• Journal of Microbiology and Biotechnology
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• 제13권2호
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• pp.256-262
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• 2003

To enhance the production of heavy chain immunoglobulin G (HC IgG) in the suspension cultures of transgenic tobacco cells (Nicotiana tabacum), the effects of adding various cytokinins (i.e., growth regulators) and organic nitrogen sources to culture media were investigated. Four different cytokinins including kinetin, isopentenyladenine (IPA), 6-benzylaminepurine (BA), and zeatin were tested with or without dichlorophenoxyacetic acid (2,4-D), which is a typical growth regulator supplemented in the standard Murashige and Skoog (MS) medium. The productivity of intracellular HC IgG was increased by 36 and $42\%$, compared to the control, especially when IPA (2 mg/l) or BA (0.2 mg/l) was added to the media in the presence of 2,4-D, respectively. In the study of organic nitrogen sources, addition of each casein hydrolysate and tryptone to the culture media at a final concentration of 0.01 and 1 g/l, respectively. increased the productivity or he IgG as much as 68 and $67\%$, respectively, in comparison with the control, which was is MS medium without supplementation of any organic nitrogen sources. This study shows that the optimization of media composition could offer significant improvements in the production of foreign proteins in the suspension cultures of transgenic plants.

• Journal of Plant Biology
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• 제15권1호
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• pp.28-32
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• 1972

Young haploid leaf derived from the anthers of tobacco plant was cultuerd and plantlets of various ploidies were obtained. When the leaf was put on the medium supplemented with kinetin as growth regulator, plantlets developed directly from the leaf, and the plants coming out in early stage of culture were all haploid. Plants developing in later stage were mostly haploids with some exception of diploid and aneuploid. Leaves were also cultured on the callus-inducing media supplemented with 2,4-D and kinetiion, and the calluses were sub-cultured for six months. Plants developed from these calluses were mostly aneuploids of various chromosome numbers. In view of the fact that the plants directly developed from the leaf were all haploid, the tissue of the original leaf explant was assumed to be uniform as far as chromosome number was concerned. On the other hand, it seemed that the occurrence of various ploidies in the plants derived from the calluses of same origin was the result of the influence of in vitro culture. Apical meristem tissues and various multicellular bodies were formed in the epidermal and inner mesophyll tissues as well as in the sub-epidermal cells.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1985년도 워크샵 및 심포지엄 북한산국립공원의 식생
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• pp.23-33
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• 1985

Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

• 한국연초학회지
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• 제16권1호
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• pp.64-68
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• 1994

The present experiments were conducted to investigate some of the factors affecting the number of haploids derived from anther culture of Nicotiana tabacum. Anther - donor plants grown under controlled environment room at 3$0^{\circ}C$ yielded more haploid than room at 18, 25 and 26-22-18$^{\circ}C$ in anther culture. Donor plants starved of fertilizer yielded more haploids as compared to those of the well fed with fertilizer in anther culture. Pretreatment of exercised flower bud at 5$^{\circ}C$ was shown to be more effective in anther culture than pretreatment at 7 and 1$0^{\circ}C$, and the optimum temperature and period of pretreatment were 4 or 6 days at 5$^{\circ}C$.

• 한국토양비료학회지
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• 제18권1호
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• pp.89-93
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• 1985

질소(窒素)와 무기양(無機陽)이온이 다른 작물(作物)보다 풍부한 연초제조부산물(煙草製造副産物)인 담배부스러기를 재료(材料)로 하여 비료물질(肥料物質)을 개발하려고 하였다. 담배부스러기를 무게감소율(減少率)로 50%되게 탄화(炭化)시킨 후 유리된 칼륨으로 인한 염기성을 pH 7.0정도 되게 질산(窒酸)과 인산(燐酸)으로 중화(中和)하면 질소(窒素) 4%{유기태(有機態) 질소(窒素)(담배부스러기에 함유(含有)된 질소) 2%, 질산태실소 2%}, 인산(燐酸) 2%, 칼륨6%이상(以上)인 비료물질(肥料物質)이 되었다. 관행(貫行)의 연초육묘시(煙草育苗時)에 시비(施肥)되는 전체(全體) 질소량(窒素量)을 위의 중화이분탄화물(中和泥粉炭化物)로서 전량기비(全量基肥)로 $1kg/110{\ell}$ 정도를 상토(床土)와 혼합시용하여 육묘(育苗)한 결과 관행(貫行)과 같이 추비(追肥)를 시용(施用)하지 않고도 양호(良好)한 생육(生育)을 보였다. 이러한 효과(效果)는 속효성인 질산태질소와 완효성인 담배가루 구성물질의 유기태질소(有機態窒素)가 탄화물(炭化物)에 함유(含有)되고 또한 다량함유(多量含有)된 칼륨이 영양원(營養源)으로 되기 때문인 것으로 생각되었다.

