Involvement of resistance induction by Penicillum oxalicum in the biocontrol of tomato wilt

Penicillium oxalicum , a biocontrol agent for Fusarium oxysporum f.sp. lycopersici , was tested for its ability to induce resistance against tomato wilt. P. oxalicum and F. oxysporum f.sp. lycopersici were placed at separate sites on tomato plants or in soil, avoiding a direct interaction between the fungi. P. oxalicum induced resistance as expressed by a reduction in disease severity, area under disease progress curve and stunting induced by the pathogen. P. oxalicum colonized the tomato rhizosphere during the experiments but it was not detected inside stems, demonstrating that P. oxalicum and Fusarium oxysporum f.sp. lycopersici remained spatially separated. Biological control was observed both in sensitive and 'resistant' cultivars, indicating the role of a general resistance mechanism. In both cultivars P. oxalicum treatment alone did not produce disease symptoms. Therefore P. oxalicum could be a suitable biocontrol agent in cases of cultivar resistance failure. These results suggest that P. oxalicum can trigger defence mechanisms in the plant.     

Induced Resistance by Penicillium oxalicum Against Fusarium oxysporum f. sp. lycopersici: Histological Studies of Infected and Induced Tomato Stems

ABSTRACT Tomato (Lycopersicon esculentum) plants of 'Lorena' were induced with a conidial suspension (10(7) conidia per ml) of Penicillium oxalicum before inoculation with Fusarium oxysporum f. sp. lycopersici, the wilt pathogen. Histological changes occurred in plants under both growth chamber and glasshouse culture conditions and there was a reduction of disease severity. In noninduced plants, the pathogen produced almost a complete loss of cambium (75 to 100% reduction), an increase in the number of bundles, and a decrease in the number of xylem vessels (20% reduction), in which the diameter also was reduced by 20 to 30% in hypocotyls and epicotyls. The percentage of vessels colonized by F. oxysporum f. sp. lycopersici was positively correlated to the area under the disease progress curve (AUDPC). However, plants induced with P. oxalicum showed less disease, did not lose the cambium, had a lower number of bundles, and had less vascular colonization by F. oxysporum f. sp. lycopersici (35 to 99%). These effects also were observed in 'Precodor', which is susceptible to races 1 and 2 of F. oxysporum f. sp. lycopersici, and partially in 'Ramón', which is resistant to both races. Renewed or prolonged cambial activity that led to the formation of additional secondary xylem could be one of the reasons for disease reduction in P. oxalicum-induced tomato plants.     

A rapid laboratory method for assessing the biological control potential of Penicillium oxalicum against Fusarium wilt of tomato

A rapid, simple and reliable procedure was developed to evaluate biological control of Fusarium wilt of tomato by Penicillium oxalicum . The method consists in growing tomato plants in flasks with nutrient solution in a growth chamber. Plants were previously treated in the seedbed with a conidial suspension (10⁷ conidia mL⁻¹) of P. oxalicum 7 days before transplanting. Fusarium oxysporum f.sp. lycopersici (race 2) was added to the Hoagland solution just before transplanting. Different concentrations and several isolates of F. oxysporum f.sp. lycopersici were tested. Using this method, plants showed typical symptoms of the disease and the effect of the biocontrol agent was clear. Consumption of nutrient solution was reduced in diseased plants, and this reduction was diminished by treatment with P.oxalicum . Consumption of nutrient solution was correlated with other disease-related parameters (AUDPC, weight of aerial parts, stunting) and was an easy and objective parameter to measure.     

Biological control of Fusarium oxysporum f. sp. lycopersici

Fungi known to produce lytic enzymes were used in an attempt to control wilt of tomato caused by Fusarium oxysporum f. sp. lycopersici (FOL). Some of the fungal species (Penicillium oxalicum, Penicillium purpurogenum and Aspergillus nidulans) damaged hyphae of FOL in vitro and reduced the numbers of microconidia in the soil. Treatments with fungi did not result in a reduction in either chlamydospores of FOL in soil or populations of FOL in the rhizosphere of tomato. P. oxalicum was the most effective agent of biocontrol, and it reduced disease severity in both non-autoclaved (20% decrease) and sterile soil. In sterile soil, P. oxalicum reduced disease with different levels of severity (27% decrease at high levels and 50% decrease at low levels). Disease control by A. nidulans and P purpurogenum was only achieved when disease severity was low in sterile soil (55% and 45%, respectively).     

