Antagonistic effects of soil bacteria onFusarium oxysporum Schlecht f. sp.dianthi (Prill and Del.) Snyd. and Hans

Summary Fusarium oxysporum f. sp.dianthi, pathogenic on carnation plants is very sensitive toBacillus subtilis M51 inhibition.Fusarium oxysporum disease (fusariosis) is prevented for a period of two months after treatment of plants withBacillus subtilis M51. The persistence ofB. subtilis M51, marked for selenomycin resistance (MZ51) and inoculated on the roots of carnation cuttings was studied. Soil used was two types: naturally infested withFusarium oxysporum and free from this pathogen. Bacterial cells presence on the roots was detected by direct plating and the presence of the pathogen in the roots was investigated by histological assays. Evidence gathered by these procedures suggest that plant protection is dependent on the physical presence ofB. subtilis M51 cells on the roots.     

Correlations between Accumulation of Dianthramides, Dianthalexin and Unknown Compounds, and Partial Resistance to Fusarium oxysporum f. sp. dianthi in Eleven Carnation Cultivars

The accumulation of dianthramides, dianthalexin and other phenolic compounds in carnation cells at different times before and after inoculation of plants with Fusarium oxysporum f. sp. dianthi, and the development of wilt symptoms were compared for eleven carnation cultivars. Untreated and water-treated controls contained only a little acetone-extractable phenolics. Upon inoculation with F. oxysporum f. sp dianthi, all cultivars accumulated the same range of phenolic compounds, including several dianthramides and dianthalexin, but in different proportions. The total amount of accumulated phenolic compounds per fresh material weight of the extracted stem segments could not be related to the disease resistance level of the different cultivars. However, the accumulation of dianthramides and dianthalexin in the cultivars was positively correlated to resistance, while the accumulation of two other, as yet unidentified, compounds was inversely related to resistance.     

In vitro selection and regeneration of carnation (Dianthus caryophyllus L.) plants resistant to culture filtrate of Fusarium oxysporum f.sp. dianthi

Callus cultures derived from internodal segments of two cultivars of carnation susceptible to Fusarium oxysporum f.sp. dianthi were successfully used for in vitro selection for resistance to this pathogenic fungus. Resistant lines were selected by culturing calli on growth medium containing various concentrations of the culture filtrate of F. oxysporum f.sp. dianthi. Resistant calli obtained after two cycles (25 days/cycle) of selection were used for plant regeneration. About 32% of the plants regenerated from the resistant calli had acquired considerable resistance against the pathogen in the field. No phenotypic variation was observed in the selected regenerates.     

A new benzoic acid derivative isolated from Piper cf. cumanense Kunth (Piperaceae)

New benzoic acid derivative (1), together with five known compounds has been isolated from the inflorescences of Piper cf. cumanense Kunth (Piperaceae). The structure was identified on basis of spectroscopic analysis and comparison with literature data. The compound (1) showed antifungal activity against Fusarium oxysporum f. sp. dianthi and Botrytis cinerea.     

Effect of Simulated Soil Solarization and Organic Amendments on Fusarium Wilt of Rocket and Basil Under Controlled Conditions

Pot trials were carried out under controlled conditions to evaluate the effectiveness against Fusarium wilt of rocket (Fusarium oxysporum f.sp. conglutinans) and basil (F. oxysporum f.sp. basilici) of soil amendments based on a patented formulation of Brassica carinata defatted seed meal and compost, combined or not with a simulation of soil solarization. The soil solarization treatment was carried out in a growth chamber by heating the soil for 7 and 14 days at optimal (55–52°C for 6 h, 50–48°C for 8 h and 47–45°C for 10 h/day) and sub‐optimal (50–48°C for 6 h, 45–43°C for 8 h and 40–38°C for 10 h/day) temperatures similar to those observed in summer in solarized soil in greenhouses in Northern Italy. Two subsequent cycles of plant cultivation were carried out in the same soil. Even at sub‐optimal temperature regimes, 7 days of thermal treatment provided very valuable results in terms of disease control on both rocket and basil. In general, the thermal treatment was more effective against F. oxysporum f.sp. basilici than against F. oxysporum f.sp. conglutinans. Control of Fusarium wilt of rocket is improved with 14 days of thermal treatment. The combination of organic amendments with a short period of soil solarization (7 or 14 days), although not providing any improvement to the level of disease management, did significantly increase biomass and positively affected yield.     

