Benomyl-resistant Fusarium-isolates in ecological studies on the biological control of fusarium wilt in carnation

Ecological properties and stability of benomyl resistance of three benomyl-resistant mutants of nonpathogenicFusarium-isolates antagonistic to fusarium wilt in carnation, and three benomyl-resistant mutants of a pathogenic isolate ofFusarium oxysporum f.sp.dianthi were evaluatedin vitro and in glasshouse experiments. The benomyl resistance of the nonpathogenic mutants was stable under all conditions tested, also after a 1000-fold increase of the population in sterilized soil. Mutants of the pathogen were stable during allin vitro tests, but after proliferation in carnation stems only part of the population was benomyl resistant.Compared to the wild type, mutants of the pathogen were less pathogenic, also if thein vitro propeties were similar. Colonization of carnation by benomyl-resistant nonpathogenicFusarium in the presence of the pathogen showed that the antagonistic effect correlated with the presence of the nonpathogenic isolates within the carnation stem. The wild types and two of the mutant nonpathogenicFusarium-isolates controlled fusarium wilt in the susceptible cultivar Lena for 50% or more.UV-induced benomyl resistance appeared to be a valuable marker to distinguish between differentFusarium isolates and to study the population dynamics, but intensive screening of the mutants is a prerequisite since alterations in antagonism and pathogenicity can occur.     

Siderophore-mediated competition for iron and induced resistance in the suppression of fusarium wilt of carnation by fluorescent Pseudomonas spp

The mechanisms of suppression of fusarium wilt of carnation by two fluorescentPseudomonas strains were studied.Treatments of carnation roots withPseudomonas sp. WCS417r significantly reduced fusarium wilt caused byFusarium oxysporum f. sp.dianthi (Fod). Mutants of WCS417r defective in siderophore biosynthesis (sid^–) were less effective in disease suppression compared with their wild-type. Treatments of carnation roots withPseudomonas putida WCS358r tended to reduce fusarium wilt, whereas a sid^– mutant of WCS358 did not.Inhibition of conidial germination of Fod in vitro by purified siderophores (pseudobactins) of bothPseudomonas strains was based on competition for iron. The ferrated pseudobactins inhibited germination significantly less than the unferrated pseudobactins. Inhibition of mycelial growth of Fod by bothPseudomonas strains on agar plates was also based on competition for iron: with increasing iron content of the medium, inhibition of Fod by thePseudomonas strains decreased. The sid^– mutant of WCS358 did not inhibit Fod on agar plates, whereas the sid^– mutants of WCS417r still did. This suggests that inhibition of Fod by WCS358r in vitro was only based on siderophore-mediated competition for iron, whereas also a non-siderophore antifungal factor was involved in the inhibition of Fod by strain WCS417r.The ability of thePseudomonas strains to induce resistance against Fod in carnation grown in soil was studied by spatially separating the bacteria (on the roots) and the pathogen (in the stem). Both WCS417r and its sid^– mutant reduced disease incidence significantly in the moderately resistant carnation cultivar Pallas, WCS358r did not.It is concluded that the effective and consistent suppression of fusarium wilt of carnation by strain WCS417r involves multiple mechanisms: induced resistance, siderophore-mediated competition for iron and possibly antibiosis. The less effective suppression of fusarium wilt by WCS358r only depends on siderophore-mediated competition for iron.     

