• Horticultural Science & Technology
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• v.29 no.4
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• pp.336-344
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• 2011

Using the abundant available information about the tomato genome, we developed DNA markers that are linked to disease resistant loci and performed marker-assisted selection (MAS) to construct multi-disease resistant lines and varieties. Resistance markers of Ty-1, T2, and I2, which are linked to disease resistance to Tomato yellow leaf curl virus (TYLCV), Tomato mosaic virus (ToMV), and Fusarium wilt, respectively, were developed in a co-dominant fashion. DNA sequences near the resistance loci of TYLCV, ToMV, and Fusarium wilt were used for primer design. Reported candidate markers for powdery mildew-resistance were screened and the 32.5Cla marker was selected. All four markers (Ty-1, T2, I2, and 32.5Cla) were converted to cleavage amplification polymorphisms (CAPS) markers. Then, the CAPS markers were applied to 96 tomato lines to determine the phenetic relationships among the lines. This information yielded clusters of breeding lines illustrating the distribution of resistant and susceptible characters among lines. These data were utilized further in a MAS program for several generations, and a total of ten varieties and ten inbred lines were constructed. Among four traits, three were introduced to develop varieties and breeding lines through the MAS program; several cultivars possessed up to seven disease resistant traits. These resistant trait-related markers that were developed for the tomato MAS program could be used to select early stage seedlings, saving time and cost, and to construct multi-disease resistant lines and varieties.

• The Korean Journal of Mycology
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• v.48 no.4
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• pp.389-396
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• 2020

Lentinula edodes is one of the most widely consumed edible mushrooms in Korea. Mating in L. edodes is regulated by a tetrapolar system, and two unlinked genetic loci, A and B, are known to be major determinants of the mating types, as reported in other heterothallic basidiomycetes. The A locus of L. edodes encodes a pair of homeodomain (HD) transcription factors. The highly variable N-termini of these HD transcription factors contribute to the diversity among the A mating types. In this study, we developed a cleaved amplified polymorphic sequence (CAPS) marker to discriminate 11 different A mating type alleles predominant among both cultivated and wild strains. Amplification of the variable region of the A locus followed by digestion with HaeIII and EcoRI restriction enzymes enabled successful discrimination among the 11 A mating type alleles. We also evaluated the applicability of this method in the identification of two A mating types of a dikaryotic strain.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.65 no.4
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• pp.447-456
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• 2020

Peanut is an allotetraploid derived from a single recent polyploidization. Polyploidization has been reported to have caused significant loss in genetic diversity during the domestication of cultivated peanuts. Single nucleotide polymorphism (SNP)-based markers such as cleaved amplified polymorphic sequences (CAPS) derived from next-generation sequencing (NGS) have been developed and widely applied for breeding and genetic research in peanuts. This study aimed to identify the genetic diversity and population structure using 30 CAPS markers and 96 peanut accessions from five different origins. High genetic dissimilarities were detected between the accessions from Korea and those from the other three South American origins generally regarded as the origin of peanuts, while the accessions from Brazil and Argentina presented the lowest genetic dissimilarity. Based on the results of the present study, accessions from Korea have unique genetic variation compared to those from other countries, while accessions from the other four origins are closely related. Our study identified the genetic differentiation in 96 peanut accessions from five different origins, and this study also showed the successful application of SNP information derived from re-sequencing based on NGS technology.

• Horticultural Science & Technology
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• v.34 no.6
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• pp.912-923
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• 2016

Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.

• Horticultural Science & Technology
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• v.29 no.2
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• pp.130-134
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• 2011

Paprika (Capsicum annuum L.), a colored bell-type sweet pepper, is one of the most important money making vegetable crops in Korea. The cultivation area, total production, and exports of paprika are gradually getting increased, but the paprika cultivars used in Korea are all imported. It was well-known that the genic male sterility (GMS) is the main way to produce paprika hybrid seeds. However, it is little known that how many and what kinds of ms genes are used for breeding of paprika $F_1$ varieties. In this study, eight paprika cultivars ('Special', 'Debla', 'Plenty', 'Fiero', 'Boogie', 'Fiesta', 'Derby', and 'Minibell'), popularly cultivated in Korea and three different genic male sterile lines ('GMSP', 'GMS3', and 'GMSK') were used. For allelism test among the $F_1$ cultivars, half diallel crosses were performed. The result demonstrated that the most of the GMS in paprika cultivars except for 'Minibell' were same allele. To identify which GMS gene(s) were used for paprika $F_1$ cultivars, top crosses between previously known GMS lines and the $F_1$ cultivars were performed. As a result, we found that the $ms_k$ and the $ms_p$ genes were alleles for the GMS of 'Minibell' and for the other cultivars, respectively. We also confirmed that the GMS gene identification using GMSK-CAPS marker linked to the $ms_k$ gene and the PmsM1-CAPS marker linked to the $ms_p$ gene in $F_2$ progenies of 'Minibell' and 'Fiesta' and 'Derby' cultivars, respectively. In addition, we developed the PmsM2-CAPS marker for 'Plenty', 'Fiero', and 'Boogie' cultivars. We expect that these markers will be very useful for breeding new maternal (male sterile) line of paprika.