Impact of compost application on Fusarium wilt disease incidence and microelements contents of basil plants

Fusarium oxysporum was isolated from stem of basil plants showing symptoms of wilt, stem blight and collar root rot. Pathogenicity tests indicated that F. oxysporum f. sp. basilici is the causal agent of this disease. This is the first report of this pathogen in Egypt. The suppressive effects of six types of composts on Fusarium wilt disease incidence in basil were evaluated under greenhouse conditions. The effectiveness of these composts and their relation to the microelements content in treated plants was also assessed. Soil treatments with Khaya and Eucalyptus composts significantly reduced the infection percentage and disease severity of basil wilt. Otherwise, the applications of Araucaria, Datura, Ficus and Azadirachta composts showed no effect on both infection percentage and disease severity. Moreover, the Khaya and Eucalyptus compost treatments increased the levels of Iron (Fe), Zinc (Zn) and Manganese (Mn) in treated basil plants than application of Araucaria, Datura, Ficus and Azadirachta composts. In the case of Copper (Cu) content, it was significantly higher only in Eucalyptus-compost-treated plants than in other compost applications. These composts not only reduced the disease incidence but also increased both fresh and dry weight (FW and DW) and microelements contented in basil treated plants. In general, although soil amendment with either Khaya or Eucalyptus compost can reduce the disease incidence of Fusarium wilt on basil plants, microelements contented; FW and DW of these effects can be variable depending on their levels added in soil. According to the results of this study, it can be concluded that the use of compost in the soil as an organic fertiliser increased exchangeable form of microelements in the soil and also the availability of these elements by basil plants.     

Grape marc compost: microbial studies and suppression of soil-borne mycosis in vegetable seedlings

Compost suppression of soil-borne diseases in horticultural crops has been attributed to the activities of antagonistic microorganisms. A great diversity of microorganisms, capable of suppressing pathogens naturally colonize compost. A large number of microbes appeared in microbiological analyses of grape marc compost. Most microorganisms were bacteria. Average percentages were 31% mesophilic and 28% thermophylic bacteria, 16% mesophilic actinomycetes and 20% thermophylic actinomycetes. Only a few mould and yeast morphologies were obtained, 4% and 1% respectively. Antagonist in vitro assays were performed with 432 microbial morphologies isolated from grape marc compost. The microbes isolated were extremely effective antagonists in in vitro assays against all the fungal pathogens tested. Seven microorganisms were selected for further bioassay with Rhizoctonia solani in radish, Fusarium oxysporum f. sp. radicis-cucumerinum in melon, and Phytophthora parasitica in tomato and two microorganisms with Pythium aphanidermatum in cucumber. Those experiments indicate that grape marc compost reduces the severity of Pythium damping-off in cucumber, but does not reduce the severity of Phytophthora root rot in tomato, Fusarium oxysporum f. sp. radicis-cucumerinum in melon and Rhizoctonia solani in radish. Better suppressive effects were not demonstrated by either compost or vermiculite amended with microbes selected from grape marc compost.     

Identification and characterization of non-pathogenic Fusarium oxysporum capable of increasing and decreasing Fusarium wilt severity

Fusarium wilt of banana is a potentially devastating disease throughout the world. Options for control of the causal organism, Fusarium oxysporum f.sp. cubense (Foc) are limited. Suppressive soil sites have previously been identified where, despite the presence of Foc, Fusarium wilt does not develop. In order to understand some aspects of this disease suppression, endophytic Fusarium oxysporum isolates were obtained from banana roots. These isolates were genetically characterized and compared with an isolate of Fusarium oxysporum previously identified as being capable of suppressing Fusarium wilt of banana in glasshouse trials. Three additional isolates were selected for glasshouse trials to assess suppression of Fusarium wilt in two different cultivars of banana, Cavendish and Lady Finger. One isolate (BRIP 29089) was identified as a potential biocontrol organism, reducing the disease severity of Fusarium wilt in Lady Finger and Cavendish cultivars. Interestingly, one isolate (BRIP 45952) increased Fusarium wilt disease severity on Cavendish. The implications of an isolate of Fusarium oxysporum, non-pathogenic on banana, increasing disease severity and the potential role of non-pathogenic isolates of Fusarium oxysporum in disease complexes are discussed.     

Kinetic of the production of polygalacturonase and pectin lyase by two closely relatedFormae speciales ofFusarium oxysporum

The kinetic of thein vitro production of polygalacturonase and pectin lyase of two closely related fungi,Fusarium oxysporum f.sp.lycopersici andF. oxysporum f.sp.radicis-lycopersici, was examined under various culture conditions such as the source of carbon, the pH, and the age of cultures. Over a 5-day period, the production of these enzymes by various isolates of the sameforma specialis (f. sp.) ofF. oxysporum was not significantly different (P ≥ 0.05). However, the amount of the enzymes produced differed markedly between both f. sp. The different carbon sources added to the culture media, such as citrus pectin, apple pectin, tomato cell wall fragments, andd-galacturonic acid, proved to be higher pectinase inducible substrates than sucrose and glucose. For both fungi, polygalacturonase and pectin lyase activities were optimal at pH 5.0 and 8.0, respectively. Furthermore, pectin lyase production had a partial Ca²⁺ requirement in contrast to polygalacturonase production which was limited by Ca²⁺. In most experiments performed, the production of polygalacturonase appeared superior withF. oxysporum f.sp.radicislycopersici than withF. oxysporum f.sp.lycopersici. On the other hand, pectin lyase production ofF. oxysporum f.sp.lycopersici was approximately 10-fold greater than that byF. oxysporum f.sp.radicis-lycopersici in media supplemented withd-galacturonic acid.     

