• Title/Summary/Keyword: Calli

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Mannose-Based Selection with Phosphomannose-Isomerase (PMI) Gene as a Positive Selectable Marker for Rice Genetic Transformation

  • Penna, Suprasanna;Ramaswamy, Manjunatha Benakanare;Anant., Bapat Vishvas.
    • Journal of Crop Science and Biotechnology
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    • v.11 no.4
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    • pp.233-236
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    • 2008
  • A positive selectable marker system was adapted for transformation of mature embryo-derived calli of Indica rice (Oryza sativa L.) utilizing the PMI gene encoding for phosphomannose-isomerase that converts mannose-6-phosphate to fructose-6-phosphate. The transformed cells grew on medium supplemented with 3% mannose as carbon source and calli were selected on media containing various concentrations of mannose. Molecular analyses showed that the transformed plants contained the PMI gene. The results indicate that the mannose selection system can be used for Agrobacterium-mediated transformation of mature embryo in rice to substitute the use of conventional selectable markers in genetic transformation.

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Root and Shoot Formation in Explant and Callus Derived from Root and Cotyledon of GinBeng(Panun ginseng C. A. Meyer) (인삼근 및 자엽 Callus의 기관분화에 관한 연구)

  • Choe, Gwang-Tae;Kim, Myeong-Won;Sin, Hui-Seok
    • Journal of Ginseng Research
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    • v.5 no.1
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    • pp.35-40
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    • 1981
  • Explants of mature root tissues and calli derived from root and cotyledon of Panax ginseng were cultured in vitro on Murashige and Skoog medium supplemented with 2, 4-dichlorophen-oxyacetic acid(3,4-D), naphthaleneacetic acid(NAA), benzyladenine, and gibberellic acid to assess their capacity to regenerate organs. Root formation at high percentage (46.2-61.1%) was obtained 20-30 days after culturing on media supplemented with combinations of NAA(5 mg/l) and kinetin (1 mg/l), And calli derived from cotyledon produced numerous embryoids in media($\frac{1}{2}$MS) containing 2,4-D(0.5 mg/l) and kinetin (0.5 mg/l). Reculture of these embryoids in media($\frac{1}{2}$MS) enriched with 1 mg/l of benzyladenine and 1 mg/l of gibberellic acid resulted in more plantlet regeneration.

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Karyological Analysis of Somaclonal Variation in Callus Cells of Allium victorialis var. platyphyllum (산마늘의 캘러스세포에서 체세포군 변이의 핵학적 분석)

  • Seo, Bong-Bo
    • Journal of Plant Biology
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    • v.38 no.4
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    • pp.321-328
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    • 1995
  • Calli obtained from basal disc explants of Allium victorialis var.platyphyllum wre grown in three kinds of nutrient media (MS, BDS, and B5), and the frequencies of mitotic index and the chromosomal aberrations were analysed. The mitotic index varied from 0.55% to 1.01% with respect to culture media and ages. The mitotic irregularities like micro-, bi- and multi-nuclei, chromosome bridge and laggards were noted in each types of calli. The chromosome number variations observed in metaphase stage were identified aneuploid and tetraploid. Structural variations such as dicentric chromosomes, centromere breakage and small chromosomes were observed. Relationship between basal medium and chromosomal variability was not observed in this study. But, in BDS medium, NAA and BA had a more effect on number variation than kinetin.

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Somatic hybridization between Nicotiana rustica and N. tabacum through protoplast fusion (Nicotiana tabacum과 N. rustica 체세포 잡종식물의 육성)

  • Choe, Sang-Ju;Lee, S. C.;Hong, B. H.
    • Journal of the Korean Society of Tobacco Science
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    • v.15 no.2
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    • pp.123-129
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    • 1993
  • Mesophyll protoplasts derived from young leaves of Nicotiana rustica and N. tabacum cv Burley 21 were fused with the aid of polyethylene glycol(PEG). Cytological examination of protoplasts after PEG treatment revealed 12.8 % heterokaryocytes. After 7 weeks culture, the hybrid calli showing greenish white with a compact appearance were selected in contrast to parental type calli tinged with white or green color. The somatic hybrid plants were verified by morphological, biochemical and cyclological analysis. A heterosis effect for plant vigor and height was observed but the shape of leaves and flower characteristics were intermediate between N. tabacum and N. rutstica. The isozyme banding patterns for peroxidase of somatic hybrid lines were compared with the parent species. A number of isozyme bands derived from both parental species were found in the hybrids. Somatic hybrid plants have been successfully backcrossed to the parental N. tabacum particularly with somatic hybrid plants as female parents. These hybrid plants yielded small seeds, only few which were germinable.

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Expression of the Glyphosate Resistant Gene, cp4-epsps, through Plastid Transformation in Rice (Oryza sativa L.) (벼 색소체 형질전환을 이용한 글리포세이트 저항성 유전자 cp4-epsps의 발현)

  • Kang, Kyung-Su;Kim, Min-Kyun
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.75-84
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    • 2006
  • Heteroplasmic rice plastid transformant was generated using suspension cells as bombardment materials. PCR analyses confirmed incorporation of aadA and cp4-epsps genes into the rice plastid genome by homologous recombination events via the flanking sequences of the trnI and trnA. Transplastomic calli were actively proliferated when cultured on AAM2 medium supplemented with various concentrations (500-3000 mg/L) of streptomycin in dark condition, and transplastomic suspension cells showed resistance to nonselective herbicide, glyphosate. Through 'agarose pie selection' method, heteroplastomic calli, containing considerably high level of transplastome and expressing the CP4 EPSPS protein, were obtained. They were further regenerated to green shoots with healthy roots.

