• Journal of Life Science
• /
• v.23 no.2
• /
• pp.299-305
• /
• 2013

Paenibacillus is a gram-positive, spore-forming aerobes that was previously classified as a Bacillus species. Paenibacillus sp. CK214 was highly motile on LB agar plates and showed typical colonial morphology of Paenibacillus. However, its motility was defective in the absence of glucose. Electron microscopic observation revealed that the cells of CK214 cultured on LB agar plates were peritrichously flagellated but not flagellated in the presence of glucose. Flagellar filaments were purified by centrifugation after shearing off from the CK214 cells with vigorous pipetting. The purified protein was composed of a single flagellin with an apparent molecular size of 29 kDa. Recognition of the protein by anti-Edwardsiella tarda flagellin protein antibody demonstrates that the protein is a flagellin protein. A decreased level of flagellin protein was detected in CK214 cells grown under glucose-supplemented media.

• Animal Bioscience
• /
• v.34 no.3_spc
• /
• pp.371-384
• /
• 2021

Objective: Wheat bran (WB) was co-fermented with Aspergillus oryzae and phytase (Phy) to determine whether co-fermentation improve WB phosphorus and fiber utilization in Isa-brown layers. Methods: A total of 112 Isa brown layer were randomly divided into 7 treatments with 8 replicates per a treatment and 2 hens per a replicate. The treatments included basal diet (control), basal diet supplemented with 250 unit/kg Phy (control+Phy), diet with 10% WB (10% WB), diet with 5% WB and 250 unit/kg Phy (5% WB+Phy) diet with 10% WB and 250 unit/kg Phy (10% WB+Phy), diet with 5% fermented WB supplemented with molasses and phy (PCFWH) and 125 unit/kg Phy (5% PCFWH), and diet with 10% PCFWH (10% PCFWH). The intestinal microbial population, intestinal morphology, serum antioxidant enzyme activities, and excreta phosphorus content were assessed. Results: In PCFWH, spore counts, protease activity, xylanase activity, and ferulic acid were 8.50 log/g dry matter (DM), 190 unit/g DM, 120 unit/g DM, and 127 ㎍/g, respectively. Xylobiose and xylotriose were released in PCFWH, while they were not detectable in WB. Antioxidant capacity was also enhanced in PCFWH compared to WB. The 10% WB+Phy and 10% PCFWH groups produced higher egg mass, but hens fed 5% WB+Phy had the lowest amount of feed intake. Eggs from 10% PCFWH had better eggshell weight, eggshell strength, and eggshell thickness. Birds fed with 10% PCFWH also had higher serum superoxide dismutase and catalase activities. Compare to control, 10% PCFWH significantly reduced excreta phosphorus content. Conclusion: Diet inclusion of 10% PCFWH improved egg quality, antioxidant status, and excreta phosphorus content of laying hens.

• Journal of Microbiology and Biotechnology
• /
• v.33 no.5
• /
• pp.607-620
• /
• 2023

The biocontrol approach using beneficial microorganisms to control crop diseases is becoming an essential alternative to chemical fungicides. Therefore, new and efficient biocontrol agents (BCA) are needed. In this study, a rhizospheric actinomycete isolate showed unique and promising antagonistic activity against three of the most common phytopathogenic fungi, Fusarium oxysporum MH105, Rhizoctonia solani To18, and Alternaria brassicicola CBS107. Identification of the antagonistic strain, which was performed according to spore morphology and cell wall chemotype, suggested that it belongs to the Nocardiopsaceae. Furthermore, cultural, physiological, and biochemical characteristics, together with phylogenetic analysis of the 16S rRNA gene (OP869859.1), indicated the identity of this strain to Nocardiopsis alba. The cell-free filtrate (CFF) of the strain was evaluated for its antifungal potency, and the resultant inhibition zone diameters ranged from 17.0 ± 0.92 to 19.5 ± 0.28 mm for the tested fungal species. Additionally, the CFF was evaluated in vitro to control Fusarium wilt disease in Vicia faba using the spraying method under greenhouse conditions, and the results showed marked differences in virulence between the control and treatment plants, indicating the biocontrol efficacy of this actinomycete. A promising plant-growth promoting (PGP) ability in seed germination and seedling growth of V. faba was also recorded in vitro for the CFF, which displayed PGP traits of phosphate solubilization (48 mg/100 ml) as well as production of indole acetic acid (34 ㎍/ml) and ammonia (20 ㎍/ml). This study provided scientific validation that the new rhizobacterium Nocardiopsis alba strain BH35 could be further utilized in bioformulation and possesses biocontrol and plant growth-promoting capabilities.

