Soybean is the one of recalcitrant legume species for shoot induction. Shoot regeneration via direct organogenesis was investigated in five soybean cultivars, ‘Dawon’, ‘Pungsan’, ‘Daewon’, ‘Taekwang’ and ‘Chongdoo 1’ by using cotyledonary node explants. Out of 5 soybean cultivars, an efficient shoot regeneration condition was developed in the two soybean cultivars, ‘Dawon’ and ‘Pungsan’. When various kinds of plant growth regulators with different concentration were estimated, the optimum medium condition for shoot induction in both soybean cultivars was MS + B5 vitamin supplemented with BA at concentration 2 ㎎/L. In addition, shoot formation efficiency was increased with 97.09% and 93.88% by the pretreatment of BA onto the explants before in vitro culture in both cultivars. Shoot induction in ‘Dawon’ cultivar was originated from epidermal tissue and sub-epidermal layers when histological changes were investigated under shoot regeneration after culturing cotyledonary node segments on shoot induction medium for 0 to 21 days. Especially, cell dedifferentiation was observed from parenchyma cells to meristematic cell in 3-day cultured segments.

An efficient shoot regeneration condition for pea cv. ‘Sparkle’ was developed by using optimum explant, plant growth regulator concentrations, and pretreatment of BA onto explant. The average shoot number per explant showed the highest on two kinds of shoot induction media (MSB5 media containing 2 ㎎/L BA and a combination of 2 ㎎/L BA and 1 ㎎/L TDZ) when cotyledonary node explants were cultured. Moreover, the pretreatment of explant in 200 ㎎/L BA solution was found to be more effective in shoot induction than that of non-pretreatment. By histological study, cell division and proto-meristem were formed near the surface of the sub-epidermal and epidermal cell layers of cotyledonary node in earlier than 3 days after culture. The analysis of genetic stability of regenerants by using thirteen ISSR markers showed that in vitro regenerated plants showed polymorphism with 8.3% compared with their mother plants.

The quality control of natural products is principal key to guarantee the Good Manufacturing Practices (GMP) and Good Clinical Practices (GCP) for the functional food, pharmaceuticals and cosmeceuticals in the industry. In this study, we examined the quantitative analysis of berberine as marker substance of Coptidis Rhizoma by high performance liquid chromatography-photodiode array detector (HPLC-DAD). The HPLC method was validated and met all the requirements for the quality control analysis recommended by FDA and ICH. The berberine was separated on a Xterra C18 column (5 ㎛, 4.6 × 250 ㎜) using mobile phase consisting of distilled water and acetonitrile with KH2PO4 (3.4 g) and Na2SO4 (1.7 g). Calibration curve of berberine has been estimated (y = 42293.47x-41589 with the correlation coefficient 0.9999). The amount of berberine was calculated as 4.25%. And berberastine, palmatine, columbamine, jatrorrhizine, epiberberine, berberine and coptisine in the Coptidis Rhizoma were identified by high performance liquid chromatography - electrospray ionization-mass spectrometer (HPLC-ESI-MS) method.

다양한 환경에서 수집한 국내 민들레속 유전자원 수집종의 엽록체 DNA 영역(trnL-trnF와 rps16-trnK) 염기서열을 이용하여 종내․ 간 변이 및 배수성을 구명하여 유전자원 육성의 기초 자료룰 제공하고자 수행하였다. 민들레속 유전자원의 배수성은 털민들레, 서양민들레, 붉은씨서양민들레가 3배체이고, 흰 민들레와 흰노랑민들레는 4배체였다. 염기서열의 길이는 trnLtrnF 영역에서 자생종류인 털민들레, 흰민들레, 흰노랑민들레 가 931 bp에서 935 bp, 서양민들레는 910 bp, 붉은씨서양민들레 는 975 bp로 종간 차이를 나타내었고, 종 특이적 염기서열 88개, 자생종 및 귀화종 특이적 염기서열 41개가 검출되었다. rps16- trnK 영역은 털민들레 882∼883 bp, 흰민들레 875∼881 bp, 흰 노랑민들레는 878∼883 bp 서양민들레 874∼876 bp, 붉은씨서 양민들레는 847∼848 bp로 37개 종특이적 염기서열이 검출되 었다. 염기서열의 유사도는 trnL-trnF 영역에서 0.860∼1.000 사이로 평균 0.949이며, rps16-trnK 영역의 유사도는 0.919∼ 1.000 사이로 평균 0.967이었다. 염기서열을 바탕으로 유연관계를 분석한 결과, trnL-trnF 영역은 크게 자생종류와 귀화종 류로 구분되었으며, 서양민들레와 붉은씨서양민들레는 같은 종 간에 유집되었고, 자생종류는 분리되지 않았으며, rps16-trnK 4개 그룹과 유집되지 않은 5개체로 나뉘었다. 흰노랑민들레는 두 영역 모두 흰민들레와 동일 계통군을 형성하였고, 염기서열 상 두 종간 뚜렷한 차이가 없었다. 유연관계에서 모두 독립적으로 존재한 흰민들레 No. 10 (조계산)과 털민들레 1번(광양)은 민들레 유전자원 육성소재로 활용이 기대된다.

