Colletotrichum acutatum J. H. Simmonds is an important fungal pathogen that causes anthracnose disease in crops. In this study, 59 C. acutatum isolates from regions in South Korea were collected and phenotypically compared. The average colony diameters of isolates from Gyeongsangbuk-do, Chungcheongbuk-do, and Gangwon-do were 4.36 cm, 4.24 cm, and 4.38 cm, respectively, following 6 days of growth on V8 juice agar. The average conidium concentrations of the isolates from each province were 14.43, 12.83, and 10.17 × 104/mL after 7 days of growth on V8A. Amplified fragment length polymorphism(AFLP) analysis using three primer pairs identified 5 polymorphic bands and 56 monomorphic bands. Two polymorphic bands were identified using the E-ACC/M-CTT primer pair, one polymorphic band was identified using the E-ACG/M-CTG primer pair, and two polymorphic bands were identified using the E-ACG/M-CTT primer pair. A dendrogram was constructed using the AFLP data, and four clusters were identified.

무당벌레(Harmonia axyridis)는 종내에서 초시색상패턴이 매우 다양하게 존재한다. 본 논문에서는 서로 다른 색상패턴의 무당벌레를 대상 으로 amplified fragment length polymorphism (AFLP)을 실시하여 무당벌레의 초시색상 패턴간 유전형질의 차이를 확인하고자 하였다. 총 28 개의 프라이머 조합으로 실험을 실시한 결과, 총 2,741 개의 밴드가 검출되었다. 그 중 20 개의 밴드(S1-S20)만이 특정 색상패턴에서 나타났다. 이들 가운데 9 개의 밴드를 색상에 연관된 AFLP 후보 지표로 선발하였다. 밴드 가운데 S1과 S2, S20은 Succinea 1, 2 변이형에 공통적으로 나타났으며, S3와 S5는 Conspicua 변이형에 특이적이었다. 또한 S13는 Spectabilis 변이형에, S15와 S18, S19는 Succinea 2 변이형에 특이적이었다. 특정 색상패턴에만 나타나는 9 개의 AFLP 지표들은 cloning을 통해 염기서열 분석을 실시하였고, GenBank를 이용해 다른 염기 서열과 비교를 해보았지만 아무런 상동성도 찾을 수가 없었다. 무당벌레 종 내 유전적 다양성을 평가한 결과, Spectabilis가 Conspicua보다 Succinea 변이형에 높은 유사성을 보였다. 색상에 연관된 AFLP 후보 지표를 기준으로 sequence characterized amplified region (SCAR) 지표로 변환하여 9 개의 AFLP 분자지표들 가운데에서 5 개만이 SCAR 지표로 전환될 수 있었으며, 이를 통해 AFLP 지표가 무당벌레의 색상과 연관되어 있는지 확인할 수 있었다.

Investigating loci compositions by conventional methods is limited in fully addressing complex gene information. We applied self-organizing map (SOM) to characterize Amplified Fragment Length Polymorphism (AFLP) of aquatic insects in six streams in Japan in responding to environmental variables. Locus band presence patterns were clustered by the trained SOM. Presence and absence data of loci were altered and cluster change through recognition was Subsequently expressed to indicate sensitivity to environmental variables. The outlier loci were determined based on the 90th percentile. Subsequently environmental responsiveness was obtained for each outlier in different species. Outlier loci were overall sensitive to pollutants and feeding material. Poly-loci like responsiveness was detected in adapting to environmental constraints. SOM training combined with recognition could be an efficient means of characterizing loci information without knowledge on population genetics a prior.

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(GST = 0.075, θB = 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.

To identify DNA markers linked to a elytra polymorphism, amplified fragment length polymorphism (AFLP) analysis was performed on DNA samples from four each colour pattern individuals (2 females and males), for example, succinea 1, succinea 2, conspicua, and spectabilis. As a result of performing AFLP analysis with the restriction endonuclease combination EcoRⅠ and Mse I, total of 2,269 AFLP fragments which were specific to succinea, conspicua and spectabilis was identified using 24 different AFLP primer combinations. Among these 2,269 fragments, 16 bands which were the most specific to one color patterns were isolated, cloned and sequenced. Subsequent UPGMA cluster analysis revealed that population of H. axyridis was divided four major group and these genetic tree showed that H. axyridis elytra colour diversity was affected by genetic polymorphism. It is considered that these genetic analyses may be facilitated the understanding of molecular genetic mechanism related with the wing colour pattern formation in this species.