• KSBB Journal
• /
• v.22 no.5
• /
• pp.297-304
• /
• 2007

For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

• Asian Journal of Turfgrass Science
• /
• v.22 no.2
• /
• pp.171-178
• /
• 2008

We have found the clusters of tiny spiny puffball-like mushrooms growing gregariously in fairy ring (arcs) rimmed by a zone of darker green grass in the golf courses. Macroscopic as well as microscopic characters were examined for the morphology of fruiting body. Exoperidium is thin and densely spiny with minute fibrillae at early stage. The connivent spines were soft and quite persistent. In age, the fibrillae scrumble away with a powdery coating, which leaves white endoperidium becoming pale brown. It's interior was white and fleshy at first, but turns into an olive-colored dust as the gleba, the spore-producing tissue, develops to maturity and loaded with olive-brown spore mass. Then, distinct apical pore developed on the endoperidium. Rudimentary subgleba(sterile base) was narrow, chambered, delineated from the gleba by a membrane in young material. These characters suggested this fungus is a Vascellum, a member of the family Lycoperdaceae. The shapes of the spores were globose, echinulate, $3{\sim}3.5{\mu}m$ in diameter, thick-walled, and olive brown. Capillitial threads were $8-9{\mu}m$ wide, mostly colorless in KOH solution and thin-walled, which designated as "paracapillitium". This is an another character that distinguishes this mushroom from Lycoperdon spp. The spines developed on exoperidium were characteristically connivent; their apices joined together in a point, leaving a space below, which gives the appearance of vault to each group of usually 5 to 6 fibrillae. Based on the above characters, this fungus is identified as Vascellum curtisii (Berkeley). The characters distinguishable this from Lycoperdon pulcherrimum, and Vascellum pretense are discussed in detail. Control trial was also attempted. Strong vertical raking(SVR) followed by applying 500x detergent solution (Spark, Aekyung Co. Seoul) resulted in excellent control over any other treatments. In this plot, fruiting body was not developed throughout the end of mushroom growing season.

• Korean Journal of Soil Science and Fertilizer
• /
• v.20 no.1
• /
• pp.77-84
• /
• 1987

The population of actinomycetes in paddy soils, with hay compost and inorganic fertilizer had been applied respectively was investigated. Actinomycetes were isolated by using selective medium and the population densities of actinomycetes in paddy soil was examined. The population of actinomycetes were reached at a range from $2.1{\times}10^6$ to $7.4{\times}10^7$ per gram of the soil. The composition of actinomycetes flora changed considerably after hay compost applied. The significant positive correlations between the organic matter content in paddy soil and the actinomycetes populations were given at 1.38 to 2.69 level. According to the result of several morphological observation, similar strains isolated were classified into 21 groups. More detailed taxonomic characterization were carried out on the isolated strains. Therefore, 15 groups of Streptomyces and 6 groups of non-Streptomyces were classified into actinomycetes isolates, percentage of streptomyces and non-Streptomyces strains were 87.2% and 12.8% in the isolated 250 actinomycetes strains respectively. Streptomyces with sporophore of the spiral chain form accounted for 80% of all the Streptomyces isolates. Surface morphology of spores were determined with the electron microscope, three species have a spiny surface, and 13 strains have a smooth spore surface.

• The Korean Journal of Mycology
• /
• v.30 no.2
• /
• pp.73-77
• /
• 2002

This study was conducted to investigate morphological characteristics of veiled lady mushroom (Dictyophora sp.). Morphological observation of mycelium showed the presence of chlamydospore-like swelling cells, and typical mycelial champ connections. Fruiting body of the veiled lady mushroom consists of cap, velum, stipe, and vulva. Oval-shaped eggs were surrounded by exodermis, gelatin layers and endodermis, and the fruiting bodies remained inside volva as liquid gelatin layer. Dark green spore layer was formed at the top of cap. The surface eggs in the cultivated veiled lady mushroom was covered with white needle-shaped mycelial projection that was oval in shape. The velum of fruiting body stretched down to the top of volva. The net was regularly shaped, and smelled like fragrance with chestnut flower. The strain KACC 50650 was identified as Dictyophora echinovolvata based on the morphology of eggs and fruiting bodies, and named as Huin-Dol-Gi-Mang-Tae mushroom in Korean.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.6
• /
• pp.1286-1298
• /
• 2005