The morphological characteristics and genetic relationships among 32 germplasms of Zanthoxylum schinifolium and Zanthoxylum piperitum collected from two farms in Korea were investigated. The traits with the most variability were seed color, leaf size, and spine size. The intraspecific polymorphism of Z. schinifolium and Z. piperitum was 96.5% and 60.3%, respectively. The genetic diversity and Shannon’s information index values ranged from 0.11 to 0.33 and 0.19 to 0.50, with average values of 0.26 and 0.42, respectively. Two ISSR primers (UBC861 and UBC862) were able to distinguish the different species. The genetic similarity matrix (GSM) revealed variability among the accessions ranging from 0.116 to 0.816. The intraspecific GSM for Z. schinifolium and Z. piperitum was 0.177–0.780 and 0.250–0.816, respectively. The GSM findings indicate that Z. schinifolium and Z. piperitum accessions have high genetic diversity and possess germplasms qualifying as good genetic resources for cross breeding. The clustering analysis separated Z. schinifolium and Z. piperitum into independent groups, and all accessions could be classified into three categories. Z. Schinifolium var. nermis belonged to independent groups. Comparison of the clusters based on morphological analysis with those based on ISSR data resulted in an unclear pattern of division among the accessions. The study findings indicate that Z. schinifolium and Z. piperitum accessions have genetic diversity, and ISSR markers were useful for identifying Z. schinifolium and Z. piperitum.

Plant regeneration via organogenesis and somatic embryogenesis was investigated in Korean soybean cultivars including Cheongja 3, Jinpumkong 2, Taekwangkong and Uram. Cotyledon, cotyledon+hypocotyl and hypocotyl segments of 7-day-old seedlings were cultured on MS medium containing various concentration (0, 1, 2 and 4 ㎎/L) of BA and TDZ. The results showed that MS medium supplemented with BA 2.0 ㎎/L yielded the highest shoot formation ratio of 83.3%. In 4 cultivars, Taekwangkong showed the highest ratio of shoot formation. When various sizes of immature cotyledons (S: 1∼ 2 ㎜, M: 3∼5 ㎜, L: 6∼8 ㎜) were tested on MS medium containing 2,4-D 40 ㎎/L for somatic embryogenesis, the optimum size for embryogenic callus induction was 3∼5 ㎜ in length of immature cotyledons. In 4 cultivars, Taekwangkong showed the highest percentage of embryogenic callus induction. The results indicate that Taekwangkong is the best soybean cultivar for plant regeneration via organogenesis and embryogenic callus induction among the 4 cultivars.

기내배양한 양란 심비디움 원괴체에 전자빔(2 MeV/n, 0.5 mA)을 10 Gy～300 Gy로 조사하여 생장 변화와 유전적 변이의 다양성을 검토하였다. 전자빔을 조사 후 배양 10주째에 선량의 증가에 따라 원괴체로부터 신초유도가 감소하였고, 200 Gy 이상의 선량을 조사한 처리구에서는 신초분화가 크게 억제되었다. 원괴체 18개체를 ISSR 분석한 결과 대조구는 12.5% 다형성을 나타내었고, 전자빔 조사구에서는 42.2%의 다형성을 나타내었다. RAPD 분석한 결과 대조구에서 13%의 다형성을 나타내었고, 전자빔 조사구에서는 43.8%의 다형성을 나타내었다. 따라서 RAPD 및 ISSR 분석 모두에서 전자빔 조사시 대조구보다 3.5배 정도 다형성이 증가한 것으로 나타났다. 유전적 유사도 지수(GSM)는 전자빔 처리가 대조구보다 낮게 나타나 전자빔 처리에 의하여 개체들 간의 유전적 다양성이 높아진 것으로 보인다. 군집분석 결과 RAPD와 ISSR 분석 모두 대조구와 전자빔 처리구가 분리되어 그룹을 이루었다. 이와 같이 전자빔 조사로 기내 배양 양란 심비디움의 유전적 다형성이 증대됨을 확인하였다.

