This study describes the identification of Panax species using mitochondrial consensus primers. Initially, a total of thirty primers were tested in ten Korean ginseng cultivars and two foreign Panax species, P. quinquefolius and P. notoginseng. In the polymerase chain reaction (PCR) amplification results, three primers (cox1, nad1/2-3 and nad2/1-2) generated co-dominant polymorphic banding patterns discriminating Korean ginseng cultivars from P. quinquefolius and P. notoginseng. However, these primers could not generated polymorphisms among the Korean ginseng cultivars, and simply represented species-specific polymorphisms for P. quinquefolius and P. notoginseng. Primers PQ91 and PN418 were designed from the consensus sequence of nad1/2-3 region. Two banding patterns (A or B) were detected in PQ91. Korean ginseng cultivars and P. notoginseng shared the same banding pattern (A type) and P. quinquefolius was identified another banding pattern (B type). In the case of PN418, two banding patterns (A or B) were detected in the Korean ginseng cultivars and two foreign Panax species. Korean ginseng cultivars and P. quinquefolius shared the same banding pattern (A type) and P. notoginseng was identified another banding pattern (B type). The combination banding patterns of three Panax species, Korean ginseng cultivars (Panax ginseng C. A. Mey.), P. quinquefolius and P. notoginseng, was identified as 'AA', 'BA' and 'AB', respectively. Consequently, PQ91 and PN418 primer sets can be used to distinguish among Panax species.

본 연구는 인삼 기내에서 생산된 재분화 식물체의 토양 순화를 위해 순화과정을 단축하고 생존율을 높일 수 있는 방법을 개발하기 위하여 실시하였다. GA3를 0.4mg l-1로 9시간을 처리하였을 때 토양에서 생존율은 59.6%로 가장 높게 나타났으며, 그 다음이 1mg l-1에서 9시간을 처리하였 때 43.7%의 생존율을 나타냈다. 재분화 식물체의 뿌리길이가 4 cm 이상일 때는 48개체가 생존하여 생존율은 53.3%로 처리 중 가장 높게 나타났으며, 뿌리 무게는 4 g 이상일때는 46.7%가 생존하여 가장 높은 비율을 나타냈다. 4년생 때 지상부 생육 특성을 조사한 결과, 초장은 35.3 cm, 경장은 18.3 cm, 엽장은 12.1 cm, 엽폭은 4.8 cm로 이식 재배한 연풍의 생육 특성과 비교해 보면 재분화 식물체의 생육이 약간 떨어짐을 볼 수 있었다. 3년생 때 지하부 특성을 보면 근장은 15.3 cm, 동장은 4.5 cm, 동직경은 2.1 cm, 그리고 근중은 15.4 g으로 지하부도 지상부와 마찬가지로 대조구인 묘삼을 이식하여 재배한 처리구에 비하여 생육이 약간 떨어졌다.

FPS (farnesyl diphosphate synthase) plays an essential role in organ development in plants. However, FPS has not previously been identified as a key regulatory enzyme in triterpene biosynthesis. In order to investigate the effect of FPS on ginsenosides biosynthesis, we over-expressed FPS of Centella asiatica (CaFPS) in Panax giseng adventitious roots. PCR analysis showed the integrations of the CaFPS and hygromycin phosphotransferase genes and we ultimately selected three lines. The result of Southern blot analysis demonstrated the introduction of the CaFPS gene into genome of ginseng. In addition, the results of RT-PCR analysis revealed that CaFPS gene overexpression induced an accumulation of its transcription in the ginseng adventitious roots. To determine whether or not the overexpression of the CaFPS gene contributes to the downstream gene expression associated with triterpene biosynthesis, the level of mRNAs was analyzed by real-time PCR. The result showed that no differences were detected in any expression of all genes. To determine quantitatively the content of ginsenosides in transgenic ginseng adventitious roots, HPLC analysis was conducted. The content of total 7 ginsenosides was increased to 1.8, 1.4, and 1.7 times than that of the controls, respectively. This indicated that the overexpression of CaFPS in ginseng adventitious roots causes an increase in ginsenoside content, although down stream genes of FPS gene were suppressed by CaFPS overexpression.

