Survival of Clavibacter michiganensis ssp. michiganensis in infected tomato stems under natural field conditions in California, Ohio and Morocco

The survival and half-life of Clavibacter michiganensis ssp. michiganensis ( C. michiganensis ), the causal agent of bacterial canker of tomato, were determined in infected plant debris under natural field conditions in California, Ohio and Morocco using a semiselective agar medium. The organism survived significantly longer in tomato stems left on the soil surface than in stems buried in the soil at all locations studied. The pathogen was recovered in high amounts from tomato stems left on the soil surface for 314 days in Ohio and California, USA, and for 194 and 132 days in Melk Zhar and Aït Melloul, Morocco, respectively; it was recovered from stems buried in the soil for up to 314 days in Ohio, up to 240 days in California, and up to 60 days in Aït Melloul and Melk Zhar. The half-life of the pathogen in stems left on the soil surface ranged from 23·2 to 24·8 days in the USA, and from 7·8 to 12·3 days in Morocco, whereas the half-life in buried stems ranged from 14·0 to 16·7 days in the USA and from 3·7 to 9·5 days in Morocco. Based on the half-life data, the predicted survival times of C. michiganensis in stems on the soil surface in Ohio, California, Melk Zhar and Aït Melloul would be up to 822, 770, 424 and 261 days, respectively, while the predicted survival times in stems buried in the soil would be 541, 497, 305 and 128 days, respectively. These results show that the survival and half-life of C. michiganensis in plant debris are relatively long and are influenced by both tissue exposure and geographic location.     

Temperature at the early stages of Clavibacter michiganensis subsp. michiganensis infection affects bacterial canker development and virulence gene expression

Clavibacter michiganensis subsp. michiganensis (Cmm), the causal agent of bacterial canker and wilt, causes severe economic losses in tomato net‐houses and greenhouses worldwide. In this study, seedlings which were transplanted and inoculated monthly over 2 years wilted and died earlier in the spring (21–24°C) and autumn (18–23°C) than in the winter (15–18°C) and summer (28–31°C): T₅₀ (the time taken for 50% of the plants to wilt or die) was 2 and 3–4 months after inoculation, respectively. A highly significant correlation was found between the average temperatures during the first month after inoculation and T₅₀; the shortest T₅₀ mortality (70 days) was observed for an average temperature of 26°C. Expression of virulence genes (pat‐1, celA, chpC and ppaA) by Cmm was higher in plants inoculated in the spring than in those inoculated in the summer. In another set of experiments, seedlings were inoculated and maintained in controlled‐environment growth chambers for 2 weeks. Subsequently, they were transplanted and maintained in commercial‐type greenhouses for 4–5 months. The temperatures prevailing in the first 48 h after inoculation were found to affect Cmm population size and virulence gene expression and to have season‐long effects on bacterial canker development.     

Clonal populations of Clavibacter michiganensis subsp. michiganensis are responsible for the outbreaks of bacterial canker in greenhouse tomatoes in Italy

Clavibacter michiganensis subsp. michiganensis (Cmm) strains, collected in greenhouses from 17 farms during tomato bacterial canker outbreaks occurring between 2005 and 2008 in Sicily, were analysed by a multiphasic approach. Population studies were conducted to investigate the possible sources of inocula. Cmm strains were characterized by PCR assays targeting virulence genes, fingerprinting techniques, metabolic profiles and virulence. These strains were comparatively analysed with Cmm strains isolated in other parts of Italy over a period of 15 years. Chromosomal genes encoding virulence determinants tomA, ppaA, chpC, and the plasmid‐encoded genes pat‐1 and celA were detected by PCR in all tested strains, except for four Sicilian Cmm strains where the pat‐1 gene was not amplified. Using BOX‐PCR, Cmm strains were differentiated into 13 haplotypes and clonal populations were identified. Cmm strains isolated from different farms in 2008 showed the same BOX‐PCR haplotype. A distinct BOX‐PCR haplotype was obtained from atypical Cmm strains lacking pat‐1 and isolated in 2006/7 from three farms. Cmm strains with two different haplotypes were detected in one farm, whereas the other farms contained strains with only a single haplotype. A new fAFLP protocol based on the amplification of ApaI/MseI fragments was developed and was able to differentiate C. michiganensis subspecies. Different populations were delineated for the multiple outbreaks occurring in Sicily, whereas similar populations were recorded in other Italian regions over a period of 12 years. The results are consistent with previous studies that demonstrate that Cmm outbreaks are associated with propagation material.     