Dandelion has been widely utilized for medicinal and edible purposes. This research was conducted to evaluate the effect of supplemental LED (light-emitting diode) light on germination, growth characteristics and anthocyanin content of dandelion (Taraxacum officinale) seedling using LED blue (460 nm), red (660 nm, R), blue + red (B:R=6:4) and fluorescent lamp light treatment. By LED illumination to T. officinale seed germination speed was delayed, and germination rate was the highest in the fluorescent light. The growth characteristics (plant height, number of leaves, root length and fresh weight) were greatly influenced by supplemental LED light compared with control treatment, and the growth promotion was the most effective in the red LED illumination. After 60 days of red and mixed LED light treatments, anthocyanin content of dandelion plants was significantly changed. The anthocyanin content was increased by 12~19 mg/100 g under the red LED and the mixed light conditions compared with the control and the blue LED. Results indicate that illumination with red and mix LEDs, compared with other light treatments, is beneficial for promotion of growth and anthocyanin content in dandelion.

Mutation breeding is characterized by its merit, creation of new mutant characters and addition of very few traits without disturbing other characters of variety. Gamma ray is generally used to induce mutation in various crops. Heading dat is one of the key factors in the regional and seasonal adaptation of rice verity. This study was carried out to evaluate agronomic characteristics, genetic variation and grain quality of early heading rice (Oryza sativa L. cv. Dongjin 1) lines derived from gamma-ray (Co60, 300 Gy) irradiation. The average heading date of the early heading lines in M7 and M8 generation was faster than that of mother verity as 11 (line γ-2), 10 Aug (line γ-5), 6 Aug (γ-1 line), 5 Aug (γ-3 and X-1), 4 (line γ-4) days, respectively. The selected lines showed shorter culm length and ear length compared with the control. Total spikelets per panicle, percent ripened grain and thousand grain weight of the lines were also mostly shorter or lower than those of the control except total spikelets per panicle of γ-4 and γ-5 lines. When genetic variations of rps16-trnK region were evaluated by nucleotide analysis, nucleotide length of the rps16-trnK region was 664 bp in all the early-heading lines and control. Out of 5 sites of nucleotide transposition detected in the region, however, 2 sites were appeared only in the early-heading lines. Amylose content of the early heading lines ranged from 18.3% to 18.4% and the content was included in high eating quality. Protein content of the lines ranged from 6.6% to 7.0%. The content belonged to Top-rice showing under 7% of protein content. Toyo taste value of the lines ranged from 76.3% to 84.8%.

Ionizing radiations have been effective mutagen to overcome the limitation of the useful genetic resources in natural environment. This study was conducted to investigate an effect of proton beam on germination, growth patterns in the irradiated dandelion (Taraxacum officinale), and genetic variation in 12 morphological mutants induced in proton-beam irradiated dandelion. Percentage germination rate was drastically decreased over 250Gy. The lethal dose 50 (LD50) of germination was estimated between 250 Gy to 500 Gy. Significant decreases in growth patterns (plant height, number of leaf and fresh weight) were observed by increase of dose (Gy) of proton beam irradiation. According to the correlation analysis between dosage and growth factors, the orders of compactness of correlation were germination, plant height, fresh weight and number of leaf, respectively. Twelve morphological variants such as, dwarf, color, plastid, growth and leaf shape were screened at 50 to 250 Gy of the beam irradiation. As a result of ISSR analysis of the 12 variants, out of 33 bands detected overall, 8 bands were identified to be polymorphic with a rate of 24.2% at the control group. While 33 bands detected overall, 21 bands were identified to be polymorphic with a rate of 63.6% at the proton beam irradiation. The result indicates that the dandelion with proton beam treatment might be promoted variation at DNA level

Dandelion has been widely used as a folkloric medicine for treatment of diverse diseases. This study was conducted to evaluate the effect of light spectrum using red (660 nm), blue (460 nm), red and blue mixed (Red : Blue = 6 : 4) LED (light emitting diodes) and fluorescent lamp on growth and functional components of dandelion (Taraxacum officinale). When LED was illuminated to T. officinale cv. Goldenboll, seed germination was delayed, and germination rate was the highest in the control (fluorescent light). The growth (plant height, root length and fresh weight) except leaf number was increased under the LED treatments compared with the control, and the growth promotion was the most effective in the red LED illumination. Total polyphenol contents in dandelion irradiated with the red and blue mixed or the red LED were 121.77 mg% or 115.36 mg%, respectively, which were greater than those in dandelion treated with blue LED and fluorescent lamp. Asparagine, proline, serine, threonine, glutamic acid and arginine were the predominant amino acids in dandelion and total amino acid was the highest under the Red LED illumination. The results indicate that application of the red and the mixed LED illumination promote growth and increase functional components during cultivation of dandelion