This study was carried out to identify Korean ginseng cultivars using peptide nucleic acid (PNA) microarray. Sixty-seven probes were designed based on nucleotide variation to distinguish Korean ginseng cultivars of Panax ginseng. Among those PNA probes, three (PGB74, PGB110 and PGB130) have been developed to distinguish five Korean ginseng cultivars. Five Korean ginseng cultivars were denoted as barcode numbers depending on their fluorescent signal patterns of each cultivar using three probe sets in the PNA microarray. Five Korean ginseng cultivars, Chunpoong, Yunpoong, Gopoong, Gumpoong and Sunpoong, were simply denoted as '111', '222', '211', '221' and '122', respectively. This is the first report of PNA microarray which provided an objective and reliable method for the authentication of Korean ginseng cultivars. Also, the PNA microarray will be useful for management system and pure guarantee in ginseng seed.

This study was carried out to research microorganisms having the antifungal activity against ginseng Alternaria blight pathogen Alternaria panax and ginseng anthracnose pathogen Colletotrichum gloeosporioides. Eleven Bacillus strains. were isolated from Korean traditional soybean paste and Kimchi. Among the 11 isolates, DJ5, KC1, KC2 and KC4 showing antagonistic activity on the mycelial growth of A. panax and C. gloeosporioides in pairing culture were finally selected as the antagonistic microorganisms. Based on 16s rRNA sequence and phylogenetic tree analysis, they were identified as Bacillus spp.. The selected microorganisms were investigated antagonistic activity by measured leaf-segment colonization in pot test. When Bacillus sp. were injected after A. panax treatment, KC1, KC2 and KC4 showed similar effect to chemical pesticides treated control. To measure preventive effect of Bacillus sp, antagonistic microorganisms were injected and C. gloeosporioides was treated in pot. When measuring the effectiveness for the prevention of Anthracnose, All Bacillus spp. showed approximately 83~90 % degree of superior preventive effect. In general, The four Bacillus spp. isolated from Korean traditional fermented foods showed therapeutic effect of Alternaria blight and preventive effect of Anthracnose.

In this study, Expressed Sequence Tag-Simple Sequence Repeat (EST-SSR) analyses were used to clarify the genetic polymorphisms among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Polymorphic and reproducible bands were produced by 14 primers out of total 30 primers used in this study. Fourteen EST-SSR loci generated a total of 123 bands. Amplified PCR products showed the highly reproducible banding patterns at 110~920 bp. The number of amplified bands for each EST-SSR primers ranged from 2 to 19 with a mean of 8.8 bands. P26 and P35 primers showed 13 and 12 banding patterns, respectively. The number of alleles for each EST-SSR locus ranged from 1.67 to 2.00 with a mean of 1.878 alleles. P34 and P60 primers showed the highest and the lowest genetic polymorphism, respectively. Cluster analysis based on genetic similarity estimated by EST-SSR markers classified Korean cultivars and breeding lines into 4 groups. Group included Gopoong and Chunpoong and 9 breeding lines (55%), group included 2 breeding lines (10%), group included 3 breeding lines (15%), group included Gumpoong and 3 breeding lines (20%). Consequently, the EST-SSR marker developed in this study may prove useful for the evaluation of genetic diversity and differentiation of Korean ginseng cultivars and breeding lines.

These studies were conducted to provide basic information on Korean ginseng cultivars and breeding lines (Panax ginseng C. A. Mey.) and to identify the variations that can be utilized in ginseng breeding programs. The agronomic characteristics was used to clarify the genetic relationships among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Angle of petiole and number of fibrous root showed a wide variation from 15.0~67.8˚ and 0~5, respectively. The average plant length was 54.2cm with a range of 37.9~64.8cm and the average stem diameter was 5.6mm with a range of 4.0~7.5mm. The average stem length was 31.9cm with a range of 21.8~37.9cm and the average root weight was 38.1 g with a range of 23.0~52.0 g. The 24 Korean ginseng cultivars and breeding lines were classified into 4 groups based on agronomic characteristics using the complete linkage cluster analysis. The I, II, III and IV groups included the 60.8%, 7.4%, 13.1% and 8.7% of the cultivars and breeding lines, respectively. The breeding lines in group I could be characterized as the group with the highest growth characters and yield components, such as plant length, stem diameter and root weight. The root weight, the yield component, had highly significant positive correlations with stem diameter, plant length and stem length.

In this study, simple sequence repeat (SSR) analyses were utilized for evaluation of genetic diversity and discrimination of 17 accessions. Five cultivars, which were developed from Korea, and 12 foreign accessions, which were collected from China, Japan, Russia and USA, were evaluated by nine markers out of 22 SSR markers. A total of 39 alleles were detected, ranging from 2 to 8, with an average of 4.3 alleles per locus. The expected heterozygosity and PIC values were 0.627 and 0.553, with a range from 0.21 (GB-PG-078) to 0.76 (GB-PG-142) and from 0.19 (GB-PG-078) to 0.70 (GB-PG-142), respectively. Four makers out of nine SSR markers, GB-PG-026, GB-PG-043, GB-PG-142 and GB-PG-177, were selected as key factors for discrimination of Korean ginseng cultivars and foreign accessions. All of Korean ginseng cultivars and foreign accessions were individually by the combination of four SSR markers. Consequently, the SSR markers developed in this study may prove useful for the evaluation of genetic diversity and discrimination of Korean ginseng cultivars and foreign accessions.