• 한국약용작물학회:학술대회논문집
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• 2011.09a
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• pp.212-213
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• 2011

• Horticultural Science & Technology
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• v.32 no.2
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• pp.227-234
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• 2014

The responses to chilling temperature of 12 Korean cucumber varieties was compared to those of two U.S.A. (previously determined cold tolerant NC76 and 'Chipper'), and Chinese and Japanese germplasms. Seedlings of each entry were exposed to $4^{\circ}C$ (Experiment 1) and $1^{\circ}C$ (Experiments 2 and 3) at the first-true leaf stage for eight and nine hours, respectively, under 80% relative humidity (RH) and $149{\mu}moles{\cdot}m^{-2}{\cdot}s^{-1}$ photosynthetic photon flux (PPF). The chilling response [damage rating (DR)] of each accession was based on visual ratings (1 to 5) after treatment, where 1 = no damage, 2 = slight, 3 = moderate, 4 = advanced, and 5 = severe damage. Predictably the cumulative average DR of chilling tolerant line NC76 and 'Chipper' after chilling w as 1 and 1.1, respectively. Korean 'Nacdongchungjang' was most sensitive to chilling temperatures [DR = 2.3] when compared to the other entries examined. The sensitivity to chilling of 'Nacdongchungjang' was followed by Chinese 'Dongguan' [DR = 1.7]. In contrast, 'Saeronchungjang' (DR = 1) and 'Janghyungnachap' (DR = 1) were the most chilling tolerant of the Korean accessions examined and equivalent to the response of line NC76 and 'Chipper'. Nevertheless, chloroplast type genotyping of these accessions with known chilling-linked sdCAPS genomic markers revealed genotypic differences between chilling tolerant lines (NC76 and 'Chipper') and all Korean lines examined.

• Horticultural Science & Technology
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• v.35 no.3
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• pp.333-343
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• 2017

Bloomless cucumber fruits are commercially produced by grafting onto the pumpkin stocks (Cucurbita moschata) to restricted silicon ($SiO_2$) absorption. Inhibition of silicon absorption in bloomless stocks is conferred by a mutant allele of the CmLsi1 homologous to Lsi1 in rice. In this study, we characterized the Lsi1 homologs in pumpkin (C. moschata) and its cold-tolerant wild relative C. ficifolia ('Heukjong') in order to develop a DNA marker for selecting a bloomless trait and to establish the molecular basis for breeding bloomless stock cultivars of C. ficifolia. A Cleaved amplified polymorphic sequence (CAPS) marker (CM1-CAPS) was designed based on a non-sysnonymous single nucleotide polymorphism (SNP, C>T) of the CmLsi1 mutant-type allele, and its applicability for Marker-assisted selection (MAS) was confirmed by evaluating three bloom and five bloomless pumpkin stock cultivars. Quantitative RT-PCR of the CmLsi1 for these stock cultivers implied that expression level of the CmLsi1 gene does not appear to be associated with the bloom/bloomless trait and may differ depending on plant species and tissues. A full length cDNA of the Lsi1 homolog [named CfLsi1($B^+$)] of 'Heukjong' (C. ficifolia), was cloned and sequence comparison between CmLsi1($B^+$) and CfLsi1($B^+$) revealed that there exists total 24 SNPs, of which three were non-synonymous. Phylogenetic analysis of CfLsi1($B^+$) and Lsi1 homologs further revealed that CfLsi1($B^+$) is closesly related to Nodulin 26-like intrinsic proteins (NIPs) and most similar to CpNIP1 of C. pepo than C. moschata.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.108-120
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• 2017

Modern watermelon cultivars (Citrullus lanatus [Thunb.] Matsum.& Nakai var. lanatus) have fruits with diverse phenotypes, including fruit shape, rind patterns, and flesh color. Molecular markers enable efficient selection of plants harboring desirable phenotypes. In the present study, publicly available DNA markers tightly linked to fruit shape, rind stripe pattern, and flesh color were evaluated using 85 watermelon accessions with diverse fruit phenotypes. For fruit shape, the dCAPS SUN - Cla011257 marker revealed an 81% of marker - trait match for accessions with elongated or round fruits. For rind stripe pattern, the SCAR wsb6-11marker was effective for selecting Jubilee-type rind pattern from other rind patterns. For flesh color, the Clcyb.600 and Lcyb markers derived from a mutation in the Lycopene ${\beta}$ - cyclase (Lcyb) gene, were effective at selecting red or yellow flesh. Forty-eight accessions possessing diverse fruit - related traits were selected as a reference array and their genetic relationships assessed using 16 SSR markers. At a coefficient of 0.11, the 48 accessions grouped into two major clades: Clade I and Clade II. Clade I subdivided further into subclades I - 1 and I - 2 at a coefficient of 0.39. All accessions with colored flesh were classified into Clade I, whereas those with white - flesh were classified into Clade II. Differences in fruit traits between subclades I - 1 and I - 2 were observed for rind pattern and fruit color; a majority of the accessions with Crimson-type striped or non-striped rind were grouped together in subclade I - 1, while most accessions in subclade I - 2 had a Jubilee - type rind stripe pattern. These results imply that reference array watermelon accessions possess distinguishable genetic structure based on rind stripe pattern. However, no significant grouping pattern was observed based on other fruit-related traits.

• Journal of Microbiology
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• v.40 no.4
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• pp.254-259
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• 2002

The Phytophthora infestans requires two mating types for sexual reproduction. Amplified fragment length polymorphism (AFLP) was used to specifically detect different mating types of P. infestans. The AFLP primers E+AA (5'-GACTGCGTACCAATTCAA-3') and M+CAA (5'-GATGAGTCCTGAG-TAAC AA-3') detected a fragment that is specific in the A2 mating type of P. infestans. This fragment was cloned and sequenced. Based on the sequence data, PHYB-1 and PHYB-2 primer were designed to detect the A2 mating type of P. infestans. A single 347 bp segment was observed in the A2 mating type of P. infestans, but not in the A1 mating type of P. infestans or other Phytophthora spp. Identification of mating type was performed with phenotype (sexual reproduction) and genotype (CAPs marker) methods. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using mating type-specific primers, a unique band was obtained within annealing temperatures of 57$^{\circ}C$-62$^{\circ}C$ and DNA levels of 10pg-100 ng (data not shown).