Genus Taraxacum has been widely used as a folkloric medicine for treatment of diverse diseases. The genetic diversity and relationship among 32 accessions belonging to five Taraxacum species (T. mongolicum T. coreanum, T. coreanum var. flavescens, T. officinale and T. laevigatum) which collected from field, mountain, island and seaside of Korea were evaluated using ISSR markers. A total of 142 ISSR loci detected in the overall species were all polymorphic loci (100%) and interspecies polymorphisms obtained from Korean native and naturalized species were 98.2% and 94.5%, respectively. The genetic similarity matrix (GSM) among 32 accessions ranged from 0.025 to 0.860 with an average of 0.303. According to the clustering analysis, the Korean native species and naturalized species were divided two major clusters. In addition, the different species were divided into independent groups except for the T. coreanum and T. coreanum var. flavescens, and all the 32 accessions could be classified into 7 categories. The study findings indicate that Taraxacum accessions have a high genetic diversity and the dandelion accessions as breeding materials can be effectively utilized for the improvement of Taraxacum breeding.

Ionizing radiations have been effective mutagen sources to overcome the limitation of the useful genetic resources in natural environment. The study was conducted to investigate an effect of electron beam on organogenesis, growth patterns and genetic variation in the irradiated orchid organs (rhizome and protocorm). The in vitro cultured rhizomes of orchids (Cymbidium goeringii, C. kanran) were irradiated with the electron beam in the dose range of 15Gy to 2240Gy under the condition of various beam energy and beam current (10MeV/n and 0.5mA, 10MeV/n and 0.05mA, 2MeV/n and 0.5mA). Significant decreases in growth and organogenesis were observed by increase of intensity of electron beam irradiation. The irradiation intensity of lethal dose 50 (LD50) of the in vitro cultured orchid was estimated as approximately 500Gy to 1000Gy under 10MeV/n, and 1000Gy to 2000Gy under 2MeV/n. While, the optimal irradiation dose was approximately 100Gy for growth and organogenesis of the cultures under 10MeV/n with 0.05mA treatment, and 300Gy under 2MeV/n and 0.5mA electron beam condition. RAPD and ISSR analyses for the electron beam irradiated organs were performed to analyze genetic variation under the electron beam condition (2MeV/n and 0.5mA).

RAPD (random amplified polymorphic DNA) analysis was examined to detect variation of in vitro cultured 30 rhizomes of Cymbidium goeringii and Cymbidium kanran, with long-term subculture, respectively. Out of 105 DNA bands detected, the 38 were polymorphic with a polymorphic rate 36.1% in the C. goeringii. Out of 126 DNA bands detected, the 58 were polymorphic with a polymorphic rate 46.0% in the C. kanran. The size of the amplified fragments of the two species ranged from about 200bp to 3,000bp. Genetic similarity matrix (GSM) shows from 0.810 to 1.00 with an average of 0.929 in the rhizomes of C. goeringii and 0.786 to 0.984 with an average of 0.915 in the C. kanran. The result indicate that the long-term in vitro cultured C. goeringii and C. kanran supplemented with growth regulators might be promoted in higher polymorphism and increased GSM. Thus, the higher polymorphism rate of in vitro cultured rhizomes might be resulted in long-term subculture and the plant growth regulators supplemented with the culture medium. The results provide as fundamental data to develop a new materials for plant breeding and resources plant.

To investigate flowering related genes in winter-type oilseed rape (Brassica napus L. cv. Tammi), differentially expressed genes were isolated from leaves of the plant after low temperature treatment which is requirements for floral induction. As a result of suppression subtractive hybridization (SSH), 288 clones were randomly selected from SSH library. Using reverse Northern blot analysis, 150 of 288 clones were identified to be differentially expressed. Out of these 150 clones, 45 clones showed very high identities with the known genes. Four clones showed very high identities over 90% with metallothionein-like gene that is related to flowering-induced genes. Of these 4 clones, the cDNA clone, rfs-13, revealed high identity with meotallothionein-like protein in Arabidopsis thaliana (98%) and Brassica compestris (89%). Furthermore, gene expressed in immature flower stages was confirmed by Northern blot analysis.

