• Mycobiology
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• 제48권2호
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• pp.122-132
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• 2020

Citric acid is a commercially valuable organic acid widely used in food, pharmaceutical, and beverage industries. In this study, 260 yeast strains were isolated from soil, bread, juices, and fruits wastes and preliminarily screened using bromocresol green agar plates for their ability to produce organic acids. Overall, 251 yeast isolates showed positive results, with yellow halos surrounding the colonies. Citric acid production by 20 promising isolates was evaluated using both free and immobilized cell techniques. Results showed that citric acid production by immobilized cells (30-40 g/L) was greater than that of freely suspended cells (8-19 g/L). Of the 20 isolates, two (KKU-L42 and KKU-L53) were selected for further analysis based on their citric acid production levels. Immobilized KKU-L42 cells had a higher citric acid production rate (62.5%), while immobilized KKU-L53 cells showed an ~52.2% increase in citric acid production compared with free cells. The two isolates were accurately identified by amplification and sequence analysis of the 26S rRNA gene D1/D2 domain, with GenBank-based sequence comparison confirming that isolates KKU-L42 and KKU-L53 were Candida tropicalis and Pichia kluyveri, respectively. Several factors, including fermentation period, pH, temperature, and carbon and nitrogen source, were optimized for enhanced production of citric acid by both isolates. Maximum production was achieved at fermentation period of 5 days at pH 5.0 with glucose as a carbon source by both isolates. The optimum incubation temperature for citric acid production by C. tropicalis was 32 ℃, with NH₄Cl the best nitrogen source, while maximum citric acid by P. kluyveri was observed at 27 ℃ with (NH₄)₂ SO₄ as the nitrogen source. Citric acid production was maintained for about four repeated batches over a period of 20 days. Our results suggest that apple and banana wastes are potential sources of novel yeast strains; C. tropicalis and P. kluyveri which could be used for commercial citric acid production.

• Journal of Microbiology and Biotechnology
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• 제22권2호
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• pp.207-214
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• 2012

Laccase- and peroxidase-free tyrosinase has commercial importance in the production of L-3, 4-dihydroxyphenylalanine (L-DOPA), which is mainly used in the treatment of Parkinson's disease. In the present study, isolation of an actinomycetes microbial strain capable of producing only tyrosinase is reported. Among all soil isolates, three individual colonies revealed black color around the colony in the presence of tyrosine. Further screening for laccase and peroxidase activities using syringaldazine denoted that one of the isolates, designated as RSP-T1, is laccase and peroxidase negative and produces only tyrosinase. The microbe was authenticated as Streptomyces antibioticus based on 16S ribotyping. Effective growth of this isolate was noticed with the use of medium (pH 5.5) containing casein acid hydrolysate (10.0 g/l), $K_2HPO_4$ (5.0 g/l), $MgSO_4$ (0.25 g/l), L-tyrosine (1.0 g/l), and agar (15 g/l). The scanning electron micrograph depicted that the microbe is highly branched and filamentous in nature. The enzyme production was positively regulated in the presence of copper sulfate. The impact of different fermentation parameters on tyrosinase production depicted that the maximized enzyme titer values were observed when this isolate was grown at 6.5 pH and at $30^{\circ}C$ temperature under agitated conditions (220 rpm). Among all the studied physiological parameters, agitation played a significant role on tyrosinase production. Upon optimization of the parameters, the yield of tyrosinase was improved more than 100% compared with the initial yield.

• Mycobiology
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• 제44권4호
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• pp.283-290
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• 2016

A double-stranded RNA (dsRNA) mycovirus was detected in malformed fruiting bodies of Pleurotus ostreatus strain ASI2792, one of bottle cultivated commercial strains of the edible oyster mushroom. The partial RNA-dependent RNA polymerase (RdRp) gene of the P. ostreatus ASI2792 mycovirus (PoV-ASI2792) was cloned, and a cDNA sequences alignment revealed that the sequence was identical to the RdRp gene of a known PoSV found in the P. ostreatus strain. To investigate the symptoms of PoV-ASI2792 infection by comparing the isogenic virus-free P. ostreatus strains with a virus-infected strain, isogenic virus-cured P. ostreatus strains were obtained by the mycelial fragmentation method for virus curing. The absence of virus was verified with gel electrophoresis after dsRNA-specific virus purification and Northern blot analysis using a partial RdRp cDNA of PoV-ASI2792. The growth rate and mycelial dry weight of virus-infected P. ostreatus strain with PoV-ASI2792 mycovirus were compared to those of three virus-free isogenic strains on 10 different media. The virus-cured strains showed distinctly higher mycelial growth rates and dry weights on all kinds of experimental culture media, with at least a 2.2-fold higher mycelial growth rate on mushroom complete media (MCM) and Hamada media, and a 2.7-fold higher mycelial dry weight on MCM and yeast-malt-glucose agar media than those of the virus-infected strain. These results suggest that the infection of PoV mycovirus has a deleterious effect on the vegetative growth of P. ostreatus.

