Identification and characterization of the <Emphasis Type="Italic">Enterobacter</Emphasis> complex causing mulberry (<Emphasis Type="Italic">Morus alba</Emphasis>) wilt disease in China

Mulberry wilt disease (MWD) was recently identified in Hangzhou, Zhejiang province, China. Typical symptoms of the disease are browning of vascular tissues, leaf wilt, defoliation, and tree decline. Unlike the symptoms of bacterial wilt disease caused by Ralstonia solanacearum, symptoms of MWD generally started from the bottom of the plants and moved upward. In inoculation experiments, four selected MWD strains caused mulberry shoot leaf wilt, discoloration, and defoliation. They also induced whole plant leaf wilt, defoliation and dark brown discoloration of vascular tissue. Based on Biolog metabolic profiles, fatty acid methyl ester analysis (FAME) and sequence analysis of the partial 16S rDNA and rpoB genes four MWD strains were identified as members of the genus Enterobacter. The 16S rDNA and rpoB gene sequences revealed a close relationship among two isolates, R2-2 and R6-2, and the E. asburiae type strain JCM6051. The isolates showed >98% similarity to E. asburiae JCM6051 in their rpoB gene. These results indicated that isolates R2-2 and R6-2 belonged to E. asburiae. No similarity in 16S rDNA sequences above 97% was found between either of the remaining isolates, R11-2 or R18-2, and any recognized Enterobacter species, suggesting that the two isolates may represent novel Enterobacter species. rpoB gene similarity values between the isolates and Enterobacter spp. type strains were <98%, providing further evidence that the two isolates may represent a novel species within the Enterobacter. The causal agent for MWD was previously reported to be E. cloacae, however, this study found that other Enterobacter spp. (E. asburiae and Enterobacter sp.) also cause MWD.     

Occurrence of blood disease of banana in Sumatra,Indonesia

Since 1991, the sudden death of cultivated banana plants has been widely observed in the southern region of Sumatra Island, Indonesia. Wilting from loss of petiole and midrib turgidity, yellowing, and necrosis of leaves was followed by death of the whole plant. Reddish brown bacterial ooze exuded from the cut surface of infected pseudostems and fruits. The colony appearance of the isolated bacterium was similar to that of Ralstonia solanacearum. The bacterium was pathogenic to banana plants but not to tomato. Its bacteriological properties agreed with those of blood disease bacterium (BDB) of banana described previously. The 16S rDNA sequence of strain Banana E had conserved bases characteristic of BDB. Based on these results, the causal agent was identified as BDB, which is a close relative of Ralstonia. The isolates have resistance against antibiotics, such as chloramphenicol and tetracycline.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB095535     

Host specificity and genetic diversity of race 4 strains of Ralstonia solanacearum

Ralstonia solanacearum race 4 isolates were obtained from Zingiberaceae plants in India during bacterial wilt outbreaks. Polyphasic phenotypic and genotypic analysis revealed intraracial diversity and dominance of biovar 3 over biovar 4. Biovar 3 strains were isolated from very severely wilted Zingiberaceae plants in the field and found to be present across diverse geographical, host and seasonal boundaries. It was hypothesized that these isolates belong to a single, ‘fast wilting’, lineage. Using one ‘fast wilting’ isolate in controlled inoculations, rapid wilt was observed in ginger within 5–7 days. Wilting was also observed in several other closely and distantly related hosts such as turmeric (Curcuma longa), aromatic turmeric (Curcuma aromatica), black turmeric (Curcuma caesia), sand ginger (Kaempferia galanga), white turmeric (Curcuma zeodaria), awapuhi (Zingiber zerumbet), greater galangal (Alpinia galanga), globba (Globba sp.), small cardamom (Elettaria cardamomum) and large cardamom (Ammomum subulatum) of the Zingiberaceae family, and in tomato (Solanum lycopersicum). Molecular analysis, including multiplex PCR‐based phylotyping, sequence analysis of 16S rDNA, 16–23S intergenic spacer and the recN gene, and multilocus sequence typing, revealed minimal differences between fast wilting isolates, confirming that almost all belong to the same lineage. Biovar 4 was isolated from plants showing slow wilt progression and self‐limiting wilting in restricted geographical locations instead, and was identified to be genetically distinct from the fast wilting biovar 3 isolates. To the authors' knowledge, this is the first report of host range and genetic analysis of R. solanacearum race 4 in India.     

