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형질전환된 포플러의 딱정벌레에 대한 저항성 유전자 ( Proteinase Inhibitor 2 ) 발현
강호덕 ( Ho Duck Kang ) 한국산림과학회 1997 한국산림과학회지 Vol.86 No.4
The resistance of a non-transgenic poplar clone, `Ogy` and three transgenic poplar lines to the cottonwood leaf beetle, Chrysomela scripta F., was evaluated by in vitro feeding. The lines were transformed with neomycin phosphotransferase II (NPT II ) as a selectable marker, proteinase inhibitor II(pin2) as a resistance gene, and CaMV 35S as a promoter. An efficient method of sterilizing the beetle eggs and introducing them into plant tissue cultures was developed. The resistance of the transgenic lines was investigated in terms of effects tin leaf area consumed, insect weight, insect developmental stages, and plantlet root dry weight after feeding. Also, leaf area consumed was examined by leaf age as measured through leaf plastochron index(LPI). The leaf area consumed and insect weight were highly significant between transformants and control, and insect development in vitro was significant among the transgenic lines. Larval infestation was the most severe around LPI 4 to 5 which were young leaves. The system provided a quick, highly controlled method to screen developing transgenic plantlets directly.
Peace 포플러(Populus koreana X P. trichocarpa)의 줄기형성에 미치는 Thidiazuron 효과
강호덕,문흥규,박인선,이민순,Kang, Ho-Duck,Moon, Heung-Kyu,Park, In-Sun,Lee, Min-Soon 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.1
기공이 잎의 앞뒤 양면에 존재하는 생리ㆍ형태적 특징을 가지고 있는 외래 교잡종인 Peace (Populus koreana ${\times}$ P. trichocarpa)포플러의 효율적인 기내증식법을 개발하고자 액아와 엽육 절편을 이용하여 기내배양을 실시하였다. 몇 종류의 배지 시험을 통해 1/2MS배지를 적정배지로 선정할 수 있었고, 이 배지에 TDZ를 처리하여 multiple shoot 및 adventitious shoot유도 효과를 조사하였다. 절간액아 및 엽육을 절편으로 TDZ의 농도별 처리로 배양 2주 후부터 multiple shoot 및 adventitious shoot가 유도되었으며 절편 및 TDZ처리 농도에 따라 차이를 나타냈다. 액아로부터는 TDZ 처리로 절편당 5-7개의 multiple shoot가 유도된 반면 엽육 절편에서는 2-5개의 adventitious shoot가 유도되었다. 한편 부정아로부터 정상적 인 줄기의 성장에 있어서도 절편에 따른 차이를 나타냈다. 이상의 결과는 저농도의 TDZ의 처리로 peace포플러의 액아 혹은 엽육 절편의 기내배양을 통해 대량증식이 가능함을 보여주는 결과이다. Shoot formation was investigated from in vitro cultivation of exotic hybrid poplar (Populus koreana ${\times}$ P. trichocarpa) with a specific stomatal character occurring both upper and lower surface of leaves. Two different explants (stem and leaf segment) of Peace poplar were cultured on half strength Murashige and Skoog (MS) basal medium supplemented with the various concentrations of thidiazuron as a plant growth regulator. Most adventitious shoots were produced from excised ends of stem or mid-veins of leaf segments. The highest average numbers of shoots were 7.1 and 5.3 with the treatments of 0.02mg/L TDZ in both explants of stem and leaf segment. The highest shooting rates were achieved to 83.3% and 47.6% with the concentrations of 0.01mg/L and 0.02mg/L TDZ by axillary bud and leaf cultures, respectively.
포플러의 미성숙 배와 캘러스에서 유전자총에 의한 GUS - gene 의 일시적 발현
강호덕,강상구,배한홍,박교수,( Ho Duck Kang,Sang Gu Kang,Han Hong Bae,Kyo Soo Park,Richard B . Hall ) 한국산림과학회 1997 한국산림과학회지 Vol.86 No.3
Excised immature ovules and calli derived from the stems of cottonwood were bombarded with microprojectiles carrying plasmid DNA containing CaMV-35S promoter and β-glucuronidase(GUS) gene. After bombarded, the expression of GUS gene was detected by the assay of 5-bromo-4-chloro-3-indolyl-β-gluconide(X-gluc). Transient gene expression was measured by counting the number of distinct regions of GUS activity per explant. As major parameters, the number of shots and the period of exposure to X-gluc after the bombardment were investigated for detecting GUS gene expression. In this experiment, the percents of GUS gene expression showing spots were 56.8 from immature ovules and 75.9 from micro-calli of cottonwood species. Among the treatments, two consecutive shots and 48 hour exposure produced abet 25.75±2.77(per ovule), 11.43±1.22(per mini petridish) spots, respectively, Microprojectile particle bombardment provides a useful method to assay transient expression in both types of explants. Furthermore, our results represent that the excised ovule and/or the calli might be stably transformed by the biolistics.
백리향 (Thymus quinquecostatus Celak.)의 기내증식에 미치는 식물생장조절물질의 영향
이금영,강호덕,Lee, Keum-Young,Kang, Ho-Duck 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.1
백리향(Thymus quinquecostatus Celak.)의 기내번식 방법을 개발하기 위해 줄기증식에 미치는 싸이토키닌 효과를 조사하였다. 시료식물은 MS배지에 thidiazuron등 5종의 싸이토키닌을 단독 또는 혼합 처리하여 광배양 또는 암배양 조건에서 식물체 증식을 유도하였다. 백리항의 대량증식에 효과적인 cytokinin종류와 농도는 명배양시 줄기수에 있어서 BA 1.0+TDZ 0.1mg/L (6.50$\pm$1.51) 였고, 신초의 길이는 zeatin 2.0+TDZ 0.1mg/L(3.27$\pm$0.7)가 최적 조건이었다. 또한 암배양시 줄기수의 경우엔 처리간에 별다른 차이 없이 비슷한 경향을 나타낸 가운데 2-ip 2.0mg/L로 처리하였을 때가 가장 뛰어났고, 줄기의 신장은 zeatin 2.0mg/L일 때가 가장 효과적이었다. 전체적으로 기내에서 대량증식율은 평균 89.58%로 비교적 대부분의 시료에서 식물체가 유도되었다. In vitro culture system was established to induce multiple shoots of Thymus quinquecostatus Celak. by investigating the effects of cytokinins. Stem explants were cultures on MS medium supplemented with either five drfferent plant growth regulators or their combinations under light or dark condition. The most effective cytokininsource was the combination of BA 1.0mg/L and TDZ 0.1mg/L for producing shoots (6.05$\pm$1.51), zeatin 2.0mg/L and TEZ 0.1mg/L for elongating shoots (3.27$\pm$0.66cm) under the light condition. In addition the most effective cytokinin was 2-ip 2.0mg/L for producing shoots(5.20$\pm$1.81), zeatin 2.0mg/L for elongating shoots(5.64$\pm$1.24cm)under the dark condition. Overall, the average percent for in vitro shooting was greater than 89.58%.