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Genetic identification of Sinomenium acutum based on chloroplast gene ndhF sequences
육진아,이혜원,고병섭 대한본초학회 2013 大韓本草學會誌 Vol.28 No.5
Objectives : This study was conducted to identify the original Sinomini Caulis et Rhizoma plant among Stephania tetrandra, Cocculus trilobus, and Aristolochiae fangchi to develop the genetic marker for Sinomini Caulis et Rhizoma. Methods : Sinomenium acutum was identified by the classification and identification committee of the National Center for Standardization of Herbal Medicines. The chloroplast ndhF gene was amplified. We performed sequences alignment analysis of Sinomenium acutum, Stephania tetrandra, C. trilobus, and A. fangchi using BioEdit program. The SFR markers designed were consisted of SF01, SR04, and SR05 primers. Results : Many variations of Sinomeni Caulis et Rhizoma are currently commercialized as herbal medicine. We compared the base sequences of the ndhF intergenic space of chloroplast DNA with Sinomenium acutum, Stephania tetrandra, C. trilobus, and A. fangchi. According to the results, it showed that the nucleotide variations were seen in 30 genes of four species. Phylogenetic analysis revealed that 4 species were classified into five groups based on an inter-group divergence in nucleotide sequence of 9%. We developed SFR marker nucleotides enough to authenticate respective species and confirmed its application on the band size at 419 base pair. These sequence differences at corresponding positions were available genetic markers to identity the Sinomeni Caulis et Rhizoma. Conclusions : Base on these results, the ndhF region was effective in distinguishing Sinomini Caulis et Rhizoma The SFR genetic marker was useful for identifying Sinomini Caulis et Rhizoma with other species.
육진아,이혜원,주영승,고병섭 한국응용생명화학회 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.2
Cynanchum wilfordii (Asclepiadaceae) is widelydistributed throughout Korea, Japan, and China. Dried roots ofthis plant have been used as a tonic to promote renal function. Dueto the morphological similarities of the dried roots of this plant tothose of Cynanchum auriculatum, which is used as a substituteherbal medicine for C. wilfordii, distinguishing these two speciesis extremely difficult. The present study was conducted to developmolecular markers to distinguish C. wilfordii and C. auriculatumby using conventional polymerase chain reaction (PCR) and realtimePCR analyses. Comparative analysis based on the sequenceof the trnL-trnF intergenic spacer revealed 4 base-pair variations,and the inter-individual sequences of the 2 species separatelyshowed 100% homology. According to these results, the variationswere divided into 2 groups. The 2 species were further distinguishedusing a sequence-characterized amplified region marker developedbased on a randomly amplified polymorphic DNA-PCR product,and then a single nucleotide polymorphism marker was designedbased on the trnL-trnF intergenic spacer for more efficientdetection in real-time PCR. The results showed that speciesspecificmolecular markers might allow accurate discrimination ofC. wilfordii and C. auriculatum.
우리나라 6개 벼 품종의 흰잎마름병 저항성 유전자 분석
육진아,최재을,강희경,최춘환 한국식물병리학회 2004 식물병연구 Vol.10 No.1
The gene analysis of resistance in rice cultivars, Daeanbyeo, Hwasunchalbyeo, Daejinbyeo, Naepungbyeo, Hwajinbyeo and Surabyeo to strains of Xanthomonas oryzae pv. oryzae was studied. F1 plants and F2 populations from the crosses between six cultivars and near isogenic lines carrying the single bacterial blight(BB) resistance gene were analyzed using Korean and Japanese BB races. Daeanbyeo, Hwasunchalbyeo, Daejinbyeo, Naepungbyeo, Hwajinbyeo and Surabyeo are alleic with IRBB101 but are non-alleic with IRBB104 and IRBB105. The allelic tests indicated that Daeanbyeo, Hwasunchalbyeo, Daejinbyeo, Naepungbyeo, Hwajinbyeo and Surabyeo have the Xa1 gene for resistance.
