종의장거리산포는집단의확산과집단간개체의흐름에중요한역할을한다. 본연구는독도에서발견된식물을형태학적특징과핵및엽록체DNA의분자마커를이용하여종을식별한결과, 참빗살나무(노박덩굴과)로확인되었다. 얕은토양층과험준한지형등입지적으로열악한대양섬인독도에서목본의분포는의미있는결과이며, 향후외부유입종의지속적인모니터링이요구된다.

Background : Kalopanacis Cortex (海桐皮) is listed in「The Korea Pharmacopoeia (KP)」as the original plant of Kalopanax septemlobus (Thunb.) Koidz. However, the dried cortex of Erythrina eariegata L (刺桐) is an adulterant, one of the most indiscriminately used herbal medicines because of its similar morphologic. Due to the morphological similarities of the dried cortex of this plant to those of K. septemlobus which is used as a substitute herbal for E. eariegata, distinguish these two species is extremely difficult. Meanwhile, K. septemlobus is a polymorphic species, its morphological characteristics showed great diversity due to the different geographical and environmental factors. For this reason, it is conducted to develop molecular markers to distinguishing these K. septemlobus with E. eariegata by using conventional polymerase chain reaction (PCR). Methods and Results : In this study, In order to clearly identify origin of K. septemlobus, E. eariegata was analyzed from four barcode regions of chloroplast DNA (psbA-trnH, rbcL, matK, atpH-atpF) and nuclear ribosomal DNA (ITS2) to evaluate the ability of discrimination for each barcode region. The present study aimed to analyze the percent of variable sites were provided the highest ITS2 (2.3%) followed by rbcL (8.2%), in oder to develop a species-specific primer that can distinguish K. septemlobus form E. eariegata. Conclusion : The INDEL markers were developed based on the divergence of each sequence, and it is possible now to identify the two species of K. septemlobus with just a single performance of PCR. This will not only prevent misused of the plant, but also to maintain the quality of the herbal medicine as well as to verify and guarantee safety for public health.

Background : In the KHP (the Korea Herbal Pharmacopoeia), the Cuscutae Semen (菟絲子) is defined as the seed of the Cuscuta chinensis Lamark (family: Convolvulaceae). Using authentic raw herbal materials is fundamental to herbal medicine quality and Cuscutae Semen is widely distributed in many asian countries. Due to having tiny bodies of seeds, it is extremely difficult to differentiate them from adulterants and closely related species by morphologic characteristics, leading to serious safety problems. For this reason, there was conducted to develop molecular markers to distinguishing these Cuscuta chinensis with Cuscuta japonica and Cuscuta pentagona by using conventional polymerase chain reaction (PCR). Method and Results : In this study we developed a clearly and efficient method to identify Cuscutae Semen on the market. These samples (C. chinensis, C. japonica and C. pentagona) were analyzed from two barcode regions of chloroplast DNA (rbcL, psbA-trnH) and nuclear ribosomal DNA (ITS2). Based on genetic distance, the precent of variable sites were provided the highest psbA-trnH value (38.7%), followed by ITS2 (23.4%), rbcL (9.9%), in order to develop a specific primer that can distinguish C. chinensis, C. japonica and C. pentagona. Conclusion : From the above results, DNA barcoding was proved to be a successful tool for authentication the three species of Cuscutae semen. The adoption of DNA barcoding as an authentication tool by food safety agencies can safeguard the interests of both consumers and traders.

