• Journal of Plant Biotechnology
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• 제35권2호
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• pp.127-132
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• 2008

In bioreactor cultures of plants, inoculum density is an important factor affecting growth and proliferation of the plantlets. To maximize shoot growth and proliferation of grape rootstock '5BB' in bioreactors, inoculum density varied at 15, 30, 45 and 60 single nodes in a 3-liter scale balloon type bioreactor, respectively and cultured for 40 days. Results suggested that the growth and the photosynthesis of the plantlet were greatly affected by inoculum density in the bioreactor. The inoculum density of 45 nodes resulted in the greatest growth (910.4 mg/shoot FW, 764.4 mg/root FW) followed by 30 nodes. $CO_2$ assimilation rate, stomatal conductance, transpiration rate of the plantlet were also highest at the inoculum density of 45 nodes. Significant reduces in shoot and root growth (426.5 mg/shoot FW, 248.4 mg/root FW) were observed at the inoculum density of 60 nodes. When the inoculum density decreased by 15 nodes, plantlets were malformed due to hyperhydricity, resulting in the highest transpiration rate and the lowest $CO_2$ assimilation rate. The plantlets stressed by the inoculum density at 15 nodes and 60 nodes showed larger number and irregular shape of stomata compared to the plantlets inoculated with 45 nodes.

• The Plant Pathology Journal
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• 제27권4호
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• pp.333-341
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• 2011

Two antifungal bacteria were selected from forest soils during the screening of microorganisms antagonistic to Cylindrocarpon destructans, a cause of ginseng root rot. The antifungal bacteria were identified as Bacillus subtilis (I4) and B. amyloliquefaciens (yD16) based on physiological and cultural characteristics, the Biolog program, and 16S rRNA gene sequencing analyses. Antagonistic activity of both bacterial isolates to C. destructans increased with increasing temperature. More rapid starch hydrolytic activity of the bacteria was seen on starch agar at higher temperatures than at lower temperatures, and in the higher density inoculum treatment than in the lower density inoculum treatment. The bacterial isolates failed to colonize ginseng root the root tissues inoculated with the bacteria alone at an inoculum density of $1{\times}10^6$ cfu/ml, but succeeded in colonizing the root tissues co-inoculated with the bacteria and C. destructans. Scanning electron microscopy showed that the pathogen was damaged by the low-density inoculum treatment with the bacterial isolates as much as by the high-density inoculum treatment. Both bacterial isolates were more effective in reducing root rot when they were treated at a concentration of $1{\times}10^6$ cfu/ml than at $1{\times}10^8$ cfu/ml. Also, only the former treatment induced prominent wound periderm formation, related to structural defense against pathogen infection. The results suggest that the bacterial antagonists may have high potential as biocontrol agents against ginseng root rot at relatively low-inoculum concentrations.

• Journal of Plant Biotechnology
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• 제6권4호
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• pp.265-268
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• 2004

Cell growth and ginseng saponin production by large-scale suspension (bioreactor) cultures of Panax ginseng were investigated under various inoculum sizes. Cell growth was low at an inoculum size of 40 g FW/L, and the maximum cell growth was obtained with increasing inoculum size up to 100 g FW/L. The cell density of 333 g FW/L and 12.7 g DW/L was obtained at inoculum size of 100 g FW/L after 30 days of cultivation. Maximum saponin production of $4.40\;\cal{mg/g}$ DW was achieved at 60 g FW/L of inoculum size. Thus, inoculum size 60 g FW/L was suitable for optimum biomass accumulation as well as saponin production during bioreactor cultivation of ginseng suspension cells.

• 한국식물병리학회지
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• 제14권6호
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• pp.594-597
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• 1998

The optimal quantity of inoculum was determined to screen resistance of garlic cultivars against Sclerotium cepivorum and 30 cultivars was tested. The growth of the pathogen in detached roots, Disease incidence was increased when the inoculum density was raised form 10 to 100 sclerotia. The optimal inoculum density to differentiate resistance or susceptibility of garlic cultivars was seemed to be 50 sclerotia. The cultivars collected from England, Japan, Nepal and Turkey, and cultivars such as common red, PI1356104 and PI135693 were less than the other cultivars in their disease incidence. The growth of S. cepivorum in detached roots varied from 23 to 33 mm according to garlic cultivars. There was no relationship between the disease incidence and the growth in detached roots. The sclerotial germination was increased significantly when root extract was extract was added. The addition of only distilled water resulted in 13% germination, but the addition of 0.25 g of root extract in 100 ml distilled water resulted in more than 85% germination. There was no difference in the stimulation of sclerotial germination among cultivars which showed different resistance.

