The aim of this study was to assess the levels of genetic diversity and relationships of Korean native black goat (n = 58), compared with the exotic breed, Boer (n = 97). For the analysis of genetic characterization 11 microsatellite markers (MAF065, INRA063, CSRD247, OarFCB20, SRCRSP5, INRA006, ILSTS008, ILSTS011, INRA005, ILSTS087, SRCRSP8) were genotyped. The number of alleles was observed 3 (INRA005) to 10 (SRCRSP8) each markers. The mean expected and observed heterozygosity (Hexp and Hobs) and polymorphism information content (PIC) for the Korean native black goat breed varied from 0.551 to 0.860, 0.517 to 0.948 and 0.464 to 0.835, respectively. Principal Components Analysis (PCoA) and FCA results showed that Korean native black goat breed was confirmed to be clearly separated from bore breed. These results were scientific evidence that Korean native black goat represents a unique and valuable animal genetic resource.
The Bemisia tabaci (Hemiptera: Aleyrodidae) is a pest that sucks a plant's phloem, excretes nectar and infects tomatoes with TYLCV (Tomato Yellow Leaf Curl Virus), causing severe economic damage. In order to explore the cause for occurrence of areas where the genetic structure of B. tabaci MED (Mediterranean) was reversed in a short period of time, this study was conducted. We collected and analyzed the B. tabaci MED populations from tomato greenhouses in some areas where their genetic structure was reversed between two years 2016 and 2017. As a result, the genetic structure of B. tabaci MED populations was found to be close between populations of the nursery and the greenhouses where tomato seedlings were supplied from that nursery, even if greenhouses were located far away each other. Further study is needed to identify the causes for occurrence of abrupt reverse of genetic structures of B. tabaci MED populations observed in some areas throughout the country.
Microsatellite SSR markers were developed and utilized to reveal the genetic diversity of 32 strains of Flammulina velutipes collected in Korea, China, and Japan. From SSR-enriched library, 490 white colonies were randomly selected and sequenced. In the 490 sequenced clones, 85 clones (17.35%) were redundant. Among the remaining 405 unique clones, 201 clones (49.6%) contained microsatellite sequences. As a result, 12 primer pairs produced reproducible polymorphic bands within diverse 4 strains and these selected markers were further characterized in 32 Flammulina velutipes strains. A total of 34 alleles were detected using the 12 markers, with an average of 3.42 alleles and the number of alleles ranged from two to seven per locus. The major allele frequency ranged from 0.42 (GB-FV-127) to 0.98 (GB-FV-166), and values for observed (HO) and expected (HE) heterozygosity ranged from 0.00 to 0.94 (mean = 0.18) and from 0.03 to 0.67 (mean = 0.32), respectively. SSR loci amplified with GB-FV-127 markers gave the highest polymorphism information content (PIC) of 0.61 and mean allele number of five, while for loci amplified with GB-FV-166 markers these values were the lowest, namely 0.03 and two. The mean PIC value (0.29) observed in the present study with average number of alleles (3.42). The genetic relationships among 32 Flammulina velutipes strains based on SSR data were generated by UPGMA cluster analysis. In conclusion, we succeeded in developing 12 polymorphic SSRs markers from SSR-enriched library of Flammulina velutipes. These SSRs are presently being used for phylogenic analysis and evaluation of genetic variations. In future, these SSR markers will be used in clarifying taxonomic relationships among the Flammulina velutipes.
근래의 육종산업에 있어서 재래 혹은 토착품종 등 고유 가축유전자원의 효율적인 보존 및 관리는 중요한 관심사이다. 본 연구는 제주재래돼지와 외래돼지 품종들간의 유전적 다양성, 집단의 구성 및 근연관계를 구명하기 위하여 수행되었다. 제주재래돼지, Landrace, Yorkshire, Berkshire 등 4품종 총 200개체를 대상으로 30개 초위성체 마커를 대상으로 대립유전자를 분석한 결과, 전체 265개의 대립유전자가 관찰되었다. 대립유전자 수의 범위는 5개(SW168)에서 22개(S0005)였으며, 전체 좌위에 대한 평균 값은 8.83개로 산출되었다. 30개 마커에 대한 기대 및 관측 이형접합도의 평균치는 0.731 및 0.615, 다형정보지수의 평균값은 0.697로 확인되었다. 제주재래돼지의 유전적 다양성은 외래 품종에 비하여 낮게 나타났다. 계통유전학적 유연관계, 요인대응분석 및 집단구조 분석 결과, 제주재래돼지는 서양유래의 상용품종과 유전적으로 명확히 구분되었다. 따라서 본 연구 결과는 제주재래돼지의 유전적 고유성 및 유전자원으로써 가치판단을 위한 과학적 근거가 될 것으로 사료된다.