• 한국자원식물학회지
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• 제17권3호
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• pp.331-338
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• 2004

본 연구에서는 카네이션과 고구마의 무병 종묘를 생산하고자 정단분열조직배양을 실시하였으며, 생산된 배양종묘의 바이러스 감염여부를 진단하기 위해 ELISA방법을 이용하여 주요 바이러스를 검정하였다. 카네이션의 정단분열조직 배양에 공시 한 'Roland' 등 4품종 중 투명화 현상이 없이 가장 정상적인 잎이 발생하고 절간신장이 이루어지며 뿌리의 발생도 양호한 생장반응을 보인 품종은 석죽계통인 'Giant Gipsy' 이었으며 MS기본배지에 NAA 0.05 ㎎/L와 BA 0.1㎎/L 혹은 kinetin 1.0㎎/L와 조합한 구에서 shoot의 재생이 양호하였다. 배양7주일 후에는 explant 1개체에서 평균 5-6개, 많은 것은 10개체 이상의 multiple shoot가 형성되었고 기내 마디삽목법에 의하여 대량증식할 수 있었다. 투명화현상이 심한 품종은 'Casha'와 'Desio'로 나타났는데 투명화 현상은 kinetin 첨가보다는 BA첨가구에서 현저하게 증가하였다. 카네이션 기내 배양종묘의 바이러스 감염여부는 ELISA방법을 이용하여 CarMV와 CarRSV를 신속하게 다량의 시료를 진단할 수 있었다. 그 결과 정단분열조직의 배양을 통해 65.2%에서 무병 주가 생산되었음을 확인할 수 있었다.

• The Plant Pathology Journal
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• 제31권3호
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• pp.278-289
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• 2015

Indigenous strains of Trichoderma species isolated from rhizosphere soils of Tea gardens of Assam, north eastern state of India were assessed for in vitro antagonism against two important tea fungal pathogens namely Pestalotia theae and Fusarium solani. A potent antagonist against both tea pathogenic fungi, designated as SDRLIN1, was selected and identified as Trichoderma viride. The strain also showed substantial antifungal activity against five standard phytopathogenic fungi. Culture filtrate collected from stationary growth phase of the antagonist demonstrated a significantly higher degree of inhibitory activity against all the test fungi, demonstrating the presence of an optimal blend of extracellular antifungal metabolites. Moreover, quantitative enzyme assay of exponential and stationary culture filtrates revealed that the activity of cellulase, ${\beta}$-1,3-glucanase, pectinase, and amylase was highest in the exponential phase, whereas the activity of proteases and chitinase was noted highest in the stationary phase. Morphological changes such as hyphal swelling and distortion were also observed in the fungal pathogen grown on potato dextrose agar containing stationary phase culture filtrate. Moreover, the antifungal activity of the filtrate was significantly reduced but not entirely after heat or proteinase K treatment, demonstrating substantial role of certain unknown thermostable antifungal compound(s) in the inhibitory activity.

• 동의생리병리학회지
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• 제18권5호
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• pp.1437-1442
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• 2004

In this study, lotus root(Nelumbo nucifera G.) and omija(Schizandrae fructus), which have been used in oriental medicine and folks remedy, were examined to apply to functional foods. We prepared yanggaeng with lotus root, sugar, oligosaccharide, agar, omija extract solution and analyzed the nutritional composition(moisture, protein, fat, ash, crude fiber, carbohydrate, free sugar and minerals). Also we investigated texture profile and evaluated sensory characteristics of developed yanggaengs and 8 commercial ones. Potassium and crude fiber contents of yanggaeng with lotus root were higher than the commercial yanggaengs. Cohesiveness, springiness and gumminess of lotus root yanggaeng were the same levels of commercial yanggaengs. Sensory evaluation with the ones, showed that the lotus yanggaeng was more desirable than the commercial ones. Yanggaeng with lotus had good scores in texture profile and sensory evaluation compared with commercial yanggaengs. These results demonstrated that lotus root hadsufficient values to use a foodstuff for yanggaeng.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.21-25
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• 2006

