다아체 형성에 의한 Philodendron Wend-imbe의 대량번식

한봉희,예병우,구대회,유희주,Han, Bong-Hee,Yae, Byeoung-Woo,Goo, Dae-Hoe,Yu, Hee-Ju 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.2

본 실험은 Philodendron wend-imbe를 기내배양하여 일시에 균일한 식물체를 대량생산하기 위하여 실시하였다. Philodendron의 경정에서 다아체 형성은 BA 5.0-10.0mg/L 또는 TDZ 0.05-0.1 mg/L가 첨가된 MS 배지에서 양호하였다. 형성된 다아체 절편체 (5-7mm)를 BA 5.0 mg/L와 sucrose 20g/L가 첨가된 MS 배지에서 배양한 결과, 신초 및 다아체의 증식이 매우 양호하였으며, 신초기부에서 callus 발생이 억제되었다 다아체 절편체에서 신초의 발생 및 발근은 IBA 1.0-2.0mg/L 또는 NAA 0.1 mg/L가 첨가된 배지가 효과적이었으며, 발근된 신초의 순화는 perlite와 peat moss의 1:1 혼합용토 또는 peat moss가 적합하였다. In order to micropropagate uniform plantlets of Philodendron wend-imbe, the shoot tips were cultured on media supplemented with 0.5-10.0mg/L BA or 0.01-1.0 mg/L thidiazuron (TDZ). The multi-bud clusters from basal part of shoots formed vigorously on media containing 5.0-10.0 mg/L BA or 0.05-0.1 mg/L TDZ. Shoot formation from the bud cluster sections (5-7mm) was achieved favorably on medium with 5.0mg/L BA and 20 g/L sucrose. Lowering of sucrose in medium to 20 g/L was effective for the inhibition of callus growth from basal part of shoots. Growth of shoots and their rooting were favorable on media containing 1.0-2.0 mg/L IBA or 0.1mg/L NAA. The rooted plantlets were acclimatizated effectively in soil mixed with perlite 1: peat moss 1 or peat moss alone.

분화용 Anthurium andreanum ′Atlanta′의 기내번식

한봉희,구대회,Han, Bong-Hee,Goo, Dae-Hoe 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.2

In order to establish micropropagation system Anthurium andreanum 'Atlanta', dwarf type, shoots of A. andreanum were cultured on medium supplemented with cytokinin. Callus was formed from the base of shoots. high frequency callus induction was obtained on medium with 10.0mg/L BA or 10.0mg/L TDZ(thidiazuron) at more than 71.8%. The shoots were cultured on media with various combinations and concentrations of TDZ, BA and 2.4-D to enhance callus induction. Callus was induced at more than 72.6% and grew vigorously on media containing 10.0mg/L BA and 0.0∼0.5mg/L 2.4-D, or 1.0mg/L TDZ. Stimulation effects of cytokinin by 2.4-D did not occur in combined treatments of cytokinin and 2.4-D. Callus was cut into sections(7${\times}$10mm), and then cultured on media with BA alone or BA and 2.4-D to regenerate shoots and to stimulate the callus growth. Shoot regeneration and callus growth were effective on media with 10.0mg/L BA alone, or 10.0mg/L BA and 0.1mg/L 2.4-D. In combined treatments of BA and 2.4-D, stmulation effects of cytocinin by 2.4-D also did not occur. Callus growth was decreased, accordiong to increasing the concentration of 2.4-D. In cimbined treatments of TDZ and 2.4-D in shoot regeneration and callus proliferation, stimulated effects of cytokinin by 2.4-D did not occur entirely. Media with 0.5∼1.0mg/L TDZ ingibited the regeneration and rooting of shoots, and callus growth from callus sections. Addition of 2.4-D on medium with TDZ ingibited the regeneration and rooting of shoots, and callus growth. Rooted plantdts were acclimatized in greenhouse. The plantlets were survived more than 98% in soil of vermiculite alone or mixed perlite 1 and vermiculite 1.