Background : Curcumae Longae Rhizoma (薑黃) is listed in「The Korea Pharmacopoeia (K P)」as the original plant of Curcuma longa L (Zingiberaceae). Meanwhile, Zeodariae Rhizoma (莪朮) is listed in 「The Korea Pharmacopoeia (KP)」as the original plant of C. phaeocaulis, C. aromatica and C. Kwangsiensi (Zingiberaceae). Due to the morphological similarities of the dried roots of this plant to those of C. phaeocaulis, C. aromatica and C. Kwangsiensis which is used as a substitute herbal for C.longa, distinguish these four species is extremely difficult. Methods and Results : A total of 90 collected samples were used in this study, In order to clearly distinguish of Curcumae Longae Rhizoma and Zeodariae Rhizoma were analysis based on sequence of the chloroplast DNA (trnK, rbcL, trnL-F, atpB-rbcL) and nuclear ribosomal DNA (ITS2). The present study aimed to analyze the percent of variable sites were provided the highest trnK (2.3%), in oder to develop a species-specific primer that can distinguish C.longa form C. phaeocaulis, C. aromatica and C. Kwangsiensis. In addition, the complete chloroplast genome of C. longa were sequenced by a 454 sequencing platform, and the structure of the obtained chloroplast genome was also analyzed. the result used that INDEL (insertion/deletion) marker for distinguish C.longa form C. phaeocaulis, C. aromatica and C. Kwangsiensis. Conclusion : The INDEL markers were developed based on the divergence of each sequence, and it is possible now to identify the four species of Curcumae Longae Rhizoma with just a single performance of PCR. This will not only prevent misused of the plant, but also to maintain the quality of the herbal medicine as well as to verify and guarantee safety for public health.

Background : Chrysanthemi Indici Flos (甘菊) is listed in 「The Korea Herbal Pharmacopoeia (KHP)」as the original plant of Chrysanthemum indicum L. C. indicum was one of the most representative medicinal plants in Asteraceae, Dried flowers of this plant have been valid chemical composition such as flavonoids, phenylpropanoids, terpenoids, and polysaccharides, possessing broad spectrum antibacterial, antiviral, antihypertensive and anti-oxidation functions. Meanwhile, C. indicum was a polymorphic species, its morphological characteristics showed great diversity due to the different geographical and environmental factors. For this reason, there was conducted to develop molecular markers to distinguishing these C. indicum with C. morifolium, C. zawadskii var. latilobum and Aster spathulifolius by using conventional polymerase chain reaction (PCR). Methods and Results : In this study, In order to clearly identify origin of Chrysanthemi Indici Flos, these samples (C. indicum, C. morifolium, C. zawadskii var. latilobum and A. spathulifolius) were analyzed from five barcoding regions of chloroplast DNA (rbcL, matK, rpoB, atpF-atpH) and nuclear ribosomal DNA (ITS2) to evaluate the ability of discrimination for each barcoding region. Based on genetic distance, the percent of variable sites were provided the highest ITS2 value (56.9%), followed by atpF-atpH (48.18%), matK (27.2%), psbK (8.2%), and rbcL (2.9%). Comparative analysis based on the complete genome sequence of the petL-petG region INDEL (insertion/deletion) that the gene annotations were registered to the GenBank (accession number: JN-867592.1, NC-020092.1, MF-034027.1, NF-279514.1). Conclusion : From the above results, we may suggest that the petL-petG region INDEL analysis were conducted for molecular authentication of four plants (C. indicum, C. morifolium, C. zawadskii var. latilobum and A. spathulifolius). The findings of results indicated that petL-petG region might be established INDEL analysis systems and hence were proved to be an effective tools for molecular evaluation and comparison of “Chrysanthemi Indici Flos” with other plants.

Background : Gentinae Macrophyllae Radix is one of the traditional medicines originated from the roots of multiple plants, Gentiana macrophylla Pall., Gentiana straminea Maxim., Gentiana crassicaulis Duthie ex Burkill and Gentiana dahurica Fisch., in Gentianaceae. Multi-origin traditional medicine usually has adulteration problem based on the morphological similarity and/or misunderstanding the species. Therefore, accurate and reliable identification criteria to ensure drug safety and quality is necessary. Methods and Results : We collected four original species of Gentinae Macrophyllae Radix from plantations and markets in China and Korea. DNA barcoding with four barcoding markers (Internal Transcribed Spacer (ITS), rbcL, trnL intron, trnL-F intergenic sapcer) was performed. Intra-specific variation was observed in ITS nucleotide sequence however, successfully distinct four original species based on the nucleotide discrepancy while trnL intron has no difference. trnL-F intergenic spacer has two transitions(T→C and A→G) sites only in G. crassicaulis and rbcL shows one transition(C→T) site in G. dahurica and G. macrophylla. Phylogenetic relationship analysis of the Gentinae Macrophyllae Radix revealed two major clades – clade I including three groups, G. macrophylla, G. straminea and G. dahurica, and clade II including G. crassicaulis. This aspects was shown more clear with multi-region combined analysis. Conclusion : DNA barcoding will be accurate and powerful criteria for the analysis the origin of Gentinae Macrophyllae Radix. However, single region analysis might be deficient such as trnL intron, rbcL and trnL-F intergenic spacer results in this research. Multi-region combined analysis based on the multiregional DNA barcode markers will be overcome the disadvantage and also increase the precision.

