• The Korean Journal of Pesticide Science
• /
• v.12 no.3
• /
• pp.291-294
• /
• 2008

The Streptomyces padanus isolate TH04, isolated from mummified peaches, showed strong antifungal activity to Monilinia fructicola. The inhibition activity of the isolate TH04 to mycelial growth and spore germination at 1% concentration of sub-antifungal powder made from culture suspension (CS) was ranged from 79.8% to 81.0% and from 73.9% to 75.8% to M. fructicola four strains, respectively. In the test of antifungal activity in mixed culture of the isolate and M. fructicola, inhibition rate was 7.5%, 86.8% and 94.0% in 0.01, 0.1, and 1% concentration of CS containing bacterial cell of the isolate, respectively. On apples (cultivar; Fuji), the control values of the isolate TH04 crude filtrates (0.1 and 1%) were 85.9% and 100%, respectively. The results suggest that the isolate TH04 indicate development possibility as biocontrol agent of brown rot caused by M. fructicola with the study on delivery method and fermentation condition to produce an antifungal compound.

• Journal of Forest and Environmental Science
• /
• v.36 no.3
• /
• pp.207-218
• /
• 2020

Ashwagandha is an important ancient medicinal crops, being affected with many diseases, among which leaf blight caused by Alternaria alternata has become the constraint resulting in huge yield losses. Continuous usage of chemical methods leads to environment, soil and water pollution. Whereas biological control of diseases is long lasting, inexpensive, eco-friendly and harmless to target organisms. In this context, it is aimed to evaluate five Trichoderma strains viz. Trichoderma virens IMI-392430, T. pseudokoningii IMI-392431, T. harzianum IMI-392432, T. harzianum IMI-392433 and T. harzianum IMI-392434 as bio-control efficacy against A. alternata and growth promoting effect in Ashwagandha. All the Trichoderma strains had varied antagonistic effects against the pathogen. In dual culture technique, the strain T. harzianum IMI-392433 showed maximum percentage inhibition of mycelial growth (54.89%) followed by T. harzianum IMI-392432 (53.83%), T. harzianum IMI-392434 (48.94%) and T. virens IMI-392430, (43.62%) against the pathogen, while the least inhibition percentage was observed with the T. pseudokoningii IMI-392431 (36.60%). The culture filtrate of the Trichoderma strain, T. harzianum IMI-392433 recorded highest inhibition on the mycelial growth (39.05%) and spore germination (80.77%) of pathogen and the lowest was recorded in T. pseudokoningii IMI-392431 (20.45 and 50%). Moreover, seeds treated with spore suspension of the strain T. harzianum IMI-392433 reduced the percentages of disease severity index significantly. The strain T. harzianum IMI-392433 also significantly increased seed germination %, seedling vigor and growth of Ashwagandha. The correlation matrix showed that root yield per plant of Ashwagandha had significant and positive correlation with plant height (r=0.726^⁎⁎), number of leaf (r=0.514^⁎⁎), number of primary branch (r=0.820^⁎⁎), number of secondary branch (r=0.829^⁎⁎), fresh plant weight (r=0.887^⁎⁎), plant dry weight (r=0.613^⁎⁎), root length (r=0.824^⁎⁎), root diameter (r=0.786^⁎⁎), root dry weight (r=0.739^⁎⁎) and fresh root weight (r=0.731^⁎⁎). The significant and negative correlation (r=-0.336^⁎⁎) was observed with the root yield and percentages of disease severity index. The study recognized that the T. harzianum IMI-392433 strain performed well in inhibiting the mycelial growth and reduced the percentages of disease severity index of pathogen as well as increased the plant growth in Ashwagandha.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.3
• /
• pp.151-156
• /
• 1994

Acifluorfen-tolerant cell lines of S, ptycanthum were isolated by stepwise selection using suspension culture. Growth of unselected line was completely inhibited at $0.5\;\mu\textrm{M}$, while some selected lines grew at $8\;\mu\textrm{M}$ acifluorfen. After subculturing on acifluorfen-free medium for 4 passages, six of the eleven cell lines screened and maintained their tolerance to $2\;\mu\textrm{M}$ acifluorfen. The regeneration capacity of selected cell lines in Solanum ptycanthum differed depending on the tell line. The acifluorfen tolerance of the somarclones regenerated from acifluorfen-tolerant cell lines differed depending on the somarclone. When plants were heated with $16\;\mu\textrm{M}$ acifluorfen, unselected control plane had over 75% phytotoxicity Many selected cell lines had less phytotoxicity than the seed-grown control plants. Tolerance to acifluorfen was inherited to the self-pollinated progenies. The inheritance patterns differed depending on the clone. Acifluorfen tolerance was inherited as a semidominant trait. Other segregation patterns were also observed. acifluorfen tolerance was recessive and acifluorfen sensitivity was dominant.