본 연구는 Cymbidium 원종 및 주요 품종을 대상으로 RAPD 분석을 이용하여 이들의 유전적 근연관계를 비교하고, 또한 유전적 구별성과 균일성을 확인하여 이들 생화학적 표지를 품종식별의 지표로 활용함으로써 교배모본을 선정할 때 품종의 기초자료를 얻기 위하여 수행하였다. 심비디움의 PCR 반응조건 구명하고자 각각의 반응액 조건을 알아본 결과 PCR tube(0.5mL)에 10ng template DNA, 100ng primer, 200μM dNTP mixture, 1unit Taq DNA polymerase, 1.5mM MgCl2, 10mM Tris-HCl을 첨가하여 25μL로 조정하여 최적 반응액을 만들었다. PCR 반응 조건은 94oC에서 5분간 예비 변성시키고, 94oC에서 3분 변성, 37oC에서 1분간 primer 접촉 및 72oC에서 5분간 증폭시키는 과정을 45회 반복했을 때가 가장 효과적이었다. 선발된 10개의 primer로부터 87개의 밴드로부터 심비디움 원종 및 품종 30종의 군집분석을 한 결과, 유사도 0.647을 기준으로 2개의 군집으로 분류할 수 있었고, I 집단은 유사도값 0.660을 기준으로 3 소군집, II집단은 유사도값 0.737을 기준으로 3 소군집이 속하였다. II집단 C. ‘Place court’ 와 C. Pure Destiny ‘Ultimate’는 유사도가 0.920으로 높게 나타났다. 중형종인 C. ‘Juulyang’과 C. ‘Tropical Yellow’도 0.908의 높은 유사도 지수를 보여 근연관계가 가깝게 나타났는데 화색은 초록색과 노란색이며 설판의 색과 화형이 비슷한 특징을 보였다. 본 연구를 통해 심비디움 원종 및 품종 30종간의 유전적 근연관계를 밝힘으로써 이후 심비디움 육종 및 유전연구에 유용한 기초자료가 될 수 있을 것으로 생각된다.
Nuclear ribosomal DNA (rDNA) was analyzed to identify inter-specific genetic relationships among 8 Cymbidium species (Cymbidium insigne, C. ensifolium, C. marginatum, C. faberi, C. gyokuchin, C. kanran, C. forrestii, and C. goeringii). Nuclear rDNA including 2 internal transcribed spacer (ITS) regions and 5.8S, was amplified using polymerase chain reaction and sequenced. The sequences were compared via pair-wise multiple alignment to determine the genetic relationships among the studied species. The lengths of the ITS1, ITS2, and 5.8S regions were 235 bp, from 255 bp to 257 bp, and from 153 bp to 165 bp, respectively. Sequence similarities in the ITS region ranged from 78.7% between C. gyokuchin and C. kanran to 96.8% between C. ensifolium and C. kanran. A phylogenetic tree was constructed from nuclear rDNA nucleotide sequence data of the 8 cymbidiums and 1 outgroup species to estimate genetic relationships. The tree revealed that cymbidiums could be classified by their ecological traits, such as their temperature preference or inflorescence pattern. The phylogenetic data is applicable for identification, classification, and breeding of cymbidiums.
Hipparchia autonoe belongs to the family Nymphalidae (Lepidoptera) and is designated as an endangered insect and national monument in Korea. It only inhabits a very restricted area on Mt. Halla but is widely distributed in several Asian countries including Mongolia. A previous study conducted to understand the genetic relationship between Mt. Halla and Mongolian H. autonoe for conservation purposes suffered from a limited number of samples. Therefore, we sequenced the DNA barcode region of an additional 36 H. autonoe individuals, combined them with previous data from 19 individuals, and performed phylogenetic and population genetic analyses. Furthermore, the internal transcribed spacer 2 (ITS2) region was also sequenced from the 36 samples as a nuclear DNA marker. The existence of independent haplotypes, sequence types, and significant FST estimates (p < 0.05) between Mt. Halla and Mongolian populations indicated hampered gene flow between the populations. Nevertheless, an absence of a reciprocal monophyletic group in Mt. Halla and Mongolian populations by cytochrome oxidase subunit I gene- and ITS2-based phylogeny suggests that the genetic isolation of the Mt. Halla population from the Mongolian populations seemed not large enough to consider them independent genetic entities.