Transmission of <Emphasis Type="Italic">Colletotrichum coccodes via</Emphasis> tomato seeds

Anthracnose of tomato caused by Colletotrichum coccodes is a devastating disease of ripe fruits. This pathogen may also infect tomato roots, stems and leaves. In the present study, C. coccodes is shown to be capable of contaminating seeds collected from artificially inoculated tomato fruits. Seedlings germinating from these infected seeds exhibited disease symptoms and therefore may transmit the pathogen to the next crop. The proportion of infected seeds ranged between 20% and 63% in all C. coccodes isolates tested and correlated with the aggressiveness of the isolates to tomato fruits. Fungicidal treatment of the collected seeds reduced, but did not eliminate, seed infection. A transgenic C. coccodes isolate expressing green fluorescent protein was used to visualize the pathogen. Mycelium was observed both on surfaces of the seed coat and within 1% of the embryos.     

Rapid detection of chitosanase activity in chitosanase gene-transformed strains of <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> by lytic infection of specific bacteriophages

 In combination with lytic infection by virulent phages, a simple method for monitoring transgenic strains of Enterobacter cloacae was developed in this study. First, 15 strains of E. cloacae were used as indicator bacteria to isolate virulent phages with different host ranges. Of the phages isolated, five isolates (EcP-22, -35, -45, -55, and -70) were used to construct a set of virulent phages corresponding to all strains of E. cloacae. Using this phage set, a rhizosphere strain (KRM-055E) of E. cloacae was effectively screened from field soil. KRM-055E was transformed with a prokaryotic chitosanase gene csnSM1 and infected with the phage EcP-03, which can lyse the strain most effectively. The lysis of KRM-055E/csn occurred 2 h after inoculation, and the chitosanase activity was simply detected by dropping the lysate onto an agar plate containing glycol chitosan. The positive signal for chitosanase activity was detected in the 2-h lysates, and the signal intensity reached a maximum in the 5-h lysate. The present assay was simple, rapid, inexpensive, easy to perform, and applicable to another strains. Received: August 2, 2002 / Accepted: October 31, 2002 Acknowledgments This work was supported in part by a grant (no. 99L01205) from the “Research for the Future” program of the Japan Society for the Promotion of Science. We are grateful to Dr. M. Sato, National Institute of Agrobiological Science, Dr. H. Okamoto, Fukui Agricultural Experiment Station, and Dr. K. Tsuda, Kyoto Prefectural Institute of Agricultural Biotechnology, for kindly providing E. cloacae strains. We thank Dr. P. Park, Kobe University, for technical support with the electron microscopic observations.     

Sequence Analysis and Detection of Ralstonia solanacearum by Multiplex PCR Amplification of 16S–23S Ribosomal Intergenic Spacer Region with Internal Positive Control

Polymerase chain reaction (PCR) methods for detection and differentiation of Ralstonia solanacearum strains were compared. The 16S–23S rRNA gene ITS sequence data revealed the two main sequence clusters (divisions I and II) of R. solanacearum and further subclusters of division II. Based on this sequence data, primers were designed which differentiated divisions I and II. Furthermore, to improve reliability of the PCR assay for routine detection of R. solanacearum in host plants, a novel multiplex PCR assay was developed in which the pathogen-specific sequences are coamplified with host plant DNA as an internal PCR control (IPC). The assay was validated during routine testing of potato samples submitted in official surveys. Of 4300 samples from 143 cultivars, 13 tested positive in both multiplex PCR and immunofluorescence (IF) assays and could be confirmed by bioassay in tomato seedlings and reisolation of the pathogen. The IPC was successfully amplified from all samples tested. A further 12 samples gave positive IF results which were not confirmed by either the multiplex PCR or tomato bioassay, indicating a greater specificity of the latter two assays.     