• Journal of Periodontal and Implant Science
• /
• v.32 no.3
• /
• pp.489-500
• /
• 2002

The purpose of this study was to evaluate newly fabricated tricalcium phosphate(TCP)/chitosan microgranuls as bone substitutes. TCP/chitosan microgranules were fabricated by dropping TCP-chitosan suspension into the NaOH/ethanol solution. The size of microgranules could be controllable via airflow rate. PDGF-BB was loaded into the fabricated granules via freeze-drying methods(300 ng/20 mg). To evaluate cell proliferation, cultured osteoblasts cell lines(MC3T3-El) was dropped on the BioOss(R), chitosan microgranules, TCP/chitosan microgranules and cultured for 1, 7 , 14, and 28 days. Scanning electron microscopic observation was done after 7 days of culture and light microscopic examination was done after 28 days of culture. PDGF-BB release from the microgranules was tested. Rabbit calvarial defects(8 mm in diameter) were formed and chitosan, TCP/chitosan, PDGF-TCP/chitosan microgranules, and BioGran(R) were grafted to test the ability of new bone formation. At SEM view, the size of prepared microgranules was 250-1000 um and TCP powders were observed at the surface of TCP/chitosan microgranules. TCP powders gave roughness to the granules and this might help the attachment of osteoblasts. The pores formed between microgranules might be able to allow new bone ingrowth and vascularization. There were no significant differences in cell number among BioOss(R) and two microgranules at 28 day. Light and scanning electron microscopic examination showed that seeded osteoblastic cells were well attached to TCP/chitosan microgranules and proliferated in a multi-layer. PDGF-BB released from TCP/chitosan microgranules was at therapeutic concentration for at least 1 week. In rabbit calvarial defect models, PDGF-TCP/chitosan microgranules grafted sites showed thicker bone trabeculae pattern and faster bone maturation than others. These results suggested that the TCP/chitosan microgranules showed the potential as bone substitutes.

• Plant Biotechnology Reports
• /
• v.2 no.2
• /
• pp.163-169
• /
• 2008

Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

• Reproductive and Developmental Biology
• /
• v.30 no.1
• /
• pp.47-52
• /
• 2006

This study was conducted to examine whether the parthenogenetic mouse embryonic stem (P-mES) cells can differentiate into functional cardiomyocytes in vitro similar to (mES) cells. p-mES04 and IVF-derived mES03 cells were cultured by suspension culture for 4 days. The formed embryoid bodies (EBs) were treated with 0.75% dimethyl-sulfoxide (DMSO) for further 4 days (4-/4+), and then plated onto gelatin coated culture dish. The appearance of contracting cardiomyocytes from the P-mES04 and mES03 cells was examined for 30 days. The highest cumulative frequency was detected at days 13 (69.83%) and 22 (61.3%), respectively. By immunocytochemistry, beating P-mES04 cells were positively stained with muscle specific anti-sarcomeric a-actinin Ab and cardiac specific anti-cardiac troponin I Ab similar to contracted mES03 cells. When the expression of cardiac muscle-specific genes was analyzed by RT-PCR, beating P-mES04 cells were expressed cardiac specific L-type calcium channel, a1C, cardiac myosin heavy chain a, cardiac muscle heavy polypeptide $7{\beta}$, GATA binding protein 4 and atrial natriuretic factor, but not expressed skeletal muscle specific L-type calcium channel, a1S, which was similar to male adult heart cells and mES03-derived beating cardiomyocytes. The result demonstrates that the P-mES cells can be used as an alternative for the study on the characteristic analysis of in vitro cardiomyocyte differentiation from the ES cells.

• Fisheries and Aquatic Sciences
• /
• v.23 no.4
• /
• pp.7.1-7.8
• /
• 2020

Background: To minimize the use of antibiotics and to obtain a more sustainable fish culture and aquaculture industry, development of alternative natural source of immunostimulant to replace antibiotic in aquafeed is highly needed. Objective: Dietary inclusion effect of yacon (YC), ginger (GG), and blueberry (BB) on growth, body composition, and disease resistance of black rockfish against Vibrio anguillarum was compared to ethoxyquin (EQ). Methods: Four hundred eighty juvenile (an initial weight of 4.2 g) fish were randomly distributed into 12 of 50 L flowthrough tanks (forty fish per tank). Four experimental diets were prepared; the control (Con) diet with 0.01% EQ inclusion, and YC, GG, and BB diets at 1% each additive inclusion. Each additive was included into the experimental diets at the expense of wheat flour. Each diet was assigned to triplicate tanks of fish and hand-fed to satiation twice daily for 8 weeks. At the end of 8-week feeding trial, 20 fish from each tank fish were artificially infected by intraperitoneal injection with 0.1 mL of culture suspension of pathogenic V. anguillarum containing 3.3 × 10⁶ cfu/mL respectively. Fish were monitored for the following 8 days after V. anguillarum infection and dead fish were removed every 6 h for the first 4 days and 12 h for the rest of the study. Results: Weight gain, specific growth rate (SGR), and feed efficiency ratio (FER) of fish fed the YC diet was higher than those of fish fed all other diets. However, feed consumption, protein efficiency ratio, and protein retention was not affected by dietary additive. Moisture, crude protein, and crude lipid content of the whole body of fish were affected by dietary additive. Analysis of the Kaplan-Meier survival curves showed that survival of fish fed the YC, BB, and GG diets was higher than the Con diet. Conclusion: Oral administration of YC can improve not only weight gain, SGR, and FER of black rockfish, but lower mortality of rockfish at occurrence of V. anguillarum.

