• 제목/요약/키워드: AFLP analysis

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AFLP fingerprinting법을 이용한 담죽엽의 감별법 연구 (The Studies on Identification of Lophatherum gracile(淡竹葉) Using AFLP fingerprinting Methods)

  • 심영훈;성락선;박주영;조창희;김지연;이종화;현성예;김선호;김동섭;장승엽
    • 생약학회지
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    • 제41권4호
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    • pp.297-302
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    • 2010
  • Lophatheri Herba is the aerial part of Lophatherum gracile Bronghiart(淡竹葉, Gramineae). 25~75 cm in length. Stem: cylindrical with nodes, empty inside, externally pale yellowish green. Leaf: dehiscent of lanceolate lamina, shrunken and rolled, 5~20 cm long, 10~35 mm wide; surface: pale green ~ yellowish green, parallel-formed with veins of square reticulate, more distinct of appearance on the lower surface. Banbusae Caulis In Taeniam is the stringy strip derived from the stem with the peeled-off epidermis of Phllostachys nigra Munro var. henosis Stapf, and Phllostachys bambusoides Siebold et Zuccarini(竹葉, Gramineae). Irregular in size and shape, thin plane ~ strip-shaped, sometimes powdery, sometimes 1~3 mm thick. Outer surface: pale green ~ yellowish green, sometimes grayish white L. gracile and P. nigra have different origins although they show similar morphologic features. We were able to distinguish between L. gracile and P. nigra which are almost indistinguishable through this study. AFLP(Amplifide Fragment Length Polymorphism) was more suitable for identifying differences between L. gracile and P. nigra in comparison with other genetic analysis using chemical analysis. Therefore. molecular biological methods are believed to be useful for discovering origins of herbal medicines.

Development of SCAR Markers for Korean Wheat Cultivars Identification

  • Son, Jae-Han;Kim, Kyeong-Hoon;Shin, Sanghyun;Choi, Induk;Kim, Hag-Sin;Cheong, Young-Keun;Lee, Choon-Ki;Lee, Sung-Il;Choi, Ji-Yeong;Park, Kwang-Geun;Kang, Chon-Sik
    • Plant Breeding and Biotechnology
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    • 제2권3호
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    • pp.224-230
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    • 2014
  • Amplified fragment length polymorphism (AFLP) is a molecular marker technique based on DNA and is extremely useful in detection of high polymorphism between closely related genotypes like Korean wheat cultivars. Six sequence characterized amplified regions (SCARs) have been developed from inter simple sequence repeat (ISSR) analysis which enabled the identification and differentiation of 13 Korean wheat cultivars from the other cultivars. We used six combinations of primer sets in our AFLP analysis for developing additional cultivar-specific markers in Korean wheat. Fifty-eight of the AFLP bands were isolated from EA-ACG/MA-CAC, EA-AGC/MA-CTG and EA-AGG/MA-CTA primer combinations. Of which 40 bands were selected to design SCAR primer pairs for Korean wheat cultivar identification. Three of 58 amplified primer pairs, KWSM006, KWSM007 and JkSP, enabled wheat cultivar identification. Consequently, 23 of 32 Korean wheat cultivars were classified by eight SCAR marker sets.

AFLP에 의한 한국산 새우난초속 식물과 그의 수종 돌연변이에 대한 분류학적 연구 (Taxonomy of Korean Calanthe species and few of its mutants based on AFLP data)

  • 스리칸스;구자춘;구자정;최경;박광우;소순구;최용국;황성수
    • 식물분류학회지
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    • 제42권3호
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    • pp.215-221
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    • 2012
  • 5개의 한국산 새우난초류 즉 새우난초, 한라새우난초, 금새우난초, 여름새우난초, 신한새우난초의 분류학적 및 유전적 유연관계가 증폭길이다형성(AFLP) 자료를 근거로 조사되었다. 위의 5개 분류군 및 노란색에 흰 설엽을 갖는 몇몇 돌엽변이체 등 16 개체가 연구에 사용되었다. AFLP를 통해서 50개의 분자마커가 동정되었다. 다변량분석의 결과는 유전적 다양성이 종내에서 보다 종간에 크게 나타났다. 유전적 비유사성은 다른 분류군에 비해서 YW 돌연변이체들 간에 가장 낮았고, 여름새우난초 개체들 간에 가장 높았다. 돌연변이체들은 주된 유집군의 밖에 유집되었다. 한라새우난초는 새우난초와 먼저 유집되어 유전적 구성이 다른 종보다 새우난초에 더 가까운 것으로 판단된다. 비록 한라새우난초가 새우난초와 금새우난초의 자연잡종에 의해 기원되었다고 하여도 분자 마커, 유집분석 그리고 비유사도 자료를 통해서 유전자이입이 새우난초 쪽으로 이루어진 것으로 판단된다. 결론적으로 한국산 새우난초류는 유전적 다양성이 있고, 한라새우난초는 새우난초 쪽으로 유전자이입이 있으며, YW 돌연변이체들은 유전적으로 금새우난초에 보다 가깝다.

