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.

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.

Root yield and quality of ginseng cultured in paddy soil was low relatively compared with that of upland soil because of moisture injury in root during rainy season. Drainage class in soils generally divided into 6 classes, and it is possible to cultivate ginseng practically in imperfectly drainage class (IDC). This study carried out to select the varieties that is suitable for paddy soil, which is easy to be generated rusty-colored root and physiological-discolored leaf. Experiment plot arranged with the condition of soil humidity contents such as poorly drainage class (PDC) and imperfectly drainage class (IDC), and upland soil. Growth characteristics and root yield were investigated in four-year-old ginseng of varieties, Cheonpoong (CP), Yeonpoong (YP), Hwangsookjong (HS), and Jakyeongjong (JK). CP among four varieties showed the highest yield in IDC and CP was the lowest ratio in leaf discoloration and rusty-colored root. HS was followed by CP in the order of root yield, but it had the weakness that the ratio of rusty-colored root was high respectively.

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.

An advanced extraction method by ultrasonic extraction with applied solid phase extraction (SPE) has been developed for the determination of simultaneous eight major ginsenosides, namely ginsenosides Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, and Rd in the root of Panax ginseng. Four extraction methods including n-BuOH reflux extraction (Method A), 70% EtOH reflux extraction (Method B), 50% MeOH reflux extraction with SPE (Method C), and 50% MeOH ultrasonication with SPE clean-up process (Method D) were investigated for the determination of eight major ginsenosides. Total contents of ginsenosides were highest by extraction of Method C as 2.408±0.011%. However, Method D was evaluated as relatively simpler and more efficient method due to short extraction time, small solvent consumption and less expensive, compared to conservative reflux method. Ginsenosides were also satisfactorily separated with good resolution and the accuracy range was between 1.05 and 4.06% as relative standard deviation (RSD) by Method D. SPE condition and HPLC condition were further optimized for determination of eight major ginsenosides by the ultrasonic extraction method. Conclusively, ultrasonic extraction of 2 g sample of ginseng using ultrasonic bath and 1 loading for SPE was evaluated as proper condition for extraction of ginseng.

해가림의 이랑방향이 인삼의 생육에 미치는 영향을 구명하고자 방위각 90˚와 270˚ (이랑방향 90˚), 120˚와 300˚ (이랑방향 120˚), 0˚와 180˚ (이랑방향 180˚)를 가로지르는 방향으로 이랑을 만들고 2, 3년생 인삼의 생육특성 및 수량성을 표준재배법의 이랑방향 120˚와 비교한 결과는 다음과 같다. 1. 이랑방향 90˚는 대조구인 120˚에 비해 오전에 투광량의 감소로 해가림 내 기온은 약간 낮았으나 15시 이후에는 투광량의 증가로 대조구보다 높은 기온을 보였다. 2. 이랑방향 180˚는 대조구인 120˚에 비해 9:00~11:00 사이에 투광량의 현저한 증가로 해가림 내 기온이 매우 높았으나 13시 이후에는 대조구와 투광량이 비슷하여 기온도 큰 차이가 없었다. 3. 이랑방향 90˚와 180˚에서 경장, 엽장 및 엽폭은 대조구인 120˚보다 모두 감소되었는데, 90˚보다 180˚에서의 감소가 현저하였다. 4. 엽소율(葉燒率)은 이랑방향 90˚에서 가장 작고 직사광선의 유입이 많았던 180˚에서 가장 컸으며 해에 따라 엽소율(葉燒率)은 큰 변이를 보였다. 5. 2004년과 같이 한발이 심한 해에는 이랑방향 180˚의 수량성이 대조구인 120˚보다 낮았으나 상대적으로 한발이 적은 2005년도에는 180˚가 대조구보다 높은 수량을 보였다.

금산 및 음성지역의 4년생 인삼재배 농가포장에서 직파재배 5개소와 이식재배 5개소를 임의로 선정하여 직파와 이식재배에 따른 생육특성 및 엑스와 조사포닌 함량을 비교한 결과는 다음과 같다. 직파재배는 4년근 생존율이 평균 48%로 이식재배의 86%보다 떨어지나 입모수가 평균 96주/3.3m2순로 이식재배의 57주보다 많고 엽면적지수가 커 수량성이 높은 반면, 주당근중은 작았다. 직파재배는 이식재배에 비해 동체의 신장이 양호하나 지근의 발달이 불량하여 동체중의 비율이 높고 지근중의 비율이 낮았으며, 직파재배는 적변 발생율이 적으나 동체와 지근부위의 엑스와 조사포닌 함량이 낮았다.