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.     

Biological Control of Fusarium-wilt in Carnation With Serratia liquefaciens and Hafnia alvei Isolated from Rhizosphere of Carnation

Serratia liquefaciens provided better protection of carnations from infection by Fusarium oxysporum f. sp. dianthi, than did Hafnia alvei. The protection occurred when the bacterial isolates were applied to the cuttings before rooting, but not when applied to the root system of rooted cuttings. S. liquefaciens was recovered from carnation stem segments along the stem up to the top after 60 days but after 120 days they were recovered only up to 2.5cm. Zones of inhibition of Fusarium-conidia sprayed on agar plates previously incubated with stem segments appeared around the bacterial colonies of S. liquifaciens after additional incubation for 24—48h.     

Suppression of Alfalfa Growth by Concommitant Populations of Pratylenchus penetrans and two Fusarium species

Growth of alfalfa (Medicago sativa cv. Vernal) seedlings was compared after inoculation with combinations of either Pratylenchus penetrans and Fusarium soloni or P. penetrans and F. oxysporum f. sp. medicaginis. A synergistic disease interaction occurred in alfalfa when F. oxysporum and P. penetrans were added simultaneously to the soil. Alfalfa growth was suppressed at all inoculum levels of P. penetrans and F. oxysporum, but not with F. solani. Seedlings inoculated with the nematode alone gave lower yields than when inoculated with either Fusarium species alone. Fusarium oxysporum, but not F. solani, was pathogenic to alfalfa under similar experimental conditions. Fusarium oxysporum did not alter the populations of P. penetrans in alfalfa roots, whereas the presence of F. solani was associated with a diminished number of P. penetrans in the roots.     

Host-specific Fusarium oxysporum Causes Wilt of Jojoba

Jojoba [Simmondsia chinensis (Link) Schneider] plantations in Israel originated from vegetative propagation, planted during 1991–92, have shown symptoms of wilting and subsequent death. Verticillium dahliae was only rarely isolated from these plants and artificial inoculation showed only mild disease symptoms. Fusarium oxysporum caused severe chlorosis, desiccation, defoliation and wilt in leaves of jojoba plants, resulting in plant death. Recovery of the fungus from artificially inoculated stem cuttings and seedlings showed for the first time that F. oxysporum was the primary pathogen. Inoculated cuttings exhibited wilt within 3 weeks, while in seedlings wilt occurred 10–24 weeks after inoculation. Seedlings and cuttings of jojoba which were inoculated with other Fusarium isolates originating from different crops (F. oxysporum f. sp. vasinfectum from cotton, F. oxysporum f. sp. dianthi from carnation, F. oxysporum f. sp. lycopersici from tomato and F. oxysporum f. sp. basilicum from basil) did not develop symptoms. Moreover, cotton, tomato, melon and cucumber seedlings inoculated with several virulent F. oxysporum isolates from jojoba did not show any symptoms of wilt or defoliation. These results indicate a high degree of specificity of the Fusarium isolates from jojoba; therefore, it is suggested that this isolate be defined as F. oxysporum f. sp. simmondsia.     

Vegetative Compatibility Grouping of the Fusarium Wilt Pathogen of Paris Daisy (Argyranthemum frutescens L.)

Fusarium oxysporum isolates, pathogenic on Paris daisy (Argyranthemum frutescens), were classified by vegetative compatibility grouping analysis. They were compared with isolates of the formae speciales chrysanthemi and tracheiphilum, and with f. sp. dianthi as a genetically distant control. The results show the uniformity of the pathogenic isolates from Paris daisy, except for one which was of a different geographic origin. They were classified as a new VC group (0052). We report also the efficacy of different media in obtaining nit mutants.     

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.     

Electrophoretic Karyotypes of Fusarium spp.

