• Horticultural Science & Technology
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• v.31 no.6
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• pp.740-747
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• 2013

Leaf mold symptoms were found on commercial tomato cultivars carrying the Cf-9, a resistance gene to leaf mold pathogen Fulvia fulva in 2012 at Buyeo, Chungnam in Korea. Fifteen-fungal isolates were obtained from four Cf-9 cultivars of tomato including 'Cutie', 'otaerangdia', 'Unicorn' and 'Rapito'. Due to their same morphological appearances and colony color, nine isolates were selected and identified as F. fulva based on molecular analysis of the internal transcribed spacer rDNA sequence. Pathogenicity of the 15 isolates on five commercial cultivars carrying Cf-4, Cf-5, and Cf-9 were tested. All the isolates showed strong pathogenicity on Cf-9 cultivars, 'Cutie' and 'Dotaerangdia', and Cf-5 cultivar, 'Yoyocaptain'. In contrast, on Cf-4 cultivar, 'Superdotaerang', five isolates were virulent and the other isolates were not. In addition, two fungal isolates, infecting Cf-9 cultivar and non-infecting Cf-4 cultivar, were selected and their pathogenicity was tested on 17 commercial cultivars reported as tomato having Cf-9 resistance gene. Among them, 15 cultivars were susceptible and 2 cultivars were resistant. It is likely that the two cultivars include other resistance gene. To our knowledge, this is the first report on the occurrence of Cf-9 infecting F. fulfva strains in Korea.

• Research in Plant Disease
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• v.15 no.3
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• pp.248-253
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• 2009

Chitosan has an antifungal activity and is widely used for control of various plant disease and plants growth in the field in Korea. Disease control efficacy of two preparations (SH-1, SH-2) of mixtures of high and low (chitooligosaccharide) molecular weight chitosan compounds against tomato leaf mold caused by Fulvia fulva was investigated under plastic greenhouse conditions. Both SH-1 and SH-2 formulations displayed potent disease control activity in two experiments. The protective activity of both preparations was comparable to synthetic thiophanate-M. The persistence activity of the formulations was sustained until 21 days after application. Effective concentration of the chtosan compounds for disease control was 1,200 mg a.i./L. In pot tests, chitosan preparations, at a concentration of 600 mg a.i./L, promoted plants growth. These results indicate that the chitosan preparations have a potential as an eco-friendly natural fungicide for the control of tomato leaf mold and plant growth regulator.

• The Plant Pathology Journal
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• v.28 no.4
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• pp.373-380
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• 2012

Burkholderia pyrrocinia CH-67 is a biocontrol bacterium with strong antifungal activity against several plant pathogenic fungi. Transposon mutagenesis was performed to identify the genes responsible for the antifungal activity of B. pyrrocinia CH-67. Of the 2,500 mutants tested using the Fulvia fulva spore screening method, a mutant deficient in antifungal activity, M208, was selected. DNA sequence analysis of the transposon-inserted region revealed that a gene encoding an adenylate kinase-related kinase was disrupted in M208. Antifungal activity was restored in M208 when a full-length adenylate kinase gene with its promoter was introduced in trans. The deduced amino acid sequence of adenylate kinase from CH-67 was 80% identical to that of B. cenocepacia MCO-3. Adenosine diphosphate supplementation or high levels of adenosine triphosphate and adenosine monophosphate together restored antifungal activity in M208, suggesting that adenylate kinase of B. pyrrocinia CH-67 is involved in antifungal activity expression.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2005.04a
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• pp.64.1-64.1
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• 2005

• The Plant Pathology Journal
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• v.27 no.1
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• pp.59-67
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• 2011

A bacterial strain CH-67 which exhibits antagonism towards several plant pathogenic fungi such as Botrytis cinerea, Fulvia fulva, Rhizoctonia solani, Sclerotinia sclerotiorum, Colletotrichum sp. and Phytophthora sp. was isolated from forest soil by a chitin-baiting method. This strain was identified as Burkholderia cepacia complex (Bcc) and belonging to genomovar IX (Burkholderia pyrrocinia) by colony morphology, biochemical traits and molecular method like 16S rRNA and recA gene analysis. This strain was used to develop a bio-fungicide for the control of tomato leaf mold caused by Fulvia fulva. Various formulations of B. pyrrocinia CH-67 were prepared using fermentation cultures of the bacterium in rice oil medium. The result of pot experiments led to selection of the wettable powder formulation CH67-C containing modified starch as the best formulation for the control of tomato leaf mold. CH67-C, at 100-fold dilution, showed a control value of 85% against tomato leaf mold. Its disease control efficacy was not significantly different from that of the chemical fungicide triflumidazole. B. pyrrocinia CH-67 was also effective in controlling damping-off caused by Rhizoctonia solani PY-1 in crisphead lettuce and tomato plants. CH67-C formulation was recognized as a cell-free formulation since B. pyrrocinia CH-67 was all lethal during formulation process. This study provides an effective biocontrol formulation of biofungicide using B. pyrrocinia CH-67 to control tomato leaf mold and damping-off crisphead lettuce and tomato.

