• Title/Summary/Keyword: Rol C.

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The Apple Rootstock Transgenic M.26 (Malus pumila) with Enhanced Rooting Ability (발근력이 향상된 사과 대목 M.26 형질전환체)

  • Kim, Jeong-Hee;Kwon, Soon-Il;Shin, Il Sheob;Cho, Kang-Hee;Heo, Seong;Kim, Hyun Ran
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.482-487
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    • 2009
  • The apple rootstock M.26 (Malus pumila) is very popular apple rootstock with semi-dwarf habit and the trees on M.26 produce high quality fruit at a young age. Since it is prone to poor prop ability to soil, however, young trees require staking in windy locations. The rolC gene was introduced into M.26 by Agrobacterium tumefaciens LBA4404 harboring pBI121 to obtain its transformants with dwarfism and enhanced rooting ability. One regenerated transgenic line was confirmed by polymerase chain reaction (PCR) analysis and Southern blot analysis of genomic DNA for the existence of rolC gene. The characteristics of transgenic line in vitro were not significantly different from non-transgenic line except for the active root formation and lateral root number. The rolC transgenic line showed reduced stem length and increased root number in vitro. Rooting ability was examined in the isolated greenhouse after mound layering. Compared to non-transgenic M.26, rolC transgenic line showed significantly higher rooting ability. The transgenic line did not show any other observable variation in shoot phenotype compared with non-transgenic line excepting increased branching

Transformation of Brassica napus via Agrobacterium Vector : Plant Regeneration and Progeny Analysis (Agrobacterium 운반체를 이용한 유채의 형질전환: 식물체재분화와 후대검정)

  • KIM, Kyung Min;SOHN, Jae Keun;CHUNG, Jae Dong
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.269-272
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    • 1997
  • Cotyledonary petioles of Brassica napus cocultivated with Agrobacterium vectors for 72 h were transferred to MS medium with 0.5 mg/L NAA, 2.0 mg/L BA, 30 mg/L kanamycin, 100 mg/L cefotaxime, 30 g/L sucrose, 3 mg/L $\textrm{AgNO}_{3}$ and 2 g/L Gelrite. The cotyledonary petioles with green shoots were selected at a frequency of 17.5% in a selection medium and then rooted. Southern blot analysis confirmed the rolC and NPT IIgenes were incorporated into the regenerated plants. The stable inheritance of rolC gene was confirmed in progeny test of transgenic plants.

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Introduction of rolC gene into Petunia hybrida (Petunia hybrida 세포내로의 rolC 유전자의 도입)

  • 정재동;김경민;남윤연;김창길;정원일
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.21-26
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    • 1999
  • These experiments were attempted to introduce rolC gene in the Petunia hybrida cv. Titan white by Agrobacterium mediated. The maximum frequency of shoot regeneration was obtained by 60% on MS medium containing 1.0 mg/L BA, 0.1 mg/L NAA, 200 mg/L kanamycin, 500 mg/L carbenicillin, 30 g/L sucrose, and 8 g/L agar. Kanamycin-resistant calli were selected from petunia leaf discs by cocultivation with Agrobacterium suspension cultures on MS medium. The addition of AgNO$_3$ and KMnO$_4$ in the medium increased the shoot regeneration by 31.3% from leaf disc as compared with non-treated leaf disc. Among clones exhibiting kanamycin resistance, only 3 clones were confirmed by southern hybridization analysis.

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Factors Affecting Introduction of rolC Gene in Lycium chinense Mill. (구기자나무(Lycium chinense Mill.)로의 rolC유전자 도입에 미치는 요인)

