• Applied Biological Chemistry
• /
• v.34 no.1
• /
• pp.54-60
• /
• 1991

Serratia marcescens CK-3, decomposing chitin which is a mar component of cell wall in phyitopathogenic fungi, was isolated from the continuous cropping rhizosphere of pepper and cucumber and its enzymatic property was examined. S. marcescens CK-3 was found tn have an tagonistic effects against, Fusarium axysporum and Rhizoctonia solani and to have complex enzyme system such as chitinase, laminarinase, and proteinase. The preferable composition of the medium for production of chitinase was fond and was as follows : colloidal chitin 1.5%, tryptone 0.5%, glucose 1.0%, peptone 0.2%, $MgSO_4{\cdot}7H_2O\;0.1%,\;K_2HPO_4\;0.1%,\;and\;NaCl\;0.1%$(w/v), pH 6.8. The maximum enzyme production was observed after culture of 72 hours at $30^{\circ}C$ using a medium containing the above chemical composition. The optimal pH and temperature for in vitro activity of chitinase from S. marcescens CK-3 were pH 7.5 and $50^{\circ}C$, respectively. The enzyme activity in-creased by metal ions such as$Ag^+$ and $Mn^{++}$.

• The Korean Journal of Mycology
• /
• v.26 no.4 s.87
• /
• pp.496-501
• /
• 1998

The mycelia of Tricholoma matsutake DGUM 26001, 26101, 26210 and FRI 91024 were used to determine the extracellular enzyme activity in mycelia. When the filtrate of culture broth after 30-day cultivation at $24^{\circ}C$ was used as a crude solution of extracellular enzyme, the average specific activity of ${\alpha}-amylase$ was 6142.3 unit/mg protein. The specific activity of xylanase was comparatively high. However, little or no enzyme activities were found for ${\beta}-glucosidase$, ligninase, CMCase, chitinase, protease, and lipase.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.10
• /
• pp.1359-1366
• /
• 2012

A strain of Streptomyces cavourensis subsp. cavourensis (coded as SY224) antagonistic to Colletotrichum gloeosporioides infecting pepper plants was isolated. SY224 produced lytic enzymes such as chitinase, ${\beta}$-1,3-glucanase, lipase, and protease in respective assays. To examine for antifungal activity, the treatments amended with the nonsterilized supernatant resulted in the highest growth inhibition rate of about 92.9% and 87.4% at concentrations of 30% and 10%, respectively. However, the sterilized treatments (autoclaved or chloroform treated) gave a lowered but significant inhibitory effect of about 63.4% and 62.6% for the 10% supernatant concentration, and 75.2% and 74.8% for the of 30% supernatant concentration in the PDA agar medium, respectively, indicative of the role of a non-protein, heat stable compound on the overall effect. This antifungal compound, which inhibited spore germination and altered hyphal morphology, was extracted by EtOAc and purified by ODS, silica gel, Sephadex LH-20 column, and HPLC, where an active fraction was confirmed to be 2-furancarboxaldehyde by GS-CI MS techniques. These results suggested that SY224 had a high potential in the biocontrol of anthracnose in pepper, mainly due to a combined effect of lytic enzymes and a non-protein, heat-stable antifungal compound, 2-furancarboxaldehyde.

• Korean Journal of Soil Science and Fertilizer
• /
• v.43 no.4
• /
• pp.490-497
• /
• 2010

A chitinolytic bacterium having a strong antagonistic activity against various pathogens including Phytophtora capsici was isolated from rhizosphere soil, and identified as Lysobacter enzymogenes (named as LE429) based on 16S rRNA gene sequence analysis. This strain produced a number of substances such as chitinase, ${\beta}-1$, 3-glucanase, lipase, protease, gelatinase and an antibiotic compound. This antibiotic compound was purified by diaion HP-20, sephadex LH-20 column chromatography and HPLC. The purified compound was identified as phenylacetic acid by gas chromatography-electron ionization (GC-EI) and gas chromatography-chemical ionization (GC-CI) mass spectrometry. In field experiment, pepper plants were treated by the strain LE429 culture (CB), neem oil solution (NO), combination (CB+NO) or control (CON). Plant height and number of branches, flowers and pods of pepper plant in CB treatment were generally highest, and followed by CB+NO, CON and NO. The fungal pathogens were strongly inhibited, while several insect pests were discovered in CB treatment. Any insect pests were not found, while all fungal pathogens tested were not suppressed in NO treatment. However, in CB+NO treatment, non incidence of fungal pathogens and insect pests were found. The strain LE429 producing secondary metabolites with neem oil should be a potential agent to control fungal diseases and insect pests.

