• Title/Summary/Keyword: 분자마커선발

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Development of PCR-based markers for selecting plastid genotypes of Solanum hjertingii (Solanum hjertingii 색소체 유전자형 선발을 위한 PCR 기반 분자마커 개발)

  • Tae-Ho Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.34-44
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    • 2023
  • The tetraploid Solanum hjertingii, a wild tuber-bearing species from Mexico is a relative of potato, S. tuberosum. The species has been identified as a potential source of resistance to blackening for potato breeding. It does not exhibit enzymatic browning nor blackspot which are physiological disorders. However, due to their sexual incompatibility, somatic hybridization between S. hjertingii and S. tuberosum must be used to introduce various traits from this wild species into potato. After somatic hybridization, molecular markers are essential for selecting fusion products. In this study, the chloroplast genome of S. hjertingii was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop specific markers for S. hjertingii. The chloroplast genome has a total sequence length of 155,545 bp, and its size, gene content, order and orientation are similar to those of the other Solanum species. Phylogenic analysis including 15 other Solanaceae species grouped S. hjertingii with S. demissum, S. hougasii, and S. stoloniferum. After detailed comparisons of the chloroplast genome sequence with eight other Solanum species, we identified one InDel and seven SNPs specific to S. hjertingii. Based on these, five PCR-based markers were developed for discriminating S. hjertingii from other Solanum species. The results obtained in this study will aid in exploring the evolutionary aspects of Solanum species and accelerating breeding using S. hjertingii.

Species-specific Marker of Phytophthora pinifolia for Plant Quarantine in Korea (국내 식물검역대상 Phytophthora pinifolia의 PCR 검출을 위한 종 특이적 마커 개발)

  • Kim, Narae;Choi, You Ri;Seo, Mun Won;Song, Jeong Young;Kim, Hong Gi
    • The Korean Journal of Mycology
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    • v.44 no.2
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    • pp.103-107
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    • 2016
  • To establish a rapid and accurate detection of Phytophthora pinifolia, which is a quarantine pathogenic fungus in Korea, a species-specific primer was developed based on the ras-related protein (Ypt1) gene. Species-specific primer based on the DNA sequences of Ypt1 gene amplified 193 bp polymerase chain reaction (PCR) product for P. pinifolia. The primer pair yielded the predicted PCR product size exactly in testing with target pathogen DNAs, but not from the other 10 species of Phytophthora and 14 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 $10pg/{\mu}L$.

Chloroplast genome sequence and PCR-based markers for S. cardiophyllum (감자 근연야생종 Solanum cardiophyllum의 엽록체 전장유전체 구명 및 이를 이용한 S. cardiophyllum 특이적 분자마커의 개발)

  • Tae-Ho Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.45-55
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    • 2023
  • The diploid Solanum cardiophyllum, a wild tuberbearing species from Mexico is one of the relatives to potato, S. tuberosum. It has been identified as a source of resistance to crucial pathogens and insects such as Phytophthora infestans, Potato virus Y, Colorado potato beetle, etc. and is widely used for potato breeding. However, the sexual hybridization between S. cardiophyllum and S. tuberosum is limited due to their incompatibility. Therefore, somatic hybridization can introduce beneficial traits from this wild species into the potato. After somatic hybridization, selecting fusion products using molecular markers is essential. In the current study, the chloroplast genome of S. cardiophyllum was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop S. cardiophyllum-specific markers. The total length of the S. cardiophyllum chloroplast genome was 155,570 bp and its size, gene content, order and orientation were similar to those of the other Solanum species. Phylogenic analysis with 32 other Solanaceae species revealed that S. cardiophyllum was expectedly grouped with other Solanum species and most closely located with S. bulbocastanum. Through detailed comparisons of the chloroplast genome sequences of eight Solanum species, we identified 13 SNPs specific to S. cardiophyllum. Further, four SNP-specific PCR markers were developed for discriminating S. cardiophyllum from other Solanum species. The results obtained in this study would help to explore the evolutionary aspects of Solanum species and accelerate breeding using S. cardiophyllum.