무당벌레(Harmonia axyridis)는 전 세계에 걸쳐 다양하게 분포하고 있으며, 여러 종류의 진딧물을 섭식하는 진딧물의 포식자로 널리 알려지면서 진딧물 의 생물적 방제 프로그램에 유용하게 사용되고 있다. 무당벌레는 같은 종 내 에서도 변이가 매우 심하여 다양한 초시 색상패턴을 나타내는데, 이러한 종이 나 개체간의 분자적 요인의 차이를 밝혀내는 Fingerprinting 방법으로는 RAPD, RFLP, AFLP방법 등이 있으며, 이 중에서 AFLP는 높은 재현성과 정확성을 나 타냄으로 가장 널리 사용되고 있다. 따라서 본 연구에서는 AFLP 분석을 통해 무당벌레의 초시 색상패턴에 따른 유전적 다양성을 알아보고자 하였다. 2007년부터 2009년까지 채집된 월동개체군 중에서 검은색 바탕에 2개의 붉 은색 점을 가진 conspicua 개체와, 검은색 바탕에 4개의 붉은색 점을 가진 spectabilis 개체, 노란색 바탕에 19개의 검은색 점을 가지고 있는 succinea1 개 체, 그리고 노란색 바탕에 검은색 점이 전혀 없는 succinea2 개체를 가지고 실 험을 실시한 결과, 8개의 AFLP primer조합을 통해 총 822개의 밴드가 확인되 었고, 그 중 37개의 밴드 특정 색상패턴에서만 확인되어, 특이적 밴드를 통해 무당벌레의 색상변이에 있어 유전적 다양성을 추측할 수 있었다.

As an effective generalist predator of aphids and other hemipteran pests, Harmonia axyridis has been a successful biological control agent. Interestingly, it was known that there were varied in color patterns on H. axyridis elytra. In fact, Seo & Youn (2007) reported that H. axyridis had five color patterns, for example, succinea 1, 2, conspicua, spectabilis, and axyridis. But there are uncertain that H. axyridis elytra colour patterns are regulated by genetic polymorphism. So we tried to what is the reason that color patterns are greatly variable. To identify DNA markers linked to a elytra polymorphism, amplified fragment length polymorphism (AFLP) analysis was performed on DNA samples from four female succinea, conspicua, spectabilis and Coccinella septempunctata which is another species in Coccinellidae. AFLP analysis with the restriction endonuclease combination EcoRⅠ and MseⅠwas performed. Using 12 AFLP primer pairs, nine AFLP fragments which is specific between succinea, conspicua, spectabilis was identified. These nine AFLP fragments were isolated, cloned and sequenced. Subsequent UPGMA cluster analysis revealed three major group of H. axyridis populations. These genetic tree showed that H. axyridis elytra colour diversity was affected by genetic polymorphism. For more genetically understanding elytra colour genes, different primer combinations may be need to be generate enough polymorphic markers. These genetic analyses may be facilitate the understanding of molecular mechanism behind wing colour pattern formation.

The multicolored Asian ladybird beetle, Harmonia axyridis, which demonstrates typical genetic polymorphism in its elytra color patterns. Early studies have color polymorphism in terms of geographical clines while a few investigated temporal populations in Coccinellidae. Nevertheless, note that geographical and temporal morph variation does not always correspond to what is expected from thermal and industrial adaption theories. A recent study of transformation and RNAi of the ladybird beetle, however, there is yet no evidence to indicate the variation is genetic or environmental factors. Here we describe a relatively new molecular fingerprinting technique, amplified fragment length polymorphism (AFLP). Because we think that color polymorphism in Coccinellidae is affected by genetic polymorphism. In total 38 markers were scored from which some markers were polymorphic. Supsequent UPGMA cluster analysis revealed 3 major group of Harmonia axyridis populations. But for strains that are more genetically similar, different primer combinations may be need to generate enough polymorphic marker.

Wheat-rye translocation lines are widely used in wheat breeding programmes by reason of biotic stress tolerances. Though there have been a number of researches regarding abiotic stress tolerance, the tolerance of the lines depends on wheat genetic background, not on rye chromosome. Here, we investigated wheat-rye translocation specific transcripts derived from cDNA-AFLP under drought stress, which may help to elucidate the reaction under the stress. ‘OK91G117’ (1BL.1RS translocation) and ‘OK91G144’ (non-translocation) were used as materials, which are near-isolines for 1RS. 25% PEG 6000 was added in culture solution to simulate drought condition and root tissues were sampled at each 0 h, 3 h, 6 h, 12 h, 24 h, and 48 h after PEG treatment for RNA extraction. As a result of cDNA-AFLP, TDFs (transcript derived fragments) that were specific to OK91G117 were sequenced. GO functions of each sequenced TDF were annotated by Blast2GO using standard parameter with cut-off level 3 and mapped to the GO term (i.e. biological process; BP, molecular function; MF, cellular component; CC). The term with “organic substance metabolic process”, “primary metabolic process”, and “cellular metabolic process” account for almost 50 % of BP. The most represented terms among probes classified to MF were “transferase activity” and most of TDF were annotated in “cell part” of CC. In addition, rye-chromatin specific markers were developed by BLAST comparing sequence of TDF with wheat and rye genome data. RT-PCR was conducted to validate expression patterns of selected TDF. Further studies will be needed to elucidate functions of the highly expressed genes under drought stress.

