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황백,趙德以,洪性式 한국식물학회 1986 Journal of Plant Biology Vol.29 No.4
Inoculation of carrot discs with Agrobacterium rhizogenes harboring Ri plasmid resulted in transformation of cells, as revealed by the tumors and hairy roots arising from them. Measurements of IAA content using HPLC indicate that it is higher in tumors and inoculated tissues than in uninoculated tissue. A lot of meristemoids and vessel elements formed in tumor tissue and the hiary roots differentiated from meristemoids. IAA content in tumor tissues is decreased with hairy root and vessel elements formation from them. Formation of wound callus in uninoculated tissues resulted on wound healing but no formation of vessel elements and hairy roots. Tumor tissues show continuous growing on hormone free medium.
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Pb가 당근 배양세포의 성장에 미치는 영향에 대한 전자현미경적 연구
황백,정상진,강영희,Hwang, Baek,Chung, Sang-Jin,Kang, Young-Hi 한국현미경학회 1979 Applied microscopy Vol.9 No.1
The present work has been carried out mainly to determine the effect of lead on the growth and cell organelles of Carrot (Daucus carota L.) callus. 1 mM of lead nitrate is added to the culture media R-2 and callus cells are cultured for 16 days. The growth rate is measured by fresh weight and structural changes of cell organelles is observed by electron microscope at every 4 days. The results show that lead inhibits the growth of Carrot callus and disturbs the structure of mitochondria remarkably.
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곡물류의 형질전환에 관한 연구.II. Electroporation에 의해 벼 원형질체로 도입된 유전자의 발현
황백,황성진,임욱빈,임형탁,강영희,Hwang, Baik,Hwang, Sung-Jin,Im, Wook-Bin,Im, Hyong-Tak,Kang, Young-Hee 한국생물공학회 1990 KSBB Journal Vol.5 No.4
Protoplasts isolated from embryogenic cell suspensions were electroporated in buffered solutions containing plasmid DNA of pBI121. Transient GUS (beta-glucuronidase) activity measurement and selection for kanamycin resistent showed that expression of foreign genes and stable loransformation were achieved. GUS transient gene expression was increased by increasing DNA concentration of pBI121 plasmid and affected by the level of the applied voltage. An optimal level of GUS activity was obtained after electroporation with a pulse of 200 voltage/1180 uF. Protoplast viability was up to the 60% at the optimal voltage. Cell colonies resistent to 200$\mu\textrm{g}$/ml kanamycin were selected in agar medium and identified by histochemical GUS assay.
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Plant Regeneration Via Organogenesis on Petiole of Centella asiatica (L.) Urban
황백 한국약용작물학회 2006 韓國藥用作物學會誌 Vol.14 No.2
An efficient plant regeneration of C. asiatica was achieved from organogenesis using petiole explants of in vitro plantlet on MS basal medium controled with different plant growth regulators (NAA, 2,4-D, IAA kinetin, and BA). Best results that 50%, efficiency of regeneration per explant for regeneration were obtained with IAA 17.13 μM and BA 8.9 μM. Formation of adventitious shoots via organogenesis from the petiole explant was verified by histological sectioning of plantlets. Regenerated plants were transplanted into soil. An efficient plant regeneration of C. asiatica was achieved from organogenesis using petiole explants of in vitroplantlet on MS basal medium controled with different plant growth regulators (NAA, 2,4-D, IAA kinetin, and BA). Best resultsthat 50%, efficiency of regeneration per explant for regeneration were obtained with IAA 17.13 μM and BA 8.9 μM. Formationof adventitious shoots via organogenesis from the petiole explant was verified by histological sectioning of plantlets. Regeneratedplants were transplanted into soil.
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Agrobacterium rhizogenes에 의하여 형질전환된 인삼(Panax ginseng C. A. Meyer)의 모상근 배양에 의한 Saponin 생산
황백,高庚珉,黃景花,黃聖振,康榮熹 한국식물학회 1991 Journal of Plant Biology Vol.34 No.4
Cultures of hairy root induced from ginseng(Panax ginseng C.A. Meyer) transformed with Agrobacterium rhizogenes (strain A_4, ATCC 15834) were established and morphologically two different hairy root strains (HB1, HB2) were obtained. To determine the optimum growth rate, the hairy root (HB2) was cultured in various liquid medium supplemented with or without plant growth hormone. The growth rate of hairy root cultured on MS medium was 1.3-3.1 times higher than those cultured on other media, and the optimum sucrose concentration and pH were 3-6%, 5.5-6.5, respectively. Also, the growth rate of hairy root was increased when 0.02 M ammonium nitrate, 1.2 mM potassium phosphate (monobasic) and 0.5 mg/l IBA were supplied to liquid medium. The saponin patterns and contents of hairy root (HB2) were determined by TLC and HPLC. The crude saponin contents were 4.67% and the total saponin contents were 1.0%, on dry weight basis.