Early Identification of Citrus Zygotic Seedlings Using Pollen-specific Molecular Markers (화분 특이적 마커를 이용한 감귤 교잡종 실생묘의 조기 동정)

  • Jin, Seong Beom;Yun, Su Hyun;Park, Jae Ho;Park, Suk Man;Koh, Sang Wook;Lee, Dong Hoon
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.598-604
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    • 2015
  • This study was carried out to develop molecular techniques to allow the selection of zygotic seedlings in the early stage of the plant development. We identified 37 pollen-specific molecular markers from RAPD analysis and successfully used them for identification of the zygotic seedlings from various hybrid crosses. Three Satsuma mandarin cultivars ('Morita unshiu', 'Nangan 20' and 'Miyagawawase') were used as mother parents and seven cultivars ('Ponkan', 'Lee', 'Kinokuni', 'Shiranuhi', 'Tamnaneunbong', 'Shinyegam', and 'Sunburst' mandarins) served as pollen parents. PCR analysis showed that 2 primers could identify zygotic hybrid seedlings. Among them, an UBC-27 primer was used to identify the zygotic seedlings from hybrid crosses of "'Nangan 20' ${\times}$ 'Kinokuni'" mandarin, "'Nangan 20 ${\times}$ Ponkan'" mandarin and "'Miyagawawase ${\times}$ Sunburst'" tangerine. In total 29 out of 40 seedlings (73%), 9 out of 47 seedlings (19%), and 13 out of 45 (29%) were identified as zygotic seedlings, respectively. These results can show that the pollen-specific markers selected in this study can be used effectively for early identification of zygotic seedlings from Citrus hybrid crosses.

A Molecular Marker Discriminating the Soybean Podworm, Matsumuraeses phaseoli and the Podborer, M. falcana (Lepidoptera: Tortricidae) (팥나방(Matsumuraeses phaseoli)과 어리팥나방(M. falcana)의 판별 분자마커)

  • Heo, Hye-Jung;Son, Ye-Rim;Seo, Bo-Yoon;Jung, Jin-Kyo;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.48 no.4
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    • pp.547-551
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    • 2009
  • Two closely related species, the soybean podworm, Matsumuraeses phaseoli, and the podborer, M. falcana, gives differential economic damages on crops. It is difficult to discriminate these potential sympatric species by morphological characters. The goal of this study was to develop a discriminating molecular marker based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). A partial genomic fragment (500 bp) of mitochondrial cytochrome oxidase I (mtCOI) was sequenced in both species, in which restriction site by Rsa I was selected as a dichotomous marker. PCR-RFLP in the mtCOI region clearly discriminated both species.

Selection and Characterization of Horticultural Traits of Tomato leaf curl virus (TYLCV)-resistant Tomato Cultivars (토마토 황화잎말림바이러스(TYLCV) 저항성 품종 선발 및 원예특성 분석)

  • Kim, Woo-Il;Kim, Kwang-Hwan;Kim, Young-Bong;Lee, Heung-Su;Shon, Gil-Man;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.328-336
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    • 2013
  • This study was conducted to evaluate imported tomato $F_1$ cultivars as breeding materials for the resistance to Tomato yellow leaf curl virus (TYLCV) by molecular markers and bioassay. From marker genotyping and disease evaluation of 40 $F_1$ cultivars, most of the cultivars declared as TYLCV-resistance carried heterozygous marker genotype for the TYLCV resistance genes Ty-1, Ty-3, or Ty-3a, and showed low disease rates. Whereas, 4 of 5 $F_1$ cultivars declared as intermediate resistance showed marker genotype for susceptibility and disease rates ranged 18.1-33.3%. However, the xx cultivars showed inconsistency in marker genotype and disease rate. Characterization of horticultural traits of the $F_1$ cultivars with TYLCV-resistance indicated that large-size fruit cultivars were higher in yield and similar in sugar contents and solid-acid ratio compared to a control cultivar preferred in the domestic market, although hardness remained to be a problem. On the other hand, cherry tomato cultivars showed lower yield and brix, but longer internode compared to a control cultivar, indicating that breeding for TYLCV-resistance using these cultivars will require more efforts and time compared to large-sized.

Transgene structures of marker-free transgenic Bt rice plants (무선발 형질전환 Bt벼의 도입유전자 구조 분석)

  • Woo, Hee-Jong;Lee, Seung Bum;Lim, Myung-Ho;Gwon, Sun-Jong;Lee, Jin-Hyoung;Shin, Kong-Sik;Cho, Hyun-Suk
    • Journal of Plant Biotechnology
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    • v.40 no.3
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    • pp.135-140
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    • 2013
  • A less simple approach developed for generation of marker-free transgenic plants is to select transformants without the use of selective marker genes. Some results about development of marker-free transgenic plants were obtained using a non-selective approach in several crops such as rice, potato and tobacco. However, the study did not provide evidence on detailed characterization of introduced gene on genome, a critical step for confirming the stable integration and transmission of a foreign gene. In this study, we evaluated structure and integration sites of transgene (mCry1Ac) in the transgenic Bt rice plants which were made via conventional Agrobacterium-mediated transformation by non-selective method. Structure and integration sites of transgene in these transgenic plants had similar fashion as those recovered under selection.