• Journal of Plant Biotechnology
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• 제4권4호
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• pp.143-150
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• 2002

A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

• The Plant Pathology Journal
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• 제29권2호
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• pp.182-192
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• 2013

Numerous root-associated bacteria (rhizobacteria) are known to elicit induced systemic resistance (ISR) in plants. Bacterial cell-density-dependent quorum sensing (QS) is thought to be important for ISR. Here, we investigated the role of QS in the ISR elicited by the rhizobacterium, Serratia marcescens strain 90-166, in tobacco. Since S. marcescens 90-166 produces at least three QS signals, QS-mediated ISR in strain 90-166 has been difficult to understand. Therefore, we investigated the ISR capacity of two transgenic tobacco (Nicotiana tabacum) plants that contained either bacterial acylhomoserine lactone-producing (AHL) or -degrading (AiiA) genes in conjunction with S. marcescens 90-166 to induce resistance against bacterial and viral pathogens. Root application of S. marcescens 90-166 increased ISR to the bacterial pathogens, Pectobacterium carotovorum subsp. carotovorum and Pseudomonas syringae pv. tabaci, in AHL plants and decreased ISR in AiiA plants. In contrast, ISR to Cucumber mosaic virus was reduced in AHL plants treated with S. marcescens 90-166 but enhanced in AiiA plants. Taken together, these data indicate that QS-dependent ISR is elicited by S. marcescens 90-166 in a pathogen-dependent manner. This study provides insight into QS-dependent ISR in tobacco elicited by S. marcescens 90-166.

• 한국작물학회지
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• 제31권2호
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• pp.231-237
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• 1986

향미종 연초 재배에서 재식밀도 및 질소 시비량에 따른 생육특성 및 수량, kg당 가격과 향미종 연초의 특징인 휘발성 유기산 함량의 관계를 분석, 재배법에 활용코자 시험한 결과, 1. 10 a당 13,885주에서 24,997주로 밀식할수록, 주당 생장량은 감소하나 LAI는 증가하였으며 수량은 21.875주까지 유의성 없이 증가하였으나 그 이하에서는 증가를 보이지 않았다. 2. 10a 당 질소 시비량을 1.5, 3.0, 4.5kg 증비 할수록 주당 생장량, 잎의 크기 및 LAI가 증가하고 수량 및 kg당 가격이 유의하게 증가하는 경향이였다. 3. 발뢰 7 일 전 엽중 질소 함량은 처리간 차를 나타냈으나, 건조엽중의 질소, 환원당, 니코딘 함량에서는 재식밀도 및 질소수준간 일정한 경향을 보이지 않았다. 4. 초장 및 엽의 크기가 작을수록 건조엽중 2-methyl butanoic, 3- methyl butanoic 및 3- methyl pentanoic acid의 함량이 높은 경향이었으며, 5. 건조엽중 휘발성 유기산 함량과 개화기의 초장(r=-0.49), 최대엽장(r=-049), 일주당 엽면적(r=-0.47)은 유의한 부의 상관을 보였다. LAI 및 단위엽면적당과는 각각 유의한 상관을 보이지 않았다.

• 한국작물학회지
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• 제28권2호
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• pp.262-266
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• 1983

인삼종자의 내과피에 부생하는 진균이 배생장 및 개압에 미치는 영향을 알기 위하여 종자와 층적장의 모래를 살균 처리한 뒤 종자를 층적하여 얻은 결과를 요약하면 다음과 같다. 1. 종자소독에 의해 종자의 내과피에 부생하는 균밀도가 현저히 저하되었으며 종자의 경도변화가 크게 억제되었고, 종자의 경도와 균밀도간에는 부(-)상관 $(r=0.984^**)$이 인정되었다. 2. 종자내 수분함량의 증가는 종자소독구가 대조구보다 20일 늦게 시작되었다. 3. 배생장과 개압은 종자소독에 의해 크게 억제되었고 종자의 경도가 낮을수록 배생장은 잘되었다. 인삼종자의 내과피에 부생하는 진균이 내과피를 경화시켜 산소, 수분 등의 공급을 쉽게 함으로써 배생장 및 개갑을 촉진시키는 것으로 생각된다.

• 생명과학회지
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• 제22권6호
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• pp.778-787
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• 2012

카드뮴에 의해 유도되는 담배의 생장, 엽록소 함량, rubisco와 rubisco activase에 미치는 SA의 영향과, 이에 대한 변성제의 효과를 연구하였다. 담배 기내생장에 대한 SA의 최적농도를 찾기 위해, $10^{-6}$ mM - $10^2$ mM SA를 처리하여 9주간 생장시킨 결과, $10^{-4}$ mM SA에서 가장 높은 생장을 보였다. SA와 카드뮴을 4개의 실험구(대조구, SA, 카드뮴, 카드뮴 + SA)로 하여 생장, 엽록소 함량 및 rubisco와 rubisco activase의 함량과 활성을 측정한 결과, 카드뮴 > 카드뮴 + SA < 대조구 카드뮴 + SA < 대조구 ${\beta}$-mercaptoethanol은 활성을 촉진시켰으며, urea, thiourea, guanidium-HCl은 억제시켰다. 이는 L-cysteine과 ${\beta}$-mercaptoethanol은 변성에 관여하지 않으며, urea, thiourea, guanidium-HCl은 변성에 관여하였음을 의미한다. Rubisco activase의 활성에 대한 변성제들의 영향을 조사한 결과, 5종 모두는 비처리구와 비교하여 현저한 영향은 나타내지 않았다.