Carnation Fusarium wilt suppression in four composts

Fusarium wilt is now a major disease of carnation crops worldwide. Methyl bromide, which is used to remedy it, is environmentally unsafe. An alternative approach integrated into biological control is to grow crops in suppressive media. Suppressiveness of seven plant growth media to Fusarium oxysporum f. sp. dianthi was evaluated in bioassays with carnation (Dianthus cariophyllus) cv. Medea. These media were: (1) grape marc compost, (2) cork compost, (3) olive oil husk + cotton gin trash composted and mixed with rice husk, (4) spent mushroom compost mixed with peat, (5) coir fibre, (6) light peat and (7) vermiculite. In order to look for carnation Fusarium wilt suppressiveness indicators, growth medium pH and β-glucosidase activity were evaluated. Furthermore, F. oxysporum populations were measured in plant growth media at the beginning and end of bioassays. The compost media showed a range of suppressiveness in comparison with peat. Grape marc compost was the most effective plant growth medium in suppressing carnation Fusarium wilt. On the other hand coir fibre, peat and vermiculite were conducive for this disease. β-glucosidase activity and pH were positively correlated with disease severity as in other reports for tomato. Therefore, these two parameters are good indicators for carnation Fusarium wilt suppressiveness, and possibly for other F. oxysporum pathosystems. All composts showed similar F. oxysporum populations at the end of the bioassays to peat and vermiculite.     

Physiological races of Fusarium oxysporum f.sp. melonis in Tunisia

Fusarium wilt of melon, caused by Fusarium oxysporum f.sp. melonis (Fom), is an important disease; races of the pathogen were identified by inoculating differential standard host cultivars. A total of ten isolates that were obtained from 23 fields located in four different geographical regions were identified as pathogenic. Results indicate that all four known Fom races, namely, 0, 1, 2 and 1.2, were found in north and middle Tunisia. Race 1.2 was the most prevalent.     

Fusarium wilt of banana: biology,epidemiology and management

Fusarium oxysporum is a soil borne hyphomycete that causes vascular wilts in several crop plants. A variety of remedial measures such as the use of fungicides, soil amendments and biological antagonists have proved insufficient in controlling F. oxysporum. Ever since it was first reported in banana crop, the only effective control strategy known is planting of resistant cultivars. However, presumably due to the high mutation rates and rapid co-evolution with its host, Fusarium wilt has surmounted host defense barriers and has already begun infecting even the resistant Cavendish varieties that dominate export markets worldwide. Transgenic banana plants showing enhanced resistance to Fusarium wilt have been developed in recent past, but they remain largely confined to the laboratory. The importance of banana as source of food and income in developing countries world over and the need to develop Fusarium wilt tolerant cultivars by novel biotechnological approaches is detailed herein. In this communication, we review the biology and management of Fusarium wilt in banana with the aim of providing the baseline of information to encourage much needed research on integrated management of this destructive banana crop disease problem.     

Back to the roots: Understanding banana below-ground interactions is crucial for effective management of Fusarium wilt

Global banana production is affected by Fusarium wilt, a devastating disease caused by the soilborne root-infecting fungus, Fusarium oxysporum f. sp. cubense (Foc). Fusarium wilt is notoriously difficult to manage because infection arises through complex below-ground interactions between Foc, the plant, and the soil microbiome in the root–soil interface, defined as the rhizosphere. Interactions in the rhizosphere play a pivotal role in processes associated with pathogen development and plant health. Modulation of these processes through manipulation and management of the banana rhizosphere provides an auspicious prospect for management of Fusarium wilt. Yet, a fundamental understanding of interactions in the banana rhizosphere is still lacking. The objective of this review is to discuss the state-of-the-art of the relatively scant data available on banana below-ground interactions in relation to Fusarium wilt and, as a result, to highlight key research gaps. Specifically, we seek to understand (a) the biology of Foc and its interaction with banana; (b) the ecology of Foc, including the role of root-exuded metabolites in rhizosphere interactions; and (c) soil management practices and how they modulate Fusarium wilt. A better understanding of molecular and ecological factors influencing banana below-ground interactions has implications for the development of targeted interventions in the management of Fusarium wilt through manipulation of the banana rhizosphere.     

