• Biotechnology and Bioprocess Engineering:BBE
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• 제7권4호
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• pp.206-211
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• 2002

Calli obtained from a shoot-tip of garlic, Allium sntivum L., were encapsulated using a calcium alginate gel. Some of the encapsulated calli were cultured on a 1/2 MS medium supplemented with 3% sucrose, 10$\^$-5/ kinetin, and 5 ${\times}$ 10$\^$-6/ M NAA whereas the remainder was stored for 40 days at 4$^{\circ}C$. All the naked calli regenerated on the solid medium, while 95% of the encapsulated calli regenerated, and 88% of the encapsulated calli regenerated after 40 days of storage at 4$^{\circ}C$. The capsule matrix delayed the germination time of the encapsulated calli, yet activated the shoot formation of the artificial garlic seeds. The shoot length of the encapsulated garlic calli was much longer than that of the naked garlic calli. The encapsulated garlic calli were dried in a laminar airflow cabinet and the conversion frequency of the dried artificial garlic seeds on a 1/2 MS medium remained at 93% with a water Loss of Less than 50%.

• Journal of Plant Biotechnology
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• 제6권4호
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• pp.229-233
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• 2004

Partial desiccation of embryogenic calli cultures or somatic embryos leads to different physiological changes and maturation of somatic embryos, leading to improved plant regeneration. Embryogenic calli was induced from immature inflorescence segments and young leaf rolls of sugarcane (Saccharum officinarum hybrids CoC-671) on Murashige and Skoog's basal medium enriched with different concentrations of 2,4-D ($1-4\;\cal{mg/l}$), L-glutamine ($100\cal{mg/l}$), malt extract ($100\cal{mg/l}$), casein hydrolysate ($1000\;\cal{mg/l}$) and coconut milk ($5\%$) and solidified with $0.2\%$ gel rite. The embryogenic calli were subjected to desiccation for 1-8 h. Desiccation of the calli for 6-7 h resulted in enhancement of plant regeneration frequency ($83-96\%$) as compared to control ($12\%$). Plantlets exhibited vigorous growth to maturity in the greenhouse. Partial desiccation of embryogenic calli offers as a simple method for improving plant regeneration frequency in sugarcane.

• 식물조직배양학회지
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• 제26권3호
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• pp.211-217
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• 1999

애기장대 (Arabidopsis thaliana) 생태종 'Columbia' 잎절편에서의 묘조, 부정근, 모용 및 캘러스 형성에 미치는 auxin과 cytokinin의 효과를 조사하였다. 광범위한 식물생장 조절물질 농도와 조합에서 묘조, 부정근, 모용 및 캘러스가 형성되었다. 묘조는 0.1mg/L IAA와 10mg/L BA에서 형성되었고, 부정근은 저농도의 IAA와 NAA에서 모두 형성되었다. 모용과 캘러스는 IAA와 NAA의 농도 증가에 따라 형성되었으나 모용은 0.5mg/L NAA와 0.1mg/L BA 혼합처리구에서 가장 많이 형성되었으며, 캘러스는 10mg/L NAA와 10mg/L BA혼합처리구에서 가장 많이 형성되었다. NAA의 단독 영향을 조사한 결과 부정근 형성은 0.1mg/L 모용은 2.0mg/L 캘러스는 10mg/L가 가장 효과적이었으며, NAA배지에 각각 일정 기간 잎 절편을 치상하였다가 NAA 무처리 배지에 옮긴 결과 부정근, 모용 및 캘러스을 형성하는 determination time은 각각 6, 7, 18일이었다.

