Water lettuce (Pistia stratiotes L.) is a free-floating perennial herbaceous plant with rosette leaves and a stem. Although this plant multiplies and has adverse effects on aquatic ecosystems, it can be used for biological purification of polluted water and production of valuable substances as a traditional medicine. In this study, we report a protocol to establish an in vitro micropropagation method based on direct shoot organogenesis from stem explants. In media comprising two types of basal medium and different growth regulators, multiple shoot organogenesis was observed on stems. The micropropagation method was most effective on Schenk and Hildebrandt (SH) medium with 1 mg/L indole-3-acetic acid and 2 mg/L 6-benzylaminopurine supplemented with 30 g/L sucrose, on which all explants produced multiple shoots. The shoots rooted spontaneously on solid half-strength Murashige and Skoog (MS) and SH media without growth regulators (1/2MSO and 1/2SHO). However, roots developed more vigorously in liquid media. Regenerated plants colonized and grew more rapidly in SH basal medium than in MS basal medium and produced 6–8 stolons within 2 weeks on 1/2SHO. In summary, we established a method for micropropagation in vitro through direct organogenesis of water lettuce, which shows the potential of water lettuce as a model aquatic plant for phytochemical and pharmacological research.

산겨릅나무(Acer tegmentosum Max)는 단풍나무과의 약용식물로 기내 대량증식 및 발근에 적합한 최적 배지와 식물생장조절물질의 농도를 구명하고, 순화조건을 검증하여 대량생산조건을 확립하기 위해 수행되었다. 산겨릅나무의 종자 소독 후 1.0mg l-1 GA3가 첨가된 MS배지에서 발아시켜 기내식물체를 확보하였고, 생장에 적합한 최적배지 구명을 위해 7종류의 다양한 식물배지에서 4주간 배양한 결과, WPM 배지가 가장 효과적이었다. 액아배양을 통한 대량증식에 최적 조건을 구명하기 위하여 4종의 cytokinin의 다양한 농도별 처리를 통해서 8주 동안 배양한 결과, 1.0mg l-1 Zeatin이 첨가된 배지에서 평균 3.3개의 다경줄기가 유도되었다. 기내발근에는 0.5mg l-1 2.4.-D가 단독 첨가된 배지에서 뿌리 수가 높았고, 다신초 개체는 기외의 토양으로 이식하여 환경적응을 유도한 후 배양묘 대량생산 조건을 확립하였다.

본 연구는 조직배양을 이용한 효율적인 블루베리의 묘 생산 체계 개발을 위하여 수행되었다. 신초형성을 위한 초대배양을 위해서는 WPM 배지가 MS 배지보다 신초형성률 , 마디수, 신초장 등에 있어서 유의적으로 우수하였다. 식물 생장 조절제가 신초형성에 미치는 식물생장조절제의 영향을 조사하기 위해서는 zeatin(1, 2mg·L-1), 2iP(10, 15mg·L-1), 그리고 BA(4, 6mg·L-1)를 처리하였으며, 신초의 증식에 미치는 영향을 확인하기 위해서는 zeatin(0.5mg·L-1), 2iP(10, 15mg·L-1), 그리고 BA(0.05mg·L-1)를 처리하였다. 신초형성 에 미치는 결과는 zeatin 처리가 신초형성률, 마디수, 신 초장 등에서 유의적으로 효과적이었다. 반면 무처리와 BA 처리에서는 신초가 형성되지 않았으며, 2iP 처리의 경우 신초형성률이 낮았다. 기내 삽목으로 생산된 블루 베리 신초의 계대배양에 있어 신초 발생 및 생장에 미치는 영향은 zeatin을 농도별로 처리한 결과, 모든 농도에서 신초의 형성률과 형성된 신초의 개수는 유의적 차이를 보이지 않은 반면 zeatin의 농도가 증가할수록 신초의 마디수는 증가하나 신초장은 짧아지는 경향을 보였다. 신초 증식을 위한 적정한 zeatin 농도는 0.5mg·L-1 였다.