신초 Cluster 형성에 의한 Lilium Asiatic Hybrid 'Hae Hwa'의 기내번식

한봉희,유희주,예병우,구대회,Han, Bong-Hee,Yu, Hee-Ju,Yae, Byeoung-Woo,Goo, Dae-Hoe 한국식물생명공학회 2002 식물생명공학회지 Vol.29 No.1

This experiment was conducted to micropropagate bulblets via shoot cluster formation and massproduce normal bulblets from the sections of proliferated shoot clusters in Lilium asiatic hybrid 'Hae Hwa'. The induction of shoot clusters from the culture of bulblet sections was more effective than that of bulb scales on MS medium with 1.0 mg/L BA and 0.5 mg/L IAA. Proliferation of shoot clusters from the formed shoot cluster sections was the most favorable on medium containing 5.0 mg/L BA and 0.5 mg/L IAA. The formation and the growth of bulblets from shoot cluster sections were achieved effectively on medium with 60∼90 g/L sucrose. The leaves derived from shoot clusters grew vigorously but the bulblets from shoot clusters grew very poor in 5L air-lift bioreactor culture. By the addition of 30 mL fresh liquid medium containing doulble strength MS salts, 250 g/L sucrose and 5 g/L activated charcoal after 8 weeks in the shoot cluster culture on MS medium with 5.0 mg/L BA and 0.5 mg/L IAA, the number of bulblets was increased in light condition, but the growth of bulblets was not affected by light. Bulblet production was possible with the bulblet product at 53 to 68 mg in fresh weight by liquid medium addition after the proliferation of shoot cluster. Lilium asiatic hybrid 'Hae Hwa'의 인편 및 자구절편 배양으로 신초 cluster를 유도하여 자구를 대량증식하고, 형성된 신초 cluster에서 정상적인 자구를 대량생산하고자 일련의 실험을 실시하였다. MS배지에 BA 1.0mg/L와 IAA 0.5 mg/L가 첨가된 배지에서 신초 cluster의 유도는 인편보다 자구절편을 배양하는 것이 효과적이었다. MS배지에 BA 5.0 mg/L와 IAA 0.5 mg/L 첨가배지에서 신초수 및 신초무게가 양호하여 형성된 신초 cluster 절편에서 신초 cluster의 증식에 적합하였다. 또한 자구의 비대는 MS 배지에 sucrose 60∼90 g/L가 첨가된 배지에서 양호하였다. MS배지에 sucrose 30∼90 g/L를 주입한 5L airlift 생물반응기에 신초 cluster 절편체를 배양한 결과, 신초만 무성하게 자랐고 자구비대는 불량하였다. 신초 cluster를 8주간 배양한 후에 2배 MS 염류와 sucrose 250g/L, 활성탄 5 g/L를 포함한 액체배지를 동일용기에 첨가하여 배양한 결과, 명배양에서 형성된 자구수는 증가하였으나 소자구의 생장은 명, 암 모두 비슷하였다. 이로써 형성된 소자구의 무게가 53∼68 mg이며, BA 5.0 mg/L와 IAA 0.5 mg/L가 첨가된 MS 배지에서 신초 cluster를 증식한 후, 액체배지 첨가방법에 의하여 효율적인 소자구의 생산이 이루어졌다.

Lilium longiflorum 'Nellie White'의 인편으로부터 Friable 배발생 캘러스를 통한 소자구 분화

한봉희,이수영,서은정,우종규,Han Bong-Hee,Lee Soo-Young,Shu Eun-Jung,Woo Jong-Gyu 한국식물생명공학회 2005 식물생명공학회지 Vol.32 No.2

