• 한국약용작물학회지
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• 제11권2호
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• pp.102-108
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• 2003

본 연구는 금산농업기술센터 인삼연구실에서 순계선발법으로 육성 중인 인삼 계통과 인삼연초연구원에서 육성한 품종을 RAPD 방법으로 계통 내의 변이와 육성계통의 순도를 검정하여 인삼의 순계선발법으로 활용하기 위한 기초자료를 얻기 위해 실시하여 얻은 결과를 요약하면 다음과 같다. 1. 10개 계통으로부터 각각 $4{\sim}5$개 개체를 임의로 수확한 49개체의 DNA를 48개의 primer를 사용하여 PCR한 결과 최소한 1개의 계통 내에서 RAPD다형성을 나타내는 4개의 primer OPA 19, OPM 11, URP 3 및 UBC 98을 선발하였다. 그중 Primer OPA 19, OPM 11 및 UBC 98은 각각 6계통, 7계통 및 1계통 내에서 개체간의 차이를 보이는 band가 증폭되었다. 2. 육성품종 천풍의 DNA를 OPA 19를 사용하여 증폭한 결과 약 1,800bp 크기의 band에서 개체간의 차이를 보였고, OPM 11을 사용하여 증폭한 경우에는 약 730bp 및 850bp 크기의 두 band에서 개체간의 차이를 나타냈으며, 육성품종 연풍은 OPM 11을 사용하여 증폭한 결과 약 730bp 크기의 band에서 개체간의 차이를 보였다. 3. 이와 같이 인삼육성계통내의 개체 간에 RAPD 다형성이 나타나는 이유는 영년작물인 인삼이 타가수정 되면 유전적으로 고정이 되는데 필요한 기간이 길어지기 때문이라고 설명할 수 있다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권9호
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• pp.1244-1249
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• 2014

Goose fatty liver is one of the most delicious and popular foods in the world, but there is no reliable genetic marker for the early selection and breeding of geese with good liver-producing potential. In our study, one hundred and twenty-four 78-day-old Landes geese bred in Shunda Landes goose breeding farm, Jiutai, Jilin, China were selected randomly. The fatty livers were sampled each week after overfeeding during a three week period. Polymerase chain reaction-single strand conformation polymorphism and DNA sequencing were used to identify single nucleotide polymorphisms (SNPs) of fatty acid synthase (FAS), which is an important enzyme involved in the synthesis of fat under both physiological and pathological conditions. Least-squares correlation was established between these SNPs and fatty liver weight, abdominal fat weight, and intestinal fat weight of the overfed Landes geese, respectively. The results showed that fatty liver weight of geese with EF and FF genotypes (amplified by primer P1) was significantly higher than that of the EE genotype (p<0.05), and liver weight of CD and DD genotypes (amplified by primer P2) was significantly higher than that of the CC genotype (p<0.05). Different genotype combinations showed different liver weights, and from highest to lowest were ABDD, DDEF, DDFF, DDEE, ABEF, ABFF, AADD, and CDEF. Further analysis of DNA sequencing showed that there were two SNPs within the 5' promoter region the FAS gene. The geese of EF and FF genotypes carried a change of T to C, and the geese of CD and DD genotypes carried a change of A to G. The changes of the bases could potentially influence the binding of some transcription factors to this region as to regulate FAS gene. To our knowledge, this is the first report of SNPs found within the 5' promoter region of the Landes goose FAS gene, and our data will provide an insight for early selection of geese for liver production.

