Background : The soil-borne ascomycete fungus Ilyonectria rdicicola species complex is commonly associated with root rot disease symptoms in ginsneg. Its virulence has been attributed, among other factors, to the activity of hydrolytic cell wall-degrading enzymes (CWDE).
Methods and Results : To establish a rapid and accurate detection of Ilyonectria rdicicola, a species-specific primer was developed based on the putative genes of cell wall–degrading enzyme (pectinase, polygalactose, xylanase, xylosidase). Species-specific primer based on the DNA sequences of gene amplified about 200 - 300 bp polymerase chain reaction (PCR) product for Ilyonectria mors-panacis.
Conclusion : The primer pair yielded the predicted PCR product size exactly in testing with target pathogen DNAs, but not from the other species of Ilyonectria and species of other phytopathogenic fungi. The primer pair also showed only the species-specific amplification curve on realtime PCR on target pathogen DNA. The detection sensitivity of real time PCR using species-specific primer pair was 10 to 100 times higher than conventional PCR, with 1 to 10 pg/㎕. The approach outlined here could be further utilized as a rapid and reliable tool for the diagnosis and monitoring of the root rot of ginseng.
Background : Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via amplification refractory mutation system (ARMS)-PCR analyses. We performed molecular authentication of twelve C. tricuspidata ecotypes from different regions using DNA sequences in the chloroplast TrnL-F intergenic region. Methods and Results : SNPs were identified based on the results of nucleotide sequence for the intergenic region of TrnL-TrnF gene (chloroplast). Molecular markers were designed for those SNPs with additional mutations on the second base from SNPs for amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). HRM pattern analyses were performed using the Mx3005P QPCR System (Agilent Technologies, CA, USA). Conclusion : We collected 12 individual lines of C. tricuspidata from various region in South Korea and China. Based on the nucleotide sequence in the trnL-trnF intergenic region of these lines, six SNPs and a deletion of 12 bps were identified and 12 individual lines were able to be grouped in one Korean ecotype and two different ecotypes of chinese lines, chinese line 1 and 2. The SNP markers developed in this study are useful for rapidly identifying these specific C. tricuspidata ecotypes collected from different regions.
Background : Medicinal crop has been used in the traditional Asian medicinal methods. From ancient times, various kinds of medicinal crop are being cultivated in East Aisa including Korea, China and Japan. In Korea, they used a variety of medicinal plants in folk medicine and oriental medicine since ancient times. Molecular markers can be widely used in a variety of settings such as effective-loci estimation, genetic-diversity characterization, allelic-effect studies, gene-flow studies, quantitative-trait locus (QTL) mapping, and evolutionary studies. The genetic analyses of crops require large numbers of useful molecular markers for genetic or QTL identification, comparative mapping and breeding. Studying the genetic diversity and genetic relationship of crops can assist breeders. Crop genetics within a breeding program enable the economic and effective cultivar development. We tried to develop a variety of molecular markers from Angelica gigas genomic sequences for genetic studies and breeding. Methods and Results : A. gigas resources cultivated in Republic of Korea were collected. Fresh leaves were ground with liquid nitrogen and gDNA was extracted using a DNeasy Plant Mini kit (Qiagen, Valencia, CA, USA). We sequenced the whole genomes of five A. gigas accessions using Illumina HiSeq 2500 platform and identified genomic Simple Sequence Repeat (SSR) and InDel markers. DNA amplification was conducted using the PCR system (Bio-Rad T-100 Thermal Cycler). PCR products were separated by capillary electrophoresis on the ABI 3730 DNA analyzer (Applied Biosystems) and Fragment analyzer automated CE system (Advanced Analytical Technologies, Ankeny, IA, USA). Conclusion : We developed novel SSR and InDel markers from A. gigas genomic sequences for further genetic studies and breeding.