Lilium longiflorum 'Nellie White'에서 부서지기 쉬운 배발생 캘러스 (FEC)를 유도하여 FEC를 통한 소자구 재분화 체계를 확립하고자 실시하였다. FEC를 통한 나리 소자구 분화는 2.0 mg/L dicamba가 첨가된 MS 기본배지에 나리 인편을 배양하여 단단한 캘러스를 유도하고, 유도된 단단한 캘러스를 동일배지에서 3번 이상 계대배양하여 단단한 일반 캘러스를 증식하였다. 증식된 단단한 캘러스는 $1{\sim}2mm$ 정도의 크기로 절단하여 2.0 mg/L dicamba와 90 g/L sucrose가 첨가된 MS배지에서 배양하여 FEC를 유도하였다. 2개월 간격으로 계대배양하면서 FEC를 유도하였으며, FEC 유도율은 단단한 캘러스를 동일 배지에 계대배양 하였을 때 증가하였다. 유도된 FEC는 $1.0{\sim}2.0\;mg/L$ dicamba와 90 g/L sucrose가 첨가된 MS배지에서 5배 이상의 증식율을 보였다. 증식된 FEC에서 소자구 분화는 0.1 mg/L BA, 1.0 g/L NAA, 30 g/L maltose가 첨가된 1/2 MS 배지에서 양호하였다. 그러나 많은 재분화된 소자구가 투명화 되었다. 건전한 소자구의 재분화는 30 g/L sucrose와 $0.5{\sim}1.0%$ 활성탄이 첨가된 MS 배지가 가장 효과적이었다. A series of experiments were performed to establish regeneration system through friable embryogenic callus (FFC) of Lilium longiflorum 'Nellie White'. Only hard and regular callus was induced from bulb scales on medium containing 2.0 mg/L dicamba and $30{\sim}90$ g/L sucrose. The induced hard callus was subcultured on medium with 2.0 mg/L dicamba and 30 g/L sucrose, and used as a material for induction of FEC. In order to induce FEC, induced hard and regular callus was chopped into $1{\sim}2\;mm$ segments, and re-cultured on medium with 2.0 mg/L dicamba and 90 g/L sucrose. FEC was induced from chopped hard calli by the subcultures of two months interval. The induction rate of FEC was enhanced when hard callus was subcultured on same medium. FEC was proliferated more than 5 times on medium with $1.0{\sim}2.0\;mg/L$ dicamba and 90 g/L sucrose. Bulblet differentiation from FEC was very favorable on MS medium supplemented with 0.1 mg/L BA, 1.0 mg/L NAA and 30 g/L maltose, but many differentiated bulblets were changed to vitrificated ones. The differentiation of normal bulblets was most effective on medium containing $0.5{\sim}1.0\%$ activated charcoal and 30 g/L sucrose.

액체배지 첨가에 의한 Anthurium andreanum ′Atlanta′의 기내생육 촉진

한봉희,구대희,Han, Bong-Hee,Goo, Dae-Hoe 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.2

In order to enhance shoot elongation and rooting of Anthurium andreanum 'Atlanta' in vitro, 15mL of liquid media containing various concentrations of activated charcoal, sucrose and MS salts were added in same vessels after small shoots were induced from the calli on mudium supplemented with 10.0mg/L BA and 0.1mg/L 2.4-D. The post-supplying of 15mL liquid medium containing MS macro and micro elements, 30g/L sucrose and 5.0∼10.0g/L activated charcoal was significantly stimulated the shoot elongation and rooting of regenerated shoots from calli. The medium addition was also resulted in the enhanced soil survival, elongation and rooting of plantlets in cultural soil mixed with perlite and vermiculite(1 : 1)

Alocasia cadieri Chantrier의 기내번식

한봉희,예병우,구대회,유희주,Han, Bong-Hee,Yae, Byeoung-Woo,Goo, Dae-Hoe,Yu, Hee-Ju 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.1

본 실험은 Alocasia cadieri Chantrier를 기내배양하여 일시에 균일한 식물체를 대량생산하기 위하여 실시하였다. Alocasia의 경정을 TDZ 0.1mg/L가 첨가된 배지에 배양하였을 때 신초증식이 가장 높았으며, 형성된 신초에서 신초의 증식은 TDZ과 NAA가 각각 0.5mg/L첨가된 배지에서 효과적이었다. 형성된 신초의 발근은 IBA또는 NAA가 첨가된 배지보다는 활성탄 2.0g/L가 첨가된 배지에서 양호하였으며, 소식물체의 순화는 perlite : vermiculite가 1:1로 혼합된 용토 또는 vermiculite가 적합하였다. In order to micropropagate uniform plantlets of Alocasia cadieri Chantrier in vitro, the shoot tips were cultured on media containing various concentrations of BA and thidiazuron (TDZ). Multiple shoot formation from shoot tips was very effective on medium containing 0.1mg/L TDZ. The formed shoots from shoot tips were separated into a shoot, and cultured on media with BA, TDZ, and NM combination for proliferation. The shoots were multiplied very vigorously on medium with 0.5mg/L TDZ and 0.5mg/L NAA. The rooting and growth of multiplied shoots were more effective on medium with 2.0g/L activated charcoal, rather than those with IBA and NAA. Rooted plantlets show high survival in soil mixed with perlite 1: vermiculite 1 or vermiculite alone.

