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식물생장조절물질과 배지첨가물질이 켄터키 블루그래스의 식물체 재분화에 미치는 영향
이상훈(Sang-Hoon Lee),이기원(Ki-Won Lee),김도현(Do-Hyun Kim),이동기(Dong-Gi Lee),원성혜(Sung-Hye Won),김기용(Ki Yong Kim),이병현(Byung-Hyun Lee) 한국초지조사료학회 2006 한국초지조사료학회지 Vol.26 No.2
To optimize tissue culture responses for genetic transformation of Kentucky bluegrass, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of a cultivar ‘Newport’ as explant tissues. The optimal concentration of 2,4-D (2.4-dichloro phenoxy acetic acid) for the induction of embryogenic callus from mature seed was 3 ㎎/L. Plant regeneration frequency was 54% when embryogenic callus was cultured on the regeneration medium supplemented with 1 ㎎/L 2,4-D and 3 ㎎/L of BA (6-benzyladenine). Addition of 1 g/L of casein hydrolysate and 500 ㎎/L of L-proline improved frequencies of embryogenic callus induction and plant regeneration up to 60.8% and 58.3%, respectively. Regenerated plants were grown normally when shoots transplanted to the soil. A rapid and efficient plant regeneration system established in this study. We suggest that the results may be useful for molecular breeding of Kentucky bluegrass through genetic transformation.
식물생장조절물질이 페레니얼 라이그래스 (Lolium perenne L.)의 캘러스 유도와 식물체 재분화에 미치는 영향
이기원(Ki-Won Lee),이동기(Dong-Gi Lee),Nagib Ahsan,원성혜(Sung-Hye Won),이상훈(Sang-Hoon Lee),김기용(Ki-Yong Kim),최기준(Gi Jun Choi),서성(Sung Seo),이병현(Byung-Hyun Lee) 한국초지조사료학회 2007 한국초지조사료학회지 Vol.27 No.4
페레니얼 라이그래스의 최적 조직배양조건을 확립하기 위하여 성숙종자로부터 배발생 캘러스 유도 및 효율적인 식물체 재분화조건을 체계적으로 확립하였다. 배발생 캘러스 유도시 첨가되는 auxin으로는 2,4-D가 가장 효율적이었으며, 9 ㎎/L 2,4-D와 0.1 ㎎/L BA가 첨가된 MS 배지에서 배발생 캘러스가 가장 높은 빈도로 유도되었다. 식물체 재분화는 배발생 캘러스를 1 ㎎/L 2,4-D와 5 ㎎/L BA가 첨가된 N6 배지에서 배양했을 때 58.3%의 높은 재분화율을 나타내었다. 본 연구를 통하여 확립된 단기간의 효율적인 재분화 시스템은 분자육종을 통한 신품종 페레니얼 라이그래스 개발에 유용하게 응용되어질 수 있을 것이다. Optimum tissue culture conditions for an efficient induction of embryogenic callus from mature seeds of perennial ryegrass (Lolium perenne L.) and regeneration of plants from callus tissues were investigated. MS medium containing 3 ㎎/L 2,4-D and 0.1 ㎎/L BA was optimal for embryogenic callus induction from mature seeds. The highest plant regeneration frequency (58.3%) was observed when the embryogenic callus tissues were cultured on N6 medium supplemented with 1 ㎎/L 2,4-D and 3 ㎎/L BA. Regenerated plants were grown normally when shoots transplanted to the soil. A short tissue culture period and high-frequency regeneration system would be helpful for molecular breeding of perennial ryegrass through Agrobacteriummediated genetic transformation.
캘러스의 형태와 항산화물질 첨가가 톨 페스큐의 식물체 재분화와 형질전환효율에 미치는 영향
이기원(Ki-Won Lee),이상훈(Sang-Hoon Lee),김도현(Do-Hyun Kim),이동기(Dong-Gi Lee),원성혜(Sung-Hye Won),이효신(Hyo-Shin Lee),이병현(Byung-Hyun Lee) 한국초지조사료학회 2006 한국초지조사료학회지 Vol.26 No.2
An efficient transformation system for the production of transgenic plants has been developed for tall fescue (Festuca arundinacea Schreb.) via Agrobacterium-mediated transformation of seed-derived callus. From the point of morphogenetic capacity, three types of callus were selected. High frequency of plant regeneration was obtained by selection of type Ⅱ callus, and the plant regeneration frequency was 52.6% when embryogenic callus were cultured on the regeneration medium. Supplementation of the media with 10 ㎎/L AgNO₃ and 40 ㎎/L cysteine enhanced frequencies of plant regeneration up to 65.3%. The highest transformation efficiency was also obtained when type Ⅱ callus were inoculated with Agrobacterium. Southern blot analysis of PCR products of transgenic plants demonstrated that transgenes were successfully integrated into the genome of tall fescue. Efficient regeneration system and transformation established in this study will be useful for molecular breeding of tall fescue through genetic transformation.
