A fungal strain of Trichoderma having strong chitinolytic activity was isolated from field soil enriched with crabshell for several years. Based on 5.8S rRNA, partial 18S, 28S rRNA genes, ITS1, ITS2 sequence analysis and morphological characteristics, the fungus was identified as Trichoderma asperellum and named as Trichoderma asperellum T-5 (TaT-5). The fungus released lytic enzymes such as chitinase and ${\beta}$-1, 3-glucanse, and produced six antifungal substances in chitin broth medium. To demonstrate the protective effect of TaT-5 against damping-off in cucumber plant caused by Rhizoctonia solani, TaT-5 culture broth (TA), chitin medium (CM) and distilled water (DW) were applied to each pot at 10 days after sowing, respectively. Then, the homogenized hyphae of R. solani were infected to each pot at 1 week after TaT-5 inoculation. During experimental period, fresh weight of shoot and root in cucumber plant more increased at TA treatment compared to other treatments. PR-proteins (${\beta}$-1, 3-glucanase and chitinase) activities in cucumber leaves markedly increased at CM and DW treatments, but the activity slightly increased and then decreased at TA treatment at 3 days after infection of R. solani. The activity of PR-proteins activities in cucumber roots at all treatments decreased with time where the degree of decrement was more alleviated at TA treatment than CM and DW. These results suggest that the lytic enzymes (chitinase and ${\beta}$-1, 3-glucanse) and antifungal substances produced by TaT-5 can reduce the pathogenic attack by R. solani in cucumber plants.
Won, Ok Jae;Kim, Young Tae;Kim, Jae Deok;Choi, Jung Sup;Ko, Young Kwan;Park, Kee Woong
Weed & Turfgrass Science
/
v.4
no.3
/
pp.219-224
/
2015
This study was conducted to evaluate the effect of herbicidin, new natural herbicidal substances, derived from soil actinomycetes Streptomyces scopuliridis. Several weed species were subjected to examine the germination inhibition and herbicidal activity at the concentration from 100 to 2,000 ppm. There was no selectivity in germination inhibition and herbicidal activity against crops. Germination of Echinochloa oryzoides, Digitaria ciliaris, Abutilon theophrasti and Amaranthus retroflexus was inhibited completely when 7.81 ppm of extract was treated in petri dish. Pre-emergence application of herbicidin in soil condition showed low inhibition against weeds. However, post application of herbicidin in green house resulted in the necrosis of weeds at the concentration of 2,000 ppm. A. retroflexus was sensitive to herbicidin at the low concentration of 62.5 ppm, whereas E. oryzoides was tolerant to lower concentration of herbicidin until it became withered at the concentration of 2,000 ppm. In conclusion, herbicidal substances derived from S. scopuliridis herbicidin, which is consisted with herbicidin A and B, have dominant effect on germination and growth inhibition. On the other hand, herbicidin was insufficient to control gramineous weeds. In future, it will be needed to develop the combination of herbicidin with other herbicide or compounds to control gramineous weeds as well.
Burkholderia sp. D5, a polyaromatic hydrocarbons(PAHs)-degrading bacterium, was isolated from oil-contaminated soil. The bacterium could utilize phenanthrene (Phe) as a sole carbon source but could not use pyrene (Pyr). However, the strain could degrade Pyr when a cosubstrate such as yeast extract (YE) was supplemented. The PAH degradation rate of the bacterium was enhanced by the addition of other organic materials such as YE, peptone and glucose. YE was a particularly effective additive in stimulating cell growth as well as PAH degradation. When 1 g-YE/L was supplemented into the basal salt medium (BSM) with 215 mg-Phe/L, the specific growth rate (0.28 h-1) and Phe-degrading rate (29.30 μmol/L/h) were enhanced approximately ten and two times more than those obtained in the BSM with 215 mg-Phe/L, respectively. Through kinetic analysis, the maximum specific growth rate (μmax) and PAH degrading rate (Vmax) for Phe were obtained as 0.34/h and 289 ${\mu}mol$/L/h, respectively. Also, μmax and Vmax for Pyr were 0.27 h-1 and 50 ${\mu}mol$/L/h, respectively. The degradation rates for each Phe (2.20 μmol/L/h) and Pyr (2.18 μmol/L/h) were lower in mixture substrates than in a single substrate (29.30 ${\mu}mol$/L/h and 9.58 ${\mu}mol$/L/h, respectively). Burkholderia sp. D5 can degrade Phe and Pyr contained in soil, and the PAH degradation rates in soil were 20.03 ${\mu}mol$/L/h for Phe and 1.09 ${\mu}mol$/L/h for Pyr.