Reduced severity and impact of Fusarium wilt on strawberry by manipulation of soil pH, soil organic amendments and crop rotation

Strawberry (Fragaria?×?ananassa) is one of the most important berry crops worldwide. Fusarium wilt poses a serious threat to commercial strawberry production worldwide and causes severe economic losses. Our previous surveys suggested that soil pH, soil amendment with organic matter and/or crop rotation could offer opportunities for improved management of strawberry disease. Studies were conducted for the first time to determine the effects of soil pH, soil amendments with manure compost and crop residue, and crop rotation on the severity and impact of Fusarium wilt on strawberry. At soil pH 6.7, plants showed the least severe disease and the lowest reductions in shoot and root dry weight (DW) of plants from disease, significantly lower than those of plants in acidic soil at pH 5.2 or 5.8. In soil amendment with manure compost at 5.0?%, plants showed the least severe disease and the lowest reductions in shoot and root DW of plants from disease, significantly lower than those of plants in the other three levels of manure compost. In soil amendment with crop residue at 2.5?% or 5.0?%, shoot and root disease of plants and reductions in shoot and root DW of plants from disease were significantly lower than those of plants in soil without crop residue or excessive crop residue amendment at 10.0?%. Plants in soil rotated with tomato not only showed the least severe disease but also showed the lowest reductions in shoot and root DW of plants from disease, significantly lower than those of plants in soil continuously planted with strawberry without rotation or rotated with capsicum. Soil pH, soil amendment with manure compost or crop residue, and crop rotation, all significantly reduced the severity and impact of Fusarium wilt on strawberry. There is great potential for manipulating soil pH, adding soil organic amendments and utilizing crop rotation, not only to successfully manage Fusarium wilt on strawberry, but to do so in a sustainable way without current reliance upon chemical fumigants.     

Antifungal activity of pomegranate peel extract against fusarium wilt of tomato

Pomegranate (Punica granatum) is an important source of bioactive compounds and has been used in folk medicine for many centuries. This paper describes the in vitro antifungal activity of pomegranate peel aqueous extract (pae) on the development of Fusarium wilt of tomato caused by Fusarium oxysporum, f. sp. lycopersici. HPLC-DAD-ESI/MS analysis was performed to identify punicalagins and ellagic acid, which are the main antifungal compounds. In vivo tests established the efficacy of pae treatments in controlling Fusarium wilt by evaluating improvements in growth variables of tomato plants. At high concentrations, pae showed allelopathic activity in tomato plants. The germination and the radicle growth of tomato seeds were significantly affected by pae. Increasing the extract concentration led to a progressive decrease in germination and in the length of the radicle. The reduction of the Fusarium population in soil and the increase in number of healthy plants obtained as a result of pae treatments indicate that this plant extract could have an important role in biologically-based management strategies for the control of Fusarium wilt in tomato crops.     

Molecular characterization of Fusarium oxysporum f.sp. cichorii pathogenic on chicory (Cichorium intybus)

Fusarium oxysporum is a ubiquitous soilborne ascomycete responsible for vascular wilt in many plant species worldwide. This species comprises more than 120 putative host-specific formae speciales capable of causing marked economic losses. In summer 2009, wilt symptoms, including chlorosis and poor development of the root system, were observed on cultivars of chicory (Cichorium intybus) in northern Italy. The causal agent isolated from symptomatic tissues in this case was identified as F. oxysporum on the basis of both morphological features and molecular analyses. In this work, we attempted to characterize the isolates of F. oxysporum from C. intybus by both biological and molecular approaches. Pathogenicity trials performed on five species of the Asteraceae family with isolates of F. oxysporum from C. intybus indicated that the pathogen has a unique host range, infecting chicory only. Neither lettuce nor endive, lawn daisy or Paris daisy developed the disease. Five cultivars within C. intybus species were tested, and the cv. ??Clio?? was the most susceptible. Phylogenetic analyses relative to the ribosomal intergenic spacer (IGS) and translation elongation factor 1-alpha (EF1-??) assigned isolates pathogenic to chicory to a single cluster, distinct from other pathogenic F. oxysporum. In light of these findings, we propose to designate this organism as Fusarium oxysporum f.sp. cichorii.     

