• Journal of Applied Biological Chemistry
• /
• v.42 no.1
• /
• pp.7-11
• /
• 1999

An extracellular chitinase of the selected strong antifungal microorganism, Serratia sp. 3095, was purified by salting out, affinity adsorption, Sepadex G-100 gel fitration, Sepadex G-75 gel fitration and DEAE Sepadex A-50 chromatography. The molecular weight of the purified chitinase was estimated to be 62,000 dalton by SDS-PAGE. Optimal pH and temperature of the chitinase were pH 7.5 and 45, respectively. The enzyme retained more than 80% of the activity between pH 5.5 and pH 10.5, and below $50^{\circ}C$ but was unstable above $60^{\circ}C$, below pH 5.0. The activity of the chitinase was inhibited about 60% by $Sn^{2+}$, 40% by $Hg^{2+}$ and $Ag^+$, 70% by AHA, 40% by iodoacetate, 35% by thiourea and p-CMB, but stabilized by SDS. $K_m$ value of the purified chitinase was 3.68 mg/ml for colloidal chitin. The chitinase from Serratia sp. 3095 showed antifungal activity to Fusariurm solani.

• Korean Journal of Environmental Agriculture
• /
• v.31 no.1
• /
• pp.68-74
• /
• 2012

BACKGROUND: Some microorganisms extant in nature have ability to suppress various plant pathogens, and also can promote plant growth. Thus microorganisms are such great source of antimicrobial agents to develop antagonistic microorganism production and eco-friendly crop management. We isolated the microorganisms in various eco-friendly formulations. The suppressive abilities against plant pathogens have been characterized in vitro level. METHODS AND RESULTS: The indigenous microorganisms have been isolated from Cooked rice, Black sugar, Rice Bran, and Red clay using dilution plating method. Population of bacteria and fungi were above 107 in the all formulations. We isolated and pure cultured the microorganisms based on morphological characteristics. Three major plant pathogens (Fusarium oxysporum, Rhizoctonia solani, Phytophthora capsici) have been used to select antagonistic microorganisms. Total 20 bacteria and 9 fungi showed the pathogen growth suppression ability in vitro condition. The selected microorganisms were identified by ITS sequence similarity. CONCLUSION: All tested eco-friendly formulations contained high-density of the microorganisms. Among the isolated microorganisms, Bacillus spp. and Streptomyces spp. showed the most effective antifungal activity against the plant pathogens such as F. oxysporum, R. solani, and P. capsici. Among the selected fungi Trichoderma sp. demonstrated antifungal activity. Our results suggest that the currently adapted eco-friendly formulations might useful for sustain agricultural system.

• Korean Journal of Organic Agriculture
• /
• v.7 no.2
• /
• pp.99-106
• /
• 1999

In order to acquire microbial agents that can be utilized for biological control of bitter rot(Glomerella cingulata), the major airborne disease to apple, the effective microorganisms were isolated, tested for antagonistic activity to the pathogen. Through the screening of more than 1,000 species of microorganisms collected in nature, 11 species of antagonists were selected. On of the 11 species, one species designated as CH141 demonstrated outstanding activity. The bacterial strain, CH1141 exerted antagonistic efficiency of 65% on Glomerella cingulata. The CH1141 was identified as a bacterial strain to Bacillus subtilis based on morphology, culture conditions, and physiobiochemical characteristics.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.15 no.1
• /
• pp.1-12
• /
• 1995

This study was to investigate an effective biological control of forage diseases and provide a basic data and a model in improving variety of antagonistic bacteria, with growth promoting effect on forage, through cell fusion. The results obtained were summarized as follows; 1. The antagonistic himbacterium against soil-borne phathogenic fungi Fusarium oxysporum and Rhizoctonia solani was isolated from continuous cropping himsphere soil of forage, and its biological and physiological characteristics were investigated. This bacterium was identified as Bacillus subrilis and named BS 101. Another strain for cell fusion was Bacillus thur ingiensis ssp. kurstaki HD-I(BT 37669) with insecticidal crystal. 2. The auxotropic mutants of BS 101 and BT 37669 were derived after mutagenesis using N-methyl-N'nitro- Nitrosoguanidine(NTG) to give amino acid requirement marker. n e s e auxotropic mutants of BS 101 and BT 37669 were named BS 1013(his-) and BT 69(asp-), respectively. 3. The best protoplast requirement was obtained using DM 3 medium, containing 5% casamino acid, 1 M $MgCI_2$ and 2% bovine semm albumin, to give Fusant 3, 7 and 8. BT toxin gene was not identified with fusants by Southern blotting. However, SDS-PAGE analysis of strains showed various protein patterns among fusants. 4. From the dark culture experiment, growth of forage in inoculated soil with antagonistic bacteria was delayed than that of non-inoculated soil with antagonistic bacteria in each continuous cropping soil and in each sterilized soil. On the other hand, growth duration of forage was different between continuous cropping soil and sterilized soil. 5. Seed germination of Alfalfa, Italian ryegrass and Orchardgrass were significantly improved by inoculation of antagonistic bacteria(p< 0.05).

