• Journal of Plant Biotechnology
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• 제50권
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• pp.190-199
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• 2023

Date palm (Phoenix dactylifera L.: Arecaceae) is a dioecious species where only female trees bear fruits. In their natural state, date palms produce dates once a year. However, in Thailand, some trees were observed to produce dates during the off-season, despite no variations in morphology. The availability of such off-season fruits can significantly increase their market value. Interestingly, most female off-season date palms investigated in this study were obtained through micropropagation. Hence, there is an urgent need for genetic markers to distinguish female offseason flowering plantlets within tissue culture systems. In this study, we aimed to develop random amplification of polymorphic DNA-sequence characterized amplified region (RAPD-SCAR) markers for the identification of female off-season flowering date palms cultivated in Thailand. A total of 160 random decamer primers were employed to screen for specific RAPD markers in off-season flowering male and female populations. Out of these, only one primer, OPN-02, generated distinct genomic DNA patterns in female off-season flowering (FOFdp) individuals compared to female seasonal flowering genotypes. Based on the RAPD-specific sequence, specific SCAR primers denoted as FOFdpF and FOFdpR were developed. These SCAR primers amplified a single 517-bp DNA fragment, predominantly found in off-season flowering populations, with an accuracy rate of 60%. These findings underscore the potential of SCAR marker technology for tracking offseason flowering in date palms. Notably, a BLAST analysis revealed a substantial similarity between the SCAR marker sequence and the transcript variant mRNA from Phoenix dactylifera encoding the SET DOMAIN GROUP 40 protein. In Arabidopsis, this protein is involved in the epigenetic regulation of flowering time. The genetic potential of the off-season flowering traits warrants further elucidation.

• Advances in Traditional Medicine
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• 제5권2호
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• pp.117-123
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• 2005

Dyers woad leaf (Daqingye) is a traditional Chinese medicine commonly used as anti-pyretic, anti-bacterial and anti-viral agent against infectious diseases. The Chinese Pharmacopoeia (2005) records that Dyers woad leaf should be derived from the leaves of Isatis indigotica Fort., but the leaves of Polygonum tinctorium Ait., Baphicacanthus cusia (Nees) Bremek. and Clerodendron cyrtophyllum Turcz. have also been used as substitutes of Dyers woad leaf in different regions of China. The leaf morphologies of these four species show a close resemblance, and based on their morphological appearance, it is difficult to identify them. Here, molecular genetic methods were developed as a target to identify different members of Dyers woad leaf. The 5S-rRNA spacer domain was amplified by polymerase chain reaction from genomic DNAs isolated from I. indigotica, P. tinctorium, B. cusia and C. cyrtophyllum, and the nucleotide sequences showed a great diversity. In addition, random amplification of polymorphic DNA analysis was also used to distinguish the members of Dyers woad leaf. These molecular methods could be used as a tool in authentic identification of Dyers woad leaf.

• 농업과학연구
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• 제48권2호
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• pp.283-297
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• 2021

Investigation of genetic diversity and molecular characterization in high yielding rice varieties is important for their identification. The experiment was conducted during 2016 - 2017 to analyse the genetic diversity of fifteen high yielding rice varieties in Bangladesh by using random amplification of polymorphic DNA (RAPD) markers. Polymorphism was revealed in 12 RAPD primers out of 30, whereas no other reaction was detected on the remaining 18 primers. The 40 out of 45 bands (88.89%) polymorphics were produced by the primers and ranged from 50 to 100%. The maximum number of polymorphic bands was produced by the primer OPB-18 whereas the lowest number of polymorphic bands belonged to OPC-12. The genetic similarity coefficients were determined with the RAPD data, which ranged from 0.47 to 0.94. The unweighted paired group of arithmetic means (UPGMA) dendrogram presented the studied rice varieties into two major clusters. Moreover, the value of Nei's genetic diversity is 0.26 and the Shanon information index is 0.41. The study produced distinct positions, suggesting that the genotypes were different from each other. The results indicated that these markers could be efficient for comparing the genetic relationships, patterns of variation, and measurement of genetic distance among rice varieties. Considering all of these results, RAPD analysis is found to be an effective tool for estimating the genetic diversity of different rice varieties. The outcomes of this research may contribute to the germplasm data of rice accessions and a future breeding program of rice genotypes.

