황벽나무 자연집단의 ISSR 유전변이

이제완,강진택,홍경낙,홍용표,안지영 한국임학회 2012 산림과학 공동학술대회 논문집 Vol.2012 No.-

Microsatellite 마커를 이용한 은행나무 천연기념물의 DNA 지문 분석

이제완,이민우,안지영,홍경낙,백승훈,김상철 한국산림과학회 2017 한국산림과학회지 Vol.106 No.4

본 연구는 국내 천연기념물 은행나무 23개체를 대상으로 8개의 microsatellite 마커를 이용하여 DNA 지문분석을 수행하였다. 그 결과, 평균 대립유전자수는 6.875개, 평균 이형접합도 관찰치와 기대치는 각각 0.711과 0.710로 나타났다. 이러한 수치는 동일한 마커로 일본 및 중국 은행나무에서 분석된 기존 연구 결과와 유사하였다. 분석된 8개 마커의 다형성 정보량(PIC) 값 및 개체식별력(PD)은 각각 0.677과 0.9999로 높은 개체식별 효율을 나타내었다. 실제 유전형을 비교한 결과에서도 모든 천연기념물 은행나무와 추가로 분석된 일반 은행나무 13개체가 모두 식별되었다. 천연기념물 은행나무에 서 계산된 PIC 값을 기준으로 상위 3개의 마커(Ging06, Gb60, Gb61)에서 계산된 개체인식력과 개체식별력이 각각 8.045×10 -5 및 99.99%로 계산되므로, 이들 3개의 마커가 은행나무 개체식별을 위한 DNA 지문 분석에 우선 적용이 가능할 것이다. 본 연구의 DNA 지문 분석 결과는 천연기념물 은행나무의 관리와 후계목 육성 및 은행나무 선발 개체의 유전적 동일성 검정에 유용한 자료로 활용될 것으로 판단된다. This study describes DNA fingerprinting analysis of twenty-three natural monument individuals of Ginkgo biloba using eight microsatellite markers. The average number of observed alleles was 6.875, and the expected heterozygosity and the observed heterozygosity were 0.711 and 0.710, respectively. This results were similar to those of the previous studies on Ginkgo trees analyzed by same markers in China and Japan. PIC value and PD were calculated at 0.677 and 0.9999 respectively, indicating a high individual identification efficiency. In fact, all of the natural monument ginkgo trees and additionally analyzed thirteen general ginkgo tress were identified by genotype comparison. PI and PD calculated in three markers (Ging06, Gb60, Gb61) with the highest PIC values calculated in natural monument ginkgo trees were 8.045×10 -5 and 99.99%, respectively. Thus, these three markers could be preferentially used in DNA fingerprinting for identifying ginkgo tree individuals. The results in this study will be useful for management of natural monument ginkgo trees, proliferation of their progeny and genetic identification of individuals selected in breeding process.

학술연구논문 구두발표 2 : 황벽나무 자연집단의 유전다양성 및 유전구조 분석

이제완 ( Jei Wan Lee ),홍경낙 ( Kyung Nak Hong ) 한국임학회 2013 산림과학 공동학술대회 논문집 Vol.2013 No.-

황벽나무 자연집단의 유전다양성 및 유전구조 분석

이제완(Jei-Wan Lee),홍경낙(Kyung-Nak Hong) 한국산림휴양복지학회 2013 한국산림휴양학회 학술발표회 자료집 Vol.2013 No.4

2분과 : 황벽나무 자연집단의 유전다양성 및 유전구조 분석

이제완 ( Jei Wan Lee ),홍경낙 ( Kyung Nak Hong ),강진택 ( Jin Taek Kang ) 한국임학회 2014 한국산림과학회지 Vol.103 No.1