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Agrobacterium rhizogense에 Hairy Root 형성에 대한 생리학적 연구. III. 당근 세포에의 A. rhizogenes의 부착
황백,황성진,안준철,조혜선,Hwang, B.,Hwang, S. J.,Ann, J. C.,Jo, H. S. 한국생물공학회 1989 KSBB Journal Vol.4 No.2
In vitro attachment experiments of bacteria to surface of host plant cell were carried out using C14 labeled cells of A. rhizogenes strain A4 and carrot protoplasts isolated from suspension culture of cells. Protoplasts were cocultivated with A. rhizogenes at various times after their isolation. Attachment kinetics showed that adherence of bacteria to protoplasts attained a maximum level within 120mins of co-cultivation. Maximum attachment occured at pH 6.0 and 24-35$^{\circ}C$. Bacterial attachment was observed at botg carrot cells with and without primary cell wall. The inhibition of transformation on the carrot root discs by A. rhizogenes was observed when non-related strain and heat inactivated bacterial strain cells were pretreated. 당근 진탕배양세포로부터 분리한 원형질체에 A.rhi-zogenes를 처리하여 bacteriad의 부착과 부착과정에 영향을 주는 여러 요인들을 조사하였다. 또한 당근뿌리 절편에 strain이 다른 bacteria와 heat-inactivated bacteria를 전처리한 후 A.rhizogenes strain A4를 접종하여 식물세포 표면에 있어서 bacterial specific receptor sites존재 유무를 살펴보았다. Bacteria의 부착은 pH 6.0, 24-35$^{\circ}C$ 조건하에서 120분동안 원형질체막에 유의적인 증가를 나타내었으며, 또한 세포벽합성에 따르는 bacteriaqnckr의 영향은 볼 수 없었다. Strain이 다른 bacteria나 heat-inactivated bacteria의 전처리에 의한 A..rhizogenes의 당근조직에서 hairy root형성 억제 현상은 식물세포 표면에 specific receptor sites가 존재 한다기보다는 부착에 있어서 전처리한 bacteria의 경쟁적 저해작용으로 형질전환이 다소 억제되는 것으로 사료되어졌다.
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Agrobacterium rhizogenes 에 의한 hairy root 형성에 대한 생리학적 연구 ; IV. Hairy root 배양 및 배양 조건에 관한 조사
황백,안준철,이재혁,Hwang, Baik,An, Jun-Cheul,lee, Jae-Hyuk 한국생물공학회 1989 KSBB Journal Vol.4 No.3
Agorobacterium rhizogenes에 의하여 유도된 당근(Daucus carota L. )의 hairy root를 배양하였으며 opine 유무에 대한 형질전환 확인 및 배지조성을 달리하여 성장률에 따른 색소함량을 비교하였고, 재분화된 식물체의 형태적 차이를 관찰하여 몇가지 결론을 얻었다. Hairy root는 균 접종 2-4주 후에 형성층 부위를 중심으로 유도되었다. 유도된 hairy root의 초기 배양에는 R.C.M 배지가 적합하였으며 MSO(2, 4-D $10^{-4} ml/ l, pH6, sucrose 5%, 질소원 0.03M 등)에서 최대 성장을 보여주었고 성장의 증가에 따른 색소의 형성은 비교적 안정하였다. 재분화된 식물체는 정상 식물체에 비하여 형태적으로 차이를 나타내었으며 형질전환된 hairy root 및 재분화된 식물체에서 mannopine 분석으로 Ri-plasmid에 의한 형질전환이 이루어졌음을 확인하였다. Hairy roots of carrot were induced by Agrobacterium rhizogenes $A_4$ strain within about 2-4 weeks after inoculated from root disc. Early axonic culture is established in RCM agar medium and following is in MS rigid medium. After 15 days culture, the hairy roots were vigrous growth in about 10 times of initial inoculum. Anthocyanin contents of hairy roots were more than of ordinary roots. 2, 4-D ($10^{-4}mg/ l$), sucrose (5%), nitrogen source (0.03M) contained medium was optimized to growth of hairy root and contents of anthocyanin. Phenotypic alterations of leaves are observed in transformed plants and determined the transformation of hairy roots and the transformed plants by opine assay.
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