Phragmites australis (reed) has received much attention as being one of the principle emergent aquatic plants for treating industrial and civil wastewater. Plant regeneration via plant tissue culture in p. australis was investigated. Three types of callus were identified from seeds on N6 medium plus 4.5 UM 2,4-dichlorophenoxyacetic acid (2,4-D). Yellow compact type showed the best redifferentiation, whereas white compact type and yellow friable were not competent to differentiate into plane. Solid medium culture was better than liquid suspension culture for enhancing callus growth when N6 medium supplemented with 4.5 ${\mu}M$ 2,4-D was used. Phytagel, as a gelling agent, was superior to agar in plant regeneration on N6 medium, supplemented with 9.4 ${\mu}M$ kinetin and 0.54 ${\mu}M$ $\alpha$-naphthaleneacetic acid (NAA). Transfer of the plantlets regenerated from kinetin and NAA-supplemented N6 medium to growth regulator-free MS medium enhanced the further development of the plantlets. Plantlets on subsequently grown to maturity when tansferred to potting soil. The regenerated plants exhibited morphologically normal. The system for plant regeneration of P. australis enables to propagate elite lines on a large scale for water purification in the ecosystem

• Current Research on Agriculture and Life Sciences
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• 제31권2호
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• pp.94-100
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• 2013

An efficient plant regeneration protocol is developed for a standard-type chrysanthemum. When petal segments derived from flower buds (4 or 8cm in diameter) were used as the culture material, the highest shoot regeneration frequency (96%) was obtained on a Murashige and Skoog (MS) medium supplemented with 0.5 mg/L IAA, 2 mg/L BA, 3% sucrose, and a 0.8% agar. Pre-culturing the explants under dark conditions for 14 days produced better results for the shoot regeneration frequency than the explants cultured under a continuous 16 h photoperiod ($40{\mu}molm^{-2}s^{-1}$). The shoot regeneration frequency ranged from 19.0% for the Shinmato cultivar to 89.1% for the Baeksun cultivar. Activated charcoal (0.2%) enhanced the root formation of the regenerated shoots in a hormone-free MS medium. The rooted plantlets were acclimatized and successfully established in a greenhouse.

• Advances in Traditional Medicine
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• 제8권4호
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• pp.395-399
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• 2008

Methanol and aqueous extract of leaves of Trema orientalis Linn. were subjected to the potential antioxidant and antibacterial activities. The pharmacological interest of this plant coupled with traditional use (antidiarrhoeal, antiseptic, analgesic etc) prompted to test for antioxidant and antibacterial activities. The antioxidant potential of the methanolic extract was determined on the basis of their scavenging activity of the stable 1,1-diphenyl-2-picryl hydrazyl free radical. $IC_{50}$ of the methanol extract of T. orientalis was $110.25\;{\mu}g/ml$ which indicated the strong antioxidant activity of the plant. However the aqueous extract showed mild antioxidant activity. In case of antibacterial activities test, the extract was subjected for its effectiveness against both Gram-positive and Gram-negative bacteria in agar diffusion method. The zones of inhibition produced by the crude methanol and aqueous extract against few sensitive strains were measured and compared with those of standard antibiotic Gentamycin. It is evident that both extracts are active against the bacteria at low concentrations. The obtained results provide a support for the use of this plant in traditional medicine and suggest its further advance investigation.

• 한국약용작물학회지
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• 제21권2호
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• pp.142-147
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• 2013

Gastrodia elata has been cultivated as an important medicinal resources to treat various human diseases. One of the major problems associated with its field production is the degeneration of seed tubers, which is mainly caused by soil-borne pathogens. This study was conducted to produce disease-free seed tubers by the development of in vitro micropropagation method. First, tubers of G. elata were treated with $HgCl_2$ prior to culturing in vitro. Among various culture medium tested, water agar (WA) and WPM medium were the most effective on the induction and growth of vegetative stems. NAA ($0.1mg/{\ell}$) or TDZ ($1.0mg/{\ell}$) in WA medium showed better growth of vegetative stems compared to other plant hormones. Finally the induction and growth of vegetative stems were better in the dark compared to the light condition. In this study, we established an in vitro micropropagation system of G. elata, which might be an efficient way to increase the yield and quality of G. elata tubers in the field production.