• 생명과학회지
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• 제13권5호
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• pp.590-595
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• 2003

The objective of this study was to assess of genetic variation within and between pumpkin species including Cucurbita maxima, C. moschata, C. pepo and C. maxima${\times}$C. moschata using RAPD markers. The 16 primers showed the amplification of 136 scorable fragments ranging from about 100 bp to 2300 bp. A total of 94 DNA fragments were polymorphic with an average 5.9 polymorphic bands per primer. A species $(C. maxima\timesC. moschata)$ has the highest number of polymorphic loci. Based on obtained data, UPGMA cluster analysis was conducted. Twenty pumpkin cultivars were classified into three large categories and identified genetic distance of cluster ranging from 0.38 and 1.00. Clustering was in accordance with the division of Curcurbitaceae into four species, C. maxima, C. moschata, C. pepo and C. $C. maxima\timesC. moschata$. Therefore, RAPD method may be essential tool for enabling discrimination of pumpkin cultivars.

• Asian-Australasian Journal of Animal Sciences
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• 제19권11호
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• pp.1556-1560
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• 2006

Six breeds of riverine buffalo viz. Murrah, Mehsana, Jaffrabadi, Nagpuri, Nili-Ravi and Bhadawari were characterized using FAO-recommended cattle specific microsatellite markers. Among the total of twenty microsatellite markers screened to explore genomic variability of six buffalo breeds, only ten were polymorphic in nature. Four out of ten polymorphic microsatellite loci were rated as informative. The numbers of alleles detected ranged from 2 to 7, with a mean of $5.5{\pm}0.07$ per microsatellite marker. The most polymorphic marker was BM1818 with a total of 7 alleles present at this locus. One breed specific marker was found in each of Mehsana (BM1818) and Bhadawari (ILSTS030) and four were found in Jaffarabadi (BM1818, ILSTS030, ILSTS054 and ILSTS011). Genetic distance (Ds) between the Mehsana and Bhadawari breed was the maximum (0.29), followed by Murrah and Mehsana (0.27), and Nili-Ravi and Bhadawari (0.26). The lowest Ds was found between the Jaffrabadi and Nagpuri breeds which was only 0.05. The highest divergence time of 1318 years was established between Mehsana and Bhadawari breeds whereas it was found to be lowest (272 years) between the Jaffrabadi and Nagpuri breeds.

• Mycobiology
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• 제47권4호
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• pp.466-472
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• 2019

For the purpose of protecting the rights of Lentinula edodes breeders, we developed a new simple sequence repeat (SSR) marker set consisting only of genetically independent tetranucleotide or longer core motifs. Using available genome sequences for five L. edodes strains, we designed primers for 13 SSR markers that amplified polymorphic sequences in 20 L. edodes cultivars. We evaluated the independence of every possible marker pair based on genotype data. Consequently, eight genetically independent markers were selected. The polymorphic information content values of the markers ranged from 0.269 to 0.764, with an average of 0.409. The markers could distinguish among 20 L. edodes cultivars and produced highly repeatable and reproducible results. The markers developed in this study will enable the precise identification of L. edodes cultivars, and may be useful for protecting breeders' rights.

• 원예과학기술지
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• 제33권5호
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• pp.722-729
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• 2015

딸기 흰가루병은 Podosphaera aphanis에 의해 발병되며 수확기에 가장 큰 피해를 주는 병으로 현재 유황, 농약으로 주로 방제 되고 있는 실정이다. 본 연구에서는 딸기 흰가루병 저항성 품종 육성을 위한 흰가루병 저항성 특이마커 개발로 내병성 육종효율을 높이고자 하였다. 흰가루병 저항성 계통 선발을 위한 분자마커를 개발하기 위해 아키히메${\times}$설향 집단을 대상으로 자가수분을 통해 후대 양성 후 병저항성을 검정하였다. 마커분석은 RAPD primer 200 세트 중 OPE10 331bp에서부터 흰가루병 저항성 특이 마커 선발하였다. 흰가루병 저항성 특이밴드만 선발하기 위하여 클로닝 후 유전자정보 분석하여 SP1F/R의 Primer를 제작하였다. 그러나 SP1F/R을 이용하여 PCR한 결과 저항성, 감수성간에 다형성이 확인되지 않아 염기서열을 정렬한 후 SNP, In/del의 다형성 유무를 확인한 결과 6개의 SNP를 확인하였다. 이들 PCR 산물을 해당 사이트와 연관된 제한효소로 절단한 결과 그 중 Eae I(Y/GGCCR)의 절단으로 231bp 위치에서 저항성과 감수성간의 다형성을 확인함으로써 흰가루병 저항성 계통선발을 위한 분자마커를 선발하였다. 이러한 과정을 통해 딸기 흰가루병 저항성 품종 육성을 위한 MAS(marker assisted selection) 체계 확립으로 내병성 육종효율 증진에 기여를 할 수 있을 것으로 기대된다.