Streptomycin resistance in Clavibacter michiganensis subsp. michiganensis strains from Chile is related to an rpsL gene mutation

Streptomycin has been used for decades in Chile to control Clavibacter michiganensis subsp. michiganensis (Cmm), the causal agent of tomato bacterial canker. The aim of this work was to evaluate streptomycin resistance and to analyse the presence of resistance-related genes in Cmm strains from Chile. A collection of 25 Cmm strains isolated from different localities in central Chile between 1996 and 2015 was analysed. Minimum inhibitory concentration (MIC) of streptomycin was determined. A search of streptomycin resistance-related genes was carried out in Cmm genomes, and the presence of these genes was studied in all Chilean strains using PCR and sequencing techniques. MIC results showed that four of 25 strains were highly sensitive to streptomycin, with MIC values <2 μg mL⁻¹. The remaining 21 strains possessed MIC of streptomycin ≥100 μg mL⁻¹. The strB gene, encoding an aminoglycoside 6-phosphotransferase that inactivates streptomycin, was detected in all Chilean strains, including sensitive and resistant strains. In the 21 resistant strains, a mutation in codon 43 of the rpsL gene was determined, conferring high streptomycin resistance. Interestingly, the four streptomycin-sensitive Cmm strains did not possess this mutation. This study proposes that the continuous use of streptomycin leads to emergence of resistant Cmm strains, challenging researchers to look for novel alternatives to control this plant pathogenic bacterium.     

Induction of the viable but nonculturable state in Clavibacter michiganensis subsp. michiganensis and in planta resuscitation of the cells on tomato seedlings

Bacterial canker of tomato is an economically important seedborne disease caused by Clavibacter michiganensis subsp. michiganensis (Cmm). Copper‐based bactericides and seed treatment with hydrochloric acid are commonly used for bacterial canker management. Recent studies have shown that some bacteria can enter a viable but nonculturable (VBNC) state, and fail to form colonies on microbiological agar media. Bacteria in the VBNC state can recover their culturability when returned to favourable conditions. This study reports the induction of the VBNC state in Cmm by CuSO₄ and low pH, and resuscitation of VBNC cells on tomato seedlings. Flow cytometry using the nucleic acid dyes SYTO 9 and propidium iodide, combined with agar plating, was used to assess VBNC cell counts. It was demonstrated that CuSO₄ and low pH induced the VBNC state in Cmm and the rate of induction increased with copper ion concentration and acidity. Pathogenicity tests showed that some of the VBNC cells induced by CuSO₄ retained their ability to colonize tomato seedlings but failed to produce typical bacterial canker symptoms by 2 months post‐inoculation. This was probably due to low levels of resuscitation of VBNC Cmm cells resulting in low levels of initial inoculum. This study has improved understanding of the VBNC state of Gram‐positive phytopathogenic bacteria. Most importantly, because copper‐based chemicals and low pH conditions are used for disease management, induction of the VBNC state and subsequent resuscitation of Cmm cells on tomato seedlings may limit pathogen detection by culture‐based assays yet present a risk for disease development in the field.     

Effect of flooding on the survival of Leptosphaeria spp. in canola stubble

This study used a versatile temperature‐control device to assess the effect of temperature (12–40°C) and duration (2–12 weeks) of flooding on the survival of Leptosphaeria spp. in canola (Brassica napus) stubble. Canola basal stems with blackleg symptoms were submerged in water in small glass jars containing 20 cm³ soil on a thermogradient plate capable of simultaneously maintaining up to 96 independent temperature regimes. Flooded stems were sampled at 2‐week intervals, surface‐sterilized, and incubated on V8‐juice agar for 10 days to recover the pathogen. Flooding for 2 weeks substantially reduced pathogen recovery relative to non‐flooded controls and the pathogen was not recovered after 6 weeks of flooding, irrespective of temperature. The pathogen was eliminated slightly more rapidly at flooding temperatures >20°C than at 12–16°C. There was no difference between Leptosphaeria maculans and L. biglobosa in their ability to survive flooding. Stem tissues degraded most rapidly during the first 2 weeks of flooding, corresponding to a quick decline in pathogen survival during the same period. These results indicate that a paddy rice crop following winter rapeseed may minimize the impact of blackleg by eradicating the inoculum of Leptosphaeria spp. in stubble.     