기상조건의 변화로 우리나라에 벼멸구의 발생이 점차 증가하며 피해가 커지고 있다. 이에 다양한 벼멸구 저항성 유전자를 가진 벼 품종의 육성이 필요하다. 국내 육성 자포니카 벼 품종 중 벼멸구 저항성 품종은 Bph18 유전자를 가진 ‘안미’를 제외하고 모두 bph2(‘화청’, ‘하남’, ‘다청’, ‘친농’, ‘친들’) 유전자를 가지고 있다. 유묘검정 결과 bph2 유전자를 가진 품종은 벼멸구 흡즙에 중도저항성을 보인 후 4주 이상 지나며 고사되는 반면, ‘안미(Bph18)’는 계속해서 강한 저항성 반응을 나타낸다. 인공교배를 통해 Bph18 유전자를 가진 계통을 육성하였다. 빠른 DNA 마커 검정을 위해 12번 염색체의 Bph18 유전자와 연관된 마커를 검색하여 PCR 후 전기영동을 수행한 결과 RM3331이 저항성 유전자별 밴드의 위치가 다른 증폭 반응을 보였다. 각 계통에 대한 밴드분석 결과 벼멸구 저항성 유전자가 없는 ‘TN1’과 ‘호평’은 위치가 다른 밴드를 보였는데 이는 인디카와 자포니카의 유전적 백그라운드의 차이로 프라이머의 증폭 부위가 달라 발생한 것으로 추측된다. Bph18 유전자를 가진 ‘익산562호’와 Bph1 유전자를 가진 ‘청청벼’, ‘Mudgo’, ‘IR09N379’의 밴드 위치가 같은 것은 두 유전자가 중복되어 위치해 있기 때문인 것으로 보인다. 따라서 RM3331로 생성된 PCR 산물의 크기는 제한효소 처리없이 2% agarose gel로 분석이 가능한 간편성을 가지고 있지만, 저항성 유전자가 중복되어 있는 Bph18과 Bph1 유전자를 구분하기는 어려우며 벼멸구 저항성 유전자를 가지고 있지 않은 계통과의 구분에 선택적으로 사용하는 것이 적합할 것으로 보인다.

This study describes the efficient method for the discrimination of 'Cheonryang' in Panax ginseng Meyer using a STS primer. A total of 208 STS primers were applied to polymerase chain reaction (PCR) amplification for discriminating Korean ginseng cultivars. Co-dominant polymorphic band patterns were generated with two primers, MFGp 0019, MFGp 0248, and successful identification of 'Cheonryang' was achieved from out of 11 Korean ginseng cultivars. Two different sizes of DNA band patterns were detected with MFGp 0019 primer. Ten Korean ginseng cultivars shared the same size of amplified DNAs (389 bp), but 'Cheonryang' showed a different size. Thus 'Cheonryang' can be efficiently distinguished from the other ten ginseng cultivars by using the MFGp 0019 primer. In the case of MFGp 0248, two different sizes of DNA band patterns were detected in the eleven ginseng cultivars. Same sized amplified DNA bands (307 bp) were shown in five cultivars (Chunpoong, Gopoong, Kumpoong, Cheongsun, Sunhyang) and 254 bp sized DNA bands were identified in the other 6 cultivars (Yunpoong, Sunpoong, Sunun, Sunone, Cheonryang, K-1). In conclusion, the two STS primers, MFGp 0019, and MFGp 0248, provide a rapid and reliable method for the specific identification of 'Cheonryang' cultivar from a large number of samples.

