Bulbophyllum auricomum Lindl. is a rare orchid and has flowers with an attractive fragrance. The present study investigated the tissue culture method for micropropagation. Capsules derived from artificial self-pollination were obtained for the best seed germination in MS basal medium. Plant growth regulators (1.0 mg·L-1 of BAP and 2.0 mg·L-1 of NAA) were affected by callus induction from subcultured pseudobulb explants. For the callus subculture, different natural plant extracts were tested in 11 treatment media. Among them, MS medium with 150 mL·L-1 of coconut water was generally effective in fresh weight (1.75 ± 0.08) and (3.01 ± 0.20) of callus proliferation and PLBs induction at 1 and 2 months, respectively, followed by an MS combination of 30 g·L-1 of banana and 20 g·L-1 of potato extract. The results of a comparative study of different MS mediums containing plant growth regulators with a natural extract combination and MS medium supplemented with natural extract only showed that MS medium supplemented with a combination of natural extracts (150 mL·L-1 of coconut water) and plant growth regulators (2.0 mg·L-1 of BAP and 1.0 mg·L-1 of NAA) obtained the highest shoot regeneration (3.37 ± 0.17) and (6.41 ± 0.68) after 1 month and 2 months of culturing, respectively.

본 연구는 스프레이 국화의 기내 대량 증식을 위한 적정 배지를 선발하고, 발근시 절편체의 부위와 순화시기가 조직배양 묘의 품질에 미치는 영향을 조사하기 위해 수행하였다. 먼저 대량증식을 위한 적정 배지를 선발하기 위해 서로 다른 조성의 식물생장조절물질을 첨가한 MS배지에 ‘Field Green’, ‘Green Diamond’, ‘Snow Pangpang’ 등 스프레이 국화 3품 종을 기내배양 하였다. 배양 6주 후의 배양체의 생육을 조사한 결과, MS+NAA 0.1 mg·L-1의 배양체의 초장이 무처리에 비해 1.5~2.0배 더 길었고, 줄기의 두께도 더 두꺼웠다. 그러나 BA 0.1 mg·L-1 이상이 첨가된 모든 배지에서는 캘러스와 다신초가 유도되었고 뿌리가 발생하지 않았으며, 신초와 마디의 길이가 짧아졌다. 따라서, MS+NAA 0.1 mg·L-1 배지가 스프레이 국화의 기내 대량증식 단계에 가장 효율적인 것으로 선발되었다. 두번째로 스프레이 국화 우량묘를 양성하기 위하 여 ‘Field Green’ 품종에서 생장점을 포함하고 있는 줄기의 선단부와 액아를 포함하는 줄기의 중단부를 각각 기내배양 한 후, 배양 2주 후부터 일주일 간격으로 순화하여 순화된 묘의 생육특성을 조사하였다. 그 결과, 선단부 배양체의 뿌리출현일수가 11.2일로 중단부 배양체보다 1.7일 더 빨랐으며, 순화율 과 생장의 균일성도 순화시기에 관계없이 중단부 배양체보다 더 좋았다. 또한 초장, 엽수, 줄기의 굵기, 생체중과 같은 조직 배양묘의 소질과 관련된 특성도 선단부 배양체에서 더 좋게 나타났다. 순화시기에 따른 묘 소질 조사에서는 기내배양 2주 후에 순화한 선단부 배양체의 경우 최종 조사일에 초장이 3.8±0.5 cm로 가장 짧았지만, 줄기의 굵기와 생체중은 1.5±0.2 mm와 1.3±0.3g으로 기내배양 3주와 4주 후에 순화한 묘에 비해 1.5~2.0배 더 두껍고 무거워 품질이 우수하였다. 이러한 결과를 바탕으로 스프레이 국화의 조직배양 우량묘를 양성하기 위해서는 기내 발근시 줄기의 선단부를 절편체로 사용 하고, 기내배양 2주 후에 짧고 굵은 뿌리가 발생하기 시작했을 때 순화하는 것이 적절하다.

This is the first report describing the seasonal conditions affecting shoot regeneration by the chrysanthemum cv. Baeksun. The shoot regeneration from petal explants was found to be more favorable from September to December, reaching the highest values in December. In addition, the quality of the shoots was also influenced according to the season of the explant collection, where healthy and uniform plants were derived from the explants collected in December. Choosing the proper season for explant collection affected the successive plant growth parameters (i.e., plant height and fresh weight). Thus, the current results strongly suggest that season plays an important role in plant tissue culturing, which is an essential tool for micropropagation and Agro-bacterium-mediated genetic transformation studies.