火龙果细菌性软腐病菌的分离与鉴定

<正>火龙果(Hylocereus undatus Britt.et Rose)属仙人掌科三角柱属植物,是集水果、花卉、蔬菜、保健为一体的热带亚热带果树,在中美洲、美国南部地区,以及以色列、越南、泰国和我国的台湾、海南、广东、福建等地均有种植。近年来,随着火龙果种植面积不断扩大,火龙果病害呈逐渐加重趋势,严重威胁火龙果的产业发展。火龙果细菌性病害主要由欧文氏菌(Erwinia sp.)、阴沟肠杆菌(En-     

Anthracnose of sweet pepper caused by Colletotrichum scovillei in Japan

Severe fruit rot of sweet pepper was found in Shimane, Hyogo, Chiba, Toyama, and Nagano prefectures, Japan from 2005 to 2011. Dark, sunken spots with concentric rings of orange conidial masses appeared on fruits. Pathogenic isolates from diseased fruits in the prefectures were identified as Colletotrichum scovillei. This species was added to the pathogens of sweet pepper anthracnose in Japan. The representative isolate was pathogenic to sweet pepper, tomato and chili pepper fruits, kidney bean pod, azuki bean, pea and strawberry leaves, but a caused no symptoms on cucumber or carrot in inoculation tests.     

Differences in virulence of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> isolates from a worldwide collection on host fruits

Phytophthora capsici causes root, crown, and fruit rot of vegetable and tropical hosts. Cucumber, zucchini, tomato, and pepper fruits were inoculated using 6-mm-diameter agar plugs of P. capsici, incubated in clear plastic boxes at room temperature (25 ± 2°C and 100% relative humidity), and virulence was estimated by measuring the lesion diameter, pathogen growth diameter, and pathogen sporulation density three (cucumber, zucchini) or four (tomato, pepper) days later. When isolates were grouped by genetic cluster, significant differences in virulence were observed on cucumber and zucchini, with isolates belonging to genetic cluster five causing larger lesions than isolates from genetic cluster six. On tomato, no significant differences were observed for isolates grouped by genetic cluster, but isolates from vegetable crops were generally more virulent than isolates from tropical hosts. Isolates from fabaceous hosts sporulated better on cucumber fruits than isolates from solanaceous hosts. Isolates from vegetable hosts sporulated better on zucchini than isolates from tropical hosts. No significant differences in lesion diameter were noted on pepper when isolates were grouped by host family of origin or genetic cluster, but differences in pathogen sporulation were apparent by host family. Our findings suggest that isolate characteristics such as host family of origin and genetic cluster membership may be used to guide initial isolate selection for cucurbit fruit resistance screening. Final isolate selection should incorporate the phenotypic and genetic diversity of P. capsici, including isolates with differing virulence to the host organ of interest.     

Meloidogyne luci,a new root‐knot nematode parasitizing potato in Portugal

In 2013, during a field survey conducted in Portugal on potato, Solanum tuberosum, an unusual esterase (EST) phenotype was detected in a root‐knot nematode (RKN) from potato roots collected in Coimbra. This Portuguese isolate was purified and maintained on tomato, S. lycopersicum, and morphological, biochemical and molecular characteristics were studied. Perineal pattern morphology was highly variable, similar to Meloidogyne ethiopica and not useful for identification. The EST phenotype, from young egg‐laying females, displayed three bands similar to the Brazilian M. luci (L3) and distinct from M. ethiopica (E3). Phylogenetic analyses of mitochondrial cytochrome oxidase subunit I and the mitochondrial DNA region between COII and 16S rRNA genes revealed that the Portuguese isolate grouped with M. luci isolates close to M. ethiopica isolates. However, considering the ITS1‐5.8S‐ITS2 region, the Portuguese isolate grouped with isolates of M. luci, M. ethiopica and M. hispanica, which limits the confidence of this region for M. luci diagnosis, and its differentiation from other species with morphological similarities. The M. luci pathogenicity to potato was also assessed in 16 commercial cultivars and compared with M. chitwoodi, considered to be a quarantine RKN species by EPPO. All potato cultivars were susceptible to both Meloidogyne species with gall indices of 5 and higher reproduction factor values ranging from 12.5 to 122.3, which suggests that M. luci may constitute a potential threat to potato production. In the present study, M. luci is reported for the first time attacking potato in Portugal.     

<Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis>: a new potential biocontrol agent of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis>, causal agent of potato brown rot

Stenotrophomonas maltophilia was isolated from the rhizosphere of eggplant in the Nile Delta of Egypt, and its antagonistic potential against Ralstonia solanacearum race 3 biovar 2, the causal agent of potato brown rot, was in vitro evaluated on KB agar medium and in vivo on potato plants. In vitro, four isolates of S. maltophilia (PD3531, PD3532, PD3533, and PD3534) appeared antagonistic. The isolate (PD3533) was screened as the most promising antagonist for the in vivo tests. In the greenhouse, the antagonist was applied directly to soil or by bacterization of potato eyepieces. Stenotrophomonas maltophilia significantly suppressed potato brown rot in Egyptian clay soil but not in Dutch clay soil. Survival of a rifampicin and chloramphenicol-resistant S. maltophilia strain PD4560 was investigated in two pairs of clay soils, conventionally and organically managed, from Egypt and the Netherlands. The survival of S. maltophilia was significantly less in Dutch than in Egyptian soils, while the converse occurred for R. solanacearum. These results are in agreement with those obtained in the in vivo biocontrol tests. In conclusion, S. maltophilia may be useful for control of brown rot in the area where it was originally isolated, the Nile Delta in Egypt.     

Alternaria Brown Spot of Minneola in Greece; Evaluation of Citrus Species Susceptibility

During the last three years, a new disease was observed in northwestern Greece on Minneola trees, hybrid of mandarin and grapefruit. On May small brown necrotic leaf spots surrounded by yellow halo areas of various sizes appeared and covered a major portion of the leaves with extension of necrosis into the veins. On young fruits small, slightly depressed black spots were the first symptoms, which later became 2–7 mm in diameter. Brown spots were observed on the leaves and fruits in several orchards in the same area, causing leaves and fruits to drop. In some orchards over 50% of the fruits were affected. From the fruit and leaf spots the typical small-spore species Alternaria alternata was isolated. Pathogenicity tests were performed by artificially inoculating fruits of Minneola, common mandarin and Clementine. The symptoms of the disease were reproduced only on fruits of Minneola hybrids by the specific strain of the fungus Alternaria alternata pv. citri. Different citrus susceptibility tests indicated that mandarins Minneola, Nova and Page were very susceptible to tested isolates while Clementine SRA and Poros Clementine were not. All lemons and lime Seedless were not susceptible. Grapefruit New Hall was not susceptible, while the Star Ruby was. Orange Lane Late, Navel Late, Oval Poros, Olinda, Navel Athos were not susceptible and only Moro showed reaction being slightly susceptible only to one isolate.     

Internal fruit rot of netted melon caused by <Emphasis Type="Italic">Pantoea ananatis</Emphasis> (=<Emphasis Type="Italic">Erwinia ananas</Emphasis>) in Japan

An internal fruit rot with a malodor was found in netted melons (Cucumis melo L.) in commercial greenhouses in Kochi Prefecture, Japan, in 1998, despite their healthy appearance and lack of water-soaking or brown spots on the surface. A yellow bacterium was consistently isolated from the affected fruits. To confirm the pathogenicity of eight representative isolates of the yellow bacterium, we stub-inoculated ovaries (immature-fruits) 5–7 days after artificial pollination, with a pin smeared with bacteria. After the melon fruits had grown for 60 more days, an internal fruit rot resembling the natural infection appeared, and the inoculated bacterium was reisolated. The melon isolates had properties identical with Pantoea ananatis, such as gram-negative staining, facultative anaerobic growth, indole production, phenylalanine deaminase absence, and acid production from melibiose, sorbitol, glycerol, and inositol. Phylogenetic analysis based on 16S rDNA sequences showed that the melon bacterium positioned closely with known P. ananatis strains. The melon bacterium had indole acetic acid (IAA) biosynthesis genes (iaaM and iaaH) and a cytokinin biosynthesis gene (etz). The bacterium could be distinguished from the other ‘Pantoea’ group strains by rep-PCR genomic fingerprinting. From these results, the causal agent of internal fruit rot was identified as a strain of P.ananatis [Serrano in (Philipp J Sci 36:271–305, 1928); Mergaert et al. in (Int J Syst Bacteriol 43:162–173, 1993)]. The nucleotide sequence data reported are available in the DDBJ database under accessions AB297969, AB373739, AB373740, AB373741, AB373742, AB373743 and AB373744.     