생물반응기를 이용한 Lilium longiflorum ′Geogil′의 대량번식

한봉희,서은정,예병우,유희주,Han, Bong-Hee,Suh, Eun-Jung,Yae, Byeoung-Woo,Yu, Hee-Ju 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.3

자구를 형성 및 비대시키기 위하여 BA 0.5 mg/L와 IAA 0.5 mg/L가 첨가된 MS 배지에서 Lilium longiflorum 'Geogia'의 신초를 증식하였다. 증식된 신초 cluster를 5 L air lift 생물반응기에 배양하였다. 자구비대는 sucrose 60 g/L 및 1∼2배 사이의 MS 배지 염류농도에서 효과적이었다. 재료 투입량은 100 g을 투입하는 것이 자구형성 및 자구비대에 적절하였다. 배양방법은 air-lift 방법이 자구의 형성 및 비대를 촉진하였으며, 주입공기량은 200∼300 mL$.$$min^{-1}$을 주입하는 것이 자구비대에 효과적이었다. Shoot clusters were induced from bulb scales of Lilium longiflorum 'Geogia', and proliferated on medium containing 0.5 mg/L BA and 0.5 mg/L IAA. Thereafter, these shoot clusters were cultured in 5 L air-lift bioreactors to form and grow normal bulblets. Number of bulblets increased on medium with 30 g/L sucrose, but growth of bulblets was effective on medium with 60 g/L sucrose. The number of bulblets from shoot clusters had no differences, though bulblet growth was very effective on medium with between full and double strength of MS salts. The inoculation of 100 g shoot clusters as a cultural material was suitable for formation and growth of bulblets in 5 L bioreactors. Air-lift type was more effective for the formation and growth of bulblets than that in ebb and flood one, and 200∼300 mL$.$min$^{-1}$ injection of air was suitable in growth of bulblets.

Agrobacterium LBA4404에 의한 국화 ′Shuho-no-chikara′에 LEAFY유전자의 도입

한봉희,예병우,이숙이,이수영,신학기,Han, Bong-Hee,Yae, Byeoung-Woo,Yi, Sook-Yi,Lee, Soo-Young,Shin, Hack-Kee 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.4

Several experiments were carried out to transfer LEAFY gene to Dendranthema grandiflora 'Shuho-no-chikara' by Agrobacterium LBA4404 carrying pSK109 encoding LEAFY gene. Kanamycin 10mg/L was used in first selection medium, and 20mg/L in the second one. Co-culture for 3 days was more effective in increasing transformation efficiency than that for 7 days. The transformation efficiency by Agrobacterium LBA4404 carrying pSK109 encoding LEAFY gene was about 2.8% until the second selection, but only 0.13% of shoots (two plants) was confirmed as a transgenic plants in Southern analysis. The escape of putative transformants was occured seriously in the process of selections, PCR analysis for confirming of neomycin phosphotransferaseII (npt II), and Southern analysis for LEAFY gene. One transgenic plant appeared 7 days'early flowering in field. pSK109를 포함하고 있는 Agrobacterium LBA4004을 통하여 국화 'Shuho-no-chikara'에 LEAFY 유전자을 도입하였다. 엽절편체의 생존율은 kanamycin 10mg/L가 첨가된 배지에서 생존율 약 22%, 절편체의 재분화율이 10%정도였다. 그러나 kanamycin 20mg/L가 첨가된 배지에서는 엽절편은 약 5%정도 생존하였으나 전혀 재분화가 나타나지 않았다. 따라서 kanamycin 10mg/L를 1차 선발배지로, 20mg/L를 2차 선발배지로 선발하였다. Agrobacterium LBA4404와 국화 엽절편체를 3일간 공동배양 하는 것이 형질전환에 효과적이었다. LEAFY유전자가 삽입된 pSK109 vector를 포함한 Agrobacterium LBA4404의 형질전환 효율은 2차 선발까지 약 2.8% 였으며, Southern blot한 결과 0.13%만이 형질 전환체로 확인되었다. 형질 전환체 한계통은 포장에서 약 1주일 정도 조기개화하였고 화형은 정상이었다.

알스트로메리아의 器內大量繁殖

한봉희(Han Bong-Hee),김영진(Kim Young-Jin),최주견(Choi Ju-Kyun) 한국원예학회 1993 한국원예학회 학술발표요지 Vol.11 No.1

나리 ‘Casa Blanca’의 인편배양에서 유기한 저반부로부터 자구의 형성과 증식에 미치는 생장조절제와 광의 영향

한봉희(Han Bong Hee),예병우(Yae Byeong Woo),구대회(Goo Dae Hoe),고재영(Ko Jae Young) 한국원예학회 1999 Horticulture, Environment, and Biotechnology Vol.40 No.4