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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.
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.
황백(Baik Hwang),조덕이(Duck Yee Cho),홍성식(Sung Sik Hong) 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.
황백 ( Baik Hwang ),김미경 ( Mee Kyung Kim ),( I . K . Vasil ) 한국식물학회 1988 Journal of Plant Biology Vol.31 No.1
Several cultivars of rice were examined for induction of embryogenic callus on a medium containing MS salts, vitamins and 2,4-D under darkness. Embryogenic callus was obtained from cultivar Cheonma with high ratio and embryo-like structures were formed from the callus on a medium with or without reduced 2,4-D. Somatic embryoids with a plumule and radicle axis surrounded by a scutellum were observed. These embryoids germinated and produced plantlets in 30 days on the same medium. Protoplasts isolated from an embryogenic cell suspension culture derived from embryogenic callus were cultured either in liquid or in agar medium and protoplast derived cell colonies were obtained in 3-4 weeks.
Agrobacterium rhizogenes 에 의하여 형질전환된 인삼 ( Panax ginseng C. A. Meyer ) 의 모상근 배양에 의한 Saponin 생산
황백(Baik Hwang),고경민(Kyeong Min Ko),황경화(Kyeong Hwa Hwang),황성진(Sung Jin Hwang),강영희(Young Hee Kang) 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.