Diversity in Six Goat Populations in the Middle and Lower Yangtze River Valley

  • Jiang, X.P.;Liu, G.Q.;Ding, J.T.;Yang, L.G.;Cao, S.X.;Cheng, S.O.
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권2호
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    • pp.277-281
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    • 2003
  • Amplified fragment length polymorphism (AFLPs) markers were used to investigate the genetic variation in six autochthonous goat populations distributed in the middle and lower Yangtze River valley. The goat populations were Chengdu Grey Goat (CGG), Chuandong White Goat (CWG), Banjiao Goat (BG), Matou Goat (MG), Hui Goat (HG) and Yangtze River Delta White Goat (YRDWG). A total of 180 individuals (30 per population) were analysed using ten selected AFLP primer combinations that produced 78 clear polymorphism loci. The variability at AFLP loci was largely maintained within populations, as indicated by the average genetic similarity, and they were ranged from 0.745 to 0.758 within populations and 0.951 to 0.970 between populations. No breed specific markers were identified. Cluster analysis based on Nei' genetic distance between populations indicated that Chengdu Grey Goat is the most distant population, while CWG and YROWG were the closest populations, followed by BG, HG and MG. Genetic diversity of the goat populations didn' confirm what was expected on the basis of their geographical location, which may reflect undocumented migrations and gene flows and identify an original genetic resource.

PCR-based Assay for the Specific Detection of Pseudomonas syringae pv. tagetis using an AFLP-derived Marker

  • Song, Eun-Sung;Kim, Song-Yi;Chae, Soo-Cheon;Kim, Jeong-Gu;Cho, Heejung;Kim, Seunghwan;Lee, Byoung-Moo
    • 식물병연구
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    • 제21권1호
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    • pp.1-5
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    • 2015
  • A PCR method has been developed for the pathovar-specific detection of Pseudomonas syringae pv. tagetis, which is the causal agent of bacterial leaf spots and apical chlorosis of several species within the Compositae family. One primer set, PSTF and PSTR, was designed using a genomic locus derived from an amplified fragment length polymorphism (AFLP) fragment produced a 554-bp amplicon from 4 isolates of P. syringae pv. tagetis. In DNA dot-blot analysis with the PCR product as probe, a positive signal was identified in only 4 isolates of P. syringae pv. tagetis. These results suggest that this PCR-based assay will be a useful method for the detection and identification of P. syringae pv. tagetis.

Genetic Relationships of Lactuca spp. Revealed by RAPD, Inter-SSR, AFLP, and PCR-RFLP Analyses

  • Yang, Tae-Jin;Jang, Suk-Woo;Kim, Won-Bae
    • Journal of Crop Science and Biotechnology
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    • 제10권1호
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    • pp.27-32
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    • 2007
  • RAPD, Inter-SSR, and AFLP markers were used to assess the genetic diversity of lettuce cultivars and the phylogenetic relationships in Lactuca spp. A total of 216 polymorphic bands from seven RAPD primers, four Inter-SSR primers, and five AFLP primer combinations were used to elucidate the genetic similarity among lettuce cultivars. Forty-four lettuce accessions were subdivided into discrete branches according to plant type: crisphead, butterhead, and stem type, with some exceptions. The leafy- and cos-type accessions were intermingled in other groups with no discrete branch indicating that these are more diverse than others. Three accessions, including the Korean cultivar 'Cheongchima', the Korean local landrace 'Jinjam', and the German cultivar 'Lolla Rossa' were classified as the most diverse accessions. Twenty bands were unique in specific cultivars. Among these, three were specific in a plant type; one in Korean leafy type, one in crisphead type, and one in cos type lettuce. In the phylogenetic analysis among Lactuca species, L. saligna, L. serriola, and L. georgica clustered in a sister branch of the L. sativa complex. Two L. virosa accessions show the highest intra-specific relationships. L. perennis outlied from all the other Lactuca species at a genetic similarity of 0.53 and clustered with two Cichorium species, C. intybus and C. endivia, with genetic similarity of 0.67. The phylogenetic tree was supported by data from polymorphism of chloroplast genome which was revealed by PCR-RFLP.