The principal objective of this study was to develop a discrimination method using SSR markers in Korean ginseng cultivars. Five cultivars--Chunpoong, Yunpoong, Gopoong, Sunpoong, and Kumpoong--were evaluated by nine markers out of 22 SSR markers. A total of 23 alleles were detected, ranging from 1 to 4, with an average of 2.6 alleles per locus, and an averages of gene diversity (GD) of 0.480. Nine markers were tested in order to distinguish among five Korean ginseng cultivars. Two markers out of nine SSR markers, GB-PG-065 and GB-PG-142, were selected as key markers for discrimination among Korean ginseng cultivars. Two genotypes were detected in GB-PG-065. Chunpoong and Kumpoong shared the same allele type, and Yunpoong, Gopoong, and Sunpoong shared another identical allele type. In the case of GB-PG-142, a specific allele type differentiated from those of other four cultivars was observed only in Sunpoong cultivar. Consequently, the SSR markers developed in this study may prove useful for the identification of Korean ginseng cultivars and the development of ginseng seed management systems, as well as tests to guarantee the purity of ginseng seeds.

This study describes an efficient approach to the development of DNA markers for use in distinguishing the Scrophularia species that have been used as useful medicinal crops. In order to distinguish Scrophularia species, DNA sequences of rpl-5 region in mitochondrial DNA of Scrophularia species were analysed for detecting sequence variations, and the PCR-RFLP method was applied for developing practicable DNA marker patterns. Several DNA variations were detected by the sequence comparison of rpl-5 region among Scrophularia species. Genetic relationship analysis of Scrophularia species was carried out based on these DNA variations. DNA variations of rpl-5 region were revealed that it was significantly efficient in genetic relationship analysis of Scrophularia species. In addition, Scrophularia species tested in this study were completely discriminated by four polymorphic genotypes by PCR-RFLP combined with Tsp509 I (^AATT) restriction enzyme. Our results suggested that DNA sequence variations of rpl-5 region were sufficiently useful for genetic relationship analysis of Scrophularia species. Polymorphic genotypes by PCR-RFLP using the Tsp509 I enzyme will be useful for discrimination of Scrophularia species as a practicable DNA markers.

This study was undertaken to develop a technique of discrimination using SSR makers in boxthorn cultivars.Forty one boxthorn cultivars, which were collected from Korea and China, were evaluated by 10 SSR markers. Total of 61alleles were detected, ranging from 3 to 13 with an average of 6.1 alleles per locus. The averages of gene diversity and PICvalues were 0.482 and 0.428, with a range from 0.25 (GB-LCM-022 and GB-LCM-087) to 0.83 (GB-LCM-167) and from0.24 (GB-LCM-022 and GB-LCM-087) to 0.81 (GB-LCM-167), respectively. Five markers out of 10 markers, GB-LCM-022, GB-LCM-075, GB-LCM-104, GB-LCM-167 and GB-LCM-217, were selected as key markers for discrimination inboxthorn cultivars. All of boxthorn cultivars were individually distinguished by the combination of five SSR markers.

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.

국내자생 쑥속의 식물을 약용작물 및 산업화에 활용하기 위한 기초자료를 얻고자 본 연구를 수행하였다. 그 결과, 수집된 쑥속 식물들은 사철쑥 (A. capillaris)을 포함한 20종 1아종 2변종 24분류군으로 분류 되었으며, 이를 바탕으로 25개의 화기형질을 이용하여 주성분 분석과 군집분석을 수행하였다. 주성분 분석결과 제1주성분은 전체 분산의 44.73%, 제2주성분은 16.86%, 제 3주성분은 8.88%, 제4주성분은 7.07%의 기여율을 보였으며, 상위 제4주성분까지의 누적 기여율이 77.56%였다. 군집분석 결과, 자방의 퇴화, 아관목, 두화의 크기 등의 주요형질에 의해 크게 3개의 군으로 구분되어졌으며, 화기구조의 식별형질로는 기발표된 Dracunculus, Abrotanum, Absinthium 3절과 완전히 일치하지는 않았으나 국내 자생쑥의 분류형질로 활용이 가능하였다.