• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.2
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• pp.142-146
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• 2005

Thirty-two germplasms of Korean adlay landraces were examined to analyse the genetic relationship through the amplified fragment length polymorphism (AFLP) approach. Total number of AFLP products generated by 12 selective primer combinations was 882. The number of polymorphic fragments by each primer combination greatly varied from 4 to 51 with a mean of 20.3, bands visible on the polyacrylamide gel. A genetic similarity coefficient was used for cluster analysis following UPGMA (unweighted pair grouping method of averages) method. The resulting clusters were represented in the form of a dendrogram. The clustering was not tight in the dendrogram. There was generally no clear grouping of the adlay according to the geographic regions in which germplasms were collected. The present AFLP analysis imply that although Korean adlay displayed a larger amount of AFLP variation within germplasms, the variation was shown independently without reflecting a clinal variation. This study demonstrated that AFLP method can be used to examine the genetic relationships among different germplasms of adlay.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.44 no.2
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• pp.112-116
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• 1999

Genetic diversity of 31 rice varieties including 25 japonica and 6 indica varieties was evaluated using a combination of 19 microsatellite or simple sequence repeats (SSRs) and 28 random decamer oligonucle-otide primers. All 19 microsatellite primer sets representing 19 loci in the rice genome showed polymorphisms among the 31 varieties and revealed 91 alleles with an average of 4.80 bands per primer. Also all 28 random decamer primers used were informative and generated 114 non-redundant bands with a mean of 4.07 bands. Microsatellite markers detected higher number of alleles than random primers .although the mean difference was not statistically significant. A cluster analysis based on Nei＇s genetic distances calculated from the 205 bands resolved the 31 varieties into two major groups that correspond to indica and japonica subspecies, which is consistent with the genealogical information. As few as six random decamer primers or a combination of one microsatellite and four random decamer primers were sufficient to uniquely differentiate all 31 varieties. These combinations would be potentially useful in rice variety protection and identification considering that 25 out of 31 varieties used in this study are japonica rices with high grain quality and have close make up.

• Research in Plant Disease
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• v.24 no.3
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• pp.233-236
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• 2018

A multiplex RT-PCR (mRT-PCR) assay was developed for the detection of the recently reported viruses, Cherry virus A (CVA), Little cherry virus 1 (LChV-1), and Little cherry virus 2 (LChV-2), in cherry plants in Korea. Eight sets of primers were designed for each virus and their specificity was tested by using various combinations of mixed primer sets. From the designed primer sets, one combination was selected and further evaluated to estimate the optimum temperature and detection limits of the mRT-PCR. A newly developed mRT-PCR assay was also tested using 20 cherry samples collected in the field. This mRT-PCR assay may be a useful tool for field surveys of diseases and the rapid detection of these three viruses in cherry plants.

• The Korean Journal of Mycology
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• v.41 no.1
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• pp.14-20
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• 2013

We already reported four groups which contains some similar strains based on URP-PCR in the previous paper. The objective of this study was to confirm those strains by the amplified fragment length polymorphism (AFLP) and vegetative compatibility group (VCG). AFLP analysis showed no difference among these strains except ASI 2595 and 2183 in Weonhyeong group and ASI 2829 in Suhan group. They showed specific DNA bands only in the result of P + AG/M + AAG and P + GT/M + ATG primer combinations out of eight different combinations. The AFLP primers produced a total of 330 fragments between 80 and 1000 bp in length for 31 Pleurotus ostreatus strains. At a genetic similarity of 0.96, the UPGMA analysis separated the isolates into four distinct clusters. Each group was classified by similar strains. Confrontation test by vegetative compatibility groups (VCGs) also showed distinct line between strains from different groups, but no line between similar strains within the cluster. Our results indicate that most of similar strains was not distinctness. Thus, similar strains are considered to be very close on the genealogy of their parent or same strain with different name.

• Molecules and Cells
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• v.25 no.2
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• pp.205-210
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• 2008

To develop molecular markers linked to the $L^4$ locus conferring resistance to tobamovirus pathotypes in pepper plants, we performed AFLP with 512 primer combinations for susceptible (S pool) and resistant (R pool) DNA bulks against pathotype 1.2 of pepper mild mottle virus. Each bulk was made by pooling the DNA of five homozygous individuals from a T10 population, which was a near-isogenic $BC_4F_2$ generation for the $L^4$ locus. A total of 19 primer pairs produced scorable bands in the R pool. Further screening with these primer pairs was done on DNA bulks from T102, a $BC_{10}F_2$ derived from T10 by back crossing. Three AFLP markers were finally selected and designated L4-a, L4-b and L4-c. L4-a and L4-c each underwent one recombination event, whereas no recombination for L4-b was seen in 20 individuals of each DNA bulk. Linkage analysis of these markers in 112 $F_2$ T102 individuals showed that they were each within 2.5 cM of the $L^4$ locus. L4-b was successfully converted into a simple 340-bp SCAR marker, designated L4SC340, which mapped 1.8 cM from the $L^4$ locus in T102 and 0.9 cM in another $BC_{10}F_2$ population, T101. We believe that this newly characterized marker will improve selection of tobamovirus resistance in pepper plants by reducing breeding cost and time.