• Plant Biotechnology Reports
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• 제3권4호
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• pp.277-283
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• 2009

One of the limitations to conducting maize Agrobacterium-mediated transformation using explants of immature zygotic embryos routinely is the availability of the explants. To produce immature embryos routinely and continuously requires a well-equipped greenhouse and laborious artificial pollination. To overcome this limitation, an Agrobacterium-mediated transformation system using explants of type II embryogenic calli was developed. Once the type II embryogenic calli are produced, they can be subcultured and/or proliferated conveniently. The objectives of this study were to demonstrate a stable Agrobacterium-mediated transformation of maize using explants of type II embryonic calli and to evaluate the efficiency of the protocol in order to develop herbicide-resistant maize. The type II embryogenic calli were inoculated with Agrobacterium tumefaciens strain C58C1 carrying binary vector pTF102, and then were subsequently cultured on the following media: co-cultivation medium for 1 day, delay medium for 7 days, selection medium for $4{\times}14$ days, regeneration medium, and finally on germination medium. The T-DNA of the vector carried two cassettes (Ubi promoter-EPSPs ORF-nos and 35S promoter-bar ORF-nos). The EPSPs conferred resistance to glyphosate and bar conferred resistance to phosphinothricin. The confirmation of stable transformation and the efficiency of transformation was based on the resistance to phosphinothricin indicated by the growth of putative transgenic calli on selection medium amended with $4mg\;1^{-1}$ phosphinothricin, northern blot analysis of bar gene, and leaf painting assay for detection of bar gene-based herbicide resistance. Northern blot analysis and leaf painting assay confirmed the expression of bar transgenes in the $R_1$ generation. The average transformation efficiency was 0.60%. Based on northern blot analysis and leaf painting assay, line 31 was selected as an elite line of maize resistant to herbicide.

• Advances in Traditional Medicine
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• 제9권2호
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• pp.174-181
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• 2009

Pterocarpus marsupium, Clitoria ternatea, and Sanseveiria cylindrica are some of the important and endangered medicinal plant species of India. Despite of medicinal properties, antibacterial potential of the plants have not yet been explored. The present study was designed to optimize the in vitro technique for micropropagation and to screen the extracts from leaves and in vitro raised calli for antibacterial properties. Excised leaf-explants from the parent plants were surface sterilized and cultivated on Murashige & Skoog's (MS) medium containing $N^6$-benzyladenine (BA) in concentrations of 1, 2, 5, and $10{\mu}M$. Optimal growth of calli was noticed at a concentration of $5{\mu}M$, therefore the extracts from calli grown at this concentration were further studied for antibacterial activity. Both alcoholic and aqueous extracts from leaves of respective plants, and their in vitro raised calli were tested for antibacterial activity by agar well diffusion method against a range of Gram-positive and Gram-negative bacteria. Aqueous extracts showed antibacterial activity against limited number of bacterial species; notably the extracts of C. ternatea which showed antibacterial activity against Streptococcus pyogenes, Bacillus subtilis and Bacillus cereus. Alcoholic extracts of all three plants showed antibacterial activity against a wider range of bacteria. Among the Gram-positive bacteria, extracts from C. ternatea showed strong antibacterial activity against Bacillus spp., whereas the extracts of S. cylindrica showed good antibacterial potential for Staphylococcus aureus, S. epidermidis and S. pyogenes. The extracts from all three plants showed antibacterial activity against Gram-negative bacteria, including, Salmonella spp. and Shigella dysenteriae; organisms causing enteric fever and dysentery. In most of the cases, the extracts from respective calli showed comparable, and in some cases better, result in comparison to the extracts from parent leaves. To the best of our knowledge this is the first preliminary report on antibacterial potential, especially through calli extracts, of these plants; and in vitro cultivation of the explants may be used to obtain phytotherapeutic compounds.