In Vitro Morphogenesis through Leaf Explants of Gypsophila paniculata L.

  • Jo, Man-Hyun;Ham, In-Ki;Song, Nam-Hyun
    • Plant Resources
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    • v.3 no.2
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    • pp.135-137
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    • 2000
  • Callus cultures from leaf explants of Gypsophila paniculata L. cv. 'Bristol Fairy' have been tested their growth and morphogenic capacity on Murashige and Skoog medium supplemented with 0.l, 0.5, 1 and 3 mg/L 2,4-D. The frequency of callus formation ranged from 43.3% to 100%. The optimal 2,4-D concentration for promoting callus formation and growth was 0.5 to 3 ㎎/L. 4.2∼ 5.6% of adventitious roots were obtained with the use of 0.1 and 0.5 mg/L 2,4-D. Calli grown well on 1.0 mg/L 2,4-D was the heaviest among the calli grown in various concentrations.

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Studies on The Anther Culture of Solanum nigrum L. II (Solanum nigrum L.의 약배양에 관한 연구 II)

  • 한창열
    • Journal of Plant Biology
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    • v.14 no.2
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    • pp.7-10
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    • 1971
  • Haploid cell obta-ined from microspores of Solanum nigrum were cultured on two kinds of medium, "Callus-inducing medium" and "Differentiation medium", in order to conduct histological studies of callus and examine differentiation of plantlets. On the callus-inducing medium the calli grew rapidly. The bulk of callus mass was light brown colored "Wet callus" covered on the surface with thin layers of rough and gleaming "White callus". The wet callus was consisted of parenchyma and meristematic tissues, while the white callus had no meristematic tissues. Large parenchyma cells, by successive divisions, became multicellular or poly nucleate cells which developed later to be meristematic tissues. The calli embedded on the differentiation medium quickly turned to dark brown color. Plantlets, however, came out later from these blackened callus mass. In the callus sectioned about ten weeks after imbedding on the differentiation medium, radially elongated tissue, concentric tissue, epidermis, tracheid-like structure, and plant jprimordia were observed.ure, and plant jprimordia were observed.

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Development of Seashore Paspalum Turfgrass with Herbicide Resistance

  • Kim, Kyung-Moon;Song, In-Ja;Lee, Hyo-Yeon;Raymer, Paul;Kim, Beom-Seok;Kim, Wook
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.4
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    • pp.427-432
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    • 2009
  • Seashore Paspalum (Paspalum vaginatum Swartz) is a warm season grass and indigenous to tropical and subtropical regions of coastal areas worldwide. The species is used as feed for cattle and horses and has been very successful for golf courses worldwide. One of the most outstanding characteristics of seashore paspalum is its tolerance to saline soils compared to other warm season turfgrasses. The development of new seashore paspalum cultivars with improved traits could be facilitated through the application of biotechnological strategies. The purpose of this study was to product for herbicide resistant seashore paspalum using Arobacterium-mediated transformation and this study is the first report on transformation and herbicideresistant transgenic plants in seashore paspalum. Embryogenic calli were induced from the seeded variety of pseashore paspalum. Embryogenic calli were transformed with Agrobacterium tumefaciens strain EHA105 carrying the binary vector pCAMBIA3301 with two genes encoding gusA and bar. Transformed calli and plants were selected on medium containing 3 mg/l PPT. PCR detected the presence of the gusA and bar gene, indicating both genes are integrated into the genome of seashore paspalum. A chlorophenol red assay was used to confirm that the bar gene was expressed. By application of herbicide BASTA, the herbicide resistance in the transgenic seashore paspalum plants was confirmed.

Studies on the Induction of Transformation in Cereal Plane V. Transformation of Wheat by Electroporation (곡물류의 형질전환 유도에 관한 연구 V. Electroporation에 의한 밀의 형질전환)

  • Song, Jung-Won;Jung, Byung-Kyun;Bae, Dong-Kyu;Im, Hyung-Tak;Nam, Back-Hee;Jung, Hyun-Sook;Hwang, Baek
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.187-192
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    • 1994
  • Wheat (Triticum aestium L.cv Cho-Kwang) explants were transformed by electrporation. Excised root segments were elechoporated with plasmid DNA of pBI121 and transferred to medium containing 300 mg/L kanamycin. Transformed calli formed within 5-7 days of culture and were selected from electroporated tissue on medium containing kanamycin after 4 weeks. The highest transformation frequency was obtained after electroporation with a pulse of 200 V/800 uF and calli formed at frequencies up to 2.5%. GUS ($\beta$-glucuronidase) assay and dot blot analysis showed that the foreign gene was capable of expressing in root explants subjected to electroporation and calli derived from the explants..

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