이탈리안 라이그래스의 성숙종자 유래 캘러스로부터 효율적인 식물체 재분화
우현숙,이상훈,이동기,김진수,원성혜,이병현,Woo, Hyun-Sook,Lee, Sang-Hoon,Lee, Dong-Gi,Kim, Jin-Soo,Won, Sung-Hye,Lee, Byung-Hyun 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.1
이탈리안 라이그래스의 최적 조직배양 조건을 확립하기 위하여 성숙종자로부터 최적 배발생 캘러스 유도조건 및 효율적인 식물체 재분화 체계를 확립하였다. 배발생 캘러스 유도시 첨가되는 auxin으로는 2,4-D가 가장 효율적이었으며, 5mg/L 2.4-D가 첨가된 MS배지에서 배발생 캘러스가 가장 높은 빈도로 유도되었다. 식물체 재분화는 배발생 캘러스를 1mg/L 2,4-D와 5mg/L BA가 첨가된 N6배지에서 배양했을 때 70%이상의 재분화율을 나타내었다. 기본배지의 종류에 따른 배양효율의 차이는 캘러스 유도에는 MS배지가, 식물체 재분화에는 N6배지가 효과적이었다. 배발생 캘러스의 형성율은 품종간에 큰 차이가 없었으나, 재분화 능력은 Jeanne 품종이 73%의 가장 높은 재분화율을 나타내어 품종간에 큰 차이를 나타내었다. 본 연구를 통하여 확립된 단기간 고효율 재분화 시스템은 분자육종을 통한 신품종 이탈리안 라이그래스의 개발에 유용하게 응용되어질 수 있을 것이다. As an initial step for future genetic manipulations to improve forage characteristics of Italian ryegrass (Lolium multiflorum Lam.), an efficient tissue culture system was established and the factors affecting plant regeneration were evaluated. MS medium containing 5mg/L 2,4-D was optimal for embryogenic callus induction from mature seed and had a strong effect on successive plant regeneration. The plant regeneration frequency was observed at above 70% when embryogenic calli induced were transferred to N6 medium supplemented with 1mg/L 2,4-D and 5mg/L BA. Among several basic media tested, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. Genotype was an important factor in plant regenerability. 'Jeanne' showed the highest regeneration frequency of 73%. A short tissue culture period and high-frequency regeneration system established in this study will be useful for molecular breeding of Italian ryegrass through genetic transformation.
식물생장조절물질이 레드 톱 밴트그래스(Agrostis alba L.)의 종자로부터 캘러스 유도와 식물체 재분화에 미치는 영향
박충훈(Choong-Hoon Park),김경희(Kyung-Hee Kim),이동기(Dong-Gi Lee),알람 이프테칼(Iftekhar Alam),이기원(Ki-Won Lee),이상훈(Sang-Hoon Lee),김기용(Ki-Yong Kim),원성혜(Sung-Hye Won),이병현(Byung-Hyun Lee) 한국초지조사료학회 2008 한국초지조사료학회지 Vol.28 No.4
레드 톱 밴트그래스의 성숙종자로부터 최적조직배양조건을 확립하기 위하여 종자로부터 배발생 캘러스 유도 및 식물체 재분화 체계를 확립하였다. 종자로부터 배발생 캘러스 유도시 첨가되는 auxin으로는 2,4-D가 가장 효율적이었으며, 2 ㎎/ℓ의 2,4-D가 첨가된 MS 배지에서 가장 높은 빈도로 배발생 캘러스가 유도되었다. 배발생 캘러스로부터 식물체 재분화는 0.5 ㎎/ℓ의 2,4-D와 2 ㎎/ℓ의 BA가 첨가된 N6 배지에서 배양했을 때 64.4%의 높은 재분화율을 나타내었다. 본 연구를 통하여 확립된 성숙종자로부터의 고효율 재분화 시스템은 유전자 형질전환기술을 이용한 신품종 분자육종기술개발에 유용하게 이용 될 수 있을 것이다. In order to develop a simple and reproducible protocol for red top bentgrass(Agrostis alba L.), effect of different growth regulators was investigated for embryogenic calli induction and subsequent plant regeneration using mature seeds. MS medium containing 2 ㎎/ℓ 2,4-D was optimal for embryogenic callus induction from mature seeds. The highest plant regeneration frequency (64.4%) was showed when the embryogenic callus tissues were cultured on N6 medium supplemented with 0.5 ㎎/ℓ 2,4-D and 2 ㎎/ℓ BA. Regenerated plantlets were grown normally when shoots transplanted to the soil. A high-frequency and efficient regeneration system from mature seeds would be helpful for molecular breeding of new variety of red top bentgrass through Agrobacterium-mediated genetic transformation.