  • 박용구;최명석;김병원;정원일;노광수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.329-334
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    • 1995
  • Transformation system of rolC gene, dwarf gene in Lycium chinenese Mill. established by using system. Pin-punctured leaves induced numerous adventious buds in abaxial side when cultured on 3/2 MS medium containing 2.0 mg/L zeatin. Survival rate and shoot regeneration frequency of leaf explants decreased as kanamycin sulfate level increased. Shoot buds were not regenerated on 3/2 MS medium containing 10 mg/L kanamycin sulfate and 2.0 mg/L zeaein. Of the level tested, 10 mg/L of kanamycin sulfate was optimum in selection of kanamycin sulfate resistant plant. Co-culture time of bacteria and leaf explants was affected at the frequency of shoot regeneration and survival of leaf explants. Leaf explants co-cultivated during above 48hr severely decreased survival rate and shooting rate. Best result on survival rate and shooting rate were obtained when exposed for 24 h. 80 explants of 105 leaf explants survived on 3/2 MS medium containing 2.0 mg/L zeatin and 10 mg/L kanamycin sulfate, and 15 shoots was regenerated on the same medium. To select kanamycin sulfate resistant plant, regenerate as cultured on 3/2 MS medium containing 10 mg/L kanamycin sulfate, and obtained 5 kanamycin resistant plants. Southern blot analysis conformed that the rolC gene was incorporated into the genomic DNA of kanamycin resistant plants.

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Growth Characteristics of Transgenic Potato Using Wild-type Agrobacterium spp. (Agrobacterium spp. 에 의하여 형질전환된 감자조직의 생장특성)

  • 양덕춘
    • Korean Journal of Plant Resources
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    • v.11 no.2
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    • pp.176-176
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    • 1998
  • This study was carried out to obtain the information for growth characteristics of crown gall tumor and hairy root transformed by Agrobacterium spp,. on the media with phytohormones, casein hydrolysate and activated charoal. Crown gall tumors and hairly roots were formed respectively on potato tuber discs infected by tumerfaciens A ch 5 and A.rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. PCR analysis of Rol C and Vir C gene fragments confirmed that crown gall root was prompted on the medium containing 2,4-D 2mg/l with casein hydrolysate lg/l. The survival ration of crown gall tumor callus derived from potato increased on medium containing the activated charcoal 0.5∼0.2mg/l because of the prevention, on the other hand, hairly roots were necrosis on the same medium. Callus derived from hairly root were excellently grown for a short time by suspension culture on liquid medium containing 2.4-d 2mg/L and casein hydrolysate lg/l.

Growth Characteristics of Transgenic Potato Using Wild-type Agrobacterium spp. (Agrobacterium spp. 에 의하여 형질전환된 감자조직의 생장특성)

  • 양덕춘
    • Korean Journal of Plant Resources
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    • v.11 no.2
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    • pp.179-187
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    • 1998
  • This study was carried out to obtain the information for growth characteristics of crown gall tumor and hairy root transformed by Agrobacterium spp,. on the media with phytohormones, casein hydrolysate and activated charoal. Crown gall tumors and hairly roots were formed respectively on potato tuber discs infected by tumerfaciens A ch 5 and A.rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. PCR analysis of Rol C and Vir C gene fragments confirmed that crown gall root was prompted on the medium containing 2,4-D 2mg/l with casein hydrolysate lg/l. The survival ration of crown gall tumor callus derived from potato increased on medium containing the activated charcoal 0.5∼0.2mg/l because of the prevention, on the other hand, hairly roots were necrosis on the same medium. Callus derived from hairly root were excellently grown for a short time by suspension culture on liquid medium containing 2.4-d 2mg/L and casein hydrolysate lg/l.

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Selection of Active Grow Hairy Root Lines in Ginseng (고생장 인삼 모상근의 선발)

  • 양덕춘;김용해;양덕조;민병훈;신성련;최광태
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.525-530
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    • 1998
  • These studies were carried out to select the active grow hairy root lines induced from various ginseng(Panax ginseng C. A. Meyer) parts. Hairy roots were induced in root explants, stem and petiole in vitro by A. rhizogenes R1000 or A. rhizogenes $A_4$. These hairy roots could be grown on the phytohormone free medium, and PCR analysis of rol C and vir C gene fragments confirmed that hairy roots were transgenic tissues. We have selected 11 hairy root lines with active growing characters among 300 hairy root lines selected based on growth and morphological characteristics on 1/2MS solid media with 250 mg/L carbenicillin. Morphological characteristics of selected 11 hairy root lines were thickness and thiness of main roots, and many projection for lateral roots, active grow of lateral roots. Among selected 11 hair root lines prominent characteristics of hairy roots with active growing characters were thiness of main roots and active grow of lateral roots. But characteristics of low growing hairy roots were thickness of main roots and low grow of lateral roots. Finally we have selected actively growing hairy roots, KGHR-1, KGHR-5, KGHR-8 among 11 hairy root lines.