• 한국수산과학회지
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• 제30권3호
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• pp.388-392
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• 1997

해조류중 우리나라 전연안에 가장 흔히 분포하는 갈파래목 녹조류인 참홑파래와 구멍갈파래, 참갈파래, 잎파래, 실파래, 가시파래등을 대상으로 RAPD방법을 이용한 각 지역 개체간의 유전적 유사도를 조사하였다. Total DNA는 엽체로부터 LiCl를 사용한 DNA추출방법으로 분리하였으며, 추출된 DNA는 $3ng/{\mu}\ell$되게 희석한후 in vitro에서 arbitrary primer와 함께 45회 PCR amplification시켰다. 그 결과 34종류 primer들로부터 240bp에서 1.5kb의 다양한 크기로 증폭된 1227개의 전기영동상의 band를 볼 수 있었으며, Jaccard공식에 따라 그 유사도를 구하였다. 파래목의 유전적 유사도는 $7\%$에서 $36\%$범위의 유사도를 보였다. 그중 참흩파래가 다른 갈파래과들로부터 가장 낮은 유사도값을 보여주었으며, 또한 primer OPB-01 (CATCCCCCTG)을 사용하였을때 갈파래과에서 공통적인 630bp크기의 생성물을 생산하지않는 특징을 나타내었다.

• 한국초지조사료학회지
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• 제18권4호
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• pp.277-284
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• 1998

재배콩과 돌콩의 유전적 다양성과 유연관계를 밝히고, 콩을 사료작물로 개량하는데 필요한 유전정보를 얻기 위하여, RAPD 분석방법을 이용한 결과는 다음과 같다. 1. 증폭된 polymorphicband의 총 수는 74개였으며, 이중 다형현상을 보인 것은 50개로 67.6%, 증폭된 band의 크기는 0.13~2.0 Kb 범위에 있었다. 2. Random primer의 sequence 5'-CAG GCC CTT C-3'를 사용하였을 때, 0.56-2.0 Kb에서 콩의 종피 색깔에 따른 변이 (누런색과 검은색)가 나타났다. 그리고 Sequence 5'-TGC TCT GCC C-3'과 5'-GTC CAC ACG G-3'인 primer를 사용하였을 때, 각각 0.63~0.94, 0.94~2.0 Kb에서 검정콩 2호에 특정적인 genetic marker가 검출되었다. 3. Genetic similarity 값에 의한 품종간 유전적 변이성은 검정콩 2호 (0.81)가 가장 낮았고 검정콩 1호 (0.46)에서 가장 높았다. 4. 콩의 품종과 돌콩의 유연관계는 황금콩과 석량붓콩이 0.52로 가장 가까웠고 검정올콩 (0.47), 검정콩 l호 (0.46), 검정콩 2호 (0.42), 돌콩 (0.38)의 순서로 멀었다.

• 대한의생명과학회지
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• 제19권3호
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• pp.213-223
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• 2013

The objectives of this study are to develop a molecular diagnosis to differentiate serotypes and mating-types of C. neoformans/C. gattii complex isolates from pigeon droppings in Korea and to elucidate molecular epidemiology of the isolates. Phenotypes and genotypes of C. neoformans/C. gattii complex isolates were identified by biochemical properties and PCR using specific CNLAC1 gene, respectively. To classify serotypes and mating-types of C. neoformans/C. gattii complex isolates, the five reference strains and thirty-three isolates in Korea were investigated by restriction fragment length polymorphism (RFLP) analysis using CNLAC1 gene for varieties, by random amplified polymorphic DNA (RAPD) for serotyping, and by PCR using specific primer sets for mating typing. All isolates in Korea were belonged to C. neoformans var. grubii (serotype A) by RFLP and RAPD patterns which showed high sensitivity and specificity. Therefore, RFLP and RFLP were available to differentiate varieties and serotypes of C. neoformans. Amplification patterns of the five reference strains by specific PCR for mating typing were differentiable, and all isolates were classified into $MAT{\alpha}$. All C. neoformans environmental isolates in Korea were Cr. neoformans serotype A and $MAT{\alpha}$ which is a more virulent pathogen. This study suggests that RFLP and RAPD are rapid and correct molecular diagnosis tools for epidemiology of C. neoformans/C. gattii complex isolates.

• 생명과학회지
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• 제28권9호
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• pp.1118-1126
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• 2018