• Korean Journal of Organic Agriculture
• /
• v.6 no.1
• /
• pp.133-149
• /
• 1997

Recently, the importance of management and cultivation of grasses has been increased in Korea. Among these cultural practices, the appropriate control of diseases is considered more important than other cultivation techniques such as fertilization and irrigation. The damages of brown patch and large patch caused by Rhizoctonia spp. and Pythium blight caused by Pythium spp. are serious in the major cultivation area of turfgrass in Korea. Since these diseases are difficult to control by agrochemicals, the damages are very serious if these are occured. The periodic spray of agrochemicals, to protect and control these diseases could make some problems of toxicity and environmental pollution as well as rising of non-target diseases. Therefore, the biological methods to control diseases have been required to decrease problems resulted from overuse of agrochemicals, to conserve natural ecosystem, and to control effectively diseases of grasses in the long period. The number of studies about biological control using antagonistic microorganisms have been increased for last half century. However, the application of biological control method has been very limited. In this study, thirteen isolates of R. cerealis, 8 isolates of R. solani and 3 isolates of Phthyn spp. have been isolated from diseased turfgrass in golf course and grass-culture area that have patch and wilting symptoms of zoysia grass and creeping bentgrass. Isolation frequency of R. cerealis and R. solani was high in especially zoysiagrass, while Pythym spp. was isolated from bent grass at low frequency but showed high pathogenicity. Totally, 205 isolates of soil microorganisms were isolated in this study as primary antagonistic microorganism by Herr's triple agar layer plate and dual culture method using rhizosphere of grasses, soil of crop field as the source of antagonistic microorganisms. Among the 205 isolates, 23 isolates were actinomycetes and 182 isolates were bacteria. All of the actinomycetes were isolated by Herr's method. Antagonistic effect of primary isolated microorganisms was tested for in vitro mycelial growth inhibition against pathogenic fungi isolated from grasses and for inhibition of disease occurrence in 24 well tissue culture plate and pot experiment. Then, four isolated of bacteria which are BG23, BG74, BG136 and BG171 were selected as antagonistic microorganisms against soil-born pathogenic fungi of bentgrass.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.18 no.3
• /
• pp.195-204
• /
• 1998

This study was designed to investigate the effects of antagonistic microorganism, Bacillus subtilis, on the growth of forage seedlings in repeated cultivation soils and unrepeated cultivation soils. The field experiment was wnducted in pots in a vinyl house using repeated and unrepeated cultivation soils. Forage types were 'Suwon 19' wrn(Zea mqs L.), 'Califbmia' white clover(Tr~oIium repens L.) and 'Fawn' tall fescue (Festuca arundianacea Schreb.). Samples of white clover and tall fescue were taken h m each pot at 36 days after seeding. Samples of wm were examined at 50 days after seeding. The most active antagonistic bacterium was isolated h m forage rhizosphere soil, and selected by reference to it's antagonistic ability on the growth of pathogenic fungi, Rhizoctonia solmi and Fusarium oxyspomm, and it was identified as Bacillus subtilis. This strain strongly suppressed the growth of fungal pathogens among isolated rhizobacteria. The dry weight of forage shoots and roots cultivated in unrepeated cultivation soils was higher than that cultivated in repeated cultivation soils. The dry weight of forage was positively affected by the inoculation of the antagonistic bacterium, Bacillus subtilis, in both repeated cultivation soils and unrepeated cultivation soils. In conclusion, the growth of forage was more affected by the inoculation of the antagonistic bacterium in unrepeated cultivation soils than that in repeated cultivation soils, and bacterization of forage with B. subtilis resulted in an inrreased yield.