Pathogenicity and vegetative compatibility grouping among Indian populations of Fusarium oxysporum f. sp. ciceris causing chickpea wilt

Thirty-nine isolates of Fusarium oxysporum f. sp. ciceri – the causal agent of chickpea (Cicer arietinum) wilt – collected from different parts of India and representing eight races of the pathogen, were analyzed for virulence and classified on the basis of vegetative compatibility grouping (VCG). The wilt incidence ranged from 24% to 100% on a highly susceptible cultivar JG 62. Six isolates, from Delhi, Gujarat, Karnataka, Punjab and Rajasthan and belonging to six different races of the pathogen, caused 100% wilt incidence. Five isolates belonging to four different races, namely, Foc 143 from Andhra Pradesh, Foc 161 from Chhattisgarh, Foc 146 from Karnataka, Foc 158 from Madhya Pradesh and Foc 50 from Rajasthan, caused low wilt incidence. For VCG analysis, nitrate non-utilizing mutants (nit) were obtained by culturing wild-type isolates on 2.5% potassium chlorate and selecting resistant sectors. Complementary nit mutants were paired in all possible combinations to determine varying degrees of heterokaryon formation within the isolates, which showed that most of the isolates were self-compatible. Pairing of all the mutants showed that the isolates included in the present study belonged to a single VCG (0280). Thus, in spite of variability in the virulence, the Indian populations of the pathogen have only one VCG.     

Colonization of lettuce cultivars and rotation crops by Fusarium oxysporum f. sp. lactucae,the cause of fusarium wilt of lettuce

The severity of fusarium wilt is affected by inoculum density in soil, which is expected to decline during intervals when a non‐susceptible crop is grown. However, the anticipated benefits of crop rotation may not be realized if the pathogen can colonize and produce inoculum on a resistant cultivar or rotation crop. The present study documented colonization of roots of broccoli, cauliflower and spinach by Fusarium oxysporum f. sp. lactucae, the cause of fusarium wilt of lettuce. The frequency of infection was significantly lower on all three rotation crops than on a susceptible lettuce cultivar, and the pathogen was restricted to the cortex of roots of broccoli. However, F. oxysporum f. sp. lactucae was isolated from the root vascular stele of 7·4% of cauliflower plants and 50% of spinach plants that were sampled, indicating a greater potential for colonization and production of inoculum on these crops. The pathogen was also recovered from the root vascular stele of five fusarium wilt‐resistant lettuce cultivars. Thus, disease‐resistant plants may support growth of the pathogen and thereby contribute to an increase in soil inoculum density. Cultivars that were indistinguishable based on above‐ground symptoms, differed significantly in the extent to which they were colonized by F. oxysporum f. sp. lactucae. Less extensively colonized cultivars may prove to be superior sources of resistance to fusarium wilt for use in breeding programmes.     

Pathogenicity of indole-3-acetic acid producing fungus <Emphasis Type="Italic">Fusarium delphinoides</Emphasis> strain GPK towards chickpea and pigeon pea

An indole-3-acetic acid (IAA) producing fungal strain was isolated from chickpea grown rhizospheric soil samples. Based on morphological and Internal Transcribed Spacer (ITS) region sequence analysis the new isolate was identified as Fusarium delphinoides. The Fusarium delphinoides strain produces and secretes IAA in-vitro as identified by HPLC and Mass spectrometry. The IAA production is dependent on tryptophan (Trp) as a nitrogen source in the medium. The IAA production is influenced by growth conditions such as pH of the medium, concentration of Trp and the nature of the carbon source. Additional nitrogen sources repress Trp dependent IAA production. Glucose and Trp served as the best carbon and nitrogen sources respectively. Pathogenicity of Fusarium delphinoides towards the plants was tested by electrolyte, nutrient leakage analysis and also by scoring the disease symptoms. Two cultivars of chickpea (ICCV-10 and L-550) and two cultivars of pigeon pea (Maruti and PT-221) were assessed for the pathogenicity by inoculating with spores of Fusarium delphinoides. The inoculation induced symptoms of Fusarium wilt as in the case of Fusarium oxysporum f. sp. ciceris (FOC), a known pathogen causing Fusarium wilt in chickpea. Electrolyte and nutrient leakage from the infected plants were used to assess the resistance, tolerance (moderately resistance) and susceptibility of the plants to the infection. Based on the results, both the pigeon pea cultivars (Maruti and PT-221) were rated as resistant, and ICCV-10 was rated as a tolerant cultivar of chickpea. However, chickpea cultivar L −550 was found to be a susceptible host for infection by Fusarium delphinoides. These results suggest that Fusarium delphinoides, which belongs to the Fusarium dimerum species group, is an IAA producing plant pathogen and causes wilt in chickpea. Further, along with pathogenicity tests, electrolyte and nutrient leakage analysis can be used to assess the pathogenicity of pathogenic fungi.     