한우 Leptin 비만 유전자와 도체 및 육질 형질과의 연관성 구명

  • Sin, Seong-Cheol;Jeong, Hwa-Cheol;Kim, Hui-Seon;Jeon, Sang-Hui;Gwon, Su-Yeon;Kim, Bo-Hyeon;Jeong, Gu-Yong;Jeong, Eui-Ryong
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2006.05a
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    • pp.112-116
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    • 2006
  • 본 연구는 인간과 동물의 식욕조절, 에너지 대사, 체지방 축적 및 지방 대사에 핵심적인 역할을 담당하는 비만 유전자 leptin의 SNP를 검색하고, 이들 SNP 마커와 한우의 도체 및 육질형질들과의 연관성을 구명하여 고급육 생산 한우 조기 선발 및 육질 진단을 위한 분자 표지 마커로 활용하기 위하여 수행하였다. 한우 leptin 유전자의 exon 2 및 3번 영역을 포함한 염기서열 분석결과 총 3개의 SNP를 검출하였고, PCR-RFLP및 SSCP기법을 이용하여 SNP유전자형을 분석한 결과 exon 2 영역 내 C1180T SNP부위가 한우의 근내 지방도 및 등지방 두께와 유의적인 연관성(p<0.05)이 있는 것으로 나타났다. 따라서, 본 연구를 통해 개발된 한우 leptin 유전자의 특정한 SNP marker는 근내 지방도가 우수한 고급육을 생산하는 한우의 조기식별 및 마블링 등 육질진단에 매우 유용한 DNA 표지인자로 활용할 수 있을 기대된다.

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Research Status for Drought Tolerance in Maize (옥수수 한발 내성에 관한 연구 현황)

  • Kim, Kyung-Hee;Moon, Jun-Cheol;Kim, Jae-Yoon;Kim, Hyo-Chul;Shin, Seung-Ho;Song, Ki-Tae;Lee, Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.4
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    • pp.401-411
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    • 2015
  • Drought stress has detrimental effects on the seedling development, vegetative/ reproductive growth, photosynthesis, root proliferation, anthesis, anthesis-silking interval (ASI), pollination and grain yield in maize. Typically, two weeks before silking through pollination are an important time in maize life. Here we reviewed the effects of drought stress on growth, physiological/ molecular researches for drought tolerance, and breeding to genomics in maize. Drought stress during kernel development increases leaf dying and lodging, decreases grain filling period and grain yield. Physiological factors of drought stress/ effects are water content, water deficits, and water potential. Nowdays molecular marker assisted breeding method is becoming increasingly useful in the improvement of new germplasm with drought stress tolerance.

Identification of White Hanwoo Breed Using Single Nucleotide Polymorphism Markers (단일염기다형성 마커를 이용한 백우 품종 식별 방법)

  • Kim, Seungchang;Kim, Kwanwoo;Roh, Heejong;Kim, Dongkyo;Kim, Sungwoo;Kim, Chalan;Lee, Sanghoon;Ko, Yeounggyu;Cho, Changyeon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.1
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    • pp.240-246
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    • 2020
  • This study was conducted to develop specific Single Nucleotide Polymorphism (SNP) markers to identify the genetic characteristics and breed of White Hanwoo (WH) using a molecular biological method. SNP genotyping was performed with an Illumina Bovine HD 777K SNP chip using DNA extracted from 48 Hanwoo and 22 WH. The minor allele frequency (MAF) difference of each SNP was calculated and the statistical significance (P-value) of the MAF difference was calculated through Fisher's Exact test (Genotype). SNPs with 100% difference in the MAF difference were selected based on marker selection criteria. The nine SNP markers with genetic differences were selected. The selected markers have different alleles as being Hanwoo- and WH- specific. Therefore, based on these results, it can be concluded that the Hanwoo and WH varieties can be clearly distinguished by using these SNPs. So, the patent of the WH breed identification markers was registered. WH is a breed that shows the characteristics of a Korean native species that is separate from the native Hanwoo. It is expected that genetic characteristics research on the WH can be used to identify the breed and as a knowledge base for enhancing the value of breeding stock.