• Korean Journal of Medicinal Crop Science
• /
• v.20 no.5
• /
• pp.339-344
• /
• 2012

This study was carried out to research microorganisms having the antifungal activity against ginseng Alternaria blight pathogen Alternaria panax and ginseng anthracnose pathogen Colletotrichum gloeosporioides. Eleven Bacillus strains. were isolated from Korean traditional soybean paste and Kimchi. Among the 11 isolates, DJ5, KC1, KC2 and KC4 showing antagonistic activity on the mycelial growth of A. panax and C. gloeosporioides in pairing culture were finally selected as the antagonistic microorganisms. Based on 16s rRNA sequence and phylogenetic tree analysis, they were identified as Bacillus spp.. The selected microorganisms were investigated antagonistic activity by measured leaf-segment colonization in pot test. When Bacillus sp. were injected after A. panax treatment, KC1, KC2 and KC4 showed similar effect to chemical pesticides treated control. To measure preventive effect of Bacillus sp, antagonistic microorganisms were injected and C. gloeosporioides was treated in pot. When measuring the effectiveness for the prevention of Anthracnose, All Bacillus spp. showed approximately 83~90 % degree of superior preventive effect. In general, The four Bacillus spp. isolated from Korean traditional fermented foods showed therapeutic effect of Alternaria blight and preventive effect of Anthracnose.

• Korean Journal of Organic Agriculture
• /
• v.8 no.3
• /
• pp.121-130
• /
• 2000

In order to screen the antagonistic bacteria which inhibit the growth of the plant pathogen, Penicillum expansum, we isolated an effective bacterial strain and investigated into the antifungal activity of the antagonist and it's identification. The eleven strains of bacteria which strongly inhibited P. expansum were isolated from the nature, and the best antagonistic bacterial strain designated as KB22, was selected. The antagonistic strain KB22 was identified to be the genus Bacillus subtilis based on morphological and biochemical characterization, The KB22 showed 55.9% of antifungal activity against the growth of P. erpansum. By the treatment of the culture broth and the heat treated culture filtrate of it, the B. subtilis KB22 showed 90% and 15% of antifungal activity, respectively.

• Microbiology and Biotechnology Letters
• /
• v.32 no.4
• /
• pp.312-316
• /
• 2004

In oder to select the powerful rhizophere-dorminatable biocontrol agent, we had isolated an indigenous antagonistic bacterium which produced antibiotic and siderophore from a disease suppressive local field soil of Gyungsan, Korea. And we could select the Pseudomosp. 4059 which can strongly antagonize against Fusarium oxysporum and Phytophthora capsici by two kinds of antifungal mechanism that can be caused by the antibiotic of Phenazin, a siderophore and a auxin like subThe selected strain was identified as Pseudomonas fluorescens (biotype A) 4059 by biochemical tests, API $\textregistered$ test, MicroLog TM system and 16S rDNA analysis. The selected antagonistic microorganism, Pseudomosp. 4059 had an antifungal mechanism of antifungal antibiotic and sidrophore. And we were confirmed the antagonistic activity of P fluorescens 4059 with in vitro antifungal test against Phytophthora capsici and in vivo by red-pepper.

• Korean Journal of Food Science and Technology
• /
• v.40 no.6
• /
• pp.669-673
• /
• 2008

For the development of a biological control method against B. cereus in cheonggukjang, 20 Bacillus spp. were isolated from the naturally fermented baektae and heuktae cheonggukjang, identified by using 16S rDNA sequences. Among the isolated strains, Bacillus sp. SC-8 was selected using the B. cereus lawn cell assay as an antagonistic microorganism against B. cereus. The culture medium of Bacillus sp. SC-8 after 24 hr of incubation at $37^{\circ}C$ also evidenced a high level of antagonistic activity. In cheonggukjang fermented with the mixed culture of Bacillus sp. SC-8 and B. cereus, antagonistic effect against B. cereus was maintained during the fermentation of cheonggukjang, while its effect was reduced during storage at $4^{\circ}C$ due to the decrement of cell population of Bacillus sp. SC-8. In Bacillus sp. SC-15, which was utilized a control, antagonistic activity against B. cereus was not demonstrated on the lawn cell plate assay and culture medium, but its effects were detected in cheonggukjang. Therefore, the production of antagonistic substances of Bacillus spp. depends on the fermentative environment.