A simple and efficient protocol was developed for somatic embryogenesis from the cotyledon explant of Paeonia lactiflora Pall. Seeds of peony obtained from fieldgrown plants were disinfested and zygotic embryos were excised. For germination, excised embryos were cultured on the Murashige and Skoog (MS) medium supplemented with 3% (w/v) sucrose, 0.8% (w/v) agar, and different concentrations of N6 benzyl-adenine (BA) and gibberellic acid (GA3). The greatest germination percentage (95%) was observed when embryos were cultured on the MS medium with 1.0 mg • L-1 BA and 0.5 mg • L-1 GA3, and maintained at 25 ± 2°C under a 16 h photoperiod. Thirty days old cotyledon explants were cultured on the MS medium supplemented with different concentrations and combinations of plant growth regulators viz., BA, GA3, 2,4-dichlorophenoxyacetic acid (2,4-D), and á-naphthalene acetic acid (NAA). After 90 days, the globular embryos were directly formed on the surface of explants. The highest frequency of somatic embryo induction (72.5) was obtained on the MS medium with 3.0 mg • L-1 BA, 1.0 mg • L-1 NAA, 1.0 mg • L-1 GA3, and 0.1% (w/v) activated charcoal (AC), with a mean number of 14 embryos per explant. Maturation of globular embryos into heart- and torpedo-shape was observed on the same medium. When the torpedo-shaped embryos were transferred onto the same MS medium supplemented with 3.0 mg • L-1 BA, 1.0 mg • L-1 NAA, 1.0 mg • L-1 GA3, and 0.1% (w/v) AC, secondary somatic embryos were observed on the surface of primary somatic embryos. When the embryos were transferred to the MS medium supplemented with 1.0mg • L-1 each of BA and GA3, all of them converted into plantlets, but their growth was very slow.

Thee kinds of explants, young spikes, immature and mature embryos, was compared in callus induction, embryogenic callus formation, differentiation and plant regeneration of Russian wildrye. Callus induction efficient of young spikes is highest. The imature embryos formed more somatic embryogenesis, and with highest frequency of embryogenic callus.

국내에 도입된 오차드그래스 27 품종과 국내 합성 품종인 Hapsung 2호의 종자로부터의 캘러스 형성율, 형성된 캘러스의 크기, 캘러스로부터의 재분화율 및 재분화 효율을 4주령 및 6주령의 캘러스에 대하여 얻은 결과는 아래와 같았다. 1. 4주령의 캘러스에 대한 종자로부터 캘러스의 형성율은 치상 종자의 50% 이상이 캘러스를 형성한 형성율 상위 품종들은 93M > Sparta > Pizza> Condor > Lidaglo > Glorus >Hapsun

Background : Glycyrrhiza uralensis Fischer is one of most important medicinal (Glycyrrhizae radix) herbs in traditional oriental medication and they are also important commercial products used worldwide in sweetening and flavoring. They contain the similar compounds, such as the triterpenoid saponins and flavonoids. Therefore, it is necessary to develop more efficient and sustainable methods for the production of G. uralensis Fischer and its medicinal constituents. This study was accomplished to investigate the effect of medium composition on in vitro propagation and plantlet regeneration from nodal explants of G. uralensis Fischer, and to establish a reliable protocol for micropropagation. Methods and Results : Young and actively growing stem segments were excised from adult plants of new cultivar ‘manju’. They were cut into a 1cm nodal segments with single node after sterilization, and cultivated in the different medium supplemented with various plant growth regulators for two weeks. For shoot multiplication, one-node stem segments, approximately 1 ㎝ in length, were taken from in vitro derived shoots and subcultured. After four weeks of culture, the efficacy of each medium on shoot proliferation and growth was determined, and these shoots were transferred onto medium with different auxin hormones for rooting. Conclusion : After seven to ten days of culture, shoots began to emerge from axillary buds. They showed a vigorous growth and elongation in multiplication medium. During culture period, in vitro cultured plantlets showed significantly different responses to the respective medium with different plant growth regulators. Our experiments confirmed that in vitro growth and multiplication of palntlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.

Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on ¼ MS for P. grandiflorum with wild and green petals and on ⅛ MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on ¼ MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, ¼ MS supplemented with 1 mg L-1 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on 0.5 mg ․ L-1 thidiazuron (TDZ) from double petal explants grown on ⅛ MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.

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.

The influences of ethylene inhibitors (AgNO3 and silver thiosulfate) and cytokinins (BAP and TDZ) on shoot regeneration from cotyledon and hypocotyl explants of B. napus cv. Youngsan were investigated. The presence of 50 μM Silver thiosulfate (STS) in shoot regeneration medium formed shoots at 60-68% after 3-4 weeks of culture, which was enhanced by 2-fold compared to that of Silver nitrate (AgNO3). Moreover, cotyledon explants were more regenerative than hypocotyls; shoots from cotyledon explants began to occur 4-5 days earlier than that of hypocotyl explants. TDZ at a concentration of 8-10 μM was effective for shoot regeneration, compared with BAP. Consequently, the optimal shoot regeneration response was observed in medium supplemented with 50 μM STS + 8 μM TDZ. In transmission electron microscopy (TEM) analysis, higher density of silver nanoparticles was shown to be accumulated widely inside the cell wall and plasmodesmata of regenerating leaf cultured in medium supplemented with AgNO3. By contrast, in the cell cultured in medium with STS, fine-grained deposits were partly observed in the surroundings of the cell wall.

Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (≥ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose- 1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

국화 ‘백마’의 잎 절편체로부터 신초 기관형성을 통한 식물체 재분화 기술을 개발하고자 배지조성과 배양기간을 달리하여 실험을 실시하였다. 잎 절편체를 식물생장조절제와 AgNO3의 농도를 달리 한 11가지 배지에서 배양하였을 때 bud는 형성되었으나 신초로의 발달은 이루어지지 않았다. Bud 유도 이후 신초 형성을 유도하고자, 서로 다른 식물생장조절제 조성을 가진 배지를 이용한 2단계의 실험을 수행하였다. Bud유도배지(BA 0.2 mg/L, IAA 1.0mg/L, 2,4-D 1.0mg/L를 첨가한 MS 배지)에서 1-4주간 배양한 후 신초 형성배지(식물생장조절제 무첨가, BA 1.0 mg/L, IBA 1.5 mg/L와 AgNO3 5.0 mg/L 또는 BA 1.0 mg/L, NAA 1.0 mg/L와 AgNO3 5.0 mg/L를 각각 첨가한 MS 배지)로 옮겨 배양하였다. Bud 유도에 이어 신초 형성을 위해서는 bud 유도 배지에서 3주 이상의 배양기간이 필요하였으며, bud 유도배지에서 4주간 배양한 후 BA 1.0 mg/L, IBA 1.5 mg/L와 AgNO3 5.0 mg/L를 첨가한 신초 형성배지로 옮겨 4주간 배양한 처리에서 신초 형성율은 50.0%, 절편체당 신초 수는 1.6개로 가장 양호한 결과를 얻을 수 있었다. 재생된 신초는 신초 형성배지에서 8주간 배양 후 신장하였으며, 1 cm 이상 신장된 신초를 분리하여 발근을 유도한 후 활착시켜 온실에서 재배하였을 때 식물체는 정상적인 표현형을 보여주었다.

본 연구는 기내에서 발아된 까마중 유식물체의 조직별로 신초 및 뿌리 유도를 통해 효율적인 재분화 조건을 확립하고자 실시하였다. 신초 형성은 식물체 부위별로 서로 달랐으며, 잎에서의 신초 형성빈도가 자엽이나 하백축 그리고 상배축보다 높았다. 절편체당 신초의 수는 잎과 자엽에서 다소 많았으며, 신초의 길이도 다른 부위에 비해 잎과 자엽에서 빠른 증식과 신장을 나타내었다. 특히, 2.0 mg L-1 BAP가 첨가된 배지에서 재분화율, 신초의 수와 길이가 증가하여 2.0 mg L-1 BAP가 고빈도 신초의 유도 및 생장에 효율적인 것으로 판단되었다. 뿌리분화는 IBA와 IAA가 NAA보다 더 효과적인 것으로 나타났으며, 0.05 mg L-1 IAA가 첨가된 배지에서 뿌리 형성율이 90%이며, 뿌리의 갯수와 길이도 각각 평균 4.0개와 7.82 cm로 가장 양호하게 나타났다. 더욱이 배양토에 이식한 재분화체는 100%의 생존율을 보였으며 정상적인 식물체로 생장하였다. 이러한 결과를 토대로 2.0 mg L-1 BAP 조건에서의 잎 절편에서 다발성 신초 유도, 그리고 0.05 mg L-1 IAA 조건에서의 뿌리형성 유도가 까마중의 대량번식을 위한 최적의 조건임을 알 수 있었다.

고감미 다수성 계통인 자가수정 스테비아의 hypocotyl 절편체를 이용하여 대량증식방법 개발에 대한 결과는 다음과 같다. 선발된 자가수정개체는 초장이 72.0~120.7 cm로 분포되어 있었고, 분지수는 4.9~56.6개로 분포되어 있었다. HPLC 분석을 통하여 rebaudioside-A(RA)성분이 평균함량 55.2%이상 함유된 자가수정 개체를 선발하였다. 스테비아의 hypocotyl 절편체를 이용하여 MS배지에 1.5 mg/L BA와 0.5 mg

Somatic embryos on growth regulator-free medium can be produced directly from cotyledon explants of Panax ginseng C. A. Meyer. When the embryo developmental stage was torpedo and cotyledon, the germination rate of embryos was quite high on MS medium supplemented with gibberellic acid (GA3). However, the percentage of plantlet formation at the cotyledon stage was higher than that at the torpedo stage. This result demonstrates that the embryo at the cotyledon stage was the most appropriate for increasing germination by GA3. Embryos cultured on medium including four levels of GA3 concentrations (3, 5, 10, or 20 mg/l) showed all quite high germination rates (87-91%). When the well-developed embryos were continuously cultured on media including high concentrations of GA3 from 10 to 20 mg/l, the percentage of formation of normal plantlets was lower than that seen under low concentrations from 3 to 5 mg/l. This treatment of high concentrations resulted in shoots with abnormal shape. The optimal GA3 treatment provides a basis for the efficient method obtaining healthy plantlets derived from ginseng somatic embryos.