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Triterpenoid production and phenotypic changes in hairy roots of Codonopsis lanceolata and the plants regenerated from them

  • Kim, Ji-Ah;Kim, Yun-Soo;Choi, Yong-Eui
    • Plant Biotechnology Reports
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    • v.5 no.3
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    • pp.255-263
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    • 2011
  • Codonopsis lanceolata (Campanulaceae) has been used in traditional medicines, as its roots contain several kinds of triterpenoid saponin with high medicinal values. In this work, we induced transgenic hairy roots of C. lanceolata and analyzed triterpenoid saponins from the hairy roots and hairy root-derived transgenic plants. Hairy roots were obtained from leaf explants by the transformation of Agrobacterium rhizogenes R1000. Transgenic hairy root lines were confirmed by the transcriptional activities of rolA, B, C, and D genes by RT-PCR. Transgenic root lines actively proliferated on hormone-free medium but not in nontransformed roots. Hairy roots contained richer triterpenoids (lancemaside A, foetidissimoside A, and aster saponin Hb) than nontransformed roots. Transgenic plants were regenerated from the hairy roots via somatic embryogenesis. They showed phenotypic alterations such as shortened shoots and an increased number of axillary buds and adventitious roots. The transgenic plants also contained higher triterpenoid levels than wild-type plants. These results suggest that hairy roots and transgenic plants of C. lanceolata could be used as medicinal materials for the production of triterpene saponins.

Effect of temperature on Antioxidant of Crude Gingerol (Gingerol이 첨가된 대두유의 산화에 미치는 온도의 영향)

  • 백숙은
    • Korean journal of food and cookery science
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    • v.10 no.2
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    • pp.121-125
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    • 1994
  • The antioxidant activity of gingerol according to temperature on soybean oil were examined by measuring peroxide value(POV). The induction period(IP) of soybean oil was 45; 276.0, 65; 17.0 and 105$^{\circ}C$ : 4.7 hours respectively by-measuring POV. The relative antioxidant effectiveness(RAE) of ginge-rol group were 45; 191, 65; 200, 65: 150, 85: 132 and 105$^{\circ}C$;106%. 'The activation energies(Ea) and temperature coefficients(Q10) for Arrhenius equations at 45∼105$^{\circ}C$, was estimated in order to find out the influence of temperature on the oxidation of soybean oil contai-ning various antioxidants. The soybean oil was more unstable at 45∼65$^{\circ}C$ than at 65∼105$^{\circ}C$ in the Ea and Q10. The soybean oils containing gingerol were more stable than the control group at 45∼105$^{\circ}C$, however, BHT group was unstable compared to gingerol group at 85∼105$^{\circ}C$.

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Overexpression of the SPP2 gene of saccharomyces cerevisiae and production of antibodiesd to Spp2p

  • Park, Kwang-Hark;Lea, Ho-Zoo;L. Woolford;Kim, Kyung-Hoon
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.201-207
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    • 1995
  • We have previously reported that SPP2 gene product of yeast Saccharomyces cerevisiae is involved in the pre-mRNA splicing. To investigate the rol ein the splicing pathway of the Spp2p protein, the SPP2 gene was overexpressed in Escherichia coli and polyclonal antibodies to Spp2p were generated from rabbits. First, a DNA fragment containing the SPP2 GENE without its promoter was subcloned into an E. coli expression vector, pKK233-3. The resulting recombinant plasmid pBQ14 contained an IPTG inducible tac promoter and the SPP2 structural gene. Overexpression of the SPP2 gene was achieved by additionof 0.1 to 1.0 mM IPTG to a logarithmic culture of E. coli JM103(pBQ14) for 90 min at 37.deg.C. Sequence of N-terminal 15 amino acids of the overproduced protein was well matched to the deduced one from the SPP2 reading frame. Then, polyclonal antibodies were generated from rabbits immunized with gel-purified SppSp protein. These antibodies reacted specifically with the Spp2p protein extracted from yeast cells expressing the SPP2 gene to a great extent. The antibodies could also block the activity of yeast splicing extracts.

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