• Journal of Forest and Environmental Science
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• 제25권1호
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• pp.57-65
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• 2009

Ficus deltoidea Jack is an important and popular medicinal plant species found in the Malaysia. Plants are being collected and used based on morphology and authentication to prevent adulteration is not in practice. In this study, twenty-six accessions of F. deltoidea Jack were collected from Kelantan and Terengganu states of Peninsular Malaysia to examine their genetic similarities and differences using randomly amplified polymorphic DNA (RAPD) technique. Out of 20 arbitrary primers, two primers (D-10 and D-11) were selected which produced reliable DNA polymorphism. D-10 and D-11 primers generated 138 RAPD bands ranging from 250 bp to 3000 bp. Ninety-nine of them were polymorphic loci (72%) and thirty-nine were nonpolymorphic loci (28%). A total of 56 bands with polymorphic loci were amplified with primer D-10 and analyzed by cluster analysis and UPGMA to present a dendrogram depicting the degree of genetic relationship among 26 accessions. Eight RAPD markers were sequenced to determine their identity. RAPD analysis showed the genetic diversity among 26 accessions of F. deltoidea Jack. The RAPD profile and RAPD marker sequences reported in this paper could be used in plant and/or plant material authentication. This study also suggested that RAPD can be a useful technique to study DNA polymorphism in F. deltoidea Jack.

• 생명과학회지
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• 제16권2호
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• pp.219-224
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• 2006

ISSR markers를 이용하여 수박 18품종의 유전적 유연관계를 분석하여 얻어진 결과를 요약하면 다음과 같다. 수박 18품종의 genomic DNA와 ISSR primer 100개를 PCR 반응시킨 결과 다형성을 뚜렷하게 나타내는 primer는 21개 이였으며, 이들 primer에 의해 증폭된 밴드는 105개 이였고 다형성을 보이는 밴드는 58개 였으며 증폭된 DNA 단편의 크기는 $0.2{\sim}5.0kb$ 사이에 위치하였다. 다형성을 나타낸 primer는 18개의 anchored primer와 3개의 non-anchored primer로 구분되었고 모든 anchored primer는 2개의 염기서열이 반복된 형태를 나타내었으며, non-anchored primer보다. 다형성 정도가 높게 나타났다. 수박 18품종은 유전적 유사도 값 0.42를 기준으로 할 때 18개 품종을 2개의 그룹 으로 구분할 수 있었으며, 국내에서 육성된 품종은 유전적 유사도가 아주 높은 것으로 분석 되었고, 이들 품종은 수박의 과형에 따라 유사하게 구분되었다.

• 한국잡초학회지
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• 제18권1호
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• pp.76-83
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• 1998

피속 잡초 수집종 33종을 대상으로 RAPD marker를 이용하여 피 수집종 간의 유전적변이를 알아보고, 수집종들을 판별할 수 있는 DNA marker를 선발하기 위하여 본 실험을 수행한 결과를 요약하면 다음과 같다. Operon사에서 제작된 74개의 10-mer RAPD primer 가운데에서 명확한 다형성을 보이는 6개 primer를 선발하였다. 이들 primer로 PCR에서 증폭된 밴드는 31개이었으며 이 가운데 다형성을 나타내는 band는 26개(83.9%)로 나타났다. 피 수집종 간의 유연 관계를 분석한 결과 공시된 피 수집종은 크게 3그룹으로 분류할 수 있었다.

• Journal of Plant Biotechnology
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• 제33권4호
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• pp.289-295
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• 2006

국내에서 유통되고 있는 토마토 품종의 판별 방법에 SSR marker의 이용 가능성에 대한 연구를 수행하여 얻어진 결과를 요약하면 다음과 같다. 토마토 28품종을 18개의 SSR marker를 이용하여 분석하였을 때 대립유전자의 수는 $2{\sim}9$개로 비교적 다양한 분포를 나타내었으며 전체 60개의 대립유전자가 분석되었다. PIC 값은 $0.476 {\sim}0.800$ 범위에 속하였으며 평균값은 0.607로 나타났다. SSR marker를 이용하여 작성된 토마토 28품종의 품종간 유전적 거리는 $0.35{\sim}0.97$의 범위로 나타났고, 유사도 지수 0.36을 기준으로 할 때 28개 품종은 체리형 토마토 그룹과 일반형 토마토 그룹으로 나눌 수 있었으며, 공시 품종 모두 SSR marker의 genotype에 의해 뚜렷이 구분되었다. 이 연구결과는 토마토의 품종식별에 기초 자료로 유용하게 이용될 수 있는 것으로 나타났다.

• Journal of Forest and Environmental Science
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• 제35권2호
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• pp.90-101
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• 2019

Biodiversity refers to the total number and variation among species of flora and fauna of an area. Due to tremendous biotic especially anthropogenic pressure these natural resources are being vanishing. In present study genetic diversity among accessions of Thamnocalamus spathiflorus was evaluated. A total of 51 vegetative characters and 42 primers (10-mer) were screened. Out of 42 screened primers, 28 polymorphic primers were selected for further analysis. A total of 263 bands were recorded as polymorphic whereas 48 bands were monomorphic. The resolving power (Rp) of 28 Randomly Amplified Polymorphic DNA (RAPD) primers ranged from 4.6 (OPE08) to 17.6 (OPA11). The polymorphic information content (PIC) value ranged from 0.21 (OPAH09) to 0.44 (OPG02). The result revealed high degree of genetic relatedness (56 to 80%). Cluster analysis revealed two major clusters both for morphology as well as RAPD. Unlike morphological characterization, the accession (D5) from Bahli, Rampur, Shimla (H.P.) was clustered separately from the others in RAPD cluster analysis. Accessions with closed locality grouped together through RAPD marker system however analogy was recorded for morphological traits. The study conducted reflects the utility of RAPD technique for species identification and phylogenetic studies in bamboo for conducting bamboo breeding program.

• Journal of Forest and Environmental Science
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• 제25권2호
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• pp.93-100
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• 2009

In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.