Lessons from 10 years of stakeholder adoption of a soil bioassay for assessing the risk of spinach Fusarium wilt

Although the maritime Pacific Northwest (PNW) is the only region of the United States suitable climatically for spinach seed production, the acidic soils are highly conducive to spinach Fusarium wilt caused by Fusarium oxysporum f. sp. spinaciae. A soil bioassay developed to quantify the risk of spinach Fusarium wilt in fields has been offered to seed growers annually since 2010. Soil sampled from growers' fields each winter was planted with highly susceptible, moderately susceptible, and partially resistant spinach inbred lines, and the plants rated weekly to calculate a Fusarium wilt severity index (FWSI) and the area under the disease progress curve (AUDPC). Results for 147 soils tested from 2010 to 2013 have been published. This study examined results for an additional 248 soils tested from 2014 to 2019 with the bioassay modified to include an option of agricultural limestone amendment to the soils tested. FWSI and AUDPC were affected significantly (p < .001) by the main effects of soil and spinach inbred line, and the interaction of these factors. Correlation analyses showed a range in degree of association of FWSI and AUDPC with spinach seed crop rotation duration and soil properties, depending on the spinach inbred line (r = −.255 to –.267, n = 172 soils with characteristics suitable for correlation analyses). Stepwise regression models for 172 soils with relevant parameters for regression analyses identified spinach seed crop rotation interval, rate of agricultural limestone amendment, soil pH, and soil Fe, Mn, and Zn concentrations as most strongly associated with FWSI and AUDPC. However, the models accounted for ≤33.4% (R²) of the variability in Fusarium wilt risk. The soil bioassay remains a primary tool for spinach seed growers to select fields with low risk of Fusarium wilt.     

Genetic variability among breeding lines and cultivars of eggplant against Fusarium oxysporum f. sp. melongenae from Turkey

Seventy isolates of Fusarium oxysporum (Schlechtend: Fr.) f. sp. melongenae Matuo and Ishigami (Fomg), the causal agent of eggplant Fusarium wilt, were tested for their interaction with different lines and cultivars to determine whether there was race-specific interaction. Also, a total of 13 cultivars were tested under greenhouse conditions to evaluate the presence of resistance to Fusarium wilt. The disease severity (%) and the area under disease progress curve (AUDPC) for each of the Fomg isolates were calculated by scale values. Results showed that neither of the resistant lines (LS1934 and LS2436) exhibited wilting symptoms, whereas susceptible local cultivars (cvs. ‘Kemer’ and ‘Hadrian’) displayed severe disease symptoms. There was no significant variation in the virulence, indicating the occurrence of a race or races among 70 Fomg isolates tested on resistant lines and susceptible cultivars. This may indicate a genetically homogeneous population structure of Fomg in Turkey. The eggplant cultivars Amadeo, Anatolia, Angela, Brigitte, Corsica, Hawk, Koksal, Nouma, Sharapova and Yula exhibited various degrees of susceptibility to three virulent Fomg isolates, but disease severity for AGR-703 was significantly different and lower among the tested eggplant cultivars, as well as rootstocks. Therefore, it is considered as the most appropriate for rootstock purposes in eggplant cultivations where Fusarium wilt is present in the soil.     

Real-time PCR for differential determination of the tomato wilt fungus, <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis>, and its races

Five primer/probe sets to identify the tomato wilt pathogen, Fusarium oxysporum f. sp. lycopersici (FOL), and its three races selectively were designed based on the rDNA-intergenic spacer and avirulence genes. Real-time PCR using genomic DNA from mycelia and soil DNA with the primer/probe sets allowed the successful identification of FOL and its races.     