Sporulation of Fusarium oxysporum f. sp. lycopersici on Stem Surfaces of Tomato Plants and Aerial Dissemination of Inoculum

ABSTRACT Plants exhibiting symptoms of wilt and xylem discoloration typical of Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici were observed in greenhouses of cherry tomatoes at various sites in Israel. However, the lower stems of some of these plants were covered with a pink layer of macroconidia of F. oxysporum. This sign resembles the sporulating layer on stems of tomato plants infected with F. oxysporum f. sp. radicis-lycopersici, which causes the crown and root rot disease. Monoconidial isolates of F. oxysporum from diseased plants were assigned to vegetative compatibility group 0030 of F. oxysporum f. sp. lycopersici and identified as belonging to race 1 of F. oxysporum f. sp. lycopersici. The possibility of coinfection with F. oxysporum f. sp. lycopersici and F. oxysporum f. sp. radicis-lycopersici was excluded by testing several macroconidia from each plant. Airborne propagules of F. oxysporum f. sp. lycopersici were trapped on selective medium in greenhouses in which plants with a sporulating layer had been growing. Sporulation on stems was reproduced by inoculating tomato plants with races 1 and 2 of F. oxysporum f. sp. lycopersici. This phenomenon has not been reported previously with F. oxysporum f. sp. lycopersici and might be connected to specific environmental conditions, e.g., high humidity. The sporulation of F. oxysporum f. sp. lycopersici on plant stems and the resultant aerial dissemination of macroconidia may have serious epidemiological consequences. Sanitation of the greenhouse structure, as part of a holistic disease management approach, is necessary to ensure effective disease control.     

Differential Colonization of Tomato Roots by Nonpathogenic and Pathogenic Fusarium oxysporum Strains May Influence Fusarium Wilt Control

ABSTRACT Histochemical staining, beta-glucuronidase (GUS) activity, or placing roots on agar were methods used to characterize interactions between the pathogenic fungus, Fusarium oxysporum f. sp. lycopersici, and the nonpathogenic biocontrol F. oxysporum strain 70T01 with respect to colonization behaviors, interaction sites, and population densities on tomato roots. Mycelia of strain 70T01, a genetic transformant expressing stable GUS activity, hygromycin B resistance, and effective disease control, were localized in epidermal and cortex cell layers of tomato roots in a discontinuous and uneven pattern. In contrast, mycelia of F. oxysporum f. sp. lycopersici were found in the vascular bundles. Thus, direct interactions between the two fungi likely happen in the root surface cell layers. Colonization density of strain 70T01 was related to the inoculation density but decreased with distance from the inoculation site. Host defense reactions, including increased cell wall thickness or papilla deposits, were adjacent to 70T01 hyphae. Experiments done in soil showed that strain 70T01 densities in roots were highest at inoculation zones and barely detectable for root segments more than 2 cm away from the inoculation sites. F. oxysporum f. sp. lycopersici densities were lowest at 70T01 inoculation zones and highest (>10 times) where strain 70T01 was not directly applied. Newly elongating roots where strain 70T01 did not reach were available for infection by the pathogen. The higher strain 70T01 density was always found when the plants were simultaneously infected by F. oxysporum f. sp. lycopersici, suggesting that F. oxysporum f. sp. lycopersici has as much influence in predisposing the plant to colonization by strain 70T01 as strain 70T01 has on providing disease protection against the pathogen.     

Suppressive effect of secondary metabolites from Streptomyces plumbeus isolate F31D against Fusarium oxysporum f. sp. lycopersici,the causal agent of Fusarium wilt of tomato

Journal of General Plant Pathology - Fusarium wilt of tomato, a disease caused by the soilborne fungus Fusarium oxysporum f. sp. lycopersici, causes major losses to tomato production. Chemical...     