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.     

The significance of guttation in the secondary spread of Clavibacter michiganensis subsp. michiganensis in tomato greenhouses

Bacterial canker, caused by Clavibacter michiganensis subsp. michiganensis (Cmm), can spread in commercial tomato greenhouses causing epidemics. Results of greenhouse experiments with Cmm‐contaminated tools demonstrated disease spread for only a limited distance (<4 plants) from infected plants. However, touching symptomless infected plants bearing guttation droplets prior to touching nearby plants spread the pathogen over longer distances within rows (>22 plants). The pathogen was exuded in large numbers in the guttation fluid of infected plants; its presence in the guttation fluid was not influenced by the inoculation procedures, leaf age or the volume of the guttation droplets. Population size of Cmm and the incidence of leaflets with epiphytic bacteria were significantly higher in plants placed in a guttation‐induction chamber than in those kept in a growth chamber with high humidity, suggesting exudation through guttation contributed to the formation of epiphytic populations on leaflets. This new knowledge may provide a simple and environmentally friendly means for decreasing the spread of the disease by avoiding contact with plants during periods when they bear guttation droplets.     

Survival of Phoma koolunga,a causal agent of ascochyta blight,on field pea stubble or as pseudosclerotia in soil

Phoma koolunga is a recently recognized pathogen in the ascochyta blight complex of field pea (Pisum sativum). Unlike the other three ascochyta blight pathogens, survival of P. koolunga is poorly understood. Survival of this fungus was examined on field pea stubble and as pseudosclerotia on the surface of, and buried in, field soil. Pseudosclerotia were formed in plates containing potato dextrose agar (PDA) mixed with sand or amended with fluorocytocin. After 1 month, P. koolunga was recovered on amended PDA from 93% of stubble sections retrieved from the soil surface, 36% of buried stubble sections and 100% of pseudosclerotia buried in field soil, pasteurized or not. The frequency of recovery of P. koolunga decreased over time and the fungus was not recovered from stubble on the soil surface at 15 months, nor was it recovered from stubble buried in soil at 11 months or later, or from pseudosclerotia buried for 18 months. In a pot bioassay, most ascochyta blight lesions developed on plants inoculated with stubble retrieved from the soil surface after 1 month. Infectivity of the inoculum decreased over time. Disease on plants inoculated with stubble that had been buried or left on the soil surface for up to 6 and 5 months, respectively, and pseudosclerotia retrieved at 14 months and later from field soil did not differ from the non‐inoculated control. These results suggest that field pea stubble may play a role in survival of P. koolunga, especially if it remains on the soil surface. In addition, pseudosclerotia were able to persist in soil and infect field pea plants into the next season.     

Clavibacter michiganensis subsp. sepedonicus

Specific scope

This standard describes a national regulatory control system for Clavibacter michiganensis subsp. sepedonicus that provides guidance on surveillance for the pathogen and its containment and eradication if found.

Specific approval and amendment

First approved in 2003–09. Revision approved in 2011–09.     

Virulence and pCM1 plasmid carriage are related to BOX-PCR fingerprint type in strains of Clavibacter michiganensis subsp. michiganensis that cause bacterial wilt and canker of tomato in Argentina

Clavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial wilt and canker in tomato, producing important economic losses worldwide. Its virulence has been related to several putative virulence factors present on a chromosomal pathogenicity island and on plasmids pCM1 and pCM2, in strain NCPPB382. We genotypically characterized a collection of Cmm isolates from the main greenhouse tomato-producing areas of Argentina by BOX-PCR fingerprinting and screened for the presence of genes and plasmids involved in pathogenicity by PCR. In addition, we evaluated in vitro cellulolytic activity and virulence in planta of selected strains. BOX-PCR fingerprinting clustered strains into four groups. Group II was dominant and included the most virulent strains, while Group III was the smallest and had the least virulent strains. All local strains exhibited similar cellulolytic activity. Most of the examined strains carry two plasmids of similar size to those of NCPPB382, although there were strains with one or three plasmids. By PCR amplification of repA, pCM1 was detected only in strains belonging to Group III, which includes local strains closely related to reference strain NCPPB382. All analysed pathogenicity genes were widespread among strains, and so in strains belonging to Groups I and II, celA found on pCM1 in NCPPB382 could be found in the chromosome or in plasmids other than pCM1. This study contributes to a better understanding of the diversity of Cmm genetic profiles and virulence of strains present in Argentina. Such information could be useful for the selection of strains for screening of host resistance and development of resistant tomato varieties.     

Development of an in vitro protocol to screen Clavibacter michiganensis subsp. michiganensis pathogenicity in different Solanum species

Clavibacter michiganensis subsp. michiganensis (Cmm) is a quarantine organism in Europe and in many other countries. It is one of the most severe bacterial pathogens affecting tomato. Screening tomato plants for their resistance level to Cmm requires a large amount of space under quarantine conditions and is therefore costly. This project developed a new inoculation protocol on in vitro tomato plants to facilitate a more economic and higher throughput disease screening. A new method using the PathoScreen system was tested to localize green fluorescent protein-tagged Cmm in planta and to quantify the pathogen based on the percentage of corrected GFP (cGFP%). The system was sensitive in detecting the GFP-tagged Cmm in the shoots, but in the roots a high autofluorescence masked detection and thus sensitivity of the assay. The in vitro protocol was tested on several wild relatives of tomato, which were previously screened in a greenhouse assay. The correlation between wilt symptoms in vitro and wilt symptoms in the greenhouse was overall moderate (r = 0.6462). The protocol worked well in differentiating the two parents that were used in the mapping studies. This study shows that the in vitro protocol can be efficiently used for resistance breeding in many tomato genotypes.     

Winter annual grassy weeds increase over‐winter mortality in autumn‐sown wheat

Over‐winter mortality, that is, winterkill, reduces cereal crop competitive ability and yield. While management and environmental variables are known to affect winterkill, the extent to which weeds contribute to increased winterkill is largely unknown. Winter annual weeds may increase winterkill through resource competition and by increasing incidence of and damage from plant pathogens that cause winterkill. We evaluated the impact of summer annual (Avena fatua) and winter annual (Bromus tectorum) weeds on the over‐winter survival rate of winter wheat over three winters, during which plots were covered with snow. Pink snow mould (Microdochium nivale), a winterkill pathogen known to infect B. tectorum and winter wheat, was common in wheat stands. In weed‐free treatments, mortality rates were initially near zero, but increased by nearly 45% in each subsequent winter, presumably due to an increase in snow mould disease in continuously cropped winter wheat. Whereas A. fatua infestation had no impact on crop survival rates, winter wheat survival in B. tectorum‐infested plots was 50% less than the weed‐free control in the second and third years of this study. Among B. tectorum‐infested plots, winter wheat over‐winter survival declined with increasing weed seed produced in the previous summer. Overall, this study demonstrated that winter annual weed infestations can reduce crop stand densities below replanting thresholds by reducing fall‐sown cereal winter survival. The effects of winter annual weeds on winter wheat may be meditated by increased proliferation of snow mould disease.     

Inoculum potential of Sclerotinia sclerotiorum sclerotia depends on isolate and host plant

The soilborne fungus Sclerotinia sclerotiorum infects many important crop plants. Central to the success of this pathogen is the production of sclerotia, which enables survival in soil and constitutes the primary inoculum. This study aimed to determine how crop plant type and S. sclerotiorum isolate impact sclerotial production and germination and hence inoculum potential. Three S. sclerotiorum isolates (L6, L17, L44) were used to inoculate plants of bean, carrot, lettuce, oilseed rape (OSR) and potato, and the number and weight of sclerotia per plant quantified. Carpogenic germination of sclerotia collected from different hosts was also assessed for L6. Production of sclerotia was dependent on both crop plant type and S. sclerotiorum isolate, with OSR and lettuce supporting the greatest number (42–122) and weight (1.6–3.0 g) of sclerotia per plant. The largest sclerotia were produced on OSR (33–66 mg). The three S. sclerotiorum isolates exhibited a consistent pattern of sclerotial production irrespective of crop type; L6 produced large numbers of small sclerotia while L44 produced smaller numbers of large sclerotia, with L17 intermediate between the two. Germination rate and percentage was greatest for larger sclerotia (4.0–6.7 mm) and also varied between host plants. Combining sclerotial production data and typical field crop densities suggested that infected carrot and OSR could produce the greatest number (3944 m^?2) and weight (73 g m^?2) of S. sclerotiorum sclerotia, respectively, suggesting these crops potentially contribute a greater increase in inoculum. This information, once further validated in field trials, could be used to inform future crop rotation decisions.     