Migheli, Q., Berio, T., and Gullino, M. L. 1993. Electrophoretic karyotypes of Fusarium spp. Experimental Mycology 17, 329-337. The electrophoretic karyotype of 17 antagonistic and pathogenic strains of Fusarium spp. has been established by using contour-clamped homogeneous electric field gel electrophoresis. Intact chromosomal DNA was prepared from fungal protoplasts with standard procedures. Up to 11 distinct chromosomal bands were resolved after 184 h of migration at 50 V. Polymorphic karyotypes were observed in different species of Fusarium, formae speciales of F. oxysporum , and races of F. oxysporum f.sp. dianthi. Using the Schizosaccharomyces pombe and Saccharomyces cerevisiae chromosomes as size standards, the size of the Fusarium genome was estimated to range from approximately 18.1 to 51.5 Mb. The suitability of electrophoretic karyotyping as a tool for strain characterization, as well as some applications in hybridization analysis of Fusarium spp., is discussed.     

Cumanensic acid, a new chromene isolated from Piper cf. cumanense Kunth. (Piperaceae)

A new chromene, cumanensic acid (1), together with eight known compounds have been isolated from the aerial part of Piper cf. cumanense Kunth (Piperaceae). Chromene structure was identified on the basis of spectroscopic analysis and comparison with literature data. The compound showed antifungal activity against Fusarium oxysporum f. sp. dianthi and Botrytis cinerea.     

Antagonistic effects of soil bacteria onFusarium oxysporum Schlecht f.sp.dianthii (Prill and Del.) Snyd. and Hans.

Summary Thirty two bacteria antagonistic to a number of phytopathogenic fungi were isolated from soil samples. One bacterial strain, designated as M 51, appeared to be particularly active towardsF. oxysporum f. sp.dianthii, in vitro andin vivo and it was inhibitoryin vitro to three otherFusarium spp. used. Tests to find if there was protection against fusarium wilt were carried out by three different methods of inoculation of the cuttings: a) dipping of cuttings for ten minutes in bacterial suspension; b) spraying of suspension on perlite where the rooted cuttings were planted; c) spraying the greenhouse bench rooting boxes, where the non-rooted cuttings were planted, with bacterial suspension. Following this all the cuttings were transplanted into soil naturally highly infested withFusarium oxysporum f. sp.dianthii (3000 units/g). Good protection against fusarium wilt was obtained for cuttings inoculated by method (b). However protection decreased gradually about 60 days after they were transplanted; both control and inoculated cuttings showed a comparable mortality rate. Method of inoculation and the development of the protective effect are discussed.     

Land use intensification affects soil microbial populations, functional diversity and related suppressiveness of cucumber Fusarium wilt in China’s Yangtze River Delta

The extent of soil microbial diversity in agricultural soils is critical to the maintenance of soil health and quality. The aim of this study was to investigate the influence of land use intensification on soil microbial diversity and thus the level of soil suppressiveness of cucumber Fusarium wilt. We examined three typical microbial populations, Bacillus spp., Pseudomonas spp. and Fuasarium oxysporum, and bacterial functional diversity in soils from three different land use types in China’s Yangtze River Delta, and related those to suppressiveness of cucumber Fusarium wilt. The land use types were a traditional rice wheat (or rape) rotation land, an open field vegetable land, and a polytunnel greenhouse vegetable land that had been transformed from the above two land use types since 1995. Results generated from the field soils showed similar counts for Bacillus spp. (log 5.87–6.01 CFU g⁻¹ dw soil) among the three soils of different land use types, significantly lower counts for Pseudomonas spp. (log 5.44 CFU g⁻¹ dw soil) in the polytunnel greenhouse vegetable land whilst significantly lower counts for Fusarium oxysporum (log 3.21 CFU g⁻¹ dw soil) in the traditional rice wheat (or rape) rotation land. A significant lower dehydrogenase activity (33.56 mg TPF kg⁻¹ dw day⁻¹) was observed in the polytunnel greenhouse vegetable land. Community level physiological profiles (CLPP) of the bacterial communities in soils showed that the average well color development (AWCD) and three functional diversity indices of Shannon index (H′), Simpson index (D) and McIntosh index (U) at 96 h incubation in BIOLOG Eco Micro plates were significantly lower in the polytunnel greenhouse vegetable land than in both the traditional rice wheat (or rape) rotation land and the open field vegetable land. A further greenhouse experiment with the air-dried and sieved soils displayed significantly lower plant growth parameters of 10-old cucumber seedlings as well as significantly lower biomass and total fresh fruit yield at the end of harvesting at day 70 in the polytunnel greenhouse vegetable soil sources. The percentages of Fusarium wilt plant death were greatly increased in the polytunnel greenhouse vegetable plants, irrespective of being inoculated with or without Fusarium oxysporum f. sp. cucumerinum. Our results could provide a better understanding of the effects of land use intensification on soil microbial population and functional diversity as well as the level of soil suppressiveness of cucumber Fusarium wilt.