• The Korean Journal of Malacology
• /
• v.28 no.1
• /
• pp.1-6
• /
• 2012

We studied that the effect of stocking density on growth and survival rate of the scallop, Chlamys farreri (initial shell height 32.97 mm and total weight 5.63 g) from June 2002 to October 2003 in the west coast of Korea. C. farreri is usually the west coast of Korea and northern China in the natural habitat of the coastal species. Range of surface water temperature in the study area was $4.3^{\circ}C$ to $25.3^{\circ}C$, salinity 29.2 psu to 32.1 psu, dissolved oxygen 5.32 mg/L to 7.51 mg/L and pH was 7.84 to 8.12, respectively. The stocking densities were 20, 30, 40 and 50 individuals per a compartment of suspension cage in culture beginning. After 16 months from initiation, ranges of shell height and mean total weight were from 64.35 mm to 76.23 mm and from 41.53 g to 64.85 g. The survival rate was from 82% to 100%. The growth rate of the scallop was negatively correlated with the stocking density. The growth of the shell height and total weight were decreased with decreasing of water temperature. Most of mortality of scallop occurred during March to April and September to October. Survival rate in the stocking density was decreased by density increase and was highest in 20 individual a compartment.

• Horticultural Science & Technology
• /
• v.31 no.5
• /
• pp.626-632
• /
• 2013

In the course of a developing screening method for resistant radish to Fusarium oxysporum f. sp. raphani, we found that the fungus produces phytotoxic compound against Raphanus sativus. The culture filtrate of F. oxysporum f. sp. raphani KR1 represented the strongest phytotoxicity when the fungus was incubated in the malt extract broth with 150 rpm at $25^{\circ}C$ for 14 days. Under bioassay-guided purification, we isolated a substance from liquid culture of F. oxysporum f. sp. raphani KR1, with phytotoxic effect against R. sativus. The compound was identified as fusaric acid by mass and nuclear magnetic resonance spectral analyses. Phytotoxicity of the compound against cruciferous vegetable crops, including radish, cabbage, and broccoli, was investigated. Fusaric acid represented phytotoxicity on radish seedlings by concentration dependant manner. And the phytotoxin demonstrated strong phytotoxicity on the resistant cultivars as well as susceptible cultivars of radish to F. oxysporum f. sp. raphani. In addition, fusaric acid isolated from the fungus also showed a potent phytotoxic efficacy against non-host Brassicaceae crops of the fungus such as cabbage and broccoli. The results demonstrate that fusaric acid produced by F. oxysporum f. sp. raphani is non-host-specific toxin and for screening of resistant radish to the fungal pathogen, spore suspension of the fungus without the phytotoxin has to be used.

• Korean Journal of Veterinary Service
• /
• v.44 no.3
• /
• pp.149-155
• /
• 2021

In virucidal efficacy testing, the chemical inactivation cannot be determined for all viruses due to the difficulties or the inability to culture sufficiently or the risk of exposure to the viruses. Therefore, disinfectants against these viruses could be evaluated by different methods and surrogate viruses are used as alternative. In this study we developed a method for efficacy testing of veterinary disinfectants using one of the candidate surrogate viruses, bacteriophage MS2, as part of the research on the selection of surrogate viruses for efficiency of efficacy testing of veterinary disinfectants. This method is based on the Animal and Plant Quarantine Agency (APQA) guidelines for efficacy testing of veterinary disinfectants. Bacteriophage and disinfectant are reacted in suspension in accordance with the APQA guidelines and then a newly established double agar layer method is applied for the efficacy test. The double agar layer method is summarized as follows: 1) The bottom agar with 1.5% agar is boiled and cooled before poured into petri dishes at volume of 20 mL, and dried under biological safety cabinet. 2) The top agar with 0.7% agar is boiled and kept at 50℃ before E. coli culture was seeded. 3) The serially diluted bacteriophage MS2-disinfectant mixtures 0.05 mL and E. coli host 0.01 mL (OD₆₀₀ 0.2~0.3) are mixed with 5 mL of top agar and incubate them at 50℃ for 5 min for reaction. 4) The resulting mixture is poured over top of a bottom agar plate and rocked sufficiently to ensure that the top agar covers the entire surface of the bottom agar. 5) The double agar layer is then placed under biological safety cabinet to allow the agar layer to solidify and subsequently incubated at 37℃ for 24 hr. 6) Following incubation, the plates may be inspected for plaques and record results.