An ultrastructural study of tomato roots inoculated with pathogenic and non-pathogenic necrotrophic fungi and a saprophytic fungus

Tomato cultivar Moneymaker was independently inoculated with Alternaria alternata, Cunninghamella elegans, Fusarium culmorum, F. oxysporum f.sp. lycopersici, F. oxysporum f.sp. pisi and Stromatinia gladioli and analysed ultrastructurally. The extent and amount of superficial fungal growth on tomato roots was similar but C. elegans , a saprophyte, was exceptional in that hyphae were not closely appressed to plant surfaces and did not adhere to plant cell walls.
In general, the type of plant responses to fungal colonization and infection were similar in all of the interactions studied, with the exception of C. elegans which did not infect tomato root tissue. The failure to penetrate tomato roots by C. elegans may have been associated with the lack of hyphal adhesion to plant cell walls. Migration of cytoplasm and wall apposition/penetration papilla formation were regularly observed in tomato root tissue beneath appressed hyphae and at sites of fungal infection. Specific cellular reactions in the exodermis, namely the formation of wall 'inclusions' and appearance of 'sensitive' cells, indicated that exodermal cells were particularly responsive to fungal challenge.
Fusarium oxysporum f.sp. lycopersici , a pathogen of tomato, invaded tomato root tissue more extensively than the other fungi inoculated onto tomato roots. Infection of tomato by the other fungi studied was variable, and the extent and success of fungal invasion was tentatively associated with their necrotrophic capability and typical host range.     

Fusarium crown and root rot of tomatoes in the UK

Fusarium crown and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici was found in the UK in 1988 and 1989 mainly in rockwool-grown tomato crops. Up to 14% of plants were affected in individual crops. In experiments, leaf and stem symptoms did not appear until the time of first fruit harvest even when the plants were inoculated at planting, first flowers or fruit set. Conidial inoculum at 10⁶ spores/plant applied at seed sowing killed 70–83% of tomato seedlings, whereas similar levels of inoculum applied to young plants caused root and basal stem decay, and eventually death but only after fruit harvest began. Disease incidence and symptom severity increased with inoculum concentration. Experimentally, the disease was more severe in peat- or compost-grown plants than in rockwool. Disease spread was only a few centimetres in 50 days in experimental rockwool-grown plants. All tomato cultivars tested were highly susceptible. Prochloraz-Mn was highly effective against the pathogen in vitro and controlled the disease in the glasshouse, but only when applied preventively. Non-pathogenic Fusarium oxysporum isolates and Trichoderma harzianum also reduced FCRR disease levels.     

Effects of soil application of fly ash on the fusarial wilt on tomato cultivars

A study was carried out in microplots to evaluate the effect of fly ash on the plant growth and yield of tomato cultivars, Pusa Ruby, Pusa Early Dwarf and New Uday, and on wilt disease caused by Fusarium oxysporum f.sp. lycopersici . Fly ash was applied to soil by broadcast or in rows at the rate of 1, 2, 3 and 4 kg ash m-2 in place of inorganic fertilizers. In control plots, NPK (about 40 : 20 : 20 kg acre-1) and compost were added in place of fly ash. Ash application greatly increased the soil contents of P, K, B, Ca, Mg, Mn, Zn, carbonates, bicarbonates and sulphates. Plants grown in the ash-treated plots, especially at 3 or 4 kg dose, showed luxuriant growth and greener foliage, and plant growth and yield of the three cultivars were significantly increased in comparison with the plants grown in plots without fly ash. The wilt fungus, F. oxysporum f.sp. lycopersici at the inoculum level of 2 g plant-1 caused significant suppression of growth and yield in all three cultivars. Application of fly ash, however, checked the suppressive effect of the fungus, leading to a significant increase in the considered variables compared with the inoculated control. Soil population of the fungus (colony-forming units g soil-1) gradually decreased with an increase in ash dose. Row application was found to be relatively more effective in enhancing the yield of tomato cultivars and suppressing the wilt disease. The greatest increase in the yield of fungus inoculated and uninoculated plants due to broadcast or row application at 3 or 4 kg ash m-2 was recorded in tomato cv. Pusa Ruby (39 - 61 and 9 - 24%), followed by Pusa Early Dwarf (31 - 61 and 17 - 34%) and New Uday (21 - 35 and 4 - 22%).     