• Journal of Ginseng Research
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• 제43권3호
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• pp.482-487
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• 2019

Background: The mixed-cultivation of different Panax ginseng cultivars can cause adverse effects on stability of yield and quality. K-1 is a superior cultivar with good root shape and stronger disease resistance. DNA markers mined from functional genes are clearly desirable for K-1, as they may associate with major traits and can be used for marker-assisted selection to maintain the high quality of Korean ginseng. Methods: Five genes encoding pathogenesis-related (PR) proteins of P. ginseng were amplified and compared for polymorphism mining. Primary, secondary, and tertiary structures of PR5 protein were analyzed by ExPASy-ProtParam, PSSpred, and I-TASSER methods, respectively. A coding single nucleotide polymorphism (SNP)-based specific primer was designed for K-1 by introducing a destabilizing mismatch within the 3' end. Allele-specific polymerase chain reaction (PCR) and real-time allele-specific PCR assays were conducted for molecular discrimination of K-1 from other cultivars and landraces. Results: A coding SNP leading to the modification of amino acid residue from aspartic acid to asparagine was exploited in PR5 gene of K-1 cultivar. Bioinformatics analysis showed that the modification of amino acid residue changed the secondary and tertiary structures of the PR5 protein. Primer KSR was designed for specific discrimination of K-1 from other ginseng cultivars and landraces. The developed real-time allele-specific PCR assay enabled easier automation and accurate genotyping of K-1 from a large number of ginseng samples. Conclusion: The SNP marker and the developed real-time allele-specific PCR assay will be useful not only for marker-assisted selection of K-1 cultivar but also for quality control in breeding and seed programs of P. ginseng.

• Plant Biotechnology Reports
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• 제2권4호
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• pp.239-243
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• 2008

Simmondsia chinensis (Link) Schneider, a multipurpose and monogeneric dioecious shrub from arid zones, has emerged as a cash crop all over the globe. Its seed propagation poses severe problems due to its male-biased population: the male:female ratio is 5:1. Investigations have been carried out to generate a sex-specific Inter-simple sequence repeat (ISSR) marker for the early detection of male and female plants. Of the 42 primers analysed with a bulk sample of pooled male DNA and a bulk sample of pooled female DNA, only one primer, UBC-807, produced a unique ~1,200 base-pair fragment in the male DNA. To validate this observation, this primer was re-tested with individual male and female samples from eight cultivars. A similar unique ~1,200 bp fragment was present in the male individuals of all eight cultivars and completely absent in the female individuals tested. This is the first report of the use of ISSR markers to ascertain sex in physiologically mature S. chinensis plants.

• 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.

• 한국자원식물학회지
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• 제17권2호
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• pp.116-121
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• 2004

고려 인삼중 폐포와 4등급 이하를 유발시키는 90％이상이 적변삼이라고 불리는 인삼의 표피 색택이 붉은 삼이 그 원인이다. 이러한 적변삼은 미국삼보다는 고려 인삼에 서 다량 발견되는 바, 적변은 유전적 요인이 있다고 사료된다. 그러므로 이 연구의 목적은 RT-PCR을 이용하여 인삼적병에 내성을 가지는 유전자를 탐색하기 위하여 실시되었다. 고려인삼 3년근 1개체 중에서 적변이 발생된 부위와 건전 부위의 RNA를 추출하여 형성된 cDNA를 여러개의 random primer를 사용하여 PCR 증폭을 한 결과 정상 부위의 cDNA에서 발견되지 않는 band가 적변삼의 부위에서 발견되었다. 따라서 band가 형성된 부위의 유전자가 적변과 관련될 가능성이 있는 것으로 사료되고 이러한 유전자는 향후 염기서열을 분석하여 어떠한 유전자인지 판명을 하여야 하며 적변관련 유전자이면 선발마커로서 사용되고 또한 형질전환을 통한 적변내성 인삼계통을 육성할 수 있으며, 만약 적변과 관련이 없는 유전자로 판명된다면 더 많은 primer를 사용하여 적변관련 유전자를 탐색해야 할 것이다.