For understanding the genetic diversity and genetic relationship between cultivated and weedy types, we evaluated genetic variation of 80 accessions of rice (O. Sativa). This included 42 cultivated accessions and 38 weedy accessions with the help of AFLP and CACTA-TD. A total of 542 loci were analyzed (255 for AFLP and 287 for CACTA-TD) of which AFLP markers exhibited 75% of polymorphism and transposon based CACTA-TD markers exhibited 93% of polymorphism. The average genetic diversity value for all 80 accessions, using AFLP markers was 0.226 (Cultivated – 0.210; Weedy 0.241) and based on CACTA-TD markers was 0.281 (Cultivated – 0.294; Weedy 0.269). A UPGMA phylogenetic tree revealed three major groups for both the marker system. The average polymorphic content value obtained with AFLP and CACTA-TD markers were 0.21 and 0.232, Effective multiplex ratio (AFLP – 47.50; CACTA-TD – 66.75), Marker Index (AFLP – 9.94; CACTA-TD – 21.13) and Resolving power (AFLP – 19.53; CACTA-TD – 34.62) indicated that the CACTA-TD markers were relatively efficient than AFLP markers.

Based on double pseudo-testcross theory, a population of 76 F1 clones, which were derived from a cross of China type tea plants (Camellia sinensis var. sinensis) with a Korean tea cultivar, ‘Kemsull’ for female parent and a Japanese tea cultivar, ‘Houshun’ for male parent, was used to construct a genetic linkage map with AFLP markers. Totally, 2,360 markers were detected by 26 pairs of primers and 90.8 markers for each pair on average. Among these, 481 markers (20.3%) were polymorphic, 392 markers (81.5%) of which showed Mendelian segregation ratio (p=0.01). Of these Mendelian segregated markers, 139 (35.5%) were segregated in 3:1 ratio and 253 (65.5%) were segregated in 1:1 ratio. The construction of AFLP molecular marker based linkage map were carried out by Joinmap 4.0 version. The linkage map of ‘Kemsull’ contained 227 markers which distributed into 18 linkage groups. The linkage map of ‘Kemsull’ covered 1,382.2 cM with the average distance between two markers of 6.0 cM. The linkage map of ‘Houshun’ contained 154 markers which were distributed into 17 linkage groups and were spanned with the total map length of 1,540.9 cM and the average distance between two markers of 10 cM. However, these AFLP markers were not distributed evenly and further even saturation is additionally required.

Amplified fragment length polymorphism (AFLP) is one of molecular marker technique based on DNA and is extremely useful in detection of high polymorphism between closely related genotypes like Korean wheat cultivars. Six Korean wheat cultivar specific marker sets have been developed from inter simple sequence repeat (ISSR) analysis and we can identify the 13 Koran wheat cultivars form other cultivars using six that (Son et al., 2013). We used four combinations of primer sets in our AFLP analysis for developing additional cultivar specific markers in Korean wheat. Twenty-one of the AFLP bands were isolated from ACG/M-CAC primer combination and 19 bands were isolated from E-AGC/M-CTG primer combination, respectively. We used forty bands to design sequence characterized amplified region (SCAR) primer pairs for Korean wheat cultivar identification. Only one of 40 amplified primer pairs, C2, were able to use for wheat cultivar identification. The DNA band of 215bp length was amplified by C2 primer pairs in ten cultivars, Eunpa, Olgeuru, Gobun, Saeol, Milsung, Sinmichal, Jokyung, Sugang, Goso, and Joah. Then C2 primer was applied to these primer sets as newly SCAR marker, six cultivars are identifying from other cultivars, additionally. Finally, to use the C2 and six primer sets, 19 Korean wheat cultivars are identified.