안선민,육진아,김영화,채병찬,김홍준,김기훈,강권규,고병섭,이미영 韓國藥用作物學會 2006 한국약용작물학회지 Vol.14 No.3
형태학적으로 구분이 어려운 황정과 위유의 범위를 구분하기 위해 황정에 속하는 층층갈고리둥굴레, 다화황정, 진황정 등 3종과 위유에 속하는 8종의 둥굴레 동속 식물 등 11종 23 개체 시료로 RAMP분석한 결과를 요약하면 다음과 같다. 1. 황정그룹과 위유그룹은 NTSYS를 통한 유연관계 분석에서 55%의 유연관계를 나타냈으며, 층층갈고리둥굴레와 층층둥굴레는 81.75%로 분석에 사용된 둥글레 동속 식물 중 가장 높은 유연관계를 나타냈다. 2. 둥굴레 동속식물에 해당하는 층층둥글레는 황정의 약재인 층층갈고리둥굴레와 외부형태학적으로, 그리고 다형성패턴결과가 매우 유사하므로 황정의 약재에 속하는 것으로 사료된다. 3. 진황정은 위유와 외부형태적으로 유사하며, 유연관계분석에서도 위유의 그룹에 속하므로 위유의 약재에 대한 구분을 새롭게 할 필요성이 있다. Two herbal medicines of the Polygonatum genus, namely Polygonati Rhizoma and Polygonati Odorati Rhizoma, are difficult to distinguish from each other through exterior morphologic aspects. Furthermore, because the standard components for physiochemical distinction have not yet been standardized, the identification of these medicines through botanic taxology is based on genetic methods of random amplified microsatellite polymorphism (RAMP). For the RAMP evaluation, we used five sets of UBC microsatellite primers 811, 818, 834, 836, 842 and random primer M1. Although no specific band that could clearly distinguish Polygonati Rhizoma from Polygonati Odorati Rhizoma was found, 11 Polygonatum plants could be divided into two groups by this method. P. sibiricum and P. stenophyllum were classified to group I and the others were to group II. As P. sibiricum and P. stenophyllum were very similar in genetic and morphologic perspective, the results suggest that P. stenophyllum belongs to the Polygonati Rhizoma family.
RAPD 방법을 이용한 국내 수집 인삼 (Panax ginseng C. A. Meyer)의 다양성 분석
서상덕,육진아,차선경,김현호,성봉재,김선익,최재을 韓國藥用作物學會 2003 한국약용작물학회지 Vol.11 No.5
본 연구는 고려인삼 재래종의 유전적 차이를 RAPD marker로 평가하였다. 재래종은 우리나라 주요 인삼재배 단지인 괴산, 금산, 남원, 포천, 양주, 연천, 영주로부터 수집한 인삼에서 10개체를 임의 선발하여 사용하였다. 9개 지역의 90개체를 대상으로 RAPD분석을 한 결과 48개의 primer 중 OPA 7, OPA 13, URP 2, URP 3, UBC 3의 5개 primer가 재현성이 있고 재래종 내 개체간에도 다형성인 band를 보였다. 재래종 집단간보다 재래종 내에서 유전적 다양성이 낮았다. 재래종 집단 간 또는 재래종 집단내의 유전적 차이가 있다는 것은 이 집단들이 헤테로라는 것을 의미한다. 이러한 결과는 우리나라에서 재배중인 고려 인삼은 유전 자원의 혼합으로 헤테로라는 것을 암시한다. 선발된 5개의 primer를 이용하여 90개체를 집괴분석한 결과 국내 재래종 인삼은 5개군으로 그리고 3개의 아군으로 구분되었다. 국내 재래종 인삼은 유전적 변이가 크므로 인삼 육종을 위한 재료로 이용될 수 있을 것이다. Genetic differences among nine land races of Korean ginseng (Panax ginseng C. A. Meyer) were examined using RAPD markers. Land races of Korean ginseng were collected from nine regions in Korea: Cheongwon, Guesan, Geumsan, Namwon, Pochun, Yangju, Yeoncheon, Yeongju. Out of 48 RAPD primers tested, 5 primers (OPA 7, OPA 13, URP 2, URP 3 and UBC 3) produced remarkable bands which showing polymorphisms among evaluated collections. Lower levels of genetic diversity were in detected same land races than among other land races. Genetic differences within and among land races indicate heterogeneity. These results indicate that cultivated ginseng in Korea is heterogeneous. Genetic similarity matrices of RAPD profiles were generated via coefficients of variation and the data were processed by the cluster analysis (UPGMA). When 90 collections were evaluated using selected 5 primers, those were clustered to 5 and 3 subgroups. These differences in genetic variation between land races of Korean ginseng implied the potential source for further breeding of Korean ginseng.
RAPD를 이용한 고려인삼 육성계통의 유전적 다양성 분석
김진희,육진아,차선경,김현호,성봉재,김선익,최재을 한국약용작물학회 2003 한국약용작물학회지 Vol.11 No.2
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 4~5개 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. This experiment was conducted to evaluate the diversity and purity of the Korean ginseng (Panax gjnseng) lines developed by the pure line selection using RAPD markers. Four primer (OPA 19, OPM 11, URP 3 and UBC 98) out of the 48 primer tested produced band which showed within-line polymorphisms at least in one line. Within-line polymorphisms were detected in six lines by OPA 19, in four lines by URP 03, in five lines by OPM 11, and in one line by UBC 98 respectively. Five plants obtained from the commercial cultivar 'Cheonpung' were differentiated using the primers OPA 