Ecological studies on entomopathogenic nematodes are required to increase control efficacy against target insect pests and to obtain basic information for mass production. Thus, effect of temperature and nematode concentration on infectivity and reproduction of Steinernema carpocapsae Pocheon and that of exposure time and soil depth on infectivity were examined using Galleria mellonella larvae. Infectivity and reproduction were examined at five temperatures, 13, 18, 24, 30 and 35$^{\circ}C$ with seven concentrations, 0, 5, 10, 20, 40, 80 and 160 infective juveniles (IJs)/larva. Temperature and nematode concentration influenced infectivity and reproduction of S. carpocapsae Pocheon. Although G. mellonella larvae were killed by S. carpocapsae Pocheon at all given temperatures and nematode concentrations, mortality was higher at 24$^{\circ}C$ than other temperatures. Lethal time of G. mellonella by S. carpocapsae Pocheon was shorter with increasing temperature and nematode concentrations. S. carpocapsae Pocheon was not established in G. mellonella at 13 and $35^{\circ}C$. Time for the first emergence from G. mellonella cadaver was longer $18^{\circ}C$ (about 20 days) than 24 and $30^{\circ}C$ (about 5 days). The highest number of progenies was obtained at $24^{\circ}C$ with 80IJs/1arva, i.e., $18.8$\times$10^4$IJs were produced from a larva. In the exposure time assay, G. mellonella death was recorded in 10 minutes when 300 IJs were inoculated per larva. When S. carpocapsae Pocheon was inoculated at the rate of $10^{9}$ IJs/ha to G. mellonella at the depth of 0, 2, 5 and 10 cm of sand columns, 100% mortality and similar sex ratio were observed but number of established IJs in cadaver was decreased with deepening the soil depth. The results indicated that optimum temperature for infectivity and reproduction of S. carpocapsae Pocheon was $24^{\circ}C$ In addition, S. carpocapsae Pocheon was effective to target insects within 5 cm from the soil surface.
With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.
Journal of Korean Society of Coastal and Ocean Engineers
/
v.30
no.6
/
pp.306-318
/
2018
In order to evaluate the seismic capacity of massive vertical type breakwaters which have intensively been deployed along the coast of South Korea over the last two decades, we carry out the preliminary numerical simulation against the PoHang, GyeongJu, Hachinohe 1, Hachinohe 2, Ofunato, and artificial seismic waves based on the measured time series of ground acceleration. Numerical result shows that significant sliding can be resulted in once non-negligible portion of seismic energy is shifted toward the longer period during its propagation process toward the ground surface in a form of shear wave. It is well known that during these propagation process, shear waves due to the seismic activity would be amplified, and non-negligible portion of seismic energy be shifted toward the longer period. Among these, the shift of seismic energy toward the longer period is induced by the viscosity and internal friction intrinsic in the soil. On the other hand, the amplification of shear waves can be attributed to the fact that the shear modulus is getting smaller toward the ground surface following the descending effective stress toward the ground surface. And the weakened intensity of soil as the number of attacking shear waves are accumulated can also contribute these phenomenon (Das, 1993). In this rationale, we constitute the numerical model using the model by Hardin and Drnevich (1972) for the weakened shear modulus as shear waves go on, and shear wave equation, in the numerical integration of which $Newmark-{\beta}$ method and Modified Newton-Raphson method are evoked to take nonlinear stress-strain relationship into account. It is shown that the numerical model proposed in this study could duplicate the well known features of seismic shear waves such as that a great deal of probability mass is shifted toward the larger amplitude and longer period when shear waves propagate toward the ground surface.