• Journal of Plant Biotechnology
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• 제7권4호
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• pp.211-218
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• 2005

Genetic transformation was carried out by using biolistic gun method. The hypocotyl derived embryogenic calli (explants) of cotton (Gossypium hirsutum L.) cv. Cocker-312 were transformed with a recombinant pGreen II plasmid, in which both, bar (selection marker) and GUS (${\beta}$-glucuronidase) reporter genes were incorporated. Explants were arranged on osmoticum-containing medium (0.5M mannitol) 4 hours prior to and 16 hours after bombardment that was resulted into an increase about >80% for GUS stable expression. 3 days after bombardment, GUS assay was performed, which exhibited, $18.36{\pm}1.00$ calli showed blue spots. The transformed embryogenic calli were cultured on selection medium (@ 6 mg/L basta) for 3 months. The putative transgenic plants were developed via selective somatic embryogenesis (@1.50 mg/L basta); maximum $27.58{\pm}1.25$ somatic embryos were obtained while $17.47{\pm}1.00$ embryos developed into plantlets (@ 0.75mg/L basta). In five independent experiments, up to 7.24% transformation efficiency was recorded. The presence of the transgenes was analyzed by using PCR and southern hybridization analysis. The transgenic plants were developed with in 6-7 months, but mostly transformants were abnormal in morphology.

• 한국자원식물학회지
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• 제24권3호
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• pp.280-285
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• 2011

In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.

• Journal of Plant Biotechnology
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• 제30권2호
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• pp.161-165
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• 2003

The effect of NAA and Benzyladenine(BA) for determination times on the formation of adventitious shoots, roots trichmoes and calli in MS basal medium was investigated in leaf segments from ecotype 'Nosses' of Arabidopsis thalliana. Adventitious shoots, roots, trichomes and calli were formed fromed from leaf segments in a wede range of NAA and BA. The optimal combination of hormones for adventitious shoots formation, 20mg/L NAA for trichome formation, 100mg/L for callus formation. Inductive times for formation of adventitious shoots, roots, trichomes and calli were determined at 14, 4, 6 and 18 days respectively by periodical transfer of leaf segments from hormines containing media to hormone free medium.

• 식물조직배양학회지
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• 제27권2호
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• pp.117-123
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• 2000

애기장대 (Arabidopsis thaliana) 생태종 'Columbia'의 잎 절편 배양시 부정근, 모용 및 캘러스 형성에 미치는 polyamine의 역할을 알아보고자 putrescine, spermidine, spermine, cyclohexylamine (CHA) 및 methylglyoxal-bis(guanylhydrazone) (MGBG)의 영향을 조사하였다 애기장대 잎 절편을 부정근 (0.1 mg/L NAA), 모용 (2.0 mg/L NAA) 및 캘러스 (10.0mg/L NAA)를 형성하는 MS배지 각각에 putrescine, spermidine 및 spermine을 1∼100 mg/L 처리하였다. Polyamine 처리시 부정근 형성구에서는 부정근과 함께 모용이 형성되었으며, 모용 형성구에서도 모용과 함께 캘러스가 형성되었다 또한 CHA와 MGBG를 1∼100mg/L 범위에서 처리한 결과 CHA는 부정근 형성구에서 다소 부정근이 증가시켰으나 모용이나 캘러스 형성구에는 영향을 미치지 않았다. MGBG는 부정근 모용 및 캘러스 모두를 억제하였고 모용 형성구에서는 고농도에서 부정근을 형성시켰다.

• Journal of Plant Biotechnology
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• 제40권3호
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• pp.163-168
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• 2013

Iris odaesanensis Y. N. Lee. is an important endangered and native plant belonging to the family Iridaceae in Korea. This study describes a method for rapid micropropagation of this species via from leaf, rhizome and root explants derived calli. Leaf, rhizome and root explants were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) for callus induction. Rhizome explants yielded calli at a frequency of 72% when cultured at 1.0 mg/l 2,4-D. Calli were maintained at 1.0 mg/l 2,4-D. These calli were transferred to MS medium supplemented with 0, 0.5, 1.0, and 2.0 mg/l 2,4-D in combination with 0, 0.5, 1.0, and 3.0 mg/l BA for adventitious shoot induction. The highest number of adventitious shoot (228.9 per petri-dish) were formed at 1.0 mg/l 2,4-D and 1.0 mg/l BA. WPM medium was the best to convert calli into plantlets, where up to 98.2% of calli were regenerated into plantlets. This in vitro propagation protocol should be useful for conservation of this endangered plant.