• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.683-692
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• 2010

Morphologically, nine different slow-growing protoclones were screened from regenerated protoplasts of heterokaryotic Agaricus bisporus. As such, the present study is the first report on differentiating homo- and heterokaryotic protoclones using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Among 80 primers tested, the seven ISSR and seven RAPD primers selected for the analysis generated a total of 94 ISSR and 52 RAPD fragments, respectively. The ISSR fingerprinting also detected more polymorphic loci (38.29%) than the RAPD fingerprinting (34.61%). A principal coordinate analysis (PCA) was employed to evaluate the resolving power of the markers as regards differentiating protoclones. As a result, the mean polymorphism information content (PIC) for each marker system (i.e., 0.787 for RAPD and 0.916 for ISSR) suggested that ISSR is more effective for determining polymorphisms. The dendrograms constructed using RAPD, ISSR, and an integrated RAPD and ISSR marker system were highly correlated with one another as revealed by a high Mantel correlation (r= 0.98). The pairwise similarity index values also ranged from 0.64 to 0.95 (RAPD), 0.67 to 0.98 (ISSR), and 0.67 to 0.98 (RAPD and ISSR), whereas the mean similarity index values of 0.82, 0.81, and 0.84 were obtained for the RAPD, ISSR, and combined data, respectively. As there was a good correspondence between the RAPD and ISSR similarity matrices, ISSR would appear to be an effective alternative to RAPD in the genetic diversity assessment and accurate differentiation of homo- and heterokaryotic protoclones of A. bisporus.

• 원예과학기술지
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• 제29권6호
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• pp.600-609
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• 2011

Allium속에 포함된 6종의 122점을 수집하고, 이 종들의 유전적 관계는 ISSR 마커를 이용하여 확인하였다. 형태적 분석은 6개의 양적 형질을 측정하고 1개의 질적 형질은 수치화하였다. SSR 분석은 17개의 primer를 사용하여 총 370개의 다형성 밴드를 얻었다. 형태적 특성 분석은 유전적 거리로 구분할 경우 3개의 그룹으로 분류되었으나, 부분적으로 몇 몇 종들은 분류에 어려움이 있었다. ISSR 결과를 바탕으로 Allium 속의 군집분석은 5개의 그룹으로 분리되었다. 형태적 분석과 SSR분석 간의 상관 관계는 유의성이 매우 낮았다(r = 0.036). 따라서 본 연구에서 개발한 ISSR 마커는 달래와 산달래의 분류와 교배 육종에 유용하게 이용될 수 있을 것이다.

• 한국자원식물학회지
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• 제19권1호
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• pp.189-195
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• 2006

본 연구는 우리나라 특산수종이며 조경 및 원예적 가치가 높은 노각나무 유전변이를 조사하기 위해 6개 천연집단을 선발하여 DNA I-SSR 표지자를 사용, 유전다양성 및 유전구조를 조사하였다. 5개의 I-SSR primer(#813, 815, 818, 820, 823)에서 총 61개의 증폭산물을 관찰할 수 있었으며, 유전 다양성을 나타내는 P(Percentage of polymorphic loci)값과S.I.(Shannon's information Index)가 남쪽에 분포하는 오봉산(P=88.5%, S.I.=0.467), 금산(P=86.9%, S.I.=0.427), 바랑산(P=83.6%, S.I.=0.425)집단이 높았으며, 북쪽(내륙)에 분포하는 소백산(P=80.3%, S.I.=0.396), 지리산(P=77.1%, S.I.=0.368), 가야산(P=75.4%, S.I.=0.358)집단은 낮았다. 전체 유전변이 중 11.8%만이 집단간에 기인하는 것으로 나타났고, 나머지 88.2%는 집단내 개체간의 차이에서 기인하였다. 유전거리를 이용하여 UPGMA법에 의한 유집분석을 실시한 결과 지리적 분포에 대한 뚜렷한 경향은 나타나지 않았다.