경기도 안성지역의 4~6년생 재래종 (자경종) 인삼포장에서 4월 28일에서 9월 30일까지 월별 지하부 생육특성과 엑스함량의 경시적 변화를 조사한 결과는 다음과 같다. 1. 동체장은 월별 수축과 회복을 반복하여 9월 수확기까지 증가되지 않았으나 동체직경은 점진적인 증가를 보였다. 2. 단위 면적당 지상부중은 4월부터 급격히 증가하여 7월에 최고를 보인 후 차차 감소되었으며, 지하부중은 5월에 감소된 후 9월 수확기까지 점차 증가되었는데, 6년생은 7월 이후 약간의 감소를 보였다. 3. 지하부의 상대생장율은 전엽기인 5월과 고온기인 8월에 감소되었고 6, 7, 9월에 다시 증가되었는데, 6년생의 지근과 세근은 4~5년생에 비해 월별 뚜렷한 변화를 보이지 않았다. 4. 상품비율 (근중 60g 이상)은 연생이 증가할수록 커서 4년생 23%, 5년생 60%, 6년생 69% 이었다. 5. 뿌리의 수분함량은 수확기까지 차차 감소되었는데, 7~9월까지 유의적인 차이를 보이지 않았으며, 9월 수확기의 수분함량은 4년생 〉 5년생 〉 6년생순 이었다. 6. 동체의 경도는 5월에 감소된 후 급격히 증가되었는데, 4~5년생은 6~9월까지 대체로 증가되었으나 6년생은 8월부터 차차 감소되어 4~5년생과 유의적인 차이를 보였다. 7. 동체와 지근의 엑스함량은 4월부터 6월까지 급격히 감소된 후 9월에 다시 증가되었는데, 4~5월의 엑스함량은 4년생이 5~6년생보다 많았으나 9월에는 연생 간에 뚜렷한 차이가 없었다.

본 연구는 고려인삼의 집단내의 유전적 변이를 작물학적 특성 및 DNA수준에서 비교하여 인삼품종육성을 위한 기초 자료를 제공하기 위하여 수행한 결과를 요약하면 다음과 같다. 1.개화기는 5월 16일부터 24일 까지 일주일에 걸쳐 개화하였고, 19일과 22일에 개화율이 가장 빈도가 높았다. 2. 줄기 색은 녹색이 13개체, 연자색이 429개체, 자색이 229개체, 진자색이 38개체로 연자색이 가장 많은 분포를 보였으며 한 집단 내에서 다양한 분포를 나타냈다. 3. 초장은 22­68cm의 넓은 범위에 분포하였으며, 20­27cm가 31개체 , 27­34cm가 88개 체, 34­41cm가 219개체 , 41­48cm가 291개체, 48­55cm가 63개체, 55­62cm가 10개체, 62­69cm가 5개체로 다양한 분포를 나타냈다. 4. 뿌리무게는 16­26g이 53개체, 26­36g이 137개체, 36­46g이 385개체, 46­56g이 94개체, 56­66g이 27개체, 66­76g이 9개체, 76­86g이 4개체, 36­46g범위에서 385(57.7%)개체로 가장 많은 분포를 나타내었으며, 16­86g의 범위로 변이 정도가 매우 컸다. 5.662개체를 대상으로 RAPD를 실시한 결과 32개의 프라이머 중 10개 가 재현성이고 다형성인 밴드를 보였다. 10개의 프라이머에서 나타난 전체 밴드수는 109개였으며 이중 103개가 다형성을 보여 다형성 비율은 94.5%였다. 6. URP5 프라이머를 이용하여 662개체를 군집 분석한 결과 밴드의 유무에 따라 16개의 그룹으로 구분하였으며, 개화기, 초장, 줄기색, 뿌리직경, 뿌리무게에 다른 분류와 일치하지 않았다.

구기자품종의 혹응애에 대한 저항성 반응 및 저항성 유전양식 을 검정하기 위하여 저항성 품종과 감수성 품종을 인공교배 하여 얻은F1집단의 포장자연 발생율에 의하여 저항성을 조사한 결과 다음과 같은 결과를 얻었다. 1. 구기자혹응애 저항성 품종인 일본 1호,중국 1호를 감수성인 CL42-56과 교배한 F1은 혹응애 저항성과 감수성이 3 ; 1로 분리되었으며, 일본 1호와 중국 1호를 교배한 F1은 혹응애 저항성과 감수성이 15 : 1로 분리 되었다. 2. 일본 1호와 중국 1호의 혹응애 저항성은 hetero 인 두 개의 우성유전자에 의해 지배되며, 저항성 유전자를 E1e1E2e2로 명명하였다. 3. 청양구기자를 CL42-56에 교배한 F1은 혹응애 저항성과 감수성의 분리비가 명확하지 않았으며 변이 의 폭이 크므로 청양구기자의 혹응애 저항성은 주동유전자와 polygenes에 의해 지배되었다. 4. 일본 1호와 중국 1호는 혹응애 저항성 품종 육성을 위한 모본으로 우수하였으며, 청양구기자도 저항성 모본으로서 가치가 인정되었다.