본 논문은 양식 어류에게 많은 경제적 피해를 입히고 있는 다양한 어류질병세균 중에서 연쇄구균증에 대한 일반적인 특성과 종류, 진단 방법에 대하여 기존에 연구 보고된 논문을 기반으로 알아보고자 한다. 대표적인 어류연쇄구균증의 원인체로는 Lactococcus garvieae, Streptococcus iniae, S. parauberis가 있다. 연쇄구균증에 감염 시 나타나는 증상으로는 체색변화, 안구이상, 아가미 퇴색, 출혈, 복부팽만, 신장과 비장의 종대 등의 보이다 폐사가 일어난다. 또한 수온이 상승하는 6월부터 10월까지 주로 일어나며 집단적으로 발병하여 폐사가 일어난다. 현재 연쇄구균증을 진단하기 위한 기술로는 16S rRNA, 16S-23S rRNA intergenic spacer region (ISR), Random Amplified polymorphic DNA (RAPD), Ribotyion (RT), Loop-mediated isothermal amplification (LAMP) 등이 있다. 이중에서 현장에서 적용 가능성이 높은 LAMP법이 각광을 받고 있지만 결과 확인 등의 문제로 인하여 현재는 어려움을 겪고 있는 실정이다. 이에 현장에서 진단부터 결과 확인까지 손쉽게 사용할 수 있도록 문제점을 보완하면 연쇄구균증에 대한 경제적 손실을 최소화 할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권9호
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• pp.1204-1209
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• 2004

Random amplification of polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out using DNA samples of 30 animals of Rathi cattle and 42 animals of Tharparkar cattle. Genomic DNA was isolated as per standard protocol and evaluated for its quality, purity and concentration. Twenty three random primers were screened out of which 15 primers yielded satisfactory amplifications and were used for further analysis. Average numbers of polymorphic fragments per primer were 7.07${\pm}$0.86 in Rathi and 6.80${\pm}$0.61 in Tharparkar cattle. The percentage of polymorphic bands in these two cattle breeds were 86 and 87%, respectively. Within breed genetic similarities for pooled over primers in the animals of Rathi and Tharparkar breeds were .577${\pm}$0.30 and 0.531${\pm}$0.02, respectively on the basis of band frequency (BF) and 0.645${\pm}$0.04 and 0.534${\pm}$0.04, respectively on the basis of band sharing (BS). Averages of between breed genetic similarities for pooled over primers were 0.97 and 0.92 according to BF and BS, respectively, which reflect higher degree of genetic similarity between Rathi and Tharparkar cattle breeds. Index of genetic distance based on BF and BS for pooled over primers was 0.030${\pm}$0.011 and 0.088${\pm}$0.031, respectively. Percentage of polymorphic bands and within-breed genetic similarities on the basis of band frequency (BF) and band sharing (BS) for pooled over primers revealed higher genetic similarity in Rathi than Tharparkar cattle population. High estimates of between breed genetic similarities for pooled over primers indicated that either Rathi is having decent from Tharparkar or both the cattle breeds are having common descent. Low value of Index of genetic distances between these two cattle breeds may be due to the fact that Rathi and Tharparkar cattle breeds are the native of Thar Desert in Northwest India. The results of between breed genetic distances also confirm the existence of high degree of genetic similarity between these two breeds of cattle.

• 한국버섯학회지
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• 제15권2호
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• pp.84-87
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• 2017

목이버섯의 고온적응성이 우수한 '현유'의 주요특성을 요약하면 다음과 같다. 균사생장적온은 $25^{\circ}C$이고 버섯발생 및 생육온도역시 $26{\sim}36^{\circ}C$로 고온성을 나타내었다. 배양기간은 40~42일, 초발이소요일수는 27일, 생육기간은 15일로 총재배기간은 82~84일이 소요되었다. 갓크기는 56.8 mm이며, 수량은 1 kg배지에서 350 g을 나타내었다. DNA다형성을 분석한 결과 OPA1, OPA7등의 primer에서 개발품종과 중국유래품종 그리고 대조구와 뚜렷한 품종간 밴드차이를 보였다.

• Journal of Ecology and Environment
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• 제36권3호
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• pp.205-210
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• 2013

Cicuta virosa L. (Apiaceae) is a perennial emergent plant designated as an endangered species in South Korea. According to the former records, only four natural habitats remain in South Korea. A former study suggested that three of four populations (Pyeongchang: PC, Hoengseong: HS, Gunsan: GS) would be classified as different ecotypes based on their different morphological characteristics and life cycle under different environmental conditions. To evaluate this suggestion, we estimated genetic diversity in each population and distance among three populations by random amplification of polymorphic DNA. Seven random primers generated a total of 61 different banding positions, 36 (59%) of them were polymorphic. Nei's gene diversity and the Shannon diversity index increased in the order of PC < HS < GS, which is the same order of population size. In the two-dimensional (2D) plot of first two principal components in principal component analysis with the presence of 61 loci, individuals could be grouped as three populations easily (proportion of variance = 0.6125). Nei's genetic distance for the three populations showed the same tendency with the geographical distance within three populations. And it is also similar to the result of discriminant analysis with the morphological or life-cycle factors from the previous study. From the results, we concluded that three different populations of C. virosa should be classified as ecotypes based on not only morphology and phenology but genetic differences in terms of diversity and distance as well.