The species status of Oeneis urda (Eversmann) and O. mongolica (Oberthür) has been argued based on morphological characters. Reexamination of their major morphological characters has shown a slight differentiation in the two species. Sequences of three mitochondrial genes (COI, ND6, and ND1) and one nuclear region (internal transcribed spacer 2, ITS2) from two O. urda populations (Yangyang and Mt. Hanla) and one O. mongolica population (Uljin) were performed for phylogenetic and population genetic inferences. Sharing of identical sequences in the ND6 gene and ITS2, minimal sequence divergence in the COI and ND1 genes, and phylogenetically undividable sequence types in all mitochondrial genes and ITS2 suggest genetic continuity between the two species. Nevertheless, significant FST estimates (p < 0.05) were found for the COI gene in comparisons between Yangyang (O. urda) and Uljin (O. mongolica), between Yangyang (O. urda) and Mt. Hanla (O. urda), and between Uljin (O. mongolica) and Mt. Hanla (O. urda) populations. These FST estimates, along with other gene-based analyses collectively suggest isolation of the two species at some point in the past, but incomplete separation between the two species on the mainland (Yangyang and Uljin) and biogeographically forced isolation of the O. urda population on Mt. Hanla collectively appear to complicate species status of these two species that were once further clearly separated.
Animal crude drugs (natural medicines derived from animal organs) have been widely used in various Chinese medicine for the therapeutic effects and for enhancement of immunologic functions. We found the specific identification methods using DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses for mitochondrial DNA (mt DNA) in order to discriminate between the animal species and organs as well as the placenta of humans. Species-specific PCR bands of D-loop mt DNA for equine, bovine, porcine, and human were 133 bp, 137 bp, 231 bp, and 240 bp, respectively. Porcine organs were identified using restriction enzyme, HphI cut into two subfragments, 36 bp and 195 bp bands in the heart, spleen, and liver, except for kidney. The heart and liver of porcine were identified using restriction enzyme, SpeI cut into two subfragments, 84 bp and 147 bp bands, except for kidney and spleen. Bovine organs were cut into 68 bp and 69 bp bands in the liver, kidney, and spleen using NalIV, except heart and placenta. Placentas of bovine and humans were easily identified using each primer. Our results suggest that sequencing of mt DNA and its PCR-RFLP methods are useful for identification and discrimination of inter- and intra-specific variations in equine, bovine, porcine, and human by routine analysis.
Phylogenetic relationships among 53 accessions of D.glomerata collected from 5 continents were investigated using simple sequence repeat (SSR) markers. 15 SSR primer pairs generated a total of 127 alleles, with an average of 8.5 alleles per locus. The average polymorphic rate (P) was 95.21 %. The genetic similarity (GS) among all accessions ranged from 0.43 to 0.94, with an average of 0.63. Analysis of molecular variance (AMOVA) indicated that larger proportions of variability existed within geographical regions (73.75%). High degree of genetic diversity was observed in Asia (P, 90.55%) and Europe (P, 86.61%) groups. Based on the cluster and principal component analysis, 53 accessions could be divided into five groups (GS=0.64) according to the nearest phylogenetic relationship.
SRAP (Sequence-related amplified polymorphism) and ISSR (Inter simple sequence repeat) molecualr markers were used to evaluate the levels and patterns of genetic diversity among 45 collections of orchardgrass from four continents. Twenty-one primer combinations were used and 480 bands were produced in SRAP, of which 405(84.38%) were polymorphic. On the other hand, twelve primers were used to generate a total of 116 bands in ISSR, of which 116(87.07%) were polymorphic. The coefficient range of genetic similarity was 0.6248-0.9686 and 0.6116-0.9231 respectively. Based on cluster and principal component analysis on the genetic characteristics, all collections could be divided into four groups and five groups in two markers, respectively. According to the analysis of genetic diversity and relationships, the appropriate strategies for collection and conservation of germplasm resources also were discussed and scientific breeding with far genetic relationship materials in orchardgrass were suggested.
