• Journal of Mushroom
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• v.16 no.1
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• pp.51-56
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• 2018

In Korea, the oak mushroom (Lentinula edodes) is highly preferred by consumers in the food industry and makes up about 97.7% of the total forest mushroom production. This indicates that the oak mushroom is an important non-timber forest product in Korea. Recently, the breeding and development of new cultivars of L. edodes have been actively initiated, and the development of molecular markers that are able to identify and discriminate the new cultivars is crucial for protecting the breeder's rights. This study was carried out to develop a cleaved amplified polymorphic sequence (CAPS) marker for the identification and discrimination of a new cultivar, Sanmaru 2ho from the 37 other oak mushroom cultivars. A single nucleotide polymorphism (SNP) was identified at the $1,803,483^{rd}$ position of scaffold2 in the genome of Sanmaru 2ho. The amplified DNA containing the SNP of Sanmaru 2ho was uniquely not cleaved by the restriction enzyme, Hha I, and thus Sanmaru 2ho was successfully distinguished from the other oak mushroom cultivars.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.4
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• pp.329-341
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• 2011

The objective of this study was to determine the genetic diversities of major rice blast resistance genes among 84 accessions of aromatic rice germplasm. Eighty four accessions were characterized by a dominant 11 set of PCR-based SNP and CAPS marker, which showed the broad spectrum resistance and closest linkage to seven major rice blast resistance (R) genes, Pia, Pib, Pii, Pi5 (Pi3), Pita (Pita-2), and Pi9 (t). The allele specific PCR markers assay genotype of SCAR and STS markers was applied to estimate the presence or absence of PCR amplicons detected with a pair of PCR markers. One indica accession, Basmati (IT211194), showed the positive amplicons of five major rice blast resistance genes, Pia, Pi5 (Pi3), Pib, Pi-ta (Pi-ta2), and Pik-5 (Pish). Among 48 accessions of the PCR amplicons detected with yca72 marker, only five accessions were identified to Pia gene on chromosome 11. The Pib gene was estimated with the NSb marker and was detected in 65 of 84 accessions. This study showed that nine of 84 accessions contained the Pii gene and owned Pi5 (Pi3) in 42 of 84 accessions by JJ817 and JJ113-T markers, which is coclosest with Pii on chromosome 9. Only six accessions were detected two alleles of the Pita or Pita-2 genes. Three of accessions were identified as the Pi9 (t) gene locus.

• Journal of Mushroom
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• v.19 no.4
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• pp.336-340
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• 2021

The cleaved amplified polymorphic sequence (CAPS) marker uses a restriction enzyme recognition site resulting from single nucleotide polymorphisms and insertions and deletions on the DNA sequence. This technique does not require expensive equipment, the process is simple, and clear results can be obtained reliably. In this study, Agaricus bisporus cultivars SaeA, SaeDo, SaeHan, SaeYeon, SaeJeong, Dodam, Seolgang, Dahyang, Hogam, and Hadam developed in Korea were discriminated using four CAPS markers. Our results indicated that it is possible to distinguish the ten cultivars and determine the genetic diversity among them.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.648-655
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• 2015

Balloon flower (Platycodon grandiflorum A. DC.) is a perennial plant of mainly Campanulaceae family, which have been widely used as a food ingredient and herbal medicine in East Asia. Although demands on related products and yearly cultivation area for balloon flower are increasing, diverse fundamental technologies and molecular breeding studies are not very well supported in Platycodons. In this study, 30 random amplification of polymorphic DNA (RAPD) primers were test in an attempt to explore genetic diversities. In addition, sequences information of the actin gene, a well conserved gene encoding a globular protein that forms microfilaments, was retrieved and analyzed. Two actin homologs were recovered; 3.4 kb fragment is a Pg-actin and 1.4 kb fragment is a Pg-actin homolog with 28.6% similarity. We have confirmed that the Pg-actin gene is configured into 4 exons and 3 introns. A single nucleotide polymorphism (SNP), G↔A, was detected on the intron 3, which served as a target for the CAPS marker development. The marker Pg-Actin-Int3 was applied to 32 balloon flower accessions. Balloon flower DNA sequence information generated in this study is expected to contribute to the analysis and molecular breeding and genetic diversity analysis of balloon flowers.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.66-66
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• 2018