一株戴尔福特菌NF83-1的鉴定及评价

为研究拮抗菌株NF83-1的分类与生物学特性以及评价其生防能力,通过生理生化特征和16S rRNA基因序列分析对菌株NF83-1进行种属鉴定,以选择性培养基检测其分泌的多种胞外酶,以结晶紫染色法检测其生物膜的形成,采用室内平板对峙生长法测定其抑菌活性,室外盆栽法测定其对番茄立枯病的防效。结果表明,菌株NF83-1被鉴定为戴尔福特菌Delftia tsuruhatensis;菌株NF83-1能产生纤维素酶和嗜铁素,且能在管壁形成较厚的生物膜。菌株NF83-1对立枯丝核菌Rhizoctonia solani、甘蓝黑斑病菌Alternaria brassicae、油菜菌核病菌Sclerotinia sclerotiorum、梨胶孢炭疽病菌Colletotrichum gloeosporioides等病原真菌,梨火疫病菌Erwinia amylovora、番茄青枯病菌Ralstonia solanacearum、水稻细菌性条斑病菌Xanthomonas oryzae pv.oryzicola等革兰氏阴性病原细菌及革兰氏阳性细菌巨大芽胞杆菌Bacillus megaterium都具有明显的抑制作用;菌株NF83-1发酵液原液和100倍稀释液对番茄立枯病的防效分别为48.78%~57.69%和26.83%~36.54%。研究表明,菌株NF83-1是一株具有较好生防应用潜力的拮抗菌株。     

Phenotypic and genetic characterization of Erwinia rhapontici isolated from diseased Asian pear fruit trees

In 1995, a serious necrotic disease appeared in Asian pear trees in the orchards of Chuncheon, South Korea. Large numbers of bacterial samples were collected, and the causative microorganism was identified as a novel pathogen, Erwinia pyrifoliae. Among the samples, four bacterial isolates with atypical characteristics were further analyzed through biochemical tests and 16S rRNA gene sequence analysis. By phenotypic and genetic analysis these isolates were identified as E. rhapontici. Phylogenetic analysis using 16S rRNA sequence data revealed that E. rhapontici forms a discrete clade with high bootstrap value close to the E. amylovora group. Pathogenicity tests on leaves of tobacco plants (Nicotiana tabacum) elicited hypersensitivity responses, but artificial inoculations on immature fruits and shoots of Asian pear (Pyrus pyrifolia) did not show any necrotic symptoms. The developed primers showed no cross reactivity when tested against other phytopathogens and were able to detect E. rhapontici from mixed culture and in planta.     

菊花绿变病植原体在中国的发生及分子鉴定

对内蒙古农业大学校园内表现花器绿变症状的菊花样品进行采集和DNA提取,应用植原体16S rRNA基因和rp基因的引物进行巢式PCR扩增,从感病样品中分别扩增得到了长度均约为1.2 kb的片段。序列一致性分析表明,菊花绿变植原体16S rRNA基因与翠菊黄化植原体匈牙利风信子株系(GenBank登录号MN080271)、印度玉米株系(KY565571)、印度繁缕株系(KC623537)和印度马铃薯株系(KC312703)的核酸一致性最高,为99.9%,rp基因序列与翠菊黄化植原体立陶宛洋葱株系(GU228514)的核酸一致性最高,为99.8%。基于16S rRNA基因和rp基因构建系统进化树时发现,菊花绿变植原体均与16SrI-B亚组成员聚为一起。16S rRNA基因相似性系数分析表明,菊花绿变植原体与洋葱黄化植原体(AP006628)的相似性系数最高为1.00,洋葱黄化植原体(AP006628)在分类上属于16SrI-B亚组。因此,我们可以确定该菊花绿变植原体属于16SrI-B亚组。这是我国首次报道菊花绿变病的发生。     