Shoot induction system was developed in the recalcitrant plant species, Brassica rapa ssp. rapifera by using optimum selection of profit organ, phytohormone combination, seedling age and kind of culture container. Out of in vitro cultured leaf segment, petiole, hypocotyl, and cotyledon with petiole, only cotyledon with petiole derived from 4 day-old seedlings induced multiple shoot. The optimum combination of auxin and cytokinin for the multiple shoot induction was MS medium containing 5mg/L BA and 0.5mg/L NAA. The major factors for multiple shoot propagation were part of plant organ, age of seedling, and ratio of auxin and cytokinin. In addition, shoot regeneration was promoted in the 100ml Erlenmeyer flask compared with the 90mm×20mm Petri-dish. The induced shoots formed roots easy on MS medium containing 0.1mg/L IBA and the whole plants were successfully cultivated in soil.

Genetic diversity of 45 tea accessions from Korea, Japan, China and Taiwan was investigated by using RAPD analysis. Out of the eighty primers screened, twenty primers generated 99 polymorphic bands with a polymorphic rate 87.0%. The size of the amplified fragments ranged from about 3,138 bp to 520 bp. By cluster analysis, all of the 45 accessions can be grouped into five groups. Over 90% of the 32 Korean accessions belonged to group II, III, IV and V. Moreover, newly developed Korean cultivars (accession no. 13, 14 and 15) belonged to very different group compared with any other Korean accessions. Among the Korean accessions, the minimum genetic similarity 0.500 was obtained between accession no. 17 and 37 and the largest genetic similarity 0.912 between no. 20 and 21.

Intraspecific genetic diversity of sixteen accessions of Mogolian Alliums including fifteen species was investigated using randomly amplified polymorphic DNA (RAPD) analysis. Twenty three out of forty primers revealed scorable polymorphism. A total of 440 RAPD markers were generated on the 16 accessions of Mongolian Alliums. Among 440 RAPDs assayed, 439 were polymorphic with a mean polymorphic rate of 99.7%. Unweighted pair-group method using an arithmetic average (UPGMA) cluster analysis using RAPD data separated the 16 Allium accessions into two broad groups at similarity index 0.70. The clustering of the species was closely related with previous classification between A. altaicum and A. fistulosum. In addition, a high genetic similarity was showed between A. cepa and A. tagar.

The effect of carbenicillin on the dedifferentiation and the regeneration efficiency of plant tissues of horseradish(Armoracia rusticana) was evaluated, Inhibition effect for callus initiation was observed when leaf blade, root and petiole segments were grown on MS medium containing 500 mg/L to 2000 mg/L carbenicillin and 0.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). The regeneration of horseradish shoots from leaf blade, root and petiole explants were decreased as the addition of carbenicillin increased from 1000 mg/L to 2000 mg/L in MS medium containing 0.5 mg/L of 6-benzylaminopurine (BAP) or kinetin. Especially, 500 mg/L carbenicillin treatment significantly inhibited shoot induction when leaf blade explants were grown on hormone-free MS medium. It was suggested that the toxic effects of combinations of carbenicillin and 2,4-D may be due to high auxin activity levels.

The concerns on the crop damage by ultraviolet (UV) radiations is increasing owing to the decrease of their absorbing stratospheric ozone in the tropospheric. Cultivar differences on early growth of UV radiation among five Korean rice cultivars, four japonica types and one Tongil type (indica-japonica cross hybrid), were studied. Pot-seeded rice plants were grown under four different radiation conditions, i.e., visible radiation only, visible radiation with supplemented with high or low dose of UV-B (280~320 nm in wavelength) and UV-C (less than 280 nm in wavelength). The inhibitory degree on plant height, shoot and root weight and length of leaf blade and leaf sheath were determined at 40 days after seeding. UV-C showed the most severe inhibitory effect on the degree of biomass gain and leaf growth in most cultivars examined, followed by high UV-B and low UV-B. Among the cultivars used, the Kuemobyeo was the most sensitive cultivar and had not repair or showed resistance ability to continued irradiation of UV radiation. However, Janganbyeo and Jaekeon showed different responses that the elongation of leaf blades was promoted on 2nd and 3rd leaves and inhibited on 4th and 5th leaves but this inhibitory degree was reduced on 6 th and 7th leaves. Such tendency on leaf growth means that both cultivars had low sensitivity and most resistant ability to continued irradiation of UV radiation. While Tongil showed different response to enhanced UV radiation, ie., low UV-B promoted leaf growth but the inhibitory was severely increased by continued irradiation of high UV-B and UV-C, which means that Tongil had high threshold of UV radiation for response as an inhibitory light of plant growth. The results of this study indicate that the differences on sensitivity or resistant to the effects of UV radiation were existed among Korean rice cultivars