Genetic groups of Clavibacter michiganensis subsp. michiganensis identified by DNA fingerprinting and the effects of inoculation methods on disease development

Clavibacter michiganensis subsp. michiganensis (CMM) strains collected in nine different prefectures in Japan were fingerprinted by inter-simple sequence repeats–polymerase chain reaction (PCR) and repetitive sequence–based PCR. Forty-three CMM strains corresponded to 22 DNA fingerprinting patterns, indicating that the CMM population in Japan has high genetic diversity. Tomato plants were inoculated with CMM by two different methods: defoliation using infected scissors and planting in soil containing infected plant debris. Defoliation resulted in a larger number of diseased plants, disease developed earlier, and plants survived for significantly fewer days. Upon planting in the infected soil, 33.3 % of plants were infected and 6.7 % were diseased, i.e., 20.1 % of infected plants developed disease symptoms. These results indicate that some plants are infected with CMM through plant debris in the soil as the primary inoculum and most infected plants maintain latent infection. Thus, growers may unwittingly carry out disbudding and defoliation of latently infected plants, resulting in secondary inoculum.     

Survival of dicarboximide-resistant strains ofBotrytis cinerea in plant debris during summer in Israel

Survival- ofBotrytis cinerea was monitored during two summer seasons. Mycelium and conidia were found dead on the surface of plant debris within 2 months of incubation, whereas a high level of viability was detected in thallus of the pathogen which was 1–2 mm inside the dry host tissue. Of the 148 samples of infected senescing cucumber female fruits, 8% survived seven warm months; half of these isolates ofB. cinerea were resistant to dicarboximides (5 (μ/ml iprodione). Of the stems of cucumber infected withB. cinerea in winter, 18% yielded the pathogen at the beginning of the following winter; 15% of the surviving isolates were resistant to dicarboximides. Cucumber seedlings artificially infected byB. cinerea did not yield the pathogen longer than 9 weeks after establishment of infection, even when incubated in the shade. Plant debris with symptoms of gray mold were kept in the shade during the summer; at the beginning of winter it was possible to establish infection ofB. cinerea from the dry debris.     

Detection of Clavibacter michiganensis ssp. michiganensis in tomato seeds by immunofluorescence microscopy and dilution plating

A method for detectingClavibacter michiganensis ssp.michiganensis in tomato seeds was evaluated. The method is based on rapid screening of tomato seed lots using indirect immunofluorescence staining (IF), followed by dilution plating of IF positive seed lots. Different polyclonal antisera, prepared againstC. michiganensis ssp.michiganensis were tested for their specificity using IF. All strains ofC. michiganensis ssp.michiganensis tested reacted with the polyclonal antisera. Two of nine saprophytic isolates from tomato seeds were positive with the antisera as well as with the control normal serum, but cells of these isolates were distinct in shape from cells ofC. michiganensis ssp.michiganensis.For extraction of the pathogen from the seed, seeds were either blended with a stomacher or soaked at 4–6 °C. The stomacher method yielded more fluorescent cells in IF than 24 h soaking of seed samples. However, soaking of seeds for 48 h generally yielded less saprophytes and overall higher numbers ofC. michiganensis ssp.michiganensis colonies in dilution plating when compared to blending by a stomacher. SCM medium was generally more selective than KBT and modified CNS medium. However, the efficacy of the medium was dependent on the seed lot and/or extraction method used. Confirmation of suspected colonies with YDC (yeast-dextrose-carbonate medium), IF and a pathogenicity test on tomato seedlings proved to be highly reliable (P>0.95). For routine testing of seed lots it is recommended to screen tomato seed lost after soaking seeds for 24 h at 4–6 °C with IF, followed by plating of IF-positive seed lots on modified CNS and SCM after soaking seeds for an additional 24 h.     