본 연구는 ISSR 표지자를 이용하여 국내 분포하는 황벽나무 7개 집단의 유전다양성과 유전구조를 분석하였다. 6개의 ISSR primer를 이용하여 분석한 결과 primer 당 평균 4.5개의 다형성 band를 확인하였고, 각집단의 다형성 유전자좌의 비율은 평균 78.8%로 나타났다. Shannon의 유전다양성 지수(I)는 0.421로 나타났고, 이형접합체 기대치(He)는 평균 0.285로 베이즈 방법을 이용한 평균 이형접합체 기대치(hs=0.287)와 유사하였다. AMOVA에서 전체 유전변이의 92.4%가 집단내 개체간 차이에 기인하며, 7.6%는 집단간 차이에 기인하였다. 베이즈 방법을 이용한 유전분화(θII)는 0.066으로 추정되었으며, 전체 집단의 근친교배율(f)은 0.479로 계산되었다. 유연관계 분석과 베이즈 군집분석결과 우리나라 황벽나무 집단은 가평, 화천, 봉평, 용평이 하나의 군집을 형성하였고, 산청 지역의 2개 집단(삼장 및 시천)이 다른 하나의 군집을 형성하였으며, 무주 집단이 산청지역의 집단과 지리적으로 근접함에도 불구하고 독립적인 군집을 나타내었다. Mantel’s test 결과 집단간 유전적 유연관계와 지리적 분포의 상관성은 나타나지 않았다. 황벽나무의 유전자원보존을 위한 대상 집단 선정 시 생태적 및 생활사적 특징과 함께 본 연구결과에서 나타난 유전다양성과 군집구조 분석결과를 고려하는 것이 효과적일 것으로 사료된다. Genetic diversity and genetic structures were estimated in seven natural populations of Phellodendronamurense Rupr in South Korea using ISSR markers. The average of polymorphic loci per primer and the proportion of polymorphic loci per population were 4.5 and 78.8% respectively with total 27 polymorphic loci from 6 ISSR primers. The Shannon`s diversity index(I) was 0.421 and the expected heterozygosity(He)was 0.285, which was similar to the heterozygosity (hs = 0.287) inferred by Bayesian method. In AMOVA,7.6% of total genetic variation in the populations was resulted from the genetic difference among populationsand the other 92.4% was resulted from the difference among individuals within populations. Genetic differentiation(θII) and inbreeding coefficient(f) for total population were estimated to be 0.066 and 0.479 by Bayesian method respectively. In Bayesian clustering analysis, seven populations were assigned into three groups. This result was similar to the results of genetic relationships by UPGMA and PCA. The first groupincluded Hwachoen, Gapyeong, Bongpyeong and Yongpyeong population, and the second included two populations in Sancheong region. Muju population was discretely assigned into the third group in spite of thegeographically short distance from the Sancheong region. There was no significant correlation between genetic relationship and geographic distribution among populations in Mantel`s test. For conservation of thephellodendron trees, it would be effective to consider the findings resulted from this study with ecological traits and life histories of this species.

◇ 2분과 : 잣나무 엽록체 Simple Sequence Repeat 표지자 개발 및 특성 분석

이제완 ( Jei Wan Lee ),백승훈 ( Seung Hoon Baek ),홍경낙 ( Kyung Nak Hong ),홍용표 ( Yong Pyo Hong ),이석우 ( Seok Woo Lee ),안지영 ( Ji Young Ahn ) 한국임학회 2015 한국산림과학회지 Vol.104 No.4

본 연구에서는 잣나무 엽록체 DNA의 전체 염기서열을 기반으로 엽록체 SSR(chloroplast simple sequence repeat) 영역을 특이적으로 증폭하는 primer를 개발하고 그 특성을 분석하였다. 잣나무 엽록체 DNA에서 총 30개의 SSR 영역을 탐색하였으며, 이들 영역을 증폭하기 위한 30개의 primer를 제작하였다. 모든 primer가 잣나무를 대상으로 PCR 증폭이 가능하였다. 근연종에 대한 primer의 종간 전환률은 잣나무와 동일한 아속(Subgenus Strobus)에 속하는 눈잣나무(100%)와 섬잣나무(97%)에서 가장 높게 나타났다. 반면 소나무아속(Subgenus Pinus)에 속하는 소나무와 구주소나무에서의 종간 전환률은 73%로 비교적 낮게 나타났다. 점봉산 잣나무 집단을 대상으로 조사한 결과 13개의 유전자좌에서 다형성이 관찰되었으며, 평균 haploid 다양도(H)는 0.512로 계산되었다. 다형적 유전자좌로부터 조합된 haplotype의 수(N)는 25개로 확인되었고, haplotype 다양도(He)는 0.992로 매우 높게 나타났다. 집단내 독특하게 관찰되는 haplotype은 22개(88%)로 전체 28개체 중에서 22개체(79%)를 식별하였다. 본 연구에서 개발한 cpSSR primer는 높은 종간 전환률을 나타냄에 따라 소나무속의 근연종, 특히 잣나무아속 수종에 활용 가능성이 높고, 잣나무 유전변 이 분석을 위한 충분한 다형성을 제공하는 유용한 표지자로 판단된다. Novel cpSSR primers were developed based on the sequence information of the Pinus koraiensis chloroplast genome. A total of 30 cpSSR loci were detected in the chloroplast genome, and a total of 30 primer sets flanking those loci were designed. All primer sets were successfully amplified for chloroplast DNA in P. koraiensis. The cross-species transferability of the 30 primer sets was considerably high in P. pumila (100%) and P. paviflora (97%) belonging to the same Subgenus (Strobus) of P. koraiensis. Meanwhile, the transferability was relatively low (73%) in P. densiflora and P. sylvestris belonging to Subgenus Pinus. A total of 13 cpSSR loci out of the 30 loci were polymorphic in the Mt. Jumbong population of P. koraiensis. The mean of haploid diversity(H) was 0.512. The number of haplotypes(N) and the haplotype diversity(He) were 25 and 0.992, respectively. Of the 25 haplotypes, 22 were unique in the analyzed population. The unique haplotypes differentiated 22 individuals (79%) from the total of 28 individuals. In conclusion, the novel cpSSR primers developed in this study would be applicable to other Pinus species, especially the subgenus Strobus, and provide a high level of polymorphism for the study of genetic variation of P. koraiensis.

