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 12 cultivars of the Jeju native Citrus are considered to have originated from China. However, the origin of the cultivar ‘Byungkyool’ (Citrus platymamma Hort. ex Tanaka) is not clearly known. We performed PCR analysis by using three primer sets designed from the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) to analyze the phylogenetic relationship between the traditional citrus cultivars and the Byungkyool cultivar. Sequence length of the nrDNA ITS1 region of JNCPCRI (Jeju Native Citrus platymamma Citrus Research Institute) cultivar was 247 bp, 8the ITS2 region was 228 bp and the total ITS region (ITS1-5.8S-ITS2) was 638 bp. Analysis of the genetic relationship based on the sequence analysis at the ITS region of the JNCPCRI cultivar revealed that the ITS1 region of the cultivar was genetically the same as that of the Byungkyool (JQ990189) cultivar, and the ITS2 region was genetically similar to the Binkyool (JQ990180), Hongkyool (JQ990178), Dangyooja (JQ990179), and Pyunkyool (JQ990181) cultivars. Moreover, the total ITS region in the 5.8S rDNA region was genetically similar to the Hongkyool (JQ990178) cultivar. In addition, the total ITS region of the JNCPCRI cultivar was the most closely related to the Cheongkyool (JQ990183) cultivar and has been reported to originate from the Binkyool (JQ990180) and Pyunkyool (JQ990181) cultivars. Although the JNCPCRI cultivar was morphologically the same as the Byungkyool (JQ990189) cultivar, the ITS region showed genetic heterogeneity. Taken together, we conclude that the genetic variation in the ITS region of JNCPCRI cultivar suggests that it was propagated through fertilization with the surrounding citrus cultivars.
Blueberry (Vaccinium spp.) is a member of the Ericaceae and eleven varieties have been registered at the Korea Seed & Variety Service for Plant Variety Protection (PVP). This study was to develop simple sequence repeat (SSR) markers next generation sequencing (NGS) analysis and to analysis genetic relationship of blueberry 31 varieties. Highbush blueberry ‘Camellia’ and rabbiteye blueberry ‘Alapaha’ varieties were used as sequencing materials. Out of total 987 SSR primers detected between ‘Camellia’ and ‘Alapaha’, 148 SSR primers were initially applied to select SSR markers for identification of blueberry varieties. Fourteen SSR markers showed polymorphism between 8 varieties. Seven SSR markers showed reproducibility and clear peak among 14 SSR markers. Genetic relationships of 31 blueberry varieties were analyzed and identified using 7 SSR markers. A total of 30 polymorphic SSR alleles were obtained and two to seven alleles were detected for each locus with an average of 4.3 alleles per locus. Average polymorphism information content was 0.556, ranging from 0.374 to 0.714. Genetic distance of clusters ranged from 0.38 to 0.93 by unweighted pair-group method with arithmetical average based on Jaccard’s distance coefficients. These newly developed SSR markers indicate usefulness for variety identification related to seed dispute and distinctness, uniformity and stability (DUS) test for blueberry.
This study was carried out to analyze correlation between heterosis of F1 hybrids and the genetic distance of parental lines. Growth characteristics such as number of leaves, plant height, bulb diameter, bulb height, bolting rate and marketable yield were examined in 15 F1 hybrids. The genetic distance among one male sterile line and fifteen restorer lines were analyzed with SSR markers. 21 primer pairs amplified polymorphic band and the average number of polymorphic allele was 3.4. As a result of UPGMA cluster analysis, the similarity coefficient value among parental lines ranged from 0.43 to 0.63. Combination of M1 and R14 is high similarity and number of leaves, plant height and yield are 10.2, 66.7cm and 6,818kg·10a-1. The case of the lowest value combination, they are 9.0 and 63.7cm and 7,728kg·10a-1, respectively. The results showed that there are no significant correlations between genetic distance and heterosis of F1 hybrids and only molecular marker does not predict growth traits and yield of onion.