Collected germplasms of five representative species belonging to Curcuma genus (C. longa, C. aromatica, C.zedoaria, C. phaeocaulis and C. kwangsiensis) were 52 samples from different farmhouse in Korea and China. To elucidatethe genetic diversity among the species, 52 samples were analyzed by genomic fingerprinting method using amplified frag-ment length polymorphism (AFLP). AFLP results of 6 primer combinations were revealed 643 total DNA fragments and349 polymorphic bands with the 54.3% ratio of polymorphism. In the analysis of coefficient similarity using unweight pairgroup method with arithmetic averages (UPGMA), 52 Curcuma germplasm lines were ranged from 0.60 to 0.99 and clus-tered distinct five groups according to the species and collected geographical levels. However, the result of principal coordi-nate analysis (PCA) by multi-variate analysis was shown significantly greater differences among species than geographicalorigins based on AFLP profiling data of these samples.

Collected germplasms of five representative dandelion species (Taraxacum ohwianum, T. platycarpum, T. platypecidum, T. officinale, and T. coreanum) were 104 lines from different habitates in Korea and China. Their genetic diversity was analyzed by genomic fingerprinting method using amplified fragment length polymorphism (AFLP). AFLP results of 6 primer combinations were revealed 1,176 total DNA fragments and 523 polymorphic bands with a 44.4% ratio of polymorphism. On the basis of similarity coefficient analysis by unweight pair group method with arithmetic averages (UPGMA), 104 dandelion germplasm lines were ranged from 0.64 to 0.99 and clustered distinct five group depending on the species. Furthermore, a principal coordinate analysis (PCA) by the application of multi-variate analysis indicated significantly greater differences among species than geographical origins.

우리나라에서 수집한 재래종 조 26계통들에 대하여 9개의 AFLP primer조합을 사용하여 유전적 다양성 및 계통유연관계를 분석한 결과는 다음과 같다. 1. 분석에 이용한 9개의 AFLP primer조합들은 국내에서 수집한 조 26계통들에서 총 170개의 DNA단편을 나타내었고, 이중에서 145개의 단편(85.3%)들은 다형성을 나타내었다. 9개의 AFLP primer조합들에서 측정된 유전적 다양성 값(Hs)은 1.84에서 6.80의 범위로 나타나, 평균 3.85값을 나타내었다. 강원지역에서 수집한 조 계통(Group 1)들은 평균 3.39의 유전적 다양성 값을 나타내었고, 경기도 등 타 지역에서 수집한 조 계통들(Group 2)은 평균 2.99의 유전적 다양성 값을 나타내었다. 2. 국내에서 수집된 재래종 조 26계통들은 유전적 유사성 73% 수준에서 크게 두 개의 그룹으로 구분되었다. Group I은 유전적 유사성 76.8% 수준에서 강원도에서 수집한 13계통들과 경기도에서 수집한 1계통을 포함하고 있었으며, Group II는 유전적 유사성 78.9% 수준에서 강원도에서 수집한 2계통들과 타 지역에서 수집한 10계통들을 포함하고 있었다. 본 연구결과는 우리나라에서 수집한 재래종 조 계통들에 대한 유전적 다양성 및 계통유연관계 이해 그리고 이들 자원의 수집 및 보존 등에 유용한 정보를 제공할 것으로 기대된다.

본 연구는 AFLP 분자마커를 이용하여 국내 및 국외에서 수집된 조(Setaria italica (L.) P. Beauv.) 37계통 (국내 26계통, 파키스탄과 중국 11계통)들에 대하여 유전적 다양성을 분석하였다. 그 결과, 분석에 이용된 9개의 AFLP primer 조합들에서 총 171개의 DNA fragment를 확인하였고, 그 중 147(85.96%)개가 다형성 band를 나타 냈다. 9개의 AFLP primer 조합들에서 나타난 다형성 band의 수는 E-AAG/M-CTT primer 조합에서 10(74.43%) 개로 가장 낮은 수의 다형성 밴드를, 그리고 E-ACT/M-CAC primer 조합에서는 17(94.44%)개로 가장 높은 수의 다형성 밴드를 각각 나타내었다. 이들 9개의 AFLP primer 조합들에서 측정된 평균 phenotypic diversity 값은 국내 수집종인 GroupⅠ에서 3.62를, 그리고 국외 수집종인 GroupⅡ에서 2.81를 각각 나타내었다. 한편 UPGMA 분석 결과에 의하면, 유전적 유사성 0.77수준에서 크게 2개의 그룹으로 나뉘어졌는데. 첫 번째 Group Ⅰ에는 파키스탄 수집종과 국내 수집종이 포함되어있었고, 두 번째 GroupⅡ에는 중국 수집종과 국내 수집종 이 각각 포함되어 있었다. 본 연구 결과는 국내 및 국외에서 수집된 조 작물의 유전적 다양성을 이해하는데 유용한 정보를 제공할 것으로 기대된다.