오대산 물황철나무(Populus koreana) 집단의 유전다양성 및 공간구조 분석

신수경,이제완,임효인,송정호 한국산림휴양복지학회 2013 한국산림휴양학회 학술발표회 자료집 Vol.2013 No.4

학술연구논문 구두발표 2 : 오대산 물황철나무(Populus koreana) 집단의 유전다양성 및 공간구조 분석

신수경,이제완,임효인,송정호 한국임학회 2013 산림과학 공동학술대회 논문집 Vol.2013 No.-

Fine-scale spatial genetic and clonal structure of Eleutherococcus senticosus populations in South Korea

홍경낙,이제완 한국산림과학회 2015 Forest Science And Technology Vol.11 No.3

For the conservation of clonal plant species, better understanding is required of the clonal recruitment process and the spatial distribution pattern of ramets and genets, especially when selecting individuals for ex situ conservation. Because of deforestation and illegal harvesting in natural populations of eleuthero (Eleutherococcus senticosus), a constant conservation effort is needed. We investigated the clonality and spatial genetic structure of two populations located in Hongcheon and Inje using 30 polymorphic markers generated by RAPD or ISSR PCR. The averages of the clonal diversity and genotypic evenness using the multilocus genotypes were 0.961 and 0.880, respectively. These were a little higher than those that have been estimated for other clonal plant species. By spatial autocorrelation analysis, the ramets in Hongcheon were randomly distributed and genetically homogeneous within a meter, while those in Inje showed slight regularity in their distribution and non-genetic patchiness. The average clone size was larger in Hongcheon, but the clonal diversity was opposite. The differences of clonality and spatial structure in both populations might be caused by the distribution of rocks in the forest floor acting as a physical barrier against rooting. There was a mild soil with small pebbles in Hongcheon, whereas the forest floor habitat in Inje was rocky. The clonality and the spatial genetic structure could be easily estimated with information on “putative” clones, verified from molecular work. The clonal recruitment process and sampling strategy for the ex situ conservation of eleuthero are also discussed.

Genetic diversity and structure of Carpinus laxiflora populations in South Korea based on AFLP markers

안지영,이제완,이민우,홍경낙 한국산림과학회 2019 Forest Science And Technology Vol.15 No.4

We applied eight primer-restriction enzyme combinations to investigate genetic diversity, genetic differentiation, and genetic structure of Carpinus laxiflora populations with AFLP markers. The average of effective alleles (Ae), the proportion of polymorphic loci (%P), Shannon’s diversity index (I), and the expected heterozygosity (He) were 1.4, 82.2%, 0.371, and 0.241, respectively. The expected heterozygosity (Hj) from Bayesian method was 0.270. The level of genetic diversity was high compared to those of Carpinus species and other species with a similar life history. The inbreeding coefficient (FIS) from approximated Bayesian method was 0.618, which was smaller than that for Acer pseudosieboldianum (FIS¼0.712). Genetic differentiation was 0.060 from AMOVA (UST) and 0.056 from Bayesian method (hII). The level of genetic differentiation was very small compared to that of Carpinus species and other species with a similar life history. According to UPGMA and Bayesian clustering, 10 populations were divided into two genetic groups. Except Mt. Chilgap and Minjuji, most of the populations were detected as weak genetic structures according to the geographical distribution such as mountain ranges. We might consider that demographic disturbance, local specific vegetation change history, and forest succession interrupted the genetic structure of C. laxiflora in South Korea.