• 원예과학기술지
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• 제29권4호
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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.

• 한국균학회지
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• 제48권4호
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• pp.389-396
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• 2020

표고는 한국에서 가장 많이 소비되는 식용버섯 중 하나이다. 표고는 사극성의 교배계를 따르며, 표고의 교배형은 자웅이주성의 다른 담자균류와 마찬가지로 서로 독립적인 두 유전자좌, A와 B에 의해 결정된다. 표고의 A 유전자좌에는 한 쌍의 homeodomain (HD) 전사인자가 암호화되어 있으며 이들의 N말단에서 나타나는 높은 변이가 A 교배형의 다양성에 중요한 것으로 알려져 있다. 본 연구에서는 표고 품종과 야생종에서 많이 발견되는 11종의 A 교배형을 구분할 수 있는 CAPS 마커를 개발하고자 하였다. A 유전자좌에서 변이가 큰 부분을 PCR을 통해 증폭한 뒤 두 가지 제한효소 HaeIII와 EcoRI로 절단하여 DNA 단편의 크기 및 양상을 살펴봄으로써 11종의 A 교배형을 서로 구별할 수 있었다. 또한 해당 방법이 이핵균주의 교배형을 확인하는 데 활용할 수 있는지도 살펴보았다.

• 한국작물학회지
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• 제65권4호
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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.

• 원예과학기술지
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• 제34권6호
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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.

• 원예과학기술지
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• 제29권2호
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• pp.130-134
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• 2011

우리나라에서 파프리카(착색단고추)는 재배면적, 생산량 및 수출량이 해마다 증가하는 추세에 있어 농가 소득에 매우 중요한 채소작물의 하나이지만 재배되고 있는 파프리카 종자는 전량 외국 종자회사에서 개발된 일대잡종종자를 수입하고 있다. 최근 파프리카 품종의 국산화를 위하여 정부의 지원을 받아 국내 종묘회사에서 파프리카 품종 개발에 힘쓰고 있다. 파프리카 일대잡종종자의 생산은 주로 유전자적 웅성불임성을 이용한다. 하지만 파프리카 품종에 사용된 유전자적 웅성불임성이 어떤 종류인지 몇 가지 종류가 사용되고 있는지 잘 알려져 있지 않다. 따라서 본 연구에서는 시판되고 있는 8개의 파프리카 품종('Special', 'Debla', 'Plenty', 'Fiero', 'Boogie', 'Fiesta', 'Derby' 및 'Minibell')을 사용하여 이들의 유전자적 웅성불임 유전자가 같은 것인지 다른 것인지를 확인하기 위하여 반 이면교잡(half diallel cross)를 수행하였는데, 'Minibell'을 제외한 나머지 7개의 파프리카 품종은 같은 유전자적 웅성불임을 사용하였다는 것을 알아냈다. 또한 어떤 종류의 유전자적 웅성불임 유전자인지를 확인하기 위하여 세 종류의 유전자적 웅성불임 계통(GMS3, GMSP 및 GMSK)에 교배하여 대립유전자 검정을 수행하였는데, 'Minibell'은 $ms_k$ 유전자를 사용하였고, 나머지 7개의 품종은 $ms_p$ 유전자를 사용하였다는 것을 알아냈다. 따라서, 'Minibell' 품종에서는 기 개발된 GMSK-CAPS 분자표지를 사용할 수 있음을 확인하였고, 'Fiesta' 및 'Derby' 품종에서는 PmsM1-CAPS 분자표지가 사용될 수 있음을 확인하였다. 또한 'Plenty', 'Fiero' 및 'Boogie' 품종 등에 사용할 수 있는 새로운 분자표지 PmsM2-CAPS를 개발하였다. 이들 분자표지는 새로운 파프리카 웅성불임(모계) 계통 및 품종개발에 큰 도움이 될 것으로 판단된다.

• 한국약용작물학회:학술대회논문집
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• 한국약용작물학회 2011년도 추계학술발표회
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• pp.212-213
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• 2011

• 원예과학기술지
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• 제32권2호
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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.

• 원예과학기술지
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• 제35권3호
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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.

• 원예과학기술지
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• 제35권1호
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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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• 제40권4호
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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).