Biological and Molecular Characterization of Fusarium oxysporum f. sp. lycopersici Divides Race 1 Isolates into Separate Virulence Groups

ABSTRACT A collection of race 1 and race 2 isolates of Fusarium oxysporum f. sp. lycopersici was screened for vegetative compatibility and characterized by random amplified polymorphic DNA (RAPD) analysis to establish the identity and genetic diversity of the isolates. Comparison of RAPD profiles revealed two main groups that coincide with vegetative compatibility groups (VCGs). In addition, several single-member VCGs were identified that could not be grouped in one of the two main RAPD clusters. This suggests that F. oxysporum f. sp. lycopersici is a polyphyletic taxon. To assign avirulence genotypes to race 1 isolates, they were tested for their virulence on a small set of tomato lines (Lycopersicon esculentum), including line OT364. This line was selected because it shows resistance to race 2 isolates but, unlike most other race 2-resistant lines, susceptibility to race 1 isolates. To exclude the influence of other components than those related to the race-specific resistance response, we tested the virulence of race 1 isolates on a susceptible tomato that has become race 2 resistant by introduction of an I-2 transgene. The results show that both line OT364 and the transgenic line were significantly affected by four race 1 isolates, but not by seven other race 1 isolates nor by any race 2 isolates. This allowed a subdivision of race 1 isolates based on the presence or absence of an avirulence gene corresponding to the I-2 resistance gene. The data presented here support a gene-for-gene relationship for the interaction between F. oxysporum f. sp. lycopersici and its host tomato.     

Origin of Race 3 of Fusarium oxysporum f. sp. lycopersici at a Single Site in California

ABSTRACT Thirty-nine isolates of Fusarium oxysporum were collected from tomato plants displaying wilt symptoms in a field in California 2 years after F. oxysporum f. sp. lycopersici race 3 was first observed at that location. These and other isolates of F. oxysporum f. sp. lycopersici were characterized by pathogenicity, race, and vegetative compatibility group (VCG). Of the 39 California isolates, 22 were in VCG 0030, 11 in VCG 0031, and six in the newly described VCG 0035. Among the isolates in VCG 0030, 13 were race 3, and nine were race 2. Of the isolates in VCG 0031, seven were race 2, one was race 1, and three were nonpathogenic to tomato. All six isolates in VCG 0035 were race 2. Restriction fragment length polymorphisms (RFLPs) and sequencing of the intergenic spacer (IGS) region of rDNA identified five IGS RFLP haplotypes, which coincided with VCGs, among 60 isolates of F. oxysporum from tomato. Five race 3 isolates from California were of the same genomic DNA RFLP haplotype as a race 2 isolate from the same location, and all 13 race 3 isolates clustered together into a subgroup in the neighbor joining tree. Collective evidence suggests that race 3 in California originated from the local race 2 population.     

Race identification of Fusarium oxysporum f. sp. lycopersici isolates obtained from tomato plants in Nova Friburgo,Brazil

European Journal of Plant Pathology - Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL) is one of the main diseases affecting tomato plants. Three races (races 1, 2 and 3) of the...     

Production, Survival, and Evaluation of Solid-Substrate Inocula of Penicillium oxalicum, a Biocontrol Agent Against Fusarium Wilt of Tomato

