• Journal of Plant Biotechnology
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• 제7권4호
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• pp.259-265
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• 2005

Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.

• Mycobiology
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• 제38권1호
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• pp.17-25
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• 2010

The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

• 한국균학회지
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• 제13권4호
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• pp.221-224
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• 1985

ATCC로 부터 구입한 11종(種)과 우리나라의 토양에서 분리 동정한 6종(種)의 Trichoderma를 실험(實驗) 재료(材料)로 하였다. 2% malt extract agar 배지(培地)에서 배양(培養)하면서 균사의 영양핵에서의 핵분열을 관찰하였다. 염색(染色)은 Giemsa staining solution으로 염색(染色)하였고 그들의 염색체(染色體) 수(數)를 세었다. 염색체의 형태(形態)는 다른 균류와 마찬가지로 점(點) 모양(dot-like)의 것이 대부분(大部分)이었다. 염색체(染色體)의 크기는 매우 작아서 $0.2{\sim}0.3\;{\mu}$정도이다. 염색체(染色體)의 수는 $5{\sim}10$개로서 n=5인 종(種)이 2종(種)(Trichoderma pseudokoningil, T. saturnisporum)이고, n=6인 종(種)은 8종(種)(T. koningil, T. viride, T. hamatum, T. longibranchiatum, T. sp. No.40, T. sp. No.58 T. sp. No.237, T. sp. No.241)으로 가장 많았다. n=7인 종(種)은 2종(種)(T. reesei, T. harzianum, T. polysporium, T. virigatum, T. sp. No.196)으로 관찰되었다. 이상의 17종(種)을 비교 검토하여 보면 Trichoderma속의 기본(基本) 염색체(染色體) 수(數)는 $4{\sim}10$개 사이에 있을 것으로 생각되며 그 중에서 가장 많은 수(數)를 나타탠 n=6을 기본(基本) 염색체수(染色體數)로 추론하였다.

• Applied Biological Chemistry
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• 제39권1호
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• pp.16-19
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• 1996

청주의 주질을 향상시키기 위한 전보의 연구에서, 유용효모로서 분리된 균주에 대하여 균학적 성질을 조사하고, 일부의 사항에 대하여는 일본청주효모와 그 특성을 비교하였다. 분리효모 KP-16, KP-21 및 KP-54 균주는 모두 saccharomyces cerevisae로 동정되었고, TTC 염색성이 pink계이었다. 맥아즙 배지에서의 피막(被膜) 형성능은 KP-16 및 KP-21 균주는 약하였으나, KP-54균주는 강하였다. 당의 발효성과 탄소원의 자화성은 KP-16과 KP-21 균주는 동일하였고, KP-54 균주는 다소 달랐다. 분리효모는 모두 ${\alpha}-methyl-D-glucoside$를 발효도 자화도 하지 못하였으며, 이는 현재 사용되고 있는 일본청주효모와 다른 점 중의 하나였다. 비타민 요구성에 있어서 분리균주는 모두 biotin과 pantothenate를 요구하였으며, biotin을 요구하는 점은 일본청주효모와는 다른 특성이었다. 내알코올성은 분리균주 모두 일본청주효모 K-7및 K-9균주보다 강하였다.

• The Plant Pathology Journal
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• 제16권1호
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• pp.25-28
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• 2000

The effect of ionic osmotic potential (Ψ$\pi$), and matric potential (Ψm) in the range of -0.2 to -4.0 Mpa on mycelial growth of three species of Pleurotus (P.florida, P.ostrenatus and P.safor-caju) were determined over a range of temperature (15-3$0^{\circ}C$) on a 2% malt extract agar medium and compared with the Ψ$\pi$ effect on growth of two strains of Trichoderma green mould. With the ionic solute KCl, optimun Ψ$\pi$for growth was -0.2 MPa for P.floreda and in the range of -0.2 to -0.5 MPa, with slight growth at -3.0 MPa and with nogrowth at -4.0 MPa. Of the species of Pleurotus, P.florida grew signigicantly slower than the other two species. Growt of the species of Pleurocus was significantly slower when water potential (Ψ$\omega$) was modified matrically with polyethylene glycol (PEG) 8000 then osmotically with KCl. They were also more sensitive to changes in Ψm than Ψ$\pi$The optimum Ψm of the Pleurotus was -0.5 Ψm, with no growth below -3.0 MPa. Of the species of Pleurotus, P.florida was most sensitive and P.sajor-caju was more tolerent to lowered Ψ$\pi$,but P.sajor-caju was most sensitive to lowered Ψm. The growth rate of the Trichoderma green mould strains was much faster than that observed for the Pleurotus spp. Optimum growth for bot strains of Trichoderma was in the range of -0.2 to -0.5 MPa. Strain CNU 503 was more tolerant to water stress than strain CNU 501. Both strains were able to grow up to 30% of optimum growth at -4.0 MPa at 25-3$0^{\circ}C$.

• 한국버섯학회지
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• 제5권2호
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• pp.81-85
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• 2007

본 연구는 번데기동충하초를 곡물배지를 이용하여 자실체를 형성하는 실험으로 액체종균제조에 적합한 조건은 맥아추출배지를 농도 0.7%, pH 5~5.5,으로 조정하였을 때 균사의 건중량은 $3.9{\sim}4.1mg/m{\ell}$로 양호 하였다. 자실체 발생 최적 환경조건은 습도 95%, 온도 $17^{\circ}C$, 조도 600 lx, 이산화탄소농도 500ppm하였을 때, 곡물배지 일반현미와 흑미에서 24.3g/병, 20.5g/병의 자실체가 형성되었다.