Stem canker and wilt of delphinium caused by Fusarium oxysporum f. sp. delphinii in Japan

Stem canker and severe wilt were observed on delphinium plants (Delphinium elatum) in Aomori Prefecture, Japan, in 2008. The fungus isolated from the diseased crown was identified as Fusarium oxysporum f. sp. delphinii on the basis of morphological characteristics, nucleotide sequences, and host range. The isolate induced similar stem canker and wilt symptoms in inoculated delphinium plants. We propose the name “stem canker and wilt” for the disease.     

A sensitive biosensor based on gold nanoparticles to detect <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> in soil

Ralstonia solanacearum, the devastating causal agent of potato bacterial wilt, is a soil-borne bacterium that can survive in the soil for a long time. The development of sensitive on-field detection methods for this pathogen is highly desirable due to its widespread host range and distribution. A novel nanobiosensor was thus developed to detect unamplified genomic DNA of R. solanacearum in farm soil. Gold nanoparticles functionalized with single-stranded oligonucleotides served as a probe to detect R. solanacearum genomic DNA. The advantages of this strategy include rapidity, facile usage and being a visual colorimetric method.     

Control of Fusarium wilt in melon by the fungal endophyte,Cadophora sp.

Two soil-borne fungal endophytes almost completely suppressed the effects of a post-inoculated and virulent strain of Fusarium oxysporum f. sp. melonis when inoculated to axenically reared melon seedlings in Petri dishes. They were identified as Cadophora sp. on the basis of ITS 1–5.8S rDNA–ITS 2 sequences and morphological characters and obtained from the roots of Chinese cabbage grown as bait plants in a mixed soil made up of samples from different forest soils from Alberta and British Columbia, Canada. Hyphae of Cadophora sp. grew along the surface of the root and colonized root cells of the cortex and reduced the ingress of the Fusarium pathogen into adjacent cells. Melon seedlings pre-inoculated with Cadophora sp. were also grown in soil amended with the different N sources, nitrate or the amino acids leucine and valine, and glucose (final C:N ratio?=?10:1). After 4 weeks, these seedlings were transplanted into the field and disease symptoms were assessed. Only the endophyte-inoculated seedlings treated with valine could effectively inhibit the development of Fusarium wilt in two plots and reduced disease symptom development by 43 and 62 %.     

Molecular analysis of Spanish populations of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">dianthi</Emphasis> demonstrates a high genetic diversity and identifies virulence groups in races 1 and 2 of the pathogen

Fusarium wilt, caused by Fusarium oxysporum f. sp. dianthi (Fod), is the most important carnation disease worldwide. The knowledge of the diversity of the soil population of the pathogen is essential for the choice of suitable resistant cultivars. We examined the genetic diversity of Fod isolates collected during the period 1998–2008, originating from soils and carnation plants in the most important growing areas in Spain. Additionally, we have included some Fod isolates from Italy as a reference. Random amplified polymorphic DNA (RAPD) fragments generated by single-primer PCR were used to compare the relationship between isolates. UPGMA analysis of the RAPD data separated Fod isolates into three clusters (A, B, and C), and this distribution was more related to aggressiveness than to the race of the isolates. The results obtained in PCR amplifications using specific primers for race 1 and race 2, and SCAR primers developed in this work, correlated with the molecular groups previously determined from the RAPD analysis, and provided new molecular markers for the precise identification of the isolates. Results from successive pathogenicity tests showed that molecular differences between isolates of the same race corresponded with differences in aggressiveness. Isolates of races 1 and 2 in cluster A (R1I and R2I isolates) and cluster C (R1-type isolates) were all highly aggressive, whereas isolates of races 1 and 2 in cluster B (R1II and R2II isolates) showed a low aggressiveness profile. The usefulness of the molecular markers described in this study has been proved in double-blind tests with Fod isolates collected in 2008. Results from this work indicate a change in the composition of the Spanish Fod population over time, and this temporal variation could be related to the continuous change in the commercial carnation cultivars used by growers. This is the first report of genetic diversity among Fod isolates in the same race.