This study was performed to analyze genetic relationship of the four major Cucurbitaceae crop. We used 120 Expressed Sequence Tag(EST)-Simple Sequence Repeat(SSR) primer sets of developed from watermelon and published in International Cucurbit Genomics Initiative (ICuGI) database. Among 120 EST-SSR primer, 51(49.17%) EST-SSR primer set successfully amplified and 49(40.8%) EST-SSR primer set showed polymorphisms among eight cultivars of Cucurbitaceae. In the first instance, amplified PCR products analysis was conducted by the agarose-gel electrophoresis then further analyzed by using Fragment Analyzer. A total 382 PCR band were producted by 49 EST-SSR primers in 24 plant panels, used the analysis of pairwise similarity and dendrogram construction. Assessment of the genetic relationships resulted in similarity index with range of 0.0103 to 0.8452. In dendrogram, 24 plant panels were formed three major groups (A, B, C) and 7 subgroups (A-1, A-2, B-1, B-2, B-3, C-1, C-2). Major group A was comprised of 2 subgroups, subgroup A-1 (6 watermelon cultivars, Citrullus lanatus var. vulgaris Schrad.) and subgroup A-2 (3 wild type watermelon, Citrullus lanatus var. citroides Mats. & Nakai). Major group B was comprised of 3 subgroups, subgroup B-1 (4 melon cultivars, Cucumis melo var. cantalupensis Naudin.), subgroup B-2 (2 oriental melon cultivars, Cucumis melo var. conomon Makino.) and subgroup B-3 (5 cucumber cultivars, Cucumis sativus L.). Major group C was comprised of 2 subgroups, subgroup C-1 [2 squash/ pumpkin cultivars, Cucurbita moschata (Duch. ex Lam.)/Duch. & Poir. and Cucurbita maxima Duch.] and subgroup C-2(2 squash/pumpkin cultivars, Cucurbita pepo L./Cucurbita ficifolia Bouche.)
무핵 포도 품종 육성 시, 주요 교배친으로 사용되는 무핵 품종 20점의 유연관계를 분석 하고, 무핵 형질과 연관된 P3_VvAGL11 마커의 이용 가능성을 검토하였다. SSR 마커 30종을 이용한 결과 218개의 대립인자가 확인되었고, 마커 당 대립인자 수는 평균 7.3개였다. PIC (Polymorphism information contents) 값은 SSR 마커에 따라 0.052에서 0.883으로 나타났고 평균 값은 0.442이었다. 또한 P3_VvAGL11 마커를 이용하여 포도 무핵 유전자원 20 점을 분석한 결과 ‘Sultanina’와 ‘Kishmish Chernyi’ 품종으로부터 유래한 18개 무핵 품종에서 무핵 특성 특이적인 밴드가 증폭되었다. 선발된 218개의 다형성 밴드를 이용하여 UPGMA 군집분석한 결과 유사도 지수 0.722를 기준으로 4개의 그룹으로 구분할 수 있었다. 각 그룹은 교배친으로 사용된 품종들의 유전적 background에 유럽종(V. vinifera), 미국종(V. labrusca), 그리고 이들의 종간교잡종인 V. labruscana의 비율에 따른 분류와 일치하였다. 품종 간 유전적 유사도는 0.592에서 0.844의 범위로 나타났으며 평균 유사도 지수는 0.715였다. 가장 높은 유사도 지수를 나타낸 것은 ‘Emerald Seedless’와 ‘Ruby Seedless’ 품종 간이었고, 가장 낮은 유사도 지수를 나타낸 것은 ‘Beauty Seedless’와 ‘Honey Seedless’ 품종 간이었다. 이러한 결과는 무핵 포도 품종 육성 시, 유전적 background가 다양한 양친 선정 및 무핵 교배 실생 조기 선발에 활용 될 수 있을 것이다.
Rice genetic resources are composed of various species and ecotypes, and each accession reveals different genetic and phenotypic characters. For the managenment of diverse rice genetic resources, seed integrity is important factor in that the individuals of one accession in self-pollinating crop might be homogeneous. To elevate the management efficiency of rice germplasm contrary to the phenotypic distinction, we focused on applicable microsatellite markers because this markers are widly used for genetic evaluation in diverse genetic resources with a character of high reproducibility and polymorphism. In this regard, we selected microsatellite markers based on genotypes; diversity set including 150 accessions using 249 SSR markers. As SSR loci with high PIC(polymorphism information content) values usually revealed multi bands in one accession, proper genotyping were difficult in these loci. Therefor, we checked the band clarity in addition to PIC values and chose 12 and 6 SSR markers finally. All accessions of rice diversity set were distinguished with the first marker set comprising 12 SSR markers, and only 3 combinations of tested accessions(0.03%, 3/11,175) showed same genotype with second marker set comprising 6 SSR markers. The tested 142 Korean bred varieties revealed 0.19%(19/10,011 combinations) and 0.69%(69/10,011 combinations) genotypic identity using first and second marker set, respectively. These newly selected markers might be useful for the analysis of genetic homogeneity and relationship in rice genetic resources.