본 연구는 복숭아 유전자원의 유전적 다양성을 분석하여 육종의 기초 자료로 활용하기 위하여 재배 품종 및 국내에서 수집한 야생 복숭아 64점을 대상으로 SRAP, AFLP와 RAPD 분석을 수행하였다. 19종과 20종의 primer조합을 이용한 SRAP와 AFLP분석에서 각각 143개(26.7%)와 193개(27.2%)의 다형성 밴드를 획득하였고, 평균 다형성 밴드 수는 7.53개와 9.65개였다. RAPD분석에서 52종의 선발 primer를 이용하여 201개(45.9%)의 다형성 밴드를 얻었으며, 평균 다형성 밴드 수는 3.90개였다. SRAP, AFLP와 RAPD분석에서 획득된 537개의 다형성 밴드를 이용하여 UPGMA(비가중 평균결합) 방식으로 유사도 및 집괴분석을 수행한 결과 유전적 유사도 0.751을 기준으로 8개의 그룹으로 분류되었다. 복숭아 유전자원 간 유전적 유사도 값은 0.370～0.978의 범위로 평균 유전적 유사도는 0.737이었다. 가장 높은 유사도 값(0.978)을 나타낸 유전자원은 PH065와 PH066 간이었고 가장 낮은 유사도 값(0.370)을 나타낸 유전자원은 Prunus davidiana와 PH020간이었다. 본 연구에서는 이용된 64종의 복숭아 유전자원은 집괴분석 결과 유전자원들이 수집된 지역은 다르지만 유전적 다양성은 낮은 것으로 추정되었다.

밀양23호/기호벼 재조합자식 유전집단을 대상으로 PCR 기반 DNA 마커들로 구성된 분자유전자지도를 만들고자, 주로 아가로스 젤 상에서 분석이 가능한 마커들을 위주로 STS, InDel, RTM, SSR 마커들을 선발하여 분석하였다. InDel 마커 37개, STS 마커 88개, RTM 마커 8개, SSR 마커 91개를 포함한 224개의 마커로 구성된 유전지도를 만들었는데, 총 유전거리는 1,425 cM이었으며, 마커간 평균거리는 6.7 cM이었다. 이들 DNA 마커들의 프라이머 시퀀스 정보를 바탕으로 e-PCR프로그램을 이용하여 각 마커들의 벼 유전체상에서의 물리적인 위치를 파악하고 물리지도를 작성하였다. 이 물리지도에서 마커간의 물리적 거리의 합은 356.8 Mbp이었으며, 총 유전거리에서 이를 나누어 구한 1 cM당 평균 물리적 거리는 250 kbp이었다. 5% 유의수준에서 분리비 편의(segregation distortion) 현상을 보인 마커는 전체 마커의 22.8%인 51개이었으며, 주로 3번 염색체의 중간부위, 6번 염색체의 거의 모든 영역, 7번 염색체의 상단부위, 8번 염색체의 하단부위, 12번 염색체의 상단부위에 분포하였다. 이 분자유전자지도는 자포니카형 품종과 통일형 품종 또는 인디카 품종간의 교배후대 집단에서 유용형질의 유전자 위치를 분석하고자 할 때 이용 가능한 마커들에 대한 정보를 제공할 것이다.

본 연구에서는 관행의 교배육종과 DNA 마커를 이용한 MAS의 접목을 통하여 벼멸구 저항성과 관련된 단점을 보완하고, 효율적으로 도열병, 줄무늬잎마름병, 흰잎마름병, 벼멸구, 끝동매미충 저항성이 집적된 복합내병충성 우량계통을 육성하고자 수행하였다. 교배모본으로는 완전미율이 높고, 끝동매미충에 저항성인 '남평'과 단간이면서 흰잎마름병에 저항성인 '주남'을 반복친으로 사용하였고, 벼멸구저항성 유전자 Bph1을 가지고 있지만 간장이 크고 재배안전성이 미흡한