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Development of AFLP and STS Markers Related to Stay Green Trait in Multi-Tillered Maize

  • Jang Cheol Seong;Lee Hee Bong;Seo Yong Weon
    • 한국작물학회지
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    • 제49권4호
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    • pp.358-362
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    • 2004
  • In order to develop molecular markers related to stay green phenotype, AFLP analysis was conducted using near-isogenic lines for either stay green or non stay green trait. Both lines have characteristics of multi-ear and tillers (MET). Two out of 64 primer combinations of selective amplification identified three reproducible polymorphic fragments in MET corn with stay green. Both of E+AGC/M+CAC and E+AAG/M+CAA primer combinations produced two and one specific polymorphic fragments linked to stay green trait, respectively. For the conversion of AFLPs to sequence tag sites (STSs), primers were designed form both end sequences of each two polymorphic fragments. One fragment, which was amplified with E+AAG/M+CAA primer combinations, possessed 298 bp long and showed a $91\%$ homology with maize retrotransposon Cinful-l. One out of two polymorphic fragments produced with E+AGC/M+CAC primer combination had 236 bp long and matched a $96\%$ homology with an intron region of 22kDa alpha zein gene cluster in Zea mays. One out of two PCR fragments amplified with MET2 primer set in the stay green MET was not produced in the non-stay green MET. The developed AFLP and STS marker could be used as an efficient tool for selection of the stay green trait in the MET inbred.

Identification of DNA Variations Using AFLP and SSR Markers in Soybean Somaclonal Variants

  • Lee, Suk-Ha;Jung, Hyun-Soo;Kyujung Van;Kim, Moon-Young
    • 한국작물학회지
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    • 제49권1호
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    • pp.69-72
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    • 2004
  • Somaclonal variation, defined as phenotypic and genetic variations among regenerated plants from a parental plant, could be caused by changes in chromosome structure, single gene mutation, cytoplasm genetic mutation, insertion of transposable elements, and DNA methylation during plant regeneration. The objective of this study was to evaluate DNA variations among somaclonal variants from the cotyledonary node culture in soybean. A total of 61 soybean somaclones including seven $\textrm{R}_1$ lines and seven $\textrm{R}_2$ lines from Iksannamulkong as well as 27 $\textrm{R}_1$ lines and 20 $\textrm{R}_2$ lines from Jinju 1 were regenerated by organogenesis from the soybean cotyledonary node culture system. Field evaluation revealed no phenotypic difference in major agronomic traits between somaclonal variants and their wild types. AFLP and SSR analyses were performed to detect variations at the DNA level among somaclonal variants of two varieties. Based on AFLP analysis using 36 primer sets, 17 of 892 bands were polymorphic between Iksannamulkong and its somaclonal variants and 11 of 887 bands were polymorphic between Jinju 1 and its somaclonal variants, indicating the presence of DNA sequence change during plant regeneration. Using 36 SSR markers, two polymorphic SSR markers were detected between Iksannamulkong and its somaclonal variants. Sequence comparison amplified with the primers flanking Satt545 showed four additional stretches of ATT repeat in the variant. This suggests that variation at the DNA level between somaclonal variants and their wild types could provide basis for inducing mutation via plant regeneration and broadening crop genetic diversity.

AFLP 마커를 이용한 단양쑥부쟁이 개체군의 유전다양성 보전을 위한 최소개체군의 크기산정 (Assessment of the Minimum Population Size for ex situ Conservation of Genetic Diversity in Aster altaicus var. uchiyamae Populations Inferred from AFLP Markers)