본 연구는 총 50개의 SSR primer를 이용하여 색소옥수수(색소종실 옥수수 12계통, 색소찰 옥수수 12계통) 및 비색소옥수수(종실옥수수 2계통, 찰옥수수 5계통) 계통들에 대하여 유전적 다양성, 계통유연관계 및 집단구조를 분석하였다.1. 그 결과 50 개의 SSR primer들은 색소 및 비색소옥수수 31계통들에서 총 282개의 대립단편을 증폭시켰으며, SSR primer들에서 증폭된 대립단편의 수는 최소 2개에서부터 최대 11개까지의 범위로 나타나 평균 5.64개가 증폭되었다. MAF(major allele frequency)는 0.23(bnlg279)에서 0.90(umc2338)의 범위로 나타나, SSR primer 당 평균 0.45개로 나타났고, 유전적 다양성(GD)값은 0.17(umc2338)에서 0.86(bnlg279)의 범위로 나타나, SSR primer 당 평균 0.67의 값을 나타내었다. 2. 31개의 색소 및 비색소옥수수 계통들의 집단구조를 분석한 결과, 이들 옥수수 계통들은 Groups I, II, III, IV, V, admixed로 구분되었다. Group I은 찰옥수수 1계통, 색소종실옥수수 1계통, 색소찰옥수수 5계통이 포함되었고, Group II는 찰옥수수 1계통, 색소찰옥수수 3계통이, Group III는 색소종실옥수수 3계통, Group IV는 색소종실옥수수 2계통이, Group V는 종실옥수수 2계통, 찰옥수수 2계통, 색소종실옥수수 5계통이 포함되어 있었다. 그리고 admixed group에는 찰옥수수 1계통, 색소종실옥수수 1계통, 색소찰옥수수 4계통 이 포함되어 있었다.3. UPGMA법에 의한 계통유연관계 분석 결과, 31개의 색소 및 비색소옥수수 계통들은 유전적 유사성 27.4%의 수준에서 크게 3개의 그룹으로 나누어졌다. Group I은 10개의 옥수수 계통(종실옥수수 2계통, 찰옥수수 2계통, 색소종실옥수수 6계통)을 포함하고 있었고, Group II는 20개의 옥수수 계통(찰옥수수 3계통, 색소종실옥수수 5계통, 색소찰옥수수 12계통)을, 그리고 Group III은 색소종실옥수수 1계통을 포함하고 있었다. 따라서 본 연구의 결과는 앞으로 강원도 농업기술원 옥수수시험장에서 색소옥수수 자식계통들에 대한 선발과 품종 육성 연구에 유용한 정보를 제공할 것으로 생각된다.
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.
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.
포도 대목 품종 육성 시, 주요 교배친으로 사용되는 대목 품종 29점 및 국내 자생 머루를 포함한 포도 속(Vitis) 야생 유전자원 13점 등 총 42점의 유전적 다양성을 분석하기 위하여 RAPD와 SSR 분석을 수행하였다. Random decamer 30 종을 분석하여 329개의 다형성 밴드(60.3%)를 얻었으며 primer 당 평균 다형성 밴드 수는 11.0개였다. SSR 마커 20종을 이용하여 분석한 결과 263개의 대립인자가 확인되었고, 마커
The genetic diversity and the genetic relationship among 30 genetic resources of T. officinale and T. coreanum collected from 20 regions in Korea were evaluated by using ISSR markers. Out of 127 loci detected overall, 122 were identified to be polymorphic with a rate of 96.0% at the 30 individuals. The intraspecific polymorphism between T. officinale and T. coreanum was 92.6% and 88.2%, respectively. The genetic similarity matrix (GSM) revealed a wide range of variablility among the 30 accessions, spanning from 0.179 to 922. According to the clustering analysis, different species T. officinale and T. coreanum, were divided into independent groups and all of the accessions could be classified into 7 categories. Especially, all of the mountain collected accessions belonged to independent groups. The study findings indicate that T. officinale and T. coreanum accessions have a high genetic diversity and accordingly carry a germ-plasm qualifying as good genetic resources for breeding.
