• 제목/요약/키워드: AFLP analysis

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Identification of Differentially Expressed Genes by cDNA-AFLP in Magnaporthe oryzae

  • Chi, Myoung-Hwan;Park, Sook-Young
    • 식물병연구
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    • 제25권4호
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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.

Genetic Variability and Phylogenetic Relationship Among Proton-Beam-Irradiated Strains of Pleurotus ostreatus

  • Kwon, Hye-Jin;Park, Yong-Jin;Yoo, Young-Bok;Park, Soon-Young;Kong, Won-Sik
    • Journal of Microbiology and Biotechnology
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    • 제17권6호
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    • pp.1041-1044
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    • 2007
  • To assess the effects of a proton beam on oyster mushrooms (Pleurotus ostreatus), the genetic diversity and phylogenetic relationships among strains induced by a proton beam were investigated based on a clustering analysis. According to an AFLP DNA polymorphism analysis, the induced strains were divided into four groups that coincided with the dose. When applying proton-beam radiation, the dissimilarity among the induced strains increased when increasing the dose. When using more than 400 Gy, the genetic dissimilarity of the irradiated strains was 46-58%. Thus, evaluating the induced strains using the AFLP technique was effective in revealing the mutation effect of the proton beam.

Differentiation and authentication of Panax ginseng (Korea and China), Panax quinquefolius, and development of genetic marker by AFLP analysis.

  • Jeong, Jae-Hun;Jung, Su-Jin;Yun, Doh-Won;Yoon, Eui-Soo;Choi, Yong-Eui
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.157.2-157.2
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    • 2003
  • Panax ginseng is one of the most important medicinal plant in the Orient. The international trade of ginseng is increasing yearly. The disguise of Chinese and American ginseng into Korean ginseng became a problem in recent years in Korea and an abroad. Obviously, an effective method of authentication of Korean ginseng from others at a DNA level, is necessary for the healthy development of the ginseng market. In order to develop convenient and reproducible methods for the identification of Korean ginseng, amplified fragment length polymorphism (AFLP) analysis was applied within Panax species (Korean cultivatied and wild ginseng, Chinese wild ginseng, American cultivatied and wild ginseng). (omitted)

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Identification of Sex-specific Expression Markers in the Giant Tiger Shrimp (Penaeus monodon)

  • Khamnamtong, Bavornlak;Thumrungtanakit, Supaporn;Klinbunga, Sirawut;Aoki, Takashi;Hirono, Ikuo;Menasveta, Piamsak
    • BMB Reports
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    • 제39권1호
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    • pp.37-45
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    • 2006
  • Bulked segregant analysis (BSA) and AFLP were used for isolation of genomic sex determination markers in Penaeus monodon. A total of 256 primer combinations were tested against 6-10 bulked genomic DNA of P. monodon. Five and one candidate female- and male-specific AFLP fragments were identified. Female-specific fragments were cloned and further characterized. SCAR markers derived from FE10M9520, FE10M10725.1, FE10M10725.2 and FE14M16340 provided the positive amplification product in both male and female P. monodon. Further analysis of these markers using SSCP and genome walk analysis indicated that they were not sex-linked. In addition, sex-specific (or differential) expression markers in ovaries and testes of P. monodon were analyzed by RAP-PCR (150 primer combinations). Twenty-one and fourteen RAP-PCR fragments specifically/differentially expressed in ovaries and testes of P. monodon were successfully cloned and sequenced. Expression patterns of 25 transcripts were tested against the first stranded cDNA of ovaries and testes of 3-month-old and broodstock-sized P. monodon (N = 5 and N = 7 - 10 for females and N = 4 and N = 5 - 7 for males, respectively). Five (FI-4, FI-44, FIII-4, FIII-39 and FIII-58) and two (M457-A01 and MII-51) derived RAP-PCR markers revealed female- and male-specific expression patterns in P. monodon. Surprisingly, MII-5 originally found in testes showed a higher expression level in ovaries than did testes of juvenile shrimps but a temporal female-specific pattern in P. monodon adults.