• 한국약용작물학회지
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• 제13권2호
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• pp.121-125
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• 2005

An analysis of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) was performed with three Angelica species (A. gigas Nakai, A. sinensis (Olive.) Diels and A. acutiloba Kitag) in an effort to distinguish between members of these three species. Two arbitrary primers (OPC02, OPD11) out of80 primers tested, produced 17 species-specific fragments among the three species. Eight fragments were specific for A. sinensis, four fragments specific for A. gigas, five specific for A. acutiloba. When primers OPC02 and OPD11 were used in the polymerase chain reaction, RAPD-PCR fragments that were specific for each of the three species were generated simultaneously. Primer OPC02 produced eight species-specific fragments: four were specific for A. sinensis, one for A. gigas, and three for A. acutiloba. Primer OPD11 produced nine speciesspecific fragments: four for A. sinensis, three for A. gigas, and two for A. acutiloba. The RAPD-PCR markers that were generated with these two primers should rapidly identify members of the three Angelica species. The consistency of the identifications made with these species-specific RAPD-PCR markers was demonstrated by the observation that each respective marker was generated from three accessions of each species, all with different origins. We also performed the RAPD-PCR analysis with the dried Angelica root samples that randomly collected from marketed and from the OPC02 primer, obtained a A. gigasspecific band and the band were cloned and sequenced.

• Journal of Plant Biotechnology
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• 제35권2호
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• pp.147-153
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• 2008

국화에 담배거세미 나방 (tobacco cutworm; Spodoptera litura)에 저항성을 나타내 국화를 육성하기 위하여 cry1Ac 유전자를 형질전환하였다. Cry1Ac 유전자는 pCAMBIA2301를 포함하는 Agrobacterium C58C1을 통하여 국화 'Linneker salmon'에 도입하였다. Agrobacterium C58C1을 접종한 후 엽절편을 10 mg/L kanamycin이 함유된 재분화 배지 (MS + 1.0 mg/L BA + 0.5 mg/L IAA)에서 1차 선발을 하였고, 재분화 배지에 20 mg/L kanamycin이 첨가된 배지에서 2차 선발을 하였으며, 20 mg/L kanamycin이 첨가된 MS 배지에서 3차 발근선발을 하였다. 3차 발근선발까지 69개의 신초 (1.6%)가 생존하여 발근하였다. NptII primer로 PCR을 한 결과 그 중 36개의 신초 (0.8%)가 putative transformant로 확인되었고, Southern 분석을 한 결과, 35개체 (0.8%)가 nptII 유전자와 cry1Ac 유전자를 가진 형질전환체로 확인되었다. Cry1Ac 유전자의 형질전환율은 0.8%로 양호하였다. 온실에서 담배거세미 나방에 대한 저항성을 검정한 결과, 3개체의 형질전환체가 저항성을 나타내는 것으로 확인되었다.

• 생명과학회지
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• 제21권12호
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• pp.1659-1665
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• 2011

감귤류 중 수술이 퇴화되어 웅성불임형질을 나타내는 '청견' 품종에 정상적인 수술의 형태를 가진 웅성가임인 '진귤' 품종을 교배하여 150개체의 $F_1$ 집단을 구축하여 수술이 퇴화되는 개체와 정상인 개체를 분리하였다. 분리된 F1 개체들을 사용하여 SRAP 기법과 집단 분리 분석법(BSA)을 조합하여 웅성 가임 연관 마커 개발에 활용하였다. $F_1$ 집단 내 150개체 중 66개체가 퇴화 수술을 갖고 있으며 웅성 가임성과 웅성 불임성의 분리비는 1:1이며 $x^2$ 값은 2.16(p=0.05)이었다. 197개의 SRAP 프라이머 조합들 중 웅성가임 특이밴드를 형성하는 3개의 SRAP 프라이머 조합(F4/R27, F39/R60, 및 F15/R37)을 선발하였으며, 이 중 F39/R60 프라이머에 특이적으로 증폭하는 DNA단편의 염기서열을 기본으로 하여 새롭게 작성한 양방향 프라이머 조합 중 웅성 가임 계통에서만 약 1.4 Kb의 특이밴드를 증폭하는 프라이머 조합, pMS 33U/pMS 1462L를 선발하여 SCAR 마커를 개발 하였다. 이러한 결과는 개발된 SCAR 마커로 무핵성 계통들의 육종 선발에 효율성을 높일 수 있을 것으로 기대된다.

• Journal of Life Science
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• 제9권1호
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• pp.14-16
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• 1999

Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.