• Microbiology and Biotechnology Letters
• /
• v.25 no.6
• /
• pp.606-611
• /
• 1997

Entomogenous fungi which attack living insects are powerful means for microbiological insecticide. The purpose of this study is to establish the culture conditions and media for mass production of Beauveria sp. C208 which has a broad host range as a potential microbiological pesticide. The temperature and pH range for optimal cultivation of this strain were 28$circ$C and pH 5.0-7.0. For Beauveria sp. C 208, 2% rice straw and 0.6% tryptone were found as the proper carbon and nitrogen sources, considering cell mass, enzyme activities such as chitinase, protease and lipase, and spore concentration.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.6
• /
• pp.919-927
• /
• 2007

Antagonistic fluorescent pseudomonads isolated from rhizospheric soil of rice were characterized by 16S rRNA amplicon and fatty acid methyl ester (FAME) analyses. Antagonistic isolates were grown in the fermentation media, and production of antibiotics was confirmed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Production of fungal cell-wall-degrading enzymes such as protease, cellulase, pectinase, and chitinase was determined. Dendrogram based on the major and differentiating fatty acids resulted into 5 clusters, viz., cluster I (P. pseudoalcaligenes group), cluster II (P. plecoglossicida group), cluster III (P. fluorescens group), cluster IV (P. aeruginosa group), and cluster V (P. putida group). Characteristic presence of high relative proportions of cyclopropane (17:0 CYCLO w7c) was observed in antagonistic bacteria. Data revealed biodiversity among antagonistic fluorescent pseudomonads associated with the rice rhizosphere. Results presented in this study will help to identify the antagonistic isolates and to determine their mechanisms that mediate antagonism against fungal pathogens of rice.

• Korean Journal of Microbiology
• /
• v.36 no.2
• /
• pp.97-102
• /
• 2000

In order to use Metarhizizmn~ anisopliae as a biological pesticide, effect of envrionmental factors on nlycelial growth, spore formation, and extracellular enzyme activity in culture broth of M. anisopliae DGUM 35001 was determined. Optimal temperature was 26^{\circ}C.$ and optimal pH ranged from 5 to 9. Among the complex media tested, MCM and SDPY media were the most favorable for mycelial growth. When Czapek-Dox agar was used as a mnimal medium, glucose and sucrose among the saccharides were very excellent source of carbohydrate. Among the biopolyners tested. chitin was the most favorable source for mycelial growth and produced high aerial inycelia. Urea and ammonium phosphate as an inorganic nitrogen source and bacto-peptone and soytone as an organic nitrogen source enhanced the mycelial growth When serine as a source of amino acid was supplemented, excellent mycelial growth was shown. Large amount of spores could be obtained from the aerial mycelia of starch medium. When the culture broth was filtrated and then the concentrate with ammonium sulfate was used as a crnde enzyme solution, high enzyme activities of amylase and protease were shown. However, lipase and chitinase activities were comparatively low.

• Research in Plant Disease
• /
• v.22 no.2
• /
• pp.81-93
• /
• 2016

Out of plant-associated bacteria, certain plant growth-promoting bacteria (PGPB) have been reported to increase plant growth and productivity and to elicit induced resistance against plant pathogens. In this study, our objective was to broaden the range of applications of leaf-colonizing PGPB for foliar parts of road tress and pepper. Total 1,056 isolates of endospore-forming bacteria from tree phylloplanes were collected and evaluated for the enzymatic activities including protease, lipase, and chitinase and antifungal capacities against two fungal pathogens, Colletotrichum graminicola and Botrytis cinerea. Fourteen isolates classified as members of the bacilli group displayed the capacity to colonize pepper leaves after spraying inoculation. Three strains, 5B6, 8D4, and 8G12, and the mixtures were employed to evaluate growth promotion, yield increase and defence responses under field condition. Additionally, foliar application of bacterial preparation was applied to the road tress in Yuseong, Daejeon, South Korea, resulted in increase of chlorophyll contents and leaf thickness, compared with non-treated control. The foliar application of microbial preparation reduced brown shot-hole disease of Prunus serrulata L. and advanced leaf abscission in Ginkgo biloba L. Collectively, our results suggest that leaf-colonizing bacteria provide potential microbial agents to increase the performance of woody plants such as tree and pepper through spray application.

• Korean Journal of Microbiology
• /
• v.40 no.2
• /
• pp.100-103
• /
• 2004

The culture condition and medium composition for the enhanced mycelial growth of Inonotus mikadoi IMSNU 32058 were investigated. The optimal temperature and pH for the mycelial growth were $27^{\circ}C$ and pH 4.5, respectively. Among the complex media tested, the malt extract medium and Phellinus igniarius medium were very good for mycelial growth of I. mikadoi. When Czapek-Dox medium was used as a minimal medium for cultivation of mycelia, xylose, raffinose and carboxymethyl cellulose were very excellent as a carbon and energy source. With respect to carbohydrate, sucrose and glucose were very good carbon sources. In general, organic complex nitrogen sources were better than other inorganic ones. As the organic complex nitrogen sources tested, yeast extract, soytone, proteose peptone and bacto peptone were the best as a source of organic nitrogen. When ammonium sulfate as an inorganic source of nitrogen was used, the enhanced mycelial growth was shown. p-Aminobenzoic acid was proved to be most appropriate source of vitamin. After the mycelia of I. mikadoi was cultivated at $27^{\circ}C$ for 5 days in MEM broth (pH 4.5), the activities of both exomycelial and endo-mycelial enzymes were determined. Among endomycelial enzymes assayed, the specific activity of laccase was much higher than those of other enzymes. When the fungus was grown in MEM broth, exomycelial specific enzyme activity of laccase was comparatively high. However, little or no enzyme activities of protease, chitinase and lipase were found.

• The Korean Journal of Mycology
• /
• v.44 no.1
• /
• pp.36-47
• /
• 2016

This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.