Development of a tomato plant resistant to Clavibacter michiganensis using the endolysin gene of bacteriophage CMP1 as a transgene

The bacteriophage CMP1 endolysin gene (lys), encoding, a peptidase that was shown to effectively reduce Clavibacter michiganensis by specifically hydrolysing its murein, was transferred into tomato plants by Agrobacterium‐mediated transformation. The presence of the gene was verified by PCR and the gene product was confirmed in immunoblots and stably expressed over three generations. Transgenic tomato plants did not show disease symptoms after infection with C. michiganensis subsp. michiganensis, despite the fact that small amounts of bacteria could still be identified in xylem sap and leaf extracts, although in significantly reduced amounts.     

Biology and control of cephalosporium stripe of wheat

Cephalosporium stripe, caused by the fungus Cephalosporium gramineum, is the only known vascular wilt disease of small grain cereals. The pathogen causes characteristic striping of leaf blades and sheaths, but can also result in seedling death, stunting, and sterile seed heads (white heads). Cephalosporium stripe is a disease of autumn (fall)‐sown wheat, especially in cool and wet production regions. The disease is further favoured by early sowing, reduced tillage practices, low pH soils, and by frost heaving that damages roots. Infections occur almost entirely from spores produced on surface crop debris that are washed into the soil, although a low level of seed transmission can also occur. The pathogen colonizes root epidermis and cortical cells, subsequently moves into the vascular tissue, and eventually spreads throughout the entire plant. Production of fungal toxin(s) and extracellular polysaccharides probably play an important role in pathogenesis. Cultural practices such as delayed sowing, crop rotation, destruction of crop debris, liming of soil and fertilizer management all have potential to reduce the incidence of cephalosporium stripe. All of these cultural practices have negative economic impacts and/or increase soil erosion, and thus there is much interest in the development of resistant cultivars. There is potential for introgression of highly effective resistance from wild species into cultivated wheat. Genes for quantitatively inherited resistance can also be accumulated within cultivated wheat to attain moderate resistance. The continued use of cultivars with moderate resistance will probably be sufficient for long‐term control of the disease.     

Application of Trichoderma harzianum Strain KABOFT4 for Management of Tomato Bacterial Wilt Under Greenhouse Conditions

Clavibacter michiganensis subsp. michiganensis is a very important pathogen that causes bacterial wilt of tomato (BWT). Biological control of plant diseases is a critical tool for protecting the environment from chemical pollution. Twenty-five isolates of the genus Trichoderma were obtained from a healthy tomato root. Of the 25 isolates, KABOFT4 showed highly antagonistic activity that controlled the growth of C. michiganensis subsp. michiganensis (Cmm7) under in vitro conditions. The 5.8S ribosomal RNA gene and internal transcribed spacer identified the isolate as Trichoderma harzianum KABOFT4. The effect of this isolate as a soil drench and/or foliar application on bacterial wilt under greenhouse conditions was studied. The germination percentage of tomato seed treated with KABOFT4 increased by 36.7% compared to infected seed treated with only the pathogen Cmm7. Under greenhouse conditions, tomato seedlings treated with KABOFT4 as a soil drench, foliar and soil treatment, and foliar treatment had a 61.3, 26.7, and 40% reduced disease severity relative to the infected control, respectively. All treatments had a positive effect on tomato plants that presented as greater vegetative growth and accumulation of dry matter. The best fresh and dry weight was recorded when plants were treated with KABOFT4 as a soil and foliar application. Tomato plants treated with KABOFT4 also had increased total phenol and flavonoid contents in inoculated and non-inoculated plants compared to untreated plants. Under greenhouse conditions, T. harzianum strains can be used as an environmentally friendly way to manage the most economically important tomato disease. The results showed that a native endophytic strain of T. harzianum was a potent biocontrol agent against C. michiganensis subsp. michiganensis. Application of this strain to tomatoes in the greenhouse resulted in a decrease in disease severity and an increase in crop biomass.