ABSTRACT Production of conidia of Penicillium oxalicum (ATCC number pending), a biocontrol agent of Fusarium oxysporum f. sp. lycopersici, was tested in liquid and solid fermentation. P. oxalicum produced 250-fold more conidia in solid than in liquid fermentation at 30 days after inoculation of substrate. Solid fermentation was carried out in plastic bags (600 cm(3)) especially designed for solid fermentation (VALMIC) containing 50 g of peat/vermiculite (PV) (1:1, wt/wt) with 40% moisture, sealed, sterilized, and then inoculated with 1 ml of a conidial suspension of P. oxalicum (10(5) conidia g(-1) dry substrate), sealed again, and incubated in darkness at 20 to 25 degrees C for 30 days. Addition of amendments to PV in a proportion of 0.5 (wt/wt) significantly increased conidial production of P. oxalicum. The best production was obtained on PV plus meal of cereal grains (barley) or leguminous seeds (lentil) (100-fold higher). Conidial production obtained after 5 days of inoculation was similar to that obtained at 30 days. However, viability of conidia produced in PV plus lentil meal was 35% higher than that of conidia produced in PV plus barley meal. Changes in proportions (1:1:0.5, wt/wt/wt; 1:1:1, wt/wt/wt; 1:0.5:0.5, wt/wt/wt; 1:1:0.5, vol/vol/vol) of components of the substrate (peat/vermiculite/lentil meal) did not enhance production or viability of conidia. Optimal initial moisture in the substrate was 30 to 40%. At lower moistures, significant reductions of production of conidia were observed, particularly at 10%. There was a general decline in the number of conidia in bags with time of storage at -80, -20, 4, and 25 degrees C, or at room temperature (range from 30 to 15 degrees C), with the highest decline occurring from 60 to 180 days. Conidial viability also was reduced with time, except for conidia stored at -20 degrees C. Fresh conidia produced in solid fermentation system or those conidia stored at -20 degrees C for 180 days reduced Fusarium wilt of tomato by 49 and 61%, respectively.     

协同增效型拮抗细菌组合CL-7和CL-8的稳定性及其对加工番茄的促生防病效果

将加工番茄根际土壤中筛选到的单一拮抗细菌进行复配，筛选获得两个具有促生拈抗作用的细菌组合CL-7和CL-8。经过20代转管连续培养检测细菌组合的稳定性，结果表明，这两个拮抗细菌组合从13代之后，其pH值基本稳定，组合内菌落种类及数目也基本稳定；且12代后抑菌活性基本保持不变。和单一拮抗菌株相比，CL-7和CL-8均能显著增加对番茄立枯病菌、辣椒疫霉病菌和枯萎病菌的抑菌效果。发芽和苗期灌根试验结果表明，拮抗细菌组合CL-7和CL-8可显著促进加工番茄幼芽及苗期的生长，苗期鲜重分别增加了147．620k和176．190k，干重分别增加了143．75％和143．75％。在温室和田间条件下，细菌组合CL-7和CL-8对加工番茄立枯病的防治效果都很显著，分别可达25．1％、40．1％和52．2％、55．5％。     

Loss of Avirulence and Reduced Pathogenicity of a Gamma-Irradiated Mutant of Fusarium oxysporum f. sp. lycopersici

ABSTRACT The tomato Fusarium resistance gene I-2 confers resistance to F. oxy-sporum f. sp. lycopersici race 2, which expresses the corresponding aviru-lence gene avrI-2. To elucidate the molecular basis of this gene-for-gene interaction, we initiated a search for the avrI-2 gene. Gamma irradiation mutagenesis, using (137)Cs, was performed to generate an avrI-2 mutant of F. oxysporum f. sp. lycopersici. To this end, a race 2 isolate was first transformed with a phleomycine resistance gene and a GUS marker gene in order to distinguish mutants from contaminating isolates. A total of 21,712 mutagenized colonies was tested for loss of avirulence on I-2-containing tomato seedlings. One mutant was selected that showed the expected loss of avirulence but, surprisingly, also showed reduced pathogenicity toward susceptible tomato plants. DNA analysis was subsequently used to visualize genomic changes in the mutant. Southern analysis on contour-clamped homogeneous electrophoretic field blots demonstrated a translocation of a 3.75-Mb chromosome in the mutant. Random amplified polymorphic DNA and amplified fragment length polymorphism analysis identified at least nine polymorphisms between the wild-type and mutant isolates. Most of these polymorphisms appeared as extra fragments in the mutant and contained repetitive DNA sequences.     

Biological Control Efficiency of Fusarium Wilt of Tomato by Nonpathogenic Fusarium oxysporum Fo-B2 in Different Environments