This study describes the efficient method for the discrimination of 'Cheonryang' in Panax ginseng Meyer using a STS primer. A total of 208 STS primers were applied to polymerase chain reaction (PCR) amplification for discriminating Korean ginseng cultivars. Co-dominant polymorphic band patterns were generated with two primers, MFGp 0019, MFGp 0248, and successful identification of 'Cheonryang' was achieved from out of 11 Korean ginseng cultivars. Two different sizes of DNA band patterns were detected with MFGp 0019 primer. Ten Korean ginseng cultivars shared the same size of amplified DNAs (389 bp), but 'Cheonryang' showed a different size. Thus 'Cheonryang' can be efficiently distinguished from the other ten ginseng cultivars by using the MFGp 0019 primer. In the case of MFGp 0248, two different sizes of DNA band patterns were detected in the eleven ginseng cultivars. Same sized amplified DNA bands (307 bp) were shown in five cultivars (Chunpoong, Gopoong, Kumpoong, Cheongsun, Sunhyang) and 254 bp sized DNA bands were identified in the other 6 cultivars (Yunpoong, Sunpoong, Sunun, Sunone, Cheonryang, K-1). In conclusion, the two STS primers, MFGp 0019, and MFGp 0248, provide a rapid and reliable method for the specific identification of 'Cheonryang' cultivar from a large number of samples.
The use of functional markers, it is expected to make direct identification about genetic diversity at DNA level and overcome the problem of recombination /linkage. These markers can be used to identify interesting alleles in a breeding program and indirectly select for the trait, saving money, time and labor. Bacterial blight of rice caused by Xanthomonas oryzaepv. Oryzae is a destructive disease in rice production worldwide. No bactericide is effective to control the bacterial blight disease yet. Xa3, which is a gene conferring resistance to BB of the rice plant has been previously characterized by map-based cloning. We have cloned and sequenced the Xa3/xa3 gene in Korean cultivar, Hwayoung, Ilmi and Goun with gene specific primers. Our work detected polymorphisms and PCR-based allele specific SNP markers were developed. Susceptible or resistant individuals from an F2 population developed from across between Milyang244 and Ilmi, Korean germplasms and near isogenic lines carrying BB resistance genes were screened with allele specific markers. We found that the genotype completely matched their phenotype to BB using ASP-primers. These markers could be effective to marker-assisted selection for the Xa3 gene in rice breeding programs.
밀가루와 면대 색이 밝은 품종의 육종효율을 증진하기 위하여 PPO 활성과 연관된 sequence-tagged site(STS) 분자표지인 PPO-05, PPO-16, PPO-18, PPO-30과 PPO-43을 이용하여 국내의 밀 품종의 PPO활성 수준과 유전자형 간의 관계를 분석하였다. Ppo-A1에 연관된 PPO-18이 국내 밀 품종의 PPO 활성 판별과 선발에 유용한 분자표지로 확인되었다. PPO 활성은 Ppo-A1b 유전자형 품종(0.446)이 P
전 세계에서 광범위하게 수집한 벼 유전자원 320개를 63개의 아종특이적마커로 분석하여 유전자원의 다양성, 유연관계 및 유전집단의 구조분석을 하여 아종특이적마커의 아종판별 효율을 검정하고 아종의 게놈 구성을 검토하고자 본 시험을 수행하였다. 1. 본 연구에서 사용한 63개의 아종특이적 마커는 벼 품종을 인디카와 자포니카 두 아종으로 구분하는데 효과적으로 이용할 수 있었다. 2. 실험에 사용한 320개의 벼 유전자원들은 자포니카군(128개)과 인디카군(1
To identify the variation of the RAPD patterns between two Atractylodes species, 52 kinds of random primers were applied to each eight of A japonica and A. macrocephala genomic DNA. Ten primers of 52 primers could be used to discriminate between the species and 18 polymorphisms among 67 scored DNA fragments (18 fragments are specific for A. japonica and A. macrocephala) were generated using these primers, 26.9% of which were polymorphic. RAPD data from the 10 primers was used for cluster analysis. The cluster analysis of RAPD markers showed that the two groups are genetically distinct. On the other hand, to identify the variation of the AFLP patterns and select the species specific AFLP markers, eight combinations of EcoRI/MseI primers were applied to the bulked A. japonica and A. macrocephala genomic DNA. Consequently, three combinations of EcoRI/MseI primers (EcoRI /Mse I ; AAC/CTA, AAC/CAA, AAG/CTA) used in this study revealed 176 reliable AFLP markers, 42.0% of which were polymorphic. 74 polymorphisms out of 176 scored DNA fragments were enough to clearly discriminate between two Atractylodes species.
The subspecies-specif ic RAPD markers were identif ied using 30 varieties (15 japonica and 15 indica) from diverse geographical origins. Of 690 random primers tested including subspecies-specif ic (SS) RAPD markers reported earlier, 55 primers generated 6