The polymorphism and the genetic relationships among 32 genetic resources of genus Nelumbo from Korea, Japan, China, USA, India, Thailand and Gabong were thoroughly investigated and extensively examined using ISSR markers. Out of 103 loci detected overall, 94 were identified to be polymorphic with a rate of 91.2%. The genetic similarity matrix revealed a wide range of variability among the 32 accessions, spanning from 0.227 to 0.833. The study findings indicate that the Nelumbo accessions have a high genetic diversity, and accordingly carry a germplasm qualifying as good genetic resources for cross breeding. According to the clustering analysis, different subspecies, N. nucifera and N. lutea, were divided into independent groups and all of the N. nucifera accessions could be classified into five categories. Compared to RAPD analysis, ISSR method showed a clearer picture of polymorphism among the accessions and exhibited a definite distinction even among the subspecies. In this respect, ISSR analysis is considered to be more effective in differentiating the accessions and subspecies of the genus Nelumbo than RAPD test.
본 연구는 배 유전자원의 유전적 변이를 DNA 수준에서 비교함으로써 육종의 기초 자료로 활용하기 위하여 유전자원 60점을 대상으로 AFLP 분석을 수행하였다. 총 20종의 AFLP 프라이머 조합을 이용하여 522개의 다형성 밴드를 얻었다. 획득된 다형성 밴드를 이용하여 UPGMA 방식으로 유사도 및 집괴분석을 수행한 결과 유전적 유사도 0.691를 기준으로 4개의 그룹으로 분류되었다. 첫 번째 그룹에는 Pyrus communis에 속하는 품종 및 P.
동아시아에 분포하는 콩과에 속하는 칡 1종의 13집단과 근연분류군 2종의 4집단 등 총 17개의 개체군에 대하여 유전적 유연관계 및 종간 특이적인 분류학적으로 유용한 분자마커를 알아보기 위해 RAPD 분석을 실시하였다. 15개의 oligo primer를 이용한 효소중합반응을 통해 증폭된 RAPD 절편들은 200 bp에서 2,800 bp 사이의 구간에서 관찰되었다. 이중 유효한 polymorphic band makers는 총 208개, monomorphic bands는 3개를 확인하였으며, 칡의 종 특이적 분자마커는 4개로 확인되었다. 이러한 자료에 근거하여 칡속의 17개 개체군 집단에 대한 UPGMA 분석을 실시하였다. 도출된 UPGMA phenogram에서 한국산 칡 9개체군과 국외산 칡 3개체군이 각각 독립적인 두 개의 작은 유집군을 형성하였으며, 이후 두 유집군이 하나로 크게 유집되어 다른 칡 근연분류군과는 뚜렷하게 구분되었다. 따라서 RAPD 분석은 칡과 근연분류군간의 유연관계 분석 및 한국산과 국외산 집단의 원산지판별에 매우 유용한 분자마커로 생각된다.
Randomly Amplified Polymorphic DNA (RAPD) was performed to define the genetic variation and relationships of Artemisia capillaris. Fifteen populations by the distributions and habitat were collected to conduct RAPD analysis. RAPD markers were observed mainly between 300bp and 1600bp. Total 72 scorable markers from 7 primers were applied to generate the genetic matrix, and 69 bands were polymorphic and only 3 bands were monomorphic. The genetic dissimilarity matrix by Nei's genetic distance (1972) and UPGMA phenogram were produced from the data matrix. Populations of Artemisia capillaris were clustered with high genetic affinities and cluster patterns were correlated with distributional patterns. Two big groups were clustered as southern area group and middle area group. The closest OTUs were GW2 and GG1 in middle area group, and GB1 from southern area group was clustered with OTUs in middle area group. RAPD data was useful to define the genetic variations and relationships of A. capillaris.