ABSTRACT Efficiency of nonpathogenic Fusarium oxysporum Fo-B2 for the biological control of Fusarium wilt of tomato, caused by F. oxysporum f. sp. lycopersici CU1, was examined in different environments: a growth chamber with sterile soil-less medium, a greenhouse with fumigated or nonfumigated soil, and nonfumigated field plots. Inoculation of Fo-B2 onto tomato roots significantly reduced the severity of disease, but the efficiency of disease suppression decreased as the experimental environment became less controlled. Relationships between the recovery of Fo-B2 from hypocotyls and the disease severity indicated that the biocontrol agent was most effective when it colonized vascular tissues intensively. Moreover, the degree of Fo-B2 colonization was greatly reduced when the seedlings were grown in nonfumigated soil. Dose-response models (negative exponential, hyperbolic saturation, and logistic) were fit to observed data collected over a range of inoculum densities of the pathogen and the antagonist; the logistic model provided the best fit in all environments. The ratios of an 50% effective dose parameter for Fo-B2 to that of CU1 increased as the environment became less controlled, suggesting that environmentally related efficiency reduction impacted the antagonist more than the pathogen. The results suggest that indigenous soil microbes were a primary factor negatively influencing the efficiency of Fo-B2. Therefore, early establishment of the antagonist in a noncompetitive environment prior to outplanting could improve the efficacy of biological control.     

The relationship between brown and corky root rot caused by Pyrenochaeta lycopersici and yield loss in tomatoes established by the use of artificially infested soils

In an experiment conducted in a commercial glasshouse, tomato plants were grown in pots containing soils infested with a range of levels of Pyrenochaeta lycopersici in a randomized block design. There was found to be a negative correlation ( r = -0.93) between total yield in kg per plant (y) and % disease severity ( x ) recorded 8 weeks after planting when the disease data were exponentiated; the best fit was obtained by the equation v = 45.7–42.3 x ^0.006 Fruit number was reduced by 0.2% for each 1% increase in the disease, but a stronger correlation was obtained by the equation y (fruit number) = 417–368 x 0.006
In a separate trial, the relationship between disease severity 8 weeks after planting and yield loss was shown to be affected by soil type. Early-season loss of potential yield was found to be lower in sandy soil than in peat and clay. By the end of the season, loss in total potential yield was greater in peat soil than in clay. The effect that soil conditions can have in modifying the relationship between disease severity and yield loss is discussed.     

短短芽孢杆菌JK-2菌株对番茄枯萎病的抑菌作用及其小区防效

对分离自土壤的短短芽孢杆菌JK-2菌株对番茄枯萎病菌的防治效果和抑制作用进行了研究。结果表明：JK-2对番茄枯萎病菌的盆栽防效和田间防效分别为83．82％、74．70％。该菌株能抑制枯萎病菌菌丝的生长，当其无菌滤液终浓度为15％时，对菌丝生长的抑制率达到81．69％。JK-2菌株对病菌孢子萌发也有较强的抑制作用。扫描电镜观察结果：JK-2菌株可造成菌丝消解、产生泡状物、破坏生长点、引起细胞内含物外溢。     

Disease Development Following Infection of Tomato and Basil Foliage by Airborne Conidia of the Soilborne Pathogens Fusarium oxysporum f. sp. radicis-lycopersici and F. oxysporum f. sp. basilici

ABSTRACT Fusarium oxysporum f. sp. radicis-lycopersici, the causal agent of Fusarium crown and root rot of tomato, and F. oxysporum f. sp. basilici, the causal agent of Fusarium wilt in basil, are soilborne pathogens capable of producing conspicuous masses of macroconidia along the stem. The role of the airborne propagules in the epidemics of the disease in tomato plants was studied. In the field, airborne propagules of F. oxysporum f. sp. radicis-lycopersici were trapped with a selective medium and their prevalence was determined. Plants grown in both covered and uncovered pots, detached from the field soil, and exposed to natural aerial inoculum developed typical symptoms (82 to 87% diseased plants). The distribution of inoculum in the growth medium in the pots also indicated the occurrence of foliage infection. In greenhouse, foliage and root inoculations were carried out with both tomato and basil and their respective pathogens. Temperature and duration of high relative humidity affected rate of colonization of tomato, but not of basil, by the respective pathogens. Disease incidence in foliage-inoculated plants reached 75 to 100%. In these plants, downward movement of the pathogens from the foliage to the crown and roots was observed. Wounding enhanced pathogen invasion and establishment in the foliage-inoculated plants. The sporulation of the two pathogens on stems, aerial dissemination, and foliage infection raise the need for foliage protection in addition to soil disinfestation, in the framework of an integrated disease management program.