중부지방 적합 자운영 (Astragalus sinicus L.) 형질 특성 및 유전적 연관성 분석에 관한 연구 (Study for Morphological and Genetic Characteristics of Chinese Milk Vetch (Astragalus sinicus L.) to Select Suitable Line in Central Area of Korea)

  • 홍선희;김재윤
    • 환경생물
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    • 제34권3호
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    • pp.169-176
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    • 2016
  • 본 연구는 중북부지방 적응성 자운영 선발 및 수집 계통간 형태적, 유전적 근연관계를 분석한 것으로서 총 수집된 16개 자운영 계통 중 파주종은 엽각이 적고, 지하부의 발육이 좋아 형태적 형질이 월동에 적합한 초형이었다. 또한 구례 수집종은 개화기가 타 수집종에 비해 일주일 가량 빨랐다. 유전적 근연관계를 검정하기 위해 8개 primer 조합으로 AFLP를 실시한 결과 총 579개의 밴드를 얻었으며, 이 중 polymorphism을 갖는 밴드는 61.54%인 336개 밴드였다. AFLP 데이터를 이용해 군집분석을 실시한 결과 계통 간 유사도 지수는 0.826~0.939 사이의 값을 가지며, 구례와 파주 수집종은 타 수집종과 유전적 유사도에서 비교적 큰 차이를 보였다.

Development of a Sequence Characteristic Amplified Region Marker linked to the L4 Locus Conferring Broad Spectrum Resistance to Tobamoviruses in Pepper Plants

  • Kim, Hyun Jung;Han, Jung-Heon;Yoo, Jae Hyoung;Cho, Hwa Jin;Kim, Byung-Dong
    • Molecules and Cells
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    • 제25권2호
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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.

중이온 빔조사 담배(Nicotiana plumbaginifolia) 식물체의 생장과 DNA 변이 (Growth and DNA Alteration of Heavy-ion Beam Irradiated Tobacco(Nicotiana plumbaginifolia) Plant)

  • 류재일;김민수;;이효연;양덕춘;배창휴
    • 한국자원식물학회지
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    • 제18권1호
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    • pp.169-178
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    • 2005
  • 중이온빔$(^{20}Ne)$을 담배 식물체에 조사(irradiation)하여 식물체의 생장과 DNA의 변이에 미치는 영향을 검토하였다. 발아율과 초장은 조사선 량이 증가함에 따라 감소하였다. 반면 $50Gy\~10Gy$의 저선량에서 추대와 개화가 촉진되어 발육은 오히려 촉진되었다. 총 100개의 primer를 이용하여 RAPD분석한 결과, 59개의 primer에서 총 336개의 DNA단편이 증폭되었고, 1개의 primer에서 중이온빔 조사처리구에서만 출현하는 DNA단편이 나타났다. AFLP분석 결과, 개체 특이적인 DNA단편은 나타난 반면 중이온빔 처리구 특이적인 DNA 단편은 관찰되지 않았다.

분자표지를 이용한 마(Dioscorea spp.)의 다양성 분석 (Genetic Diversity among Dioscorea, spp. Using Molecular Markers)

  • 장광진;최익영;박주현;박종인;윤병성;김남수
    • 현장농수산연구지
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    • 제4권1호
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    • pp.45-54
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    • 2002
  • 마(Dioscorea spp.) 계통 중 품질이 우수하고 이용 가치가 기대되는 51개 계통에 대한 특성을 조사하고 도입재배 가능성을 시험했다. 또한 종간, 계통 간을 AFLP 분석하였고 생육 패턴을 시험하였다. 1. AFLP분석의 분자마커를 이용한 도입마의 유사성 분석에서 62% 수준에서 도입마는 다른 종간에 유의성이 나타났고 주성분 분석에서도 구분이 가능하였다. 그 중 D. alata는 일부 수집 지역별로 그룹이 형성되었다. 2. 일본의 쿠마모도(Kumamoto)지방에서 수집된 D. alata의 계통들의 유사성이 높은 것으로 보아, 그 지역의 마 유전자원의 이동이 적은 것으로 보인다. 반면, Indonesia, palau Is., 파푸아뉴기니아에서의 수집종들의 유연성이 높은 것은 한 지역의 계통이 여러지역으로 이동된 것으로 사료되다. 3. D. nipponica에 속하는 부채마는 다른 종과 변이가 많은 것으로 조사되어 유전적 가치가 높은 것으로 인정된다.