초록 ▼

○ 연구결과
- 분화용 및 실내 조경용 소재로 활용가치가 높은 식충식물 13속 86개의 종 및 품종을 수집하여 기내에 도입하였다.
- Dionaea속 1종, Drosera속 7종, Byblis속 1종, Sarracenia속 1종, Darlingtonia속 1종, Heliamphora속 1종 등 12종의 기내배양에 적합한 배지 종류와 생장조절물질의 처리조건, 적정질소원, sucrose 농도 그리고 agar 첨가량 등을 구명하였다.
- Dionaea속 1종, Drosera속 11종, Byblis속 1종, Brochina

○ 연구결과
- 분화용 및 실내 조경용 소재로 활용가치가 높은 식충식물 13속 86개의 종 및 품종을 수집하여 기내에 도입하였다.
- Dionaea속 1종, Drosera속 7종, Byblis속 1종, Sarracenia속 1종, Darlingtonia속 1종, Heliamphora속 1종 등 12종의 기내배양에 적합한 배지 종류와 생장조절물질의 처리조건, 적정질소원, sucrose 농도 그리고 agar 첨가량 등을 구명하였다.
- Dionaea속 1종, Drosera속 11종, Byblis속 1종, Brochina속 1종, Cephalotus속 1종, Sarracenia속 2종, Darlingtonia속 1종, Heliamphora속 1종, Pinguicula속 3종, Nepenthes속 1종 등 총 23종의 기내배양묘를 재료로 하여 기외순화 시 적정 배양토 및 차광율, 관수방법, 피복처리 및 발근촉진제 처리의 효과 등을 구명하였다.
- Dionaea속 1종, Drosera속 4종, Pinguicula속 1종, Sarracenia속 1종의 유식물체 재배에 적합한 배양토, 차광조건, 관수방법 및 hyponex의 시비효과를 구명하였다.
- Dionaea속 1종, Drosera속 5종, Byblis속 1종, Sarracenia속 1종 및 Pinguicula속 1종 등에서 기내배양중인 식물체의 엽 절편에 EMS, NMU, NaN3 및 colchicine 등과 UV 처리의 적정조건을 구명하였고, 기외이식 후 형태적 특성을 분석하여 변이주를 선발하였다.

Abstract ▼

Subtitle Ⅰ : Selection of promising insectivorous plants for pot and indoor garden.
Eighty-six species or varieties belonging to 13 genera in promising insectivorous plants for pot and indoor garden were collected and introduced in vitro. They are 1 species of Brocchinia, 3 species or varieties o

Subtitle Ⅰ : Selection of promising insectivorous plants for pot and indoor garden.
Eighty-six species or varieties belonging to 13 genera in promising insectivorous plants for pot and indoor garden were collected and introduced in vitro. They are 1 species of Brocchinia, 3 species or varieties of Byblis, 1 species of Cephalotus, 1 species of Darlingtonia, 19 varieties of Dionaea, 25 species or varieties of Drosera, 2 species or varieties of Genlisea, 2 species of Heliamphora, 1 species of Ibicella, 8 species of Nepenthes, 5 species of Pinguicula, 17 species or varieties of Sarracenia, and 1 species of Utricularis.
Subtitle Ⅱ : Development of mass-propagation methods using tissue culture.
This study was performed to investigate the effect of materials, culture media, medium components (growth regulators, nitrogen, sucrose, activated charcoal, anti-oxidizer, agar), and medium pH on shoot regeneration in order to establish the in vitro mass-propagation system.
1. Dionaea muscipula
The ideal material for in vitro shoot regeneration of Dionaea muscipula was the upper part of petiole. Modified Parliman medium was suitable for shoot multiplication. The highest number of shoot regeneration (39.8/explant) was observed on modified Parliman medium supplemented with 5 μM kinetin and 1 μM IAA among the various combinations of growth regulators and media tested, and the shoot growth was relatively good. The best shoot regeneration was observed on the medium solidified with 0.6% agar, but growth was better on the lower concentration of agar.
