareduced lesion length and extent of pathogen colonization of shoots by 90 and 80% (P ≤ 0.05), respectively, comparable to the level of disease control obtained with the fungicide captan. Disease severity in tomato plants inoculated withF. oxysporumf. sp.lycopersiciwas decreased by 30% (P ≤ 0.05) with the biological treatment. The fungusP. purpurogenumproduced β-1,3-glucanase and chitinase activities in liquid culture that were inducible by cell walls and live mycelium ofM. laxabut not ofF. oxysporumf. sp.lycopersici.Crude filtrates or crude enzyme preparations ofP. purpurogenumcultures with lytic enzyme activities produced lysis of hyphae and spores ofM. laxaandF. oxysporumf. sp.lycopersici.These lytic effects were strong inM. laxaand ended in complete dissolution of mycelium. The induction of lytic enzymes byM. laxaand the effects of lytic enzymes on mycelia of the pathogens in relation to the different degrees of biological control obtained are discussed.     

Spatial Peculiarities in the Colonization of the Plant Rhizoplane by Microscopic Fungi

Spatial peculiarities in the colonization of the tomato, cucumber, and barley rhizoplanes by microscopic fungi were studied. The apical zone of roots was colonized with a limited number of Rstrategists (the order Mucorales, Fusariumsp., Aspergillus niger, and Mycelia sterilia). The fungal population of the root hairs and the basal zone of roots was 2- to 3-fold denser due to the prevalence of Kstrategists. Fusaria, Fusarium oxysporumin particular, colonized roots in earlier terms than the genera Trichoderma, Penicillium, Gliocladium, and others. The F. oxysporumpopulation was at a maximum in the rhizoplane zone nearest the root tip.     

Induction of defence-related enzymes in onion by combined application of fungal and bacterial biocontrol agents against Fusarium oxysporum f. sp. cepae

Basal rot disease of onion is a major problem in different onion growing regions of Tamil Nadu, India. Fungal and bacterial cultures were isolated and tested their efficiency against Fusarium oxysporum f. sp. cepae under in vitro conditions. Effective bacterial and fungal antagonists were tested alone and in combinations for the control of F. oxysporum f. sp. cepae in glasshouse experiments. Defence-related enzymes such as peroxidase, polyphenol oxidase and phenylalanine ammonia-lyase were induced and accumulated in onion treated with fungal and bacterial antagonists. Defence-related enzymes were significantly higher in onion pretreated with consortial formulation of Pf12 + Pf27 + TH3 at 5 days after the challenge inoculation with F. oxysporum f. sp. cepae and gave resistance to onion against basal rot disease.     

Studies on the rhizosphere mycoflora of broad bean and cotton

Investigations were made on the effect of species of fungi isolated from therhizosphere of broad bean (Vicia faba Linn.) variety Giza I, and cotton (Gossypium barbadense Linn.) variety Giza 47, on plant growth. Broad bean rhizosphere fungi differently affected plant growth.Fusarium moniliforme, Penicillium martensii, Rhizopus stolonifer, andCladosporium sphaerospermum stimulated both root and shoot growth.Aspergillus niger andAlternaria tenuis have an inhibitory effect on plant growth. On the other hand, the rhizosphere fungi of cotton namely,Penicillium cyclopium, Aspergillus terreus, Cephalosporium sp.,Aspergillus niger, Cladosporium sphaerospermum, andFusarium oxysporum stimulated plant growth.     

Studies on the mycoflora associated with sugarcane factory waste and pollution of River Nile in upper Egypt

Sixty-nine species and four varieties that belong to 28 genera of terrestial fungi were recovered from polluted and non-polluted water and mud samples on glucose and cellulose-Czapek's agar at 28°C. The most common species from the two substrates on the two types of media were Aspergillus flavus, A. fumigatus, A.niger, Cladosporium cladosporioides, Fusarium oxysporum, Mycosphaerella tassiana and Penicillium chrysogenum. Twenty-six species belonging to 14 genera were isolated from polluted (26 species and 14 genera) and non-polluted (17species and 10 genera) mud samples on Sabouraud's dextrose agar at 28°C. The most prevalent species were Acremonium retiulum, Alternaria alternata, Aspergillus flavus, Aphanoascus fulvescens, A. terreus, Aphanoascus sp., Penicillium funiculosum and Stachybotrys chartarum.     