A bacterial strain producing extracellular mannanases was isolated from soil of chestnut tree farm located in Gongju city of Korea by enrichment culture using Avicel as a carbon source. 16S rDNA sequence of the isolate YB-43 was highly homologous to those of genus Cellulosimicrobium strains with sequence similarities of above 99.6%. Mannanase productivity was significantly increased when the Cellulosimicrobium sp. YB-43 was grown in the presence of locust bean gum (LBG) or konjac. The mannanases were partially purified to be mannanase A (ManA) and mannanase C (ManC) by DEAE-Sepharose column and Q-Sepharose column chromatography from the culture filtrate of Cellulosimicrobium sp. YB-43 grown in LB medium supplemented with 0.7% LBG for 24 h. The partially purified ManA showed the highest activity at $55^{\circ}C$ and pH 6.5, while ManC activity was optimal at $65^{\circ}C$ and pH 7.5. ManA was stable up to $40^{\circ}C$ for 1 h, but ManC activity decreased significantly even after 1 h at $20^{\circ}C$. ManA and ManC showed difference from each other according to their substrate specificities and predominant products resulting from the mannanase hydrolysis for mannooligosaccharides. As a result, Cellulosimicrobium sp. YB-43 was found to produce two different kinds of mannanases.
1) The purpcse of the present study is to investigate the effects of culture filtrates of Fuarsium oxysporum f. vasinfectum which is known to produce wilt toxin (fusaric acid) on the germination of host plants (sesame, cotton) and non-host plants (wheat, rice). 2) The experiment on the germination of sesame, cotton, wheat and rice seeds in the seed beds separately added with culture filtr ates of 10 differential strains of Fusarium oxysporom f. vasinfectum demonstrated that culture filtrates of most strains of the fungus inhibit or retard the germination of seeds of 4 plants used in this study while those of a few strains do not give notable influence on the germination of seeds of those plants. a) Culture filtrates of strain 201 of the fungus strongly inhibited the germination of seeds of those plants in nearly same degree, but culture filtrates of the other strains, 281, 321, etc., showed remarkable differences in the toxicity inhibiting or retarding the germination of the seeds of those plants. b) In general, sesame seeds are greatly susceptible, wheat and cotton seeds are moderately susceptible and rice seeds are resistant to the toxicity of culture filtrates of the fungus. 3) In the soil containing a number of differential strains of Fusarium oxysporum f. vasinfectum, the germination of seeds and also the growth of seedlings of non-host plants are possibly checked by the toxic substance, fusaric acid produced by the fungus.
Cyclodextrin glucanotransferase (CGTase) of Bacillus sp. isolated from soil was purified and its enzymological characteristics were investigated. It was found that the production of CGTase reached to the maximum when the strain was cultured in the broth containing 0.1 % albumin, 2% $NH_4Cl$, 2% soluble starch and 0.2% $NH_2PO_4$ for 72 hrs at $37^{\circ}C$. The purity of CGTase was increased by 9.7 folds through purification procedures by the following column chromatography DEAE-cellulose ion exchange chromatography and Sephadex G-100, G-150 gel filtration and its specific activity was 528.0 unit/mg. The optimum pH and temperature for the CGTase activity were 8.0 and $80^{\circ}C$, respectively. The enzyme was stable in pH $8.0{\sim}11.0$ at $60{\sim}80^{\circ}C$. The activity of purified enzyme was inhibited by $Pb^{2+},\;Hg^{2+}$ and $Zn^{2+}$. When CGTase was treated with each 20.5 unit, 41 unit, 205 unit and 410 unit to investigate the transglucosylation to stevioside by purified cyclodextrin glucanotransferase, transglucosylation rate to stevioside was 74.9%, 75.7%, 68.7% and 57.9%. Brown effect was observed above the concentration amounting to 205 unit of our CGTase.
In order to produce clear and favorable citrus wine from Citrus unshiu produced in Cheju island, chemical and microbiological processes for alcoholic fermentation were investigated. The ratio of pressed juice passed below 100 mesh sieve and peel of mandarin orange were 55.9% and 25.6% respectively. Orange juice for fermentation source contained 8.85% total sugar, 1.43% total acid and 0.056% volatile acid. Pressed juice was adjusted to 24 degree Brix with cane sugar, and was fermented at $20^{\circ}C$ for one month. Starter screened and selected was Saccharomyces cerevisiae IAM 4274. As principal fermentation proceeded for one week, suspended solids began to precipitate slowly after then. After fermentation, clear citrus wine consisted of about 8 degree Brix residual sugar, $13.3{\sim}14.4%$ ethanol, $0.78{\sim}1.11%$ total acid, $0.05{\sim}0.07%$ methanol and $2.25{\sim}3.29%$ extract, was obtained. Color, flavor and taste of citrus wine found good with panel test. Citrus wine which was treated with fungal enzyme derived from Aspergillus niger CCM-4 was cleared much faster, and could be filtered more rapidly than the untreated. The enzyme-producing strain was isolated from field soil of Cheju island and identified.
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