• Journal of Plant Biotechnology
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• 제36권1호
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• pp.7-12
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• 2009

본 실험은 음나무 배발생 세포의 증식 및 체세포배 발달을 위한 액체 현탁 배양조건의 확립을 위해 수행되었다. 배발생 세포의 생장율은 접종 밀도가 증가할수록 감소하였다. 배발생 세포의 증식에 가장 효과적인 접종 밀도는 0.1 g/100 ml 로서 이 농도에서 세포의 생장율이 가장 높았다. 배양기간에 따른 배발생 세포의 생장 패턴 및 세포 주기 (G1, S, G2/M) 분석 결과, 세포의 생장은 배양 5일 후부터 증가가 시작되어 15일 까지 급격히 생장하였으며 그 이후에는 점차 감소하였다. 배양 기간 별 세포 주기 (cell cycle)의 변화가 명확하게 관찰되어 배양 5일째 5기는 초기의 5.5% 에서 11.7%로 두 배정도 증가하였으며, 배양 15일 이후부터는 다시 초기의 세포 주기로 되돌아가면서 안정화되는 것으로 나타났다. 따라서 음나무 배발생 세포의 현탁배양은 15일의 주기로 배양하는 것이 증식에 가장 효과적인 것으로 생각되었다. 한편 배발생 세포에서 체세포배의 유도는 배양초기의 접종 밀도가 중요한 것으로 나타났다. 0.5 g/L 의 낮은 밀도로 접종 시에는 65% 이상의 어뢰형 배가 유도된 반면 접종 밀도가 높아질수록 어뢰형으로의 배발달은 급격히 감소하였다. 초기의 접종 밀도가 증가할수록 특히 어뢰형 배의 발달은 지연되었으나 구형 및 심장형 배의 유도는 초기 접종밀도에 영향을 받지 않았다. 이상의 실험결과로 음나무 액체 배양 시 초기 접종 밀도를 조절함으로써 배발생 캘러스의 증식 및 체세포배를 효과적으로 유도할 수 있었으며 이는 체세포배 생산을 위한 배양 기간의 단축이 가능함을 보여주는 결과이다.

• 생명과학회지
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• 제14권4호
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• pp.689-695
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• 2004

2003년 11월 경상남도 부림면 신반리의 결구상추 재배 플라스틱 하우스내에서 늦가을부터 밑둥썩음병이 발생하여 수확기인 11월에 평균 5.3%의 발병율이 조사되었다. 이는 비교적 낮은 수치이나, 2차적으로 균핵병과 잿빛곰팡이병이 유발되어 큰 경제적 피해를 야기하였다. 이때의 병징은 아랫잎의 밑둥 부위에 수침상의 원형 내지 타원형의 담갈색 또는 갈색점무늬가 나타나고 진전되면 윗잎쪽으로 병반이 부정형으로 크게 확대되어 결국 잎이 무르게 썩고 누렇게 변하며 종이처럼 말라죽는 증상이 관찰되었다. 이에, 밑둥썩음병이 발생한 지역의 병든 식물체와 토양으로부터 병원균을 분리하였으며 이 중 병원성이 가장 강한 PY-1 균주가 Rhizoctonia solani AG1 (IB)로 동정되었다. 균사생육 최적온도는 25∼30$^{\circ}C였으며, 병원성 검정을 위해 가장 효과적인 접종원으로 검정 방법이 용이하면서도 효율적인 균사조각 부유액 접종원을 생육상내 폿트 검정용으로 선발하였고, 토양내 부생적 착생을 위한 플라스틱 하우스내 토경 재배 검정용으로는 WRSP배지 접종원을 선발하였다. 또한, 선발된 각 접종원의 최적 처리농도 및 처리량에서는 각각 균사조각 부유액 접종원의 경우, 평균 주당 51.1%의 발병도를 보인 $A_{550}$=1.0농도를 가장 적합한 처리 농도로 선발하였으며, WRSP 배지 접종원의 처리량 비교에서는 평균 주당 61.6%의 발병도를 보인 40 $m\ell$를 최적 처리량으로 선발하였다.