• Journal of Crop Science and Biotechnology
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• v.10 no.2
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• pp.98-105
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• 2007

Phylogenetic relationships in Korean watermelons were evaluated by genetic similarity coefficients using 15 SSR(simple sequence repeat), 14 SCAR(sequence characterized amplified region) and 14 CAPS(sequence characterized amplified region) markers. The SSR markers were selected from previously reported melon and watermelon SSRs through testing polymorphisms within a set of commercial $F_1$ varieties. The SCAR and CAPS markers were developed from polymorphic AFLP(amplified fragment length polymorphism) markers between inbred lines 'BN4001' and 'BN4002'. From the AFLP analysis, 105 polymorphic fragments were identified between the inbred lines using 1,440 primer combinations of EcoRI+CNNN and XbaI+ANNN. Based on the sequencing data of these polymorphic fragments, we synthesized sequence specific primer pairs and detected clear and reliable polymorphisms in 27 primer pairs by indels(insertion/deletion) or RFLP(restriction fragment length polymorphism). A total of 43 sequence-based PCR markers were obtained and polymorphic information content(PIC) was analyzed to measure the informativeness of each marker in watermelon varieties. The average PIC value of SCAR markers was 0.41, which was similar to that of SSR markers. Genetic diversity was also estimated by using these markers to assess the phylogenetic relationships among commercial varieties of watermelon. These markers differentiated 26 Korean watermelon varieties into two major phylogenetic groups, but this grouping was not significantly correlated with their morphological and physiological characteristics. The mean genetic similarity was 66% within the complete set of 26 commercial varieties. In addition, these sequence-based PCR markers were reliable and useful to identify cultivars and genotypes of watermelon.

• Journal of Radiation Industry
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• v.5 no.4
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• pp.347-352
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• 2011

In order to identify the genetic relationship analysis by acute and chronic gamma irradiation, Arabidopsis (Arabidopsis thaliana L.) were irradiated with 200 Gy of gamma-rays using gamma-irradiator (3,000 Ci; Nordion, Canada) and gamma-phytotron (400 Ci; Nordion, Canada) for acute and chronic irradiation, respectively. Genetic relationship among two acute gamma-irradiated plants (A1 and A24) and three chronic gamma-irradiated plants (C1W, C2W, C3W) were analyzed using the amplified fragment length polymorphism (AFLP) technique compared with each non-irradiated plant. A total of 28 EcoRI and MseI primer combinations were used to screen 8 treatments by the ABI3130 capillary electrophoresis system. Amplified products by 28 primer sets showed 1,679 bands with an average of 51 bands per primer combination. Out of the total bands scored, 1,164 fragments were polymorphic bands, with different alleles existing among the treatments. The cluster analysis was performed using the UPGMA (Unweighted Pair Group Method using Arithmetic) in the computer program NTSYS-pc. In clustery analysis, acute gamma-irradiation showed higher genetic variation compared with chronic gamma-irradiation.

• BMB Reports
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• v.36 no.5
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• pp.427-432
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• 2003

Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

• Research in Plant Disease
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• v.25 no.4
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• pp.205-212
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• 2019

Analysis of differentially expressed genes has assisted discovery of gene sets involved in particular biological processes. The purpose of this study was to identify genes involved in appressorium formation in the rice blast fungus Magnaporthe oryzae via analysis of cDNA-amplified fragment length polymorphisms. Amplification of appressorial and vegetative mycelial cDNAs using 28 primer combinations generated over 200 differentially expressed transcript-derived fragments (TDFs). TDFs were excised from gels, re-amplified by PCR, cloned, and sequenced. Forty-four of 52 clones analyzed corresponded to 42 genes. Quantitative real-time PCR showed that expression of 23 genes was up-regulated during appressorium formation, one of which was the MCK1 gene that had been shown to be involved in appressorium formation. This study will be providing valuable resources for identifying the genes such as pathogenicity-related genes in M. oryzae.

• Genomics & Informatics
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• v.1 no.2
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• pp.108-112
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• 2003

We have constructed a molecular linkage map of pepper (Capsicum spp.) in an interspecific $F_2$ population of 107 plants with 320 RFLP, 136 AFLP, and 46 SSR markers. The resulting linkage map consists of 15 linkage groups covering 1,720 cM with an average map distance of 3.7 cM between framework markers. Most RFLP markers ($80\%$) were pepper-derived clones and these markers were evenly distributed all over the genome. Genes for defense and biosynthesis of carotenoids and capsaicinoids were mapped on this linkage map. By using 30 primer combinations, AFLP markers were generated in the $F_2$ population. For development of SSR markers in Capsicum, microsatellites were isolated from two small-insert genomic libraries and the GenBank database. This combined map provides a starting point for high-resolution QTL analysis, gene isolation, and molecular breeding.