Actinomycetes isolated from medicinal plant rhizosphere soils: diversity and screening of antifungal compounds,indole-3-acetic acid and siderophore production

A total of 445 actinomycete isolates were obtained from 16 medicinal plant rhizosphere soils. Morphological and chemotaxonomic studies indicated that 89% of the isolates belonged to the genus Streptomyces, 11% were non-Streptomycetes: Actinomadura sp., Microbispora sp., Micromonospora sp., Nocardia sp, Nonomurea sp. and three isolates were unclassified. The highest number and diversity of actinomycetes were isolated from Curcuma mangga rhizosphere soil. Twenty-three Streptomyces isolates showed activity against at least one of the five phytopathogenic fungi: Alternaria brassicicola, Collectotrichum gloeosporioides, Fusarium oxysporum, Penicillium digitatum and Sclerotium rolfsii. Thirty-six actinomycete isolates showed abilities to produce indole-3-acetic acid (IAA) and 75 isolates produced siderophores on chrome azurol S (CAS) agar. Streptomyces CMU-PA101 and Streptomyces CMU-SK126 had high ability to produced antifungal compounds, IAA and siderophores.     

Allergic potential of someAspergilli on saw mill workers in Lucknow,India

Fifty fungal types were isolated from the indoor atmosphere of saw mills by exposing Petri plates containing Czapek-dox Agar, Potato-dextrose Agar and Sabouraud Agar media for 5 min. The fungal flora of the outdoor surroundings was also studied for comparison. Species ofAspergilli dominated in the saw mills, being represented by 16 species including one ascosporic form. Other fungi were species ofCladosporium, Alternaria, Curvularia, Penicillium, Fusarium, etc. Variations in the fungal population in different months were also observed. Fungal spores recovered using the Rotorod Sampler wereAlternaria, Curvalaria lunata, Curvularia tetramera, Cladosporium, Dreschslera sp.,Epicoccum sp.,Pithomyes sp.,Nigrospora, Stemphylium sp. andTorula sp. Mycelial fragments and unidentifiable spores were also seen in abundance. Varying allergic responses of patients were also recorded by testing intradermally, the antigens of nineAspergilli, vizAspergillus flavus, A. fumigatus, A. japonicus, A. melleus, A. nidulans, A. niger, A. niveus, A. tammarii and A. terreus.     

Transgenic tomato plants expressing a wheat endochitinase gene demonstrate enhanced resistance to <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis>

Various chitinases have been shown to inhibit the growth of fungal pathogens in in vitro as well as in planta conditions. chi194, a wheat chitinases gene encoding a 33-kDa chitinase protein, was overexpressed in tomato plants (cv. Pusa Ruby) under the control of maize ubiquitin 1 promoter. The integration of transgene in tomato plants was confirmed with polymerase chain reaction (PCR) and Southern blot analysis. The inheritance of the transgene in T₁ and T₂ generations were shown by molecular analysis and the hygromycin sensitivity test. The broad range of chitinase activity was observed among the transgenic lines in T₀ and a similar range was retained in the T₁ and T₂ generations. Most importantly, the transgenic tomato lines with high chitinase activity were found to be highly resistant to the fungal pathogen Fusarium oxysporum f. sp. lycopersici. Thus, the results demonstrated that the expression of the wheat endochitinase chi194 in tomato plants confers resistance against Fusarium wilt disease caused by the fungal pathogen Fusarium oxysporum f. sp. lycopersici.     

Allergic potential of someAspergilli on saw mill workers in Lucknow,India

Fifty fungal types were isolated from the indoor atmosphere of saw mills by exposing Petri plates containing Czapek-dox Agar, Potato-dextrose Agar and Sabouraud Agar media for 5 min. The fungal flora of the outdoor surroundings was also studied for comparison. Species ofAspergilli dominated in the saw mills, being represented by 16 species including one ascosporic form. Other fungi were species ofCladosporium, Alternaria, Curvularia, Penicillium, Fusarium, etc. Variations in the fungal population in different months were also observed. Fungal spores recovered using the Rotorod Sampler wereAlternaria, Curvalaria lunata, Curvularia tetramera, Cladosporium, Dreschslera sp.,Epicoccum sp.,Pithomyes sp.,Nigrospora, Stemphylium sp. andTorula sp. Mycelial fragments and unidentifiable spores were also seen in abundance. Varying allergic responses of patients were also recorded by testing intradermally, the antigens of nineAspergilli, vizAspergillus flavus, A. fumigatus, A. japonicus, A. melleus, A. nidulans, A. niger, A. niveus, A. tammarii and A. terreus.     