국화과(Compositae) 다년생 초본인 산국속(Dendranthema)은 국내 약 13여종이 자생하는 것으로 알려져 있으며, 이 중 감국(D. indicum (L.) Des Moul.)과 산국(D. boreale (Makino) Ling ex Kitam.), 구절초(D. zawadskii var. latilobum (Maxim.) Kitam.)가 주로 차 또는 한약재 등의 원료로 이용되고 있다. 차로 이용되는 꽃은 산국이 감국에 비해 상대적으로 작아서 구분이 가능하지만 시중에는 건조된 형태로 가공 유통되므로 육안으로 구분이 쉽지 않고, 산국 유래 제품들은 국내에서 감국 또는 국화로 혼용해서 표기되어 유통되고 있어 그 기원을 명확히 정립할 필요가 있다. 이에 본 연구는 감국과 산국의 분자유전학적 판별을 위해 DNA 바코드 후보 유전자를 활용하여 염기서열분석으로 확보된 SNP 및 InDel 정보를 바탕으로 CAPS 마커를 개발하고자 수행되었다. 감국과 산국 모두 trnL-trnF intergenic spacer 구간에서 약 1kb의 PCR 산물이 확인되었고, 이들 염기서열에서 분석한 2 SNP 및 3 InDel을 대상으로 CAPS 마커 개발을 위한 제한효소 사이트를 탐색하였다. Gap을 포함한 774bp (감국/산국=A/G) 위치의 SNP에서 BstUI(GC^GC)처리로 CAPS 마커로 전환 가능함이 확인되었고, 이에 감국과 산국의 PCR 산물에 제한효소를 처리한 결과, 제한효소 인식 사이트가 존재하는 산국에서 두 개의 DNA 단편이 확인되었다. 위 결과는 다양한 형태로 가공 유통되는 감국과 산국의 판별을 위한 마커로 활용될 수 있으며, 본 연구에 활용된 기술은 추후 건강기능식품 개발을 위한 원료표준화 확립 연구에 유용할 것으로 판단된다.

• Horticultural Science & Technology
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• v.33 no.4
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• pp.566-574
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• 2015

Yellow or transparent fruit peel color is caused by the accumulation or lack of naringenin chalcone (NG, C) in fruit peel and determines the red or pink appearance of tomato fruit, respectively. NGC biosynthesis is regulated by the SlMYB12 gene of the Y locus on chromosome 1, and DNA markers derived from SlMYB12 would be useful for marker-assisted selection (MAS) of tomato fruit color. To develop a gene-based marker, 4.9 kb of the SlMYB12 gene including a potential promoter region was sequenced from the red-fruited (YY) line 'FCR' and pink-fruited (yy) line 'FCP'. Sequence alignment of these SlMYB12 alleles revealed no sequence variations between 'FCR' and 'FCP'. To identify SlMYB12-linked single nucleotide polymorphisms (SNPs), 'FCR' and 'FCP' were genotyped using a SolCAP Tomato SNP array and CAPS markers (CAPS-456, 531, 13762, and 38123) were developed from the four SNPs (solcap_snp_sl_456, 531, 13762, and 38123) most closely flanking the SlMYB12. These CAPS markers were mapped using $F_2$ plants derived from 'FCR' ${\times}$ 'FCP'. The map positions of the fruit peel color locus (Y) were CAPS-13762 (0 cM) - 456 (11.09 cM) - Y (15.71 cM) - 38123 (17.82 cM) - 531 (30.86 cM), and the DNA sequence of SlMYB12 was physically anchored in the middle of CAPS-456 and CAPS-38123, indicating that fruit peel color in domesticated tomato is controlled by SlMYB12. A total of 64 SolCAP tomato germplasms were evaluated for their fruit peel color and SNPs located between solcap_snp_sl_456 and 38123. Seven SNPs that were detected in this interval were highly conserved for pink-fruited accessions and specific to transparent fruit peel traits, as depicted by a phenetic tree of 64 accessions based on the seven SNPs.

• Horticultural Science & Technology
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• v.33 no.5
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• pp.722-729
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• 2015

Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

• 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.

• The Korean Journal of Mycology
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• v.45 no.2
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• pp.114-120
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• 2017

Lentinula edodes is an edible mushroom that is mainly cultivated in Asian countries. Recently, new cultivars of this mushroom have been developed in Korea; variety protection is very important, so the development of efficient molecular markers that can distinguish each variety is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanmaru 1ho and Chunjang 3ho). These markers were developed from whole genomic sequencing data from L. edodes monokaryon strain B17 and resequencing data from 10 dikaryon strains. A single nucleotide polymorphism changed in scaffold 9 POS 1630048 in Sanmaru 1ho($G{\rightarrow}T$), and in scaffold 13 POS 920681 in Chunjang 3ho ($G{\rightarrow}A$). The restriction enzymes TspR I and Xho I distinguished Sanmaru 1ho and Chunjang 3ho, respectively, from other strains. Thus, we developed 2 CAPS markers for the identification of the L. edodes cultivars Sanmaru 1ho and Chunjang 3ho.

• The Korean Journal of Mycology
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• v.49 no.1
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• pp.33-44
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• 2021

Lentinula edodes (Berk.) Pegler, the most produced mushroom in the world, is an edible mushroom with very high nutritional and pharmacological value. Currently, interest in the protection of genetic resources is increasing worldwide, and securing the distinction between new cultivars is very important. Therefore, the development of efficient molecular markers that can discriminate between L. edodes cultivars is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanbaekhyang and Sulbaekhyang). These markers were developed from whole genome sequencing data from L. edodes monokaryon strain B17 and resequencing data from 40 cultivars. A nucleotide deletion existed in scaffold 19 POS 214449 in Sanbaekhyang (GT→G), and a single nucleotide polymorphism changed in scaffold 7 POS 215801 in Sulbaekhyang (G→A). The restriction enzymes Hha I and HpyCH4IV distinguished Sanbaekhyang and Sulbaekhyang, respectively, from other cultivars. Thus, we developed two CAPS markers for the identification of the L. edodes cultivars Sanbaekhyang and Sulbaekhyang.