• 한국환경농학회지
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• 제26권1호
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• pp.36-41
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• 2007

PDA-calcium phosphate 평판배지를 이용하여 경상남북도 일대의 토양으로부터 인산가용화능이 우수한 사상균을 분리하고 배양특성을 조사하였다. 최종적으로 분리한 인산가용화능이 우수한 사상균 PS-104 균주를 선발하여 형태적 특성을 조사한 결과 이 균주의 분생자는 비가지형으로 끝이 돌출되어 있으며, 균사가 격막으로 형성되는 등의 특징을 갖는 Aspergillus sp. PS-104 균주로 동정되었다. 이 균주의 인광석 분해능이 최대가 되는 최적 배양온도와 pH는 각각 $30^{\circ}C$와 pH 7.0이었다. 이와 같은 배양조건에서 tricalcium phosphate, aluminium phosphate, hydroxyapatite, 인광석 등의 난용성 인산염에 대한 이 균주의 인산가용화능을 살펴보면 최대 인산가용화능은 tricalcium-phosphate 처리구(1,900 ppm)에서, 최소 인산가용화능은 hydroxyapatite 처리구(320 ppm)에서 확인되었다. 또한 이 균주의 인산가용화능은 질소원으로 첨가한 ammonium 염보다 nitrate 염에서 높게 나타났다.

• The Plant Pathology Journal
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• 제15권3호
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• pp.152-157
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• 1999

An antibiotic compound was isolated from the culture of an antagonist against Fusarium oxysporum f. sp. sesami, Bacillus polymyxa strain KB-8, and tested for the control of Fusarium wilt of sesame in greenhouse conditions. Optimum conditions for culturing the antagonist to obtain the maximum antibiotic activity were determined using different culture media, initial medium acidity, and incubation periods for which yeast -malt extract agar with the initial acidity of pH 5 and over 13 days culture were best. Antibiotic substances extracted by methanol had 2 main fractions, KB-8A and KB-8B, in thin layer chromatography (OLC) with Rf values of 0.35 and 0.67 in a solvent system of chloroform : methanol = 7 : 3. The fraction KB-8A wa purified further by XAD-2, silica gel and Sephadex LH-20 column chromatography, and crystalization. Its minimum inhibitory concentrations (MICs) were $12.8\mu\textrm{g}$/ml for F. oxysporum and Alternaria mali, $6.4\mu\textrm{g}$/ml for Colletotrichum gloeosporioides and Rhizoctonia solani, and $3.2\mu\textrm{g}$/ml for Phytophthora capsici. Soil drenching of antibiotic KB-8A in the concentrations of $13.0\mu\textrm{g}$/ml and $26.0\mu\textrm{g}$/ml effectively inhibited the Fusarium wilt of sesame in a greenhouse test, which appeared to be comparable to the fungicide benlate of $6.5\mu\textrm{g}$ a. i./ml.

• Mycobiology
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• 제37권1호
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• pp.28-30
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• 2009

This study was aimed at evaluating the growth promoting effect of symbiotic algal polyol (ribitol) and plant hormones on the lichen-forming fungi (LFF), Ramalina farinacea (CH050010 and 40403) and Ramalina fastigiata. The addition of ribitol to basal (malt-yeast extract) medium enhanced the relative growth rates of all three LFF. R. farinacea (CH050010), R. farinacea (40403) and R. fastigiata (H06127) showed 35.3%, 29.0% and 29.3% higher growth rates, respectively, compared to the control. IBA (indole-3-butyric acid) and TIBA (2,3,5-tridobenzoic acid) also increased growth rates of the LFF by 34 to 64% and 7 to 28%, respectively, compared to the control. The combination of ribitol with IBA or TIBA synergistically increased the growth of all LFF. For example, ribitol and IBA treatments increased growth rates of R. farinacea (CH050010), R. farinacea (40403) and R. fastigiata (H06127) by 79.4%, 40.3% and 72.8% in, respectively, compared to those grown on the basal medium. The stimulating effect of ribitol and IBA on the LFF growth induced vertical development of the fungal mass in culture. We suggest that lichen-forming fungal growth of Ramalina lichens can be stimulated aposymbiotically by supplementing polyols and plant hormones to the basal medium in the mass production of lichen secondary metabolites under large scale culture conditions.

• 한국균학회지
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• 제40권2호
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• pp.93-97
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• 2012

본 연구는 큰느타리버섯(Pleurotus eryngii)에 감마 방사선을 조사하여 형태적, 생리적 특성이 향상된 새로운 큰느타리버섯 품종을 개발하기 위하여 수행되었다. 큰느타리버섯에서 분리한 원형질체에 0.25-1.25 KGy의 감마 방사선을 조사하였다. 원형질체는 YPMGA 배지에서 80%의 치사율을 나타냈고 이들 중 무작위로 500개의 변이체를 선발하여 PDA 배지에 배양하였다. 그 결과, 대부분의 변이체가 대조구와 비슷한 형태와 생장률을 나타냈으며, 이 중 100개의 변이체가 대조구와 약간의 차이를 보였다. 67개 변이체에 대해 cellulase 활성과 laccase 활성을 조사하였다. 그 결과, 대체적으로 대조구와 유사한 효소 활성을 나타내었고, 5개의 변이체가 대조구에 비해 더 높은 cellulase 활성을 나타내었다. 또한 감마 방사선 조사 변이체의 유전적 변이를 조사하기 위하여 UFPF-PCR을 이용하여 다형성을 조사하였다.