• 생명과학회지
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• 제6권2호
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• pp.94-103
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• 1996

To understand effect of inoculum size, cell density, sucrose concentration and concentrations of MS basal on suspension culture and protoplast isolation of wild viola(Viola patrinii DC.) callus from petiole segments this experiment was conducted. In the lot of 30 mesh inoculum size, two observations were; One was that a considerable increase in the fresh and dry weight of callus was determined. Another was that the callus mass was relatively compact compared with others. A recommendable cell density was 0.4g for 20ml culture medium and the higher sucrose concentration, the higher fresh and dry weight were obtained. The dilution of MS basal salt was differently affected on fresh and dry weight; the highest fresh weight was found in 1x MS salt, while the higest dry weight was in 1/3x dilution.The addition of casein hydrolysate(3g/L) was more effective to increase of both fresh and dry weight. THe contents of protein was great in the inoculum lots with larger inoculum sizeand higher concentration of MS basal salts contenting 3g/L of casein hydrolysate and higher sucrose compared with others. The greatest protoplasts were isolated from the lot of 10 mesh size treated with 1%pectinase SE-150 and 2% cellulase YC. In general, for optimal protoplast isolation the followingconditions were recommended; 1) Use of smaller cell size cultured for 2-5 weeks, and 2) more than 5 hours incubation using the combined mixture of the enzymes with proper concentrations.

• KSBB Journal
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• 제28권6호
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• pp.400-407
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• 2013

The growth characteristics and production of color pigments by Monascus strains were investigated during liquid culture, and production of monacolin K in red mold rice was carried out by solid state fermentation. Four different Monascus ruber strains were cultured in potato dextrose yeast extract broth (PDYB) media at $25^{\circ}C$ for 15 days. The high producing strain for red pigment was not corresponded to the strain for yellow pigment. Production of red pigment was high in the strain causing the fast pH change in culture broth. Production of monacolin K in red mold rice by solid state fermentation was influenced by a combination of wet cell weight and spore density in inoculum by liquid culture. Most strains showed the high production of monacolin K in red mold rice, when submerged fermentation was carried out for 5 days as inoculum for solid state fermentation. These results suggest that submerged fermentation period of inoculum have an effect on the production of monacolin K in red mold rice by solid state fermentation, and monacolin K in red mold rice could be increased by controlling the condition of submerged fermentation for inoculum.

• KSBB Journal
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• 제11권5호
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• pp.600-605
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• 1996

Taxus brevifolia와 Taxus cuspidata 현탁세포배 양에서 taxa!의 생산에 가장 적합한 초기 접종농도 를 결정하기 위해 접종농도를 세포생체중량으로 2.5, 5 5, 7.5, 10 g/flask가 되도록 변화시켜 세포의 생장 과 taxol의 생산을 측정하였다. Taxus brevifolia의 경우 초기 접종농도가 높을수록 lag phase가 감소되 는 경향을 보였으며, 두 가지 세포주(Taxus bre­1 vifolia와 Taxus cuspidata)에서 모두 초기 접종농도 를 5 g/flask가 taxol의 생산에 최척 임을 확인할 수 있었다. Taxol 생산에 가장 척합한 초기 접종농도로 생장배지와 생산배지에 세포를 접종시켰을 때 생장 배지에서는 taxal이 검출되지 않았지만 6% 자당을 이용한 생산배지에서는 9일째 taxol이 최대로 생산 되었다가 이후 다시 줄어드는 경향을 보였다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.103.1-103
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• 2003

An antagonistic bacteria, Stenotrophomonas maitophilia BW-13 strain which was effectively inhibited mycerial growth of Bottom rot pathogen, Rhizoctonia solani PY-1 strain was isolated from the rhizosphere of the lettuce in Uiryeong-Gun, Gyeongsangnam-Do from 2002 to 2003. For the biological control, the most suitable inoculum and its density of pathogen, PY-1 strain ware tested prior biological control test, For the pathogenicity test, A inoculum (wheat bran)sawdust+rice bran+PDB) showing disease incidence of 100％ was selected as the most suitable inoculum, which showed more effective than B inoculum (sawdust＋rice bran＋DW) and mycelial disc. also, In selection of the amount of inoculum (40g, 50g, 60g, 70g, 80g), most suitable amount of inoculum of pathogen determined as 40g showing disease incidence of 80％. For the selection of effective microorganism to control bottom rot on lettuce, about 200 isolates were isolated from the diseased soil and lettuce leaves, and examined their antifungal activity to the pathogen on PDA. As the pots assay, BW-13 strain showed the highest control value as 90％, and followed by R-13 and R-26 strain as 80％ and 60％, respectively. Selected BW-13 isolates identified as 5. maltophilia (GeneBank accession no. AJ293473.1, 99％) by 16S rRNA sequencing. This is the first report on the biological control using by S. maltophilia to the bottom rot pathogen, Rhizoctonia solani PY-1 strain.