Fungicides and some biological controller agents effects on the growth of fusarium oxysporum causing paprika wilt: Wirkung von fungiziden und einigen biologischen kontrollwirkstoffen auf das wachstum des paprikawelke verursachenden fusarium oxysporum

Soil samples from both healthy and diseased paprika roots were tested to identify their mycoflora. Thirty-one species belonging to 16 genera were collected from rhizosphere and rhizoplane samples. The most frequently isolated fungi were Aspergillus flavus, A. niger, A. terreus, Fusarium oxysporum, Penicillium jensenii and Trichoderma harzianum. Fusarium oxysporum was the most common Fusarium species in the rhizoplane samples of diseased roots and identification was confirmed by RAPD-PCR technique. Trichoderma harzianum, T. pseudokoningii and Glioclaium roseum were chosen to study their biological control efficiency against Fusarium oxysporum. These fungal species reduced the percentage of seedling infection to 25, 40 and 50%, respectively. With the increasing of fungicide (Folicur and Ridomil) doses the dry weight of F. oxysporum decreased. Also, the increasing of fungicide dose lead to a slight decrease in the dry weight of T. harzianum, T. pseudokoningii and Glioclaium roseum.     

Biological Control Activity of Three Trichoderma Isolates Against Fusarium Wilts of Cotton and Muskmelon and Lack of Correlation with their Lytic Enzymes

The enzymatic activity and the biocontrol ability of two new isolates of Trichoderma spp. (T-68 and Gh-2) were compared in laboratory and glasshouse experiments with a previously studied T. harzianum strain (T-35). In dual culture tests with Fusarium oxysporum f. sp. melonis and F. oxysporum f. sp. vasinfectum, isolates T-68 and Gh-2 overgrew the colonies of Fusarium, whereas T-35 failed to parasitize both wilt pathogens. Under glasshouse conditions, the three isolates of Trichoderma were effective in controlling Fusarium wilt of cotton but only T-35 was effective against F. oxysporum f. sp. melonis on muskmelon. When the three Trichoderma isolates were grown on liquid media containing laminarin, colloidal chitin or F. oxysporum f. sp. melonis cell walls as sole carbon sources, maximum β-1,3-glucanase and chitinase specific activity in the culture filtrates of all fungi was reached after 72h of incubation. When culture filtrates of the three Trichoderma isolates were incubated with freeze-dried mycelium of F. oxysporum f. sp. melonis or F. oxysporum f. sp. vasinfectum, different concentrations of glucose and N-acetyl-D-glucosamine were released. Overall no correlation was found between enzymatic activity and the biocontrol capability against Fusarium wilt on muskmelon and cotton.     

Genetic Diversity of Fusarium oxysporum Populations Isolated from Tomato Plants in Tunisia

Fusarium crown and root rot of tomato (Lycopersicon esculentum) caused by Fusarium oxysporum f. sp. radicis‐lycopersici is a new devastative disease of tomato greenhouse crops in Tunisia. Nothing is known neither about the population of this pathogen in this region, nor about the population of F. oxysporum f. sp. lycopersici the causal agent of Fusarium wilt of tomato. In order to examine the genetic relatedness among the F. oxysporum isolates by intergenic spacer restriction fragment length polymorphism (IGS‐RFLP) analysis and to elucidate the origin of the formae specialesradicis‐lycopersici in Tunisia by looking for genetic similarity of Tunisians isolates with isolates from a foreign source, the genetic diversity among F. oxysporum f. sp. radicis‐lycopersici and F. oxysporum f. sp. lycopersici populations was investigated. A total of 62 isolates of F. oxysporum, obtained from symptomless tomato plants, were characterized using IGS typing and pathogenicity tests on tomato plants. All Fusarium isolates were highly pathogenic on tomato. Fusarium oxysporum f. sp. radicis‐lycopersici isolates were separated into five IGS types. From the 53 F. oxysporum f. sp. radicis‐lycopersici isolates, 34 isolates have the same IGS types (IGS type 25), and the remaining 19 isolates were distributed into four IGS types. However, the only nine isolates of F. oxysporum f. sp. lycopersici have six different IGS types. This difference of diversity between the two formae speciales suggests that F. oxysporum f. sp. radicis‐lycopersici isolates have a foreign origin and may have been accidentally introduced into Tunisia.