The addition of MnSO4․4H2O and CuSO4․5H2O was favorable for shoot proliferation, with the optimum concentrations, 22.3 mg․L^-1 and 0.025 mg․L^-1 respectively. Anti-oxidizers, such as PVP, DTT and DTE, inhibited the regeneration and growth of shoots. Ascorbic acid and citric acid were favorable for shoot regeneration with the best results on 200 mg․L^-1 ascorbic acid. Activated charcoal had great influence on the shoot regeneration and growth. The maximal shoot formation, 88.5 shoots/explant, was achieved on high concentration (2%) of activated charcoal. Dark pretreatment, up to 4 weeks, enhanced shoot multiplication and growth.
2. 6 species of Drosera
Six species belonged to Drosera had a different response to culture condition. The best shoot proliferation from leaf sections of Drosera species was achieved, when cultured on MS medium containing 0.02∼0.05 μM kinetin and 0.005 μM IAA.
Drosera species showed generally low nutrient requirement. The highest shoot production was obtained with 1/4 strength of MS medium for D. binata, D.capensis, D. burmanni, and with 1/2 strength of MS medium for D. spatulata, D.tokaiensis, than with any other strength of MS medium tested. The nitrogen requirement of Drosera species were also relatively low. The maximum shoot regeneration was achieved when cultured in 1/4 strength of nitrogen contained MS medium, except D. tokaiensis and D. burmanni required 1/2 strength of nitrogen.
The optimum sucrose concentration for each species was different, such as 4% for D. binata, D. burmanni and D. spatulata; 3% for D. capensis; 2% for D. tokaiensis; 1% for D. rotundifolia. In general, activated charcoal had positive effect on shoot proliferation, but had negative effect on root formation of Drosera species.
The best shoot regeneration was achieved on MS medium containing 0.05% activated charcoal for D. binata, D. burmanni, D. spatulata, and 0.01% for D. capensis, D. tokaiensis, D. rotundifolia.
Shoot regeneration of Drosera species was better in MS liquid medium, except D. capensis and D. spatulata showing best results on 0.6% agar. The maximal shoot proliferation was achieved on the media adjusted to pH 4.5 for D. spatulata and D. burmanni, pH 5.0 for D. binata, pH 5.5 for D. tokaiensis respectively.
3. Byblis liniflora
In order to establish and optimize an in vitro micropropagation method of Byblis liniflora the effects of plant growth regulator types and concentrations on shoot proliferation and callus formation were investigated, using node explants. The highest number of shoots (16.7/explant) was observed on medium supplemented with 10 μM BA and 2 μM IAA. The callus formation were most effective on medium supplemented with 10 μM BA.
4. Sarracenia purpurea
In this work, in vitro clonal propagation of Sarracenia purpurea was obtained from seedlings germinated in vitro. The shoot proliferation was most effective on medium supplemented with 1 mg․L^-1 kinetin and 1 mg․L^-1 NAA. The sucrose concentration had a dramatic effect, with 5% being optimal, while varying pH level of media had only a moderate effect. Shoot proliferation was promoted by the addition of ascorbic acid and citric acid, and optimum content of activated charcoal was 5%. The regeneration capacity of explants was influenced by medium type and agar content, the highest number of plants was obtained on Parliman liquid medium.
5. Drosera alelea
The highest number of plants regenerating from leaf explants of Drosera alelea was obtained on medium supplemented with 0.02 mg·L^-1 or 0.05 mg·L^-1 BA. Shoot proliferation was more effect on modified Parliman medium than any other medium type, but root formation was improved on 1/4MS medium. The highest shoot production was obtained in 1/4 strength of nitrogen contained MS medium, and optimum ratio of NH₄Cl:KNO₃ was 15:15 mM. The regeneration capacity of explants was promoted by addition of 0.3 0.4% sucrose. Liquid culture medium significantly increased regeneration capacity of leaf tissue, but plant growth improved in 0.6 0.7% agar. The most effective temperature for plantlet regeneration was 25℃.
6. Darlingtonia californica
Shoot proliferation from plantlets of Darlingtonia californica was most effective on 1/2MS medium supplemented with 2 mg·L^-1 BA. While varying concentration of total nitrogen source had only a moderate effect, optimum ratio of NH₄Cl:KNO₃ was 10:20 mM. The highest number of plants was obtained in 3% sucrose and the shoot proliferation was most effective on medium without activated charcoal. Liquid culture medium significantly increased regeneration capacity of leaf tissue and fresh weight.
7. Heliamphora minor
In this work, in vitro clonal propagation of Heliamphora minor was obtained from seedlings germinated in vitro. Shoot proliferation was most effective on medium supplemented with 1 mg․L^-1 BA and 2 mg․L^-1 NAA. The shoot regeneration capacity was influenced by medium type, the highest number of plants was obtained on MS basal medium, but plant growth was more improved on 1/4MS medium. The shoot regeneration was most effective in NH₄Cl:KNO₃ =5:25 mM, but plant growth was most vigorous in NH₄Cl:KNO₃ = 15:15 mM. Generally, low concentration of sucrose (1 2%) was more effective on shoot regeneration. Optimum agar content of medium was 0.6%, Liquid culture medium significantly increased regeneration capacity of leaf tissue and fresh weight.
Subtitle III : Establishment of acclimatization conditions of shoots cultured in vitro.
Experiments were conducted to find the optimal acclimatization condition for plantlets of insectivorous plant cultured in vitro, various acclimatizing conditions were compared regarding both survival rate and growth of the plantlets.
1. Dionaea muscipula and Drosera tokainsis
Growth of Dionaea muscipula plantlets was not influenced by soil media, and also differences of anthocyanin or chlorophyll contents were significant among plantlets grown in different soil types. The highest grow rate was observed when Dionaea muscipula plantlets were acclimatized at 25℃. Plantlet growth of Drosera tokainsis was most effective in pots filled with a mixture of peatmoss:vermiculite (2:1), and anthocyanin content was highest in peatmoss: vermiculite (3:1). Plantlets acclimatized at 25℃ exhibited the further growth and chlorophyll content of leaf was also highest at 25℃.
2. 5 species of Drosera
Independent of species, the best growth of plantlets was observed when plantlets were acclimatized in pots filled with peatmoss only at 25℃, and plant growth and shoot regeneration was more effective on 50% shading than any other shading conditions.
3. Byblis filifolia
Survival rate and plantlets growth exhibited the best result when plantlets were acclimatized in compost with fertilizer with 70% shading. Among the various watering method, the further growth of plantlets was observed in spray irrigation 2 time/day. When Byblis filifolia plantlets transplanted to pots, wrapping was effective, with 4 weeks being optimal, but soaking in rooting promoters, such as Ruton, IAA (100, 200, 400 mg.$L^{-1}$) or IBA(100, 200, 400 mg.$L^{-1}$) inhibited the survival and growth of plantlets.
4. Nepenthes ventricosa
The best growth of plantlets was observed when plantlets were acclimatized in pots filled with compost without fertilizer. Among the various watering method, survival rate and plantlet growth, such as leaf length and plant width, exhibited the best result in flooding 2 times/day. When Nepenthes ventricosa plantlets transplanted to pots, wrapping for 1 week was improved plantlet growth.
5. Brochinia reducta and 16 species
The optimum soil medium for acclimatization was so different according to species, such as peatmoss:perlite (5:1) for Brochinia reducta, Drosera burmanni, Drosera lanata, Drosera petiolarlis ‘all red’, Drosera occidentalis ‘white flower’, Pinguicula ehlersae, Pinguicula moranensis; peatmoss:perlite (3:1) for Byblis filifolia, Heliamphora minor; peatmoss only for Cephalotus forlicularis, Dionaea muscipula ‘E’, Drosera occidentalis ‘pink flower’, Drosera regia, Darlingtonia californica, Pinguicula primuiflora, Sarracenia leucophylla f. red tubes, Sarracenia purpurea. Among the various watering method, survival rate of Darlingtonia californica plantlets was highest in spray irrigation 1 time/day, and survival rate and growth of Darlingtonia californica plantlets exhibited the best results in flooding 2 times/day for 45 min.
Subtitle Ⅳ : Establishment of growing process to excellent plantlets.
This study was performed to investigate the effect of soil media, shading, watering, and fertilization method on growth of insectivorous plants.
1. Dionaea muscipula
Plant growth of Dionaea muscipula was excellent in peatmoss only, peatmoss: vermiculite (2:1) or peatmoss:perlite (2:1) treatmemt with 30% shading. Among the various watering methods, the further growth of plants was observed in spray irrigation 2 times/day or 2 days flooding-1 day ebbing treatment. Survival ratio and plant growth were influenced by Hyponex concentration and fertilization method, the best results observed in foliar application with 1.0 g・L^-1.