한국꿩 (Korean ring-necked Pheasant, Phasianus colchicus karpowi)과 외국 아종의 유전적 유연관계를 파악하기 위해 야생 한국꿩, 사육 한국꿩, 사육 한국꿩과 외국꿩간의 잡종꿩, 외국꿩 4아종(중국 링넥, 흑 뮤탄트, 백 뮤탄트, 녹치)을 대상으로 ISSR 표지자 분석과 AMOVA 분석을 수행하였다. 야생 한국꿩의 전체 유전 다양성중 94.08%가 서식지 내 개체간 유전적 차이에 기인하고, 5.9% (Φ
본 연구는 파파리반딧불이 (Hotaria papcrinsis), 애반딧불이 (Luciola lateralis) 및 늦반딧 불이 (Pyrocoelia fufa)등 국내 주요 반딧불이 종의 유전적 분화 및 계통분류학적 관련을 파악하고자 하였다. 이를 위하여 mtDNA의 COI유전자 및 16S rRNA유전자 일부의 염기서열 (각 403bp 및 490bp~504bp)을 분석하였으며 아울러 GenBank에 등록된 일본 반딧불이 29종(반딧불이과 27종, 홍반딧과 1종 및 Rhagophthalmus과 1종)의 16S rRNA유전자의 동일부위 염기서열을 사용하였다. 국내 세 종간의 COI및 16S rRNA유전자의 염기서열 그리고 COI유전자의 아미노산 분화정도를 비교한 결과, 반딧불이아과(Lampyrinae)의 늦반딧불이는 애반딧불이아과(Luciolinae)에 공통적으로 속해있는 애반딧불이 및 파파리반딧불이와 다소 큰 유전적 차이를 나타냄으로 기존의 분류학적 위치를 확인하였다. 16S rRNA유전자의 염기서열을 이용, PAUP과 PHYLIP에 의한 계통분류학적 분석 결과, 우리 나라 애반딧불이는 일본 애반딧불이와 강력한 단일그룹을 형성하였으나 이들간 상당한 유전적 차이 (2.9%의 16S rRNA유전자 염기분화율)를 보였다. 국내 두 지역의 파파리반딧불이는 일본 대마도 고유종인 H. tsushimana와 같은 계통그룹을 형성하였으므로 Hotaria란 속명의 사용이 타당해 보이나 파파리반딧불이는 지역 개체간 자매분류군을 형성하지 않으므로 이에 대한 추가 연구가 요망되는 실정이다. 마지막으로, 국내 늦반딧불이 지역 개체가 일본 늦반딧불이와 강력한 단일 계통그룹을 형성한 점으로 미루어 Pyrocoelia란 속명의 사용은 타당해 보이나 다른 모든 늦반딧불이로부터 큰 유전적 거리론 보인 제주도 개체에 대한 추가적인 연구가 요망되는 실정이다. 결론적으로, 국내 반딧불이 종들은 일본에서 공통적으로 발생하는 반딧불이종 또는 속과 아주 강력한 계통그룹을 형성하였으므로 기존의 계통관련 연구를 지지하고 있는 실정이다.
Eleven Italian ryegrass cultivars were examined for their genetic polymorphisms and phylogenetic relationships using randomly amplified polymorphic DNA (RAPD) markers. In RAPD analysis of 34 random primers, 96 of total 162 bands obtained from 16 primers w
연초에 채집하여 Blackman(1987)의 기주선호성과 형태적인 특징을 이용하여 분류한 방법에 의해 두가지 타입을 (M. persicae Sulzer와 M. nicotianae Blackman)으로 구분된 무시성충 복숭아 혹진딧물 8 클론의 유전적 특성을 분석을 위하여 RAPD-PCR방법을 이용하였다. 사용된 random primer(10-mer) 100개 중에서 20개의 primer을 선발하였는데 GC content가 70, 80, 90%인 primer에서 각각 26.9%, 50.5% 및 66.^%로 GC content가 높아질수록 PCR결과 band의 양상이 좋게 나타났다. simple matching coefficient를 구하여 matrix를 작성해 본 결과 유사계수(similarity coefficient)의 범위는 0.414~0.808사이이었다. 복숭아혹진딧물 clone 간에 유사계수가 가장 높은 것은 PG2와 PG3 클론으로 0.808로 나타났으며, DBR클론을 기준으로 하여 볼 때 PG2 클론간의 유사계수를 0.414로 유사도가 가장 낮았다. 유사계수를 이용하여 8가지 클론의 진딧물들에 대한 유전적 근연관RP를 살펴보면 M. persicae typer에 속하는 PG1, PG2, PG3 클론들과 M. nicotianae type에 속하는 RED 클론이 유사도 0.643에서 한그룹, M. nicotianae type에 속하는 GR1, GR2, BRN크론들이 유사도 0.636에서 유연관계가 있었으며 그리고 M. persicae type에 the하는 DBR클론 등 세 개의 그룹으로 구분되었다. 따라서 연초에서 발생한 복숭아혹진딧물 형태형 2종 (M. persicae와 M. nicotianae)에 대하여 RAPD 기법을 이용하여 분석해본 결과 뚜렷한 유전적 유연관계는 발견하지 못하였다.