• 생명과학회지
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• 제17권1호
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• pp.24-30
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• 2007

꽃치자(Gardenia jasminoides var. radicans)와 열매치자(C. jasminoides for. grandiflora)는 Gardenia속으로는 우리나라에 두 분류군밖에 없으며 열매치자는 약용, 식용, 꽃치자는 방향제로 쓰인다. 그런데 꽃이 지고 나면 형태적으로 구분이 거의 되지 않는다 ISSR분석으로 이들 종의 유전적 다양도와 집단구조를 실시하였다. 88개의 DNA 분절에서 한 분류군에만 나타나는 특이밴드가 탐지되었다. 한국의 세 야생 집단은 분리되어 있고 패치 분포를 보이지만 재배종에 비해 높은 유전적 다양성을 유지하친 있었다. 열매치자가 꽃치자보다 유전적 다양성이 높았으며 ISSR 마커로 이들 분류군이 잘 분리되었다. 또한 야생 집단이 재배 집단보다 다양성이 약간 높으나 유의성은 없었다. 이는 재배화과정에서 유전적 다양성의 일부 상실이 있었으나 인위적인 채취와 식재로 야생 집단과 재배 집단의 유전적 교류가 존재하였음을 시사한다.

• 식물분류학회지
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• 제47권1호
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• pp.11-22
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• 2017

Prangos fedtschenkoi (Regel et Schmalh.) Korovin (Apiaceae) is an endemic species for mountainous Middle Asia, which is both a rare and useful plant. Organic extractions from this species are being used in pharmaceutics and cosmetology. In recent years, P. fedtschenkoi distribution area has considerably decreased, presumably, resulting from human activities such as agriculture, construction works, overgrazing and collection from wild for pharmaceutic purposes. Six populations were found in Uzbekistan and their genetic divergence and differentiation were studied with 10 inter-simple sequence repeat (ISSR) markers, selected out of 101. Totally 166 amplified ISSR fragments (loci) were revealed, of which 164 were polymorphic. Relatively moderate level of polymorphism was found at population level with polymorphic bands ranging from 27.71% to 47.59%. Mean P = 39.05%, $N_a=1.40$, $N_e=1.25$, S.I. = 0.21, and $H_e=0.14$ were revealed for all loci across six populations. AMOVA showed higher variation among populations (62%) than within them (38%). The Bayesian model determined 5 clusters, or genetic groups. The posteriori distribution of the Theta II estimator detected full model identifying high inbreeding, intensified by low gene flow (Nm = 0.3954). Mantel test confined population 6 as distinct cluster corresponding to geographic remoteness (R = 0.5137, $p{\leq}0.005$). Results were used as the bases for developing conserve measures to restore populations.

• 한국자원식물학회지
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• 제28권6호
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• pp.697-703
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• 2015

Twenty apple germplasm accessions from the Korean Genebank were successfully cryopreserved using two-step freezing to back up genetic resources maintained by field collections. This study examined the morphological and genetic stability of cryopreserved dormant apple buds that were stored in liquid nitrogen, and then rewarmed and regrown. Whole plants were regenerated directly from dormant buds through budding without an intermediary callus phase. The cryopreserved buds produced high levels of shoot formation (76.2-100%), similar to those of noncryopreserved buds (91.3-100%), with no observed differences between cryopreserved and noncryopreserved materials. Three of the twenty cryopreserved apple germplasm accessions were used to assess morphological and genetic stability. No differences in morphological characteristics including shoot length, leaf shape, leaf width/length ratio, and root length were observed between controls (fresh control and noncryopreserved) and cryopreserved plantlets. The genetic stability of regenerants (before and after cryopreservation) was investigated using inter simple sequence repeat (ISSR) markers. The ISSR markers produced 253 bands using four primers, ISSR 810, SSR 835, ISSR 864, and ISSR 899. These markers showed monomorphic banding patterns and revealed no polymorphism between the mother plant and regenerants before and after cryopreservation, suggesting that cryopreservation using two-step freezing does not affect the genetic stability of apple germplasm. These results show that two-step freezing cryopreservation is a practical method for long-term storage of apple germplasms.

• 생명과학회지
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• 제17권11호
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• pp.1482-1487
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• 2007

ISSR분자 마커를 이용하여 왕대속 대나무의 유연관계를 분석 실시하였다. 전세계적으로 왕대속에 속하는 4종은 경제학적, 생태학적으로 중요한 식물 자원중 하나이다. 총 11개의 ISSR 프라이머 중 9개의 프라이머에서 증폭이 일어 났으며 총 64개의 밴드를 확인 할 수 있었다. ISSR분석결과 왕대속 4종의 대나무에서 70.49%인 43개의 다형현상이 나타났다. 4개의 분류군으로 분류된 왕대속 대나무의 집단내 다양성(Hs)은 0.092,집단간 다양성(Gst)은 0.499로 나타났다. 각 종에 대한 유전자 유동(Nm)은 0.559로 나타났다. 따라서 왕대속에 속하는 4종의 유전자 유동(Nm)은 아주 낮음을 알 수 있었다 4종의 분류군에 대한 유전자 다양성(Ht)은 0.291로 낮게 나타났으며 ISSR 마커로 명확하게 그들은 분류가 되었다. 또한 왕대속의 4종은 단일계통임을 확인할 수 있었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.32-32
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• 2019