The genus Paenibacillus is a new group of bacilli separated from the genus Bacillus, and most of species have been isolated from soil. In the present study, we collected 450 spore-forming bacilli from the roots of winter crops, such as barley, wheat, onion, green onion, and Chinese cabbage, which were cultivated in the southern part of Korea. Among these 450 isolates, 104 Paenibacillus-like isolates were selected, based on their colony shape, odor, color, and endospore morphology, and 41 isolates were then finally identified as Paenibacillus spp. by 16S rDNA sequencing. Among the 41 Paenibacillus isolates, 23 were classified as P. polymyxa, a type species of the genus Paenibacillus, based on comparison of the 16S rDNA sequences with those of 32 type strains of the genus Paenibacillus from the GenBank database. Thirty-five isolates among the 41 Paenibacillus isolates exhibited antagonistic activity towards plant fungal and bacterial pathogens, whereas 24 isolates had a significant growth-enhancing effect on cucumber seedlings, when applied to the seeds. An assessment of the root-colonization capacity under gnotobiotic conditions revealed that all 41 isolates were able to colonize cucumber roots without any significant difference. Twenty-one of the Paenibacillus isolates were shown to contain the nifH gene, which is an indicator of $N_{2}$ fixation. However, the other 20 isolates, including the reference strain E681, did not incorporate the nifH gene. To investigate the diversity of the isolates, a BOX-PCR was performed, and the resulting electrophoresis patterns allowed the 41 Paenibacillus isolates to be divided into three groups (Groups A, B, and C). One group included Paenibacillus strains isolated mainly from barley or wheat, whereas the other two groups contained strains isolated from diverse plant samples. Accordingly, the present results showed that the Paenibacillus isolates collected from the rhizosphere of winter crops were diverse in their biological and genetic characteristics, and they are good candidates for further application studies.

• The Korean Journal of Mycology
• /
• v.22 no.4
• /
• pp.378-385
• /
• 1994

Interspecific somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus and Pleurotus sajor-caju. The fusion products between incompatible strains did not form clamp connections. Fruiting body of the clampless fusants was induced by light-dark cycle on saw-dust-rice bran substrate in glass bottles. Out of them, seven somatic hybrids produced fruiting bodies of intermediate morphology of the two species. Light and low temperature were the initiating factors for the development of clamped hyphae from the clampless mycelial colonies. All of these basidiocarps had clamp connections. Eight fusants from the six crosses were analysed with the segregation of genetic characters by random spore isolates. In the three combinations, unexpected alleles were shown. Somatic hybrid between P188 (P. ostreatus 2-1 + P. sajor-caju 2-53) and P. florida 2-3 by triple cross produced fruiting bodies similar to those of fusant between P. ostreatus and P. florida. All the genetic charaters from the three strains were shown to segregate and recombine.

• Journal of Food Hygiene and Safety
• /
• v.23 no.4
• /
• pp.343-347
• /
• 2008

Recently, Sunshik has been issued because of easy-cook and well-being food. Sunshik basically was made of the heated cereals. Amount of spore-forming Bacillus cereus was detected and it has been caused some problem of food safety. B. cereus was isolate from 57 out of 161 Sunshik samples resulting in the isolation rate of 35.4%. Quantitative analysis of 57 samples showed that 21 samples were less than 100 CFU/g, 33 samples were between 100 and 1,000 CFU/g and distinctively even 3 (1.9%) samples had over 1,000 CFU/g. Typical morphology of B. cereus isolated from Sunshik was observed on MYP agar and then further characteristics was identified by using VITEK 2 (Biomeriux, France). 53 strains out of 57 strains isolated from Sunshik (about 93.0%) produced diarrheal enterotoxin in brain heart infusion broth which was detected by the Bacillus cereus enterotoxin reversed passive latex agglutination test kit (Oxoid England). The D-values of the B. cereus spores were $75^{\circ}C$ (37.1mim), $80^{\circ}C$ (22.5mim), $85^{\circ}C$ (4.9mim), and $90^{\circ}C$ (3.1mim) respectively. The Z-value was calculated $12.8^{\circ}C$ in Sunshik sample inoculated with B. cereus. Therefore, the management of B. cereus in Sunshik is required for the food-safety.