An efficient method for the rapid propagation of Smilax china from axillary buds was established. Plants with thick leafage were selected from Korea native S. china population. Axillary buds of S. china collected from selected plant and were cultured in various culture media (2MS, MS, 1/2MS, WPM, B5 and SH medium). Shoot was induced from axillary bud on MS basal medium after 4 weeks of culture. 1/2MS medium showed a higher growth rate than those of the others, while the lowest shoot growth was obtained in 2MS medium. Among the sucrose concentrations, 5% sucrose was the optimum level for shoots growth from axillay buds. Among cytokinins, 0.5 mg L -1 6-benzylaminopurine (BAP) treatment showed the best performance on shoot multiplication, yielding average shoot multiplication forming about 2.4. Rooting was induced directly near the base of the shoot on 1/2MS medium containing with three-auxins α-napthalene acetic acid (NAA), indole acetic acid (IAA) and β-indolebutyric acid (IBA) (0.5 and 1.0 mg L -1 ). The 1.0 mg L -1 IBA treatments induced earliest rooting with maximum of root number and root growth. These rooted plantlets were successfully transferred to pots for 4 weeks hardening process, and were transferred to soil with above 90% survival rate.

이전 실험에서 결정된 생육 단계별 최적 환경조건을 평가하기 위한 4가지 처리는 다음과 같았다: 생육 단계별 최적 환경 조건을 사용한 광독립 영양배양(photoautotrophic optimum condition with growth stage (POG)), 생육 단계별 평균 광합성 광량자속 밀도(photosynthetic photon flux density(PPFD))와 CO2 농도를 사용한 광독립 영양배양(photoautotrophic constant condition with average PPFD and CO2 of POG(PCA)), 생육 단계별 최대 PPFD와 CO2농도를 사용한 광독립 영양배양(photoautotrophic constant condition with maximum PPFD and CO2 of POG(PCM)) 그리고 대조군으로 3%의 당을 포함한 광혼합 영양배양(photomixotrophic conventional condition with 3% sucrose(PMC)). 실험 결과 각 생육 단계별 환경제어(POG)는 기내에서 배양된 감자 소식물체의 모든 생육 관련 항목에서 유의적 증진을 유도하였다. 또한 단위 건물중 당 소비된 전력과 CO2는 모든 처리 중 POG에서 가장 낮았다. 기외 이식 이후에도 POG에서 생산된 감자 묘는 PMC에서 자란 감자 묘와 전체적으로 큰 차이 없이 왕성한 생육을 유지하였다. 특히 POC는 기존 광혼합 영양방식(PCM)과 비교했을 때 기외 이식전과 이식 후 20일째 각각 4.7배와 3.8배 높은 건물중을 기록하였다. 따라서 POG와 같은 생육 단계별 환경 조절을 통한 광독립 영양 미세 증식 방법은 에너지 절감 효과와 함께 무균의 건강한 감자 묘의 생산에 효과적이었다.

본 연구는 묘의 생육을 최대화하기 위하여 생육 단계를 임의로 구분하고 각 단계 별 적정 환경 조건을 확립함에 목적을 두었다. 생육 단계는 총 20일의 배양기간을 6일(1단계), 7일(2단계), 7일(3단계)의 3단계로 구분하였다. 첫 번째 단계는 활착기로서 환경 처리 별 생육에 큰 차이가 나타나지 않았다. 높은 환경 조건에 의한 잎의 장해를 고려하였을 때, 80μmmol·m-2·s-1 의 PPFD 및 대기 중의 CO2 농도가 적합하였다. 두 번째 단계에서는 PPFD 및 CO2 조건이 높아짐에 따라 건물 중을 중심으로 부분적으로 향상되었다. 에너지 효율과 생육을 고려할 때, 160μmmol·m-2·s-1 의 PPFD와 700μmmol·mol-1의 CO2가 적합할 것으로 생각되었다. 세 번째 단계에서는 PPFD 및 CO2 농도가 높아짐에 따라 유의적으로 생육이 향상되었으며, 잎 및 마디의 발달상태도 현저히 향상되었다. 따라서 보다 적극적으로 생육증진을 고려할 때, 320μmmol·m-2·s-1 PPFD와 1800μmol·mol-1의 CO2가 적합할 것으로 생각되었다. 생육 단계별 환경 조절은 초기단계에 상대적으로 낮은 조건을 유지하고 후기단계에서 충분한 조건을 제공함으로써 건전한 묘를 생산할 수 있고 에너지 및 물질의 투입량을 절약할 수 있다.