The aim of this study was to provide basic guidelines for conservation and management of endangered plants in the national parks of Korea. Iris dichotoma Pall. (Iridaceae), which is a popular garden plant, is considered a second-class endangered species by Korean government and it is listed as a EN (Endangered) species in Red Data Book of Korea. We analyzed ecological conditions of I. dichotoma habitats based on vegetation properties and soil characteristics. This species which is known to inhabit in grassland adjacent to the ocean of lowlands slope and its population was located at an elevation of 8 m to 11 m. In the study sites, the mean of soil organic matter, total nitrogen and soil pH were 6.16%, 0.234% and 5.39 respectively. Additionally, the genetic variation and structure of three populations were assessed using ISSR (Inter Simple Sequence Repeat) markers. The genetic diversity of I. dichotoma (P = 59.46%, H = 0.206, S = 0.310) at the species level was relatively high. Analysis of molecular variance (AMOVA) showed 82.1% of the total genetic diversity was occurred in within populations and 17.9% variation among populations. Lastly, we developed predicted distribution model based on climate and topographic factors by applying SDMs (Species Distribution Models). Consequently, current status of I. dichotoma habitats is limited with natural factors such as the increase of the coverage rate of the herbs due to ecological succession. Therefore, it is essential to establish in situ and ex situ conservation strategies for protecting natural habitats and to require exploring potential and alternative habitats for reintroduction.

• Journal of Forest and Environmental Science
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• 제38권3호
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• pp.152-158
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• 2022

A cryopreservation is an essential tool for preservation of germplasm. In this study, the possibility for cryopreservation of embryogenic cells of Siberian ginseng (Eleutherococcus senticosus) in liquid nitrogen (-196℃) was evaluated. The effects of glycerol and dimethyl sulfoxide (DMSO) at different concentrations (5%, 10% and 20%) as cryoprotectants on regrowth of cryopreserved E. senticosus embryogenic cells were tested. There was significant effect of cryoprotectants on regrowth of embryogenic cells (p=0.0019). The highest and lowest fresh mass gain were achieved when embryogenic cells were frozen with 10% DMSO and 5% glycerol (138.2±5.9 and 61.3±14.6, respectively). The effect of the cryoprotectants on the frequency embryo germination was tested. There was no significant difference between glycerol and DMSO (p=0.846). Three different concentrations of cryoprotectants did not significantly affect the frequency embryo germination (p=0.534). Finally, the genetic fidelity of the plantlets regenerated from non-cryopreserved and cryopreserved embryogenic cells was tested by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis. RAPD and ISSR analysises showed that there was no genetic variation among regenerants.

• Journal of Plant Biotechnology
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• 제47권4호
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• pp.298-308
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• 2020

Genetic diversity among Thaumatococcus daniellii populations in the southwestern region of Nigeria were assessed using morphometric and molecular markers to determine the population structure and existing genetic relationship for its improvement, conservation and sustainable utilisation. Populations from five locations in each of the six states were used for the study. Morphometric data were collected on folia characters and analysed for variability. Genome DNA was isolated from the plant leaf and amplified by polymerase chain reaction with inter-simple sequence repeat markers (ISSR) to determine the allelic polymorphism, marker effectiveness and genetic relationship of the population. The results showed significant variations in petiole length and leaf dimensions of the populations within and across the states. These morphometric traits are the major parameters that delimit the populations and they correlated significantly at P≤0.05. Analysis of the electrophoregram showed that the ISSR markers are effective for the diversity study. A total of 136 loci were amplified with an average of 7.16 loci per marker, 63.2% of the loci were polymorphic. The Principal Coordinate Analysis revealed that seven factors accounted for 81.6% of the variation and the dendrogram separated the populations into two major groups at a genetic distance of 10 (about 90% similarity) with sub-groups and clusters. Most populations within the state had a high degree of similarity, nonetheless, strong genetic relationship exists among populations from different states. The close relationship between populations across the states suggests a common progenitor, which are likely separated by ecological or geographical isolation mechanisms.