  • 김창균;김호준;최홍근
    • 한국환경생태학회지
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    • 제25권4호
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    • pp.470-478
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    • 2011
  • 본 연구는 멸종위기식물인 단양쑥부쟁이(Aster altaicus var. uchiyamae)의 개체군을 대상으로 유전다양성을 유지하는데 필요한 최소개체수를 산정하기 위하여 수행되었다. 단양쑥부쟁이가 분포하고 있는 네 지역에서 각각 유전다양성 및 유전적 분화도를 분석하였다. AFLP(amplified fragment length polymorphism) 마커를 이용한 유전적 변이의 분석결과, 총 4개의 프라이머 조합에 대해서 936개의 밴드가 확인되었으며, 그 중 934개의 밴드(99.8%)가 다형성을 보여주었다. 단양쑥부쟁이 개체군 내에서 유전다양성(PPB = 45.3%, h = 0.104, I = 0.168, hs = 0.108)은 높은 수준으로 나타났으며, 개체군 간 유전적 분화도($G_{ST}$ = 0.075, ${\theta}^B$ = 0.079)는 낮은 수준이었다. AMOVA(Analysis of molecular variance)분석 결과에서도 전체 유전적 변이 중 91%가 개체군 내에서 보이는 반면, 9%는 개체군 간 변이에 기인한 것으로 나타났다. 단양쑥부쟁이 개체군에서 보이는 유전적 특성은 개체군 간의 빈번한 유전자 이동에 기인한 것으로 사료된다. 최대화 전략법에 의하여 경기도 여주일대의 3개 개체군을 대상으로(굴암, 도리섬, 삼합) 개체군 내 최소개체수를 산정한 결과 도리섬개체군에서는 17개체, 삼합개체군에서는 16개체, 굴암개체군에서는 11개체로 파악되었다. 단양쑥부쟁이 개체군의 최소개체수에 대한 정보는 효율적인 현지 외 보전을 위한 가이드라인을 제시해 줄 수 있다.

Genetic Variation and Polymorphism in Rainbow Trout, Oncorhynchus mykiss Analysed by Amplified Fragment Length Polymorphism

  • Yoon, Jong-Man;Yoo, Jae-Young;Park, Jae-Il
    • 한국양식학회지
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    • 제17권1호
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    • pp.69-80
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    • 2004
  • The objective of the present study was to analyze genetic distances, variation and characteristics of individuals in rainbow trout, Oncorhynchus mykis using amplified fragment length polymorphism (AFLP) method as molecular genetic technique, to detect AFLP band patterns as genetic markers, and to compare the efficiency of agarosegel electrophoresis (AGE) and polyacrylamide gel electrophoresis (PAGE), respectively. Using 9 primer combinations, a total of 141 AFLP bands were produced, 108 bands (82.4%) of which were polymorphic in AGE. In PAGE, a total of 288 bands were detected, and 220 bands (76.4%) were polymorphic. The AFLP fingerprints of AGE were different from those of PAGE. Separation of the fragments with low molecular weight and genetic polymorphisms revealed a distinct pattern in the two gel systems. In the present study, the average bandsharing values of the individuals between two populations apart from the geographic sites in Kangwon-do ranged from 0.084 to 0.738 of AGE and PAGE. The bandsharing values between individuals No.9 and No. 10 showed the highest level within population, whereas the bandsharing values between individuals No.5 and No.7 showed the lowest level. As calculated by bandsharing analysis, an average of genetic difference (mean$\pm$SD) of individuals was approximately 0.590$\pm$0.125 in this population. In AGE, the single linkage dendrogram resulted from two primers (M11+H11 and M13+H11), indicating six genetic groupings composed of group 1 (No.9 and 10), group 2 (No. 1, 4, 5, 7, 10, 11, 16 and 17), group 3 (No. 2, 3, 6, 8, 12, 15 and 16), group 4 (No.9, 14 and 17), group 5 (No. 13, 19, 20 and 21) and group 6 (No. 23). In AGE, the genetic distances among individuals of between-population ranged from 0.108 to 0.392. In AGE, the shortest genetic distance (0.108) displaying significant molecular differences was between individuals No.9 and No. 10. Especially, the genetic distance between individuals No. 23 and the remnants among individuals within population was highest (0.392). Additionally, in the cluster analysis using the PAGE data, the single linkage dendrogram resulted from two primers (M12+H13 and M11+H13), indicating seven genetic groupings composed of group 1 (No. 15), group 2 (No. 14), group 3 (No. 11 and 12), group 4 (No.5, 6, 7, 8, 10 and 13), group 5 (No.1, 2, 3 and 4), group 6 (No.9) and group 7 (No. 16). By comparison with the individuals in PAGE, genetic distance between No. 10 and No. 7 showed the shortest value (0.071), also between No. 16 and No. 14 showed the highest value (0.242). As with the PAGE analysis, genetic differences were certainly apparent with 13 of 16 individuals showing greater than 80% AFLP-based similarity to their closest neighbor. The three individuals (No. 14, No. 15 and No. 16) of rainbow trout between two populations apart from the geographic sites in Kangwon-do formed distinct genetic distances as compared with other individuals. These results indicated that AFLP markers of this fish could be used as genetic information such as species identification, genetic relationship or analysis of genome structure, and selection aids for genetic improvement of economically important traits in fish species.