• Research in Plant Disease
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• v.16 no.1
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• pp.27-34
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• 2010

This study was performed to develop a formulation using an antagonistic bacterium Bacillus amyloliquefaciens A-2 to control tomato leaf mold caused by Fulvia fulva. B. amyloliquefaciens A-2 was grown in a medium with rice oil and mixed with various carrier and additives. One of the formulations, A2-MP, showed the best disease control value among the tested formulations. The disease control value of A2-MP at 100-fold and 500-fold diluted treatment was not significantly different from that of chemical fungicide triflumizole in a growth chamber. Although disease control effect was decreased by serial diluted treatment of the prepared A2-MP, 1,000-fold diluted treatment of A2-MP still showed high disease control value of 72.0%. For the green house experiments, the disease control values of A2-MP was indicated as 79.4% which is similar to that of chemical fungicide, triflumizole showing 79.6%. When the disease control activity of the formulation A2-MP was compared in tomato production conditions, disease control values of 100-fold diluted A2-MP and 3,000 fold diluted triflumizole exhibited 60%, 81.6%, respectively. The disease control efficiency by A-2MP was 73% of the disease control value of chemical fungicide. The formulation A-2MP maintained the stable bacterial viability and disease control activity when stored at $4^{\circ}C$. This result suggested that A-2MP develped from B. amyloliquefaciens A-2 could be used to control tomato leaf mold.

• Research in Plant Disease
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• v.17 no.2
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• pp.142-147
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• 2011

In order to develop a environmentally friendly control protocol for managing tomato leaf mold disease in the field, we employed bacteria- and fungi-based commercially available microbial preparations. The field experiment was conducted from April to July in 2010. Average incidence rates tomato leaf mold caused by Fulvia fulva were 13.1% at the two plastic houses located in Jangsung, Jeonnam area. Initially 11 microbial preparations were tested for antifungal activity against F. fulva in vitro. Among them, 7 selected preparations showed to be inhibited the mycelial growth of the fungal pathogen over 50%. Four microbes suppressed disease incidence as much 50% under greenhouse condition. Eventually in the field two microbial products including Bacillus subtilis GB-0365 and B. subtilis KB-401 respectively were showed control value up to 71.8% for four times sprays from 20 days to 70 days after transplanting. Furthermore, the control value of three times spray program demonstrated 79.3%. Efficacy of the three and four spray programs was more effective than that of non-spray control treatment. Our results indicated that adjustment of application method of commercially available microbial preparation could be used to control a target plant disease as an effective and efficient crop protection system for organic farming.

• The Plant Pathology Journal
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• v.25 no.4
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• pp.344-351
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• 2009

A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

• Journal of Bio-Environment Control
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• v.31 no.4
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• pp.376-383
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• 2022

The current study, which consisted of two independent studies (laboratory and greenhouse), was carried out to project the hypothesis fungi-spray scheduling for leaf mold and gray leaf spot in tomato, as well as to evaluate the effect of temperature and leaf wet duration on the effectiveness of different fungicides against these diseases. In the first experiment, tomato leaves were infected with 1 × 10⁴ conidia·mL^-1 and put in a dew chamber for 0 to 18 hours at 10 to 25℃ (Fulvia fulva) and 10 to 30℃ (Stemphylium lycopersici). In farm study, tomato plants were treated for 240 hours with diluted (1,000 times) 30% trimidazole, 50% polyoxin B, and 40% iminoctadine tris (Belkut) for protection of leaf mold, and 10% etridiazole + 55% thiophanate-methyl (Gajiran), and 15% tribasic copper sulfate (Sebinna) for protection of gray leaf spot. In laboratory test, leaf condensation on the leaves of tomato plants were emerged after 9 hrs. of incubation. In conclusion, the incidence degree of leaf mold and gray leaf spot disease on tomato plants shows that it is very closely related to formation of leaf condensation, therefore the incidence of leaf mold was greater at 20 and 15℃, while 25 and 20℃ enhanced the incidence of gray leaf spot. The incidence of leaf mold and gray leaf spot developed 20 days after inoculation, and the latency period was estimated to be 14-15 days. Trihumin fungicide had the maximum effectiveness up to 168 hours of fungicides at 12 hours of wet duration in leaf mold, whereas Gajiran fungicide had the highest control (93%) against gray leaf spot up to 144 hours. All the chemicals showed an around 30-50% decrease in effectiveness after 240 hours of treatment. The model predictions in present study could be help in timely, effective and ecofriendly management of leaf mold disease in tomato.