Serratia marcescens associated with bell pepper (Capsicum annuum L.) soft-rot disease under greenhouse conditions

Soft-rotting bacteria affecting bell peppers crops represent an economically destructive disease of growing importance worldwide. In Venezuela since 2006, soft-rot symptoms have been occasionally observed in bell pepper fruits grown under greenhouse conditions. Affected fruits presented water-soaked lesions that progressed to complete fruit maceration. Bacteria were isolated from water-soaked lesions in order to identify the causal agent of this disease. Of 13 bacterial isolates recovered from affected fruits, only isolate AGPim1A was able to produce a hypersensitive reaction in tobacco plants and to reproduce soft-rot symptoms in bell peppers fruits. Several methods, including classical bacteriological tests and carbon utilization profiling, alongside with sequence analysis of 16S rRNA and housekeeping genes gyrB and groES-groEL, allowed identification of the soft-rotting bacterium as Serratia marcescens. To our knowledge, this is the first report showing a S. marcescens strain associated with soft-rot disease in bell pepper fruits.     

Susceptibility of different plant species and tomato cultivars to two isolates of <Emphasis Type="Italic">Pepino mosaic virus</Emphasis>

As Pepino mosaic virus has become a pathogen of major importance in worldwide tomato production, information is needed on possible differences between the sensitivity of cultivars towards infection. Furthermore, it is important what hosts other than Solanaceae may be virus reservoirs and are, therefore, threats for tomato cultivation. Two PepMV isolates (PepMV-Sav, E397, a European tomato isolate and PV-0554, a Peruvian pepino isolate) differing in their origin and virulence were used for several experiments to investigate these issues. The response to mechanical inoculation with PepMV was studied using 25 tomato cultivars, seven indicator plant species, and nine other possible horticultural host plants. Symptom development after infection with PepMV was monitored and the virus was detected by DAS-ELISA and IC-RT-PCR. Garlic and broad bean were shown to be additional hosts of PepMV depending on the virus isolate. Nicotiana benthamiana seems to be the most sensitive indicator among all tested indicator plants developing symptoms. Both PepMV isolates infected all tested tomato cultivars. Development of disease symptoms depended on the cultivar and the virus isolate but symptoms were not visible in all cases. None of the cultivars showed tolerance against the two isolates but two responded with a lower susceptibility at an absorbance level of 0.2 (healthy control 0.09). It was observed that some cultivars grown hydroponically showed also lower losses in biomass and yield. Data indicated a correlation between absorbance level in DAS-ELISA and reduction in total tomato growth.     

Phylogenetic analyses of plant-growth-promoting rhizobacteria isolated from tomato,lettuce, and Japanese pepper plants in Hyogo Prefecture,Japan

Approximately 30,000 fluorescent bacterial strains isolated from tomato, lettuce, eggplant, Chinese cabbage, and Japanese pepper plants at seven different locations in Hyogo Prefecture, were screened for plant-growth-promoting (PGP) activity to induce disease resistance against bacterial wilt in tomato. The 37 strains that had higher PGP activity were subjected to molecular phylogenetic analyses using the sequences of the 16S rRNA, gyrB and rpoD genes. Most of the strains were identified as Pseudomonas fluorescens or its close relative, P. putida, while a few strains were grouped with more distantly related bacterial species such as Enterobacter and Stenotrophomonas. The phylogenetic relationships among tomato and lettuce isolates mostly coincided with the source locality and host plants, with a few exceptions. In contrast, isolates from Japanese pepper plants did not form their own cluster but represented several different bacterial species.     

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.