2. Drosera aliciae
Plant growth of Drosera aliciae was excellent in peatmoss only or mixture peatmoss and other soil. Survival rate and plant growth exhibited the best result when plants were cultivated with 30% shading. Among the various watering methods, the best result was observed in 1 day flooding-2 days ebbing treatment. Survival ratio and plant growth were influenced by Hyponex concentration and fertilization method, the best survival rate and further growth were exhibited in foliar application with 0.5 g・L^-1 or soil application with 0.1 g・L^-1.
3. Drosera capensis
Plant growth of Drosera capensis was excellent in sphagnum moss only or peatmoss:perlite (2:1) or peatmoss:vermiculite (2:1) treatment. Survival rate and plant growth exhibited the best result when plants were cultivated with 50% shading. Among the various watering methods, the best result was observed in 1 day flooding-2 days ebbing treatment. Survival ratio and plant growth were influenced by Hyponex concentration and fertilization method, the best survival rate and further growth were exhibited in foliar application with 0.5 1.0 g・L^-1 or soil application with 0.2 g・L^-1.
4. Drosera rotundifolia
The optimum soil medium for cultivation of Drosera rotundifolia was peatmoss:perlite (2:1) treatment. Survival rate and plant growth exhibited the best result when plants were cultivated with 30% shading. Among the various watering methods, the best result was observed in 3 days flooding-1 day ebbing treatment. Survival ratio and plant growth were influenced by Hyponex concentration and fertilization method, the best survival rate and further growth were exhibited in foliar application with 0.5 g・L^-1 or soil application with 0.2 g・L^-1.
5. Drosera tokaiensis
The optimum soil media for cultivation of Drosera tokaiensis were sphagnum moss only or peatmoss:vermiculite:perlite (2:1:1) treatment. Survival rate and plant growth exhibited the best result when plants were cultivated with 30% shading. Among the various watering methods, the best result was observed in 1 day flooding-1 day ebbing treatment. Survival rate and plant growth were influenced by fertilization method of Hyponex, with foliar application being more effective than soil application
6. Pinguicula moranensis
The optimum soil media for cultivation of Pinguicula moranensis were sphagnum moss olny or compost only treatment. Survival rate was excellent when plants were cultivated with 0 30% shading, but plant growth was vigorous at 50% shading. Among the various watering methods, the best result was observed in spray irrigation 2 times/day. Survival rate and plant growth were influenced by Hyponex concentration and fertilization method, foliar application with 0.5 g・L^-1 was most effective.
7. Sarracenia purpurea
The optimum soil media for cultivation of Sarracenia purpurea were peatmoss olny or peatmoss:vermiculite (2:1) only treatment. Survival rate and plant growth exhibited the best result when plants were cultivated with 30% shading. Among the various watering methods, the further growth of plants was observed in spray irrigation 1 time/day or flooding 1day-ebbing 3day treatment. Survival rate and plant growth were influenced by Hyponex concentration and fertilization method, the best survival rate and further growth were exhibited in foliar application with 1.0 g・L^-1 or soil application with 0.2g・L^-1.
Subtitle Ⅴ : Breeding of new varieties by induced mutation.
To induce mutation from explants of insectivorous plants, the effects of chemical mutagen and UV ray treatments on survival rate of explants and mutant induction was investigated according to the treatment time and concentration.
1. Investigation of optimum condition for induced mutation in vitro.
In most species, the optimum condition of EMS for mutation was determined to be 20 mM for 1 or 3 hours. In case of NMU, LD50 was close to 2 mM for 1 or 3 hours. When NaN₃ treated on explants, The 0.5 mM NaN₃ treatment for 1 or 3 hours resulted in about 50% survival rate to non-mutagenized. In most species, the optimum condition of colchicine for mutation was determined to be 0.05% for 1 or 2 days. Among the various treatment periods of UV ray for mutation, survival rate to non-mutagenized was 46.2% in explants of Drosera rotundifolia treated with UV ray for 10 min.
2. Variant selection and characteristics analysis
The frequencies of variant induction varied according to mutagen type and plant species, generally low in most of Drosera spp. and Sarracenia purpurea. The best result was observed in Pinguicula moranensis, which total 177 lines were obtained from EMS, colchicine and NaN₃ treatments. Especially, 116 lines were selected from EMS treatment. Selected lines had various leaf colors (variegated white, yellow, red or dark green) and leaf shapes (spatulate, crispate or legginess).