Cryopreservation is one of the ideal and suitable methods for long-term storage of plant germplasm. The plant contaminated with diseases and pathogens are decreased the multiplication rate, survival rate and high quality of plants after cryopreservation. The aim of this work was to improve a micropropagation method for lily in Korea, which is indigenous plant. In the last process of rinsing scales after surface-sterilization, we tried to control the diseases and pathogens lived within the tissue by rinsing in 0.03% sodium hypochlorite (NaClO) instead of sterile distilled water. Bulb scales of Lilium were cultured in vitro on MS medium supplemented with Plant Preservative Mixture (PPM). The results showed that L. tsingtauense accessions were observed ranged from 53.9 to 100% with a mean value of 76.8% and L. hansonii accessions were checked from 84.5 to 85.5% with a mean of 85% survival rate. The newly small bulb formed from bulb-scales was transferred to MS medium. We checked the presence of microorganisms and survival rate after 3 weeks in culture after examination of bacterial incidences. The results indicated that the non-contamination rate were shown ranged from 75.0 to 94.1% with mean value of 83.2% in L. tsingtauense species, and that L. hansonii were observed 85.1 to 91.7% with mean value of 88.4%. This study will provide a valuable basis for establishment of effective axenic cultures for in vitro micropropagation of Korean native lily species.

Background : Cnidium officinale Makino is a perennial herb of the family Umbelliferae, and an important traditional oriental medicinal plant in China, Japan and Korea. Cnidii Rhizoma, the dried rhizome of C. officinale have been used as traditional oriental medicine in Korea. It has been shown that the cnidii rhizomes are used in the treatment of pain, inflammation, menstrual disturbance, and anti-vitamin deficiency disease, and also act as a blood pressure depressant. In this study, we anticipated to establish the mass propagation system of C. officinale, which is a high economic value as medicinal herb, by plant tissue culture to solve the problem of root stocks contamination. Methods and Results : The whole study was carried out in the department of Herbal crop research, Eumseong, RDA. In this study, C. officinale root bud was used as a explant and it was surface sterilized by 1% sodium hypochlorite for 3 minutes, then several times washed with ddH2O. Multiple shoots were induction them on MS, B5, SH media with 0.1 - 2 ㎎/ℓ auxin (NAA, IBA) and cytokine (BA, Zeatin). In this study we obtained, 7.4 multi-shoots per an explant, and the shoot growth was also favorable in the presence of 0.2 ㎎/ℓ Zeatin. Subsequent transfer of these regenerated shoots on 1/2 SH media resulted in root formation. Rooted plantlets were able to grow in soil after a short period of acclimatization. Conclusion : This experiment was comducted to identify the optimal in vitro propagation condition of C. officinale.

We micropropagated pear (Pyrus species) using shoot tips and nodal explants from three pear genotypes. The ability to establish shoot tip cultures, proliferate shoots, induce rooting, and acclimatize the resulting plantlets are all elements of in vitro micropropagation. Shoots were induced from shoot tips on Murashige and Skoog medium (MS) with five different plant growth regulator combinations. The highest shoot formation rates were achieved for the three genotypes using MS supplemented with 1.0 ㎎/L N 6 -benzyladenine (BA) and 0.1 ㎎/L gibberellic acid (GA3). The maximum shoot number and shoot length for the three cultivars were recorded with 2.0 ㎎/L BA and 0.2 ㎎/L indole-3-butyric acid (IBA) in multiplication medium using nodal explants produced from microshoots. Nodal explants with one or two axillary buds cultured for three weeks initiated roots on medium supplemented with various concentrations of 1-naphthaleneacetic acid (NAA) or/and IBA in half-strength MS medium for adventitious rooting. The highest rooting response was with the combination of 0.2 ㎎/L NAA and 0.2 ㎎/L IBA. A combination of NAA and IBA resulted in a significant increase in the rooting ratio over NAA or IBA alone. In this medium, the root formation rate according to ranged from 68.9% for the BaeYun No. 3 genotype to 51.8% for the Hwanggeum genotype. We also investigated the influence of the concentration the polyamine phloroglucinol in rooting medium. For all three genotypes, the highest rooting ratio, longest root length, and greatest root number were observed in the treatments with 75-150 ㎎/L phloroglucinol. Most rooted plants were acclimatized successfully.

Shoot tips of chinese yam (Dioscorea opposita Thunb.) were cultured on MS medium containing 0.5 mg/L BA to produce micro-tubers in vitro. To stimulate the formation of shoots and micro-tubers, and produce large micro-tubers, the sections of micro-tubers were cultured on MS media with BA and IAA. The shoot multiplication, and the micro-tuber formation and growth were very effective on the media containing 2.0 mg/L BA and 0.5~1.0 mg/L IAA. Sucrose added to MS medium with 2.0 mg/L BA and 0.5 mg/L IAA to stimulate more micro-tuber growth. The medium added 50 g/L sucrose was very effective in the increase of plant fresh weight and micro-tuber growth. After 4 weeks' culture of micro-tuber sections on the medium with 2.0 mg/L BA, 0.5 mg/L IAA and 50 g/L sucrose, the liquid media were added into the same vessels. The micro-tuber growth was stimulated remarkably by the addition of liquid medium. The addition of 25 ml liquid medium containing 10 g/L activated charcoal, 3x MS salts and 250 g/L sucrose was the most effective in micro-tuber growth.

할미꽃의 정단 분열조직배양에 의한 기내 효율적 미세번식에 미치는 식물생장조절물질의 효과를 조사하기 위해 2,4-D, NAA, kinetin, TDZ와 BA를 혼용처리한 MS배지에 정단분열조직을 배양하였다. 배지내 2,4-D와 kinetin, 2,4-D와 TDZ, NAA와 TDZ 혼용처리는 캘러스 유도에 효과적이지 못하였으나 NAA와 BA가 혼용 처리되었을 때 배발생적 또는 기관분화성 캘러스가 유도되었다. 특히 0.5mg/L NAA와 1.0mg/L BA 혼용처리된 MS 배지에서는 캘러스로부터 고빈도로(62%) 신초가 형성되어 캘러스 유도 및 신초형성에 적합하였다. 적합배지에 10% coconut water를 첨가하여 30일마다 계대배양 하므로 캘러스 증식률, 신초 분화율 및 신초 신장이 현저히 증가되었는데, 그 결과 절편체 당 평균 12.9개의 shoot bud를 분리할 수 있었다. 할미꽃 기내 유도된 식물체에서 뿌리는 쉽게 분화되지 못하였으나 9.0mg/L NAA 단용 또는 0.5mg/L NAA와 1.0mg/L BA 혼용처리된 MS배지에서 캘러스를 통해서만 분화되었다. 이들 뿌리는 동일 배지에서 캘러스유도 및 식물체 분화를 반복적으로 실시하는 시원체가 되기도 하였다.

The effect of plant growth regulators was investigated on in vitro shoot proliferation from axillary bud explants of Hypericum erectum. To determine the optimal cytokinin for proliferation of axillay buds, we carried out screening four cytokinins (BA, kinetin, 2iP, TDZ). When nodal segments were cultured on MS medium supplemented with 4.5 μM TDZ (thidiazuron), a number of shoots were induced. Our results indicated that the addition of TDZ to culture medium resulted in the induction of significantly more axillary buds than in the addition of other cytokinins. The optimal concentration of TDZ for proliferation of axillary buds was 10 μM. 92% of shoots spontaneously rooted without any plant growth regulator (PGR) and formed whole plantlets within one month. More than 95% of these regenerants survived and they did not show any detectable variation in morphology or growth characteristics compared to their donor plants.

기관배양을 통한 대량증식 방법의 일환으로 액아배양을 이용한 multiple shoot 유도 조건을 조사하였다. 기내 배양 중인 식물체로부터 액아를 절취하여 TDZ와 BA를 각각 농도 별로 첨가한 MS배지와 1/2 MS배지에서 6주간 배양한 결과, MS기본배지에 0.005 mg·L-1 TDZ가 첨가된 처리구에서 효과가 가장 양호하였다. 한편 발생된 신초를 1 mg·L-1 GA3와 0.5 mg·L-1 IBA가 첨가된 1/2 MS배지에 배양했을 때 뿌리의 발생과 신장이 가장 양호하였다. 발근된 식물체를 포트에서 순화시켜 온실로 이식하였을 때 90%에 가까운 생존율을 보였으며 형태적인 변이 없이 생장하였다. 이상의 결과는 액아유도를 통한 대량번식의 가능성을 제시한다.

유색 칼라 세 품종(Sunlight, Chiante, Pink Persuation)의 기내 급속증식체계를 확립코자 정단분 열조직을 배양함에 있어 callus, 신초 및 뿌리 분화에 미치는 식물생장조절물질의 효과와 sucrose가 기내 괴경 형성 및 비대에 미치는 효과를 조사하였다. 식물체는 캘러스를 통하거나 또는 치상체에서 직접 분화되는 두 경로로 형성되었으며 2.0 mg/L BA 단용 처리에서 Sunlight품종은 53.3% 캘러스를, Chiante 품종은 56.7% 직접 신초를 형성하였다. 2.0-3.0 mg/L BA 단용 처리는 품종별로 식물체재 분화 빈도가 20-70%로 차이가 있었으나 세 품종 모두 3.0 mg/L. BA 단용 처리에서 신초 분화가 양호하였다. 다아체 형성에 미치는 cytokinin의 효과는 Sunlight품종은 1.0 mg/L 2ip 처리로 16개체를, Chiante 품종은 5.0 mg/L BA를 처리로 14개체를, Pink Persuasion품종은 1.0 mg/L BA처리로 12개체를 형성하였다. NAA는 세품종 뿌리분화에 효과가 없었으며 Sunlight 와 Chiante 품종은 1.0 mg/L IAA에서, Pink Persuasion 품종은 2.0 mg/L IBA가 효과적이었다. 괴경형성에 미치는 sucrose는 Sunlight 품종은 90 g/L, Chiante 및 Pink Persuasion 품종은 70 g/L 가 괴경 형성 및 비대에 효과적이었다.

The establishment of an efficient protocol for plant regeneration and micropropagation from leaf explant cultures of Chicory, Cichorium intybus L. var. sativus. is reported. Callus formation rate appeared 100% from explant in all growth regulators, but calli formed in the prensence of naphthaleneacetic acid (NAA) were appeared very compact and non-embryogenic state. The regenerated shoots were obtained from leaf explant cultures on solid MS medium containing different concentrations of cytokinins and auxin. The highest number of shoots (5.7) per explant and shoot growth (2.8cm) was obtained on MS medium containing 0.1 mg BAP L-1 and 0.1 mg NAA L-1 . Indole acetic acid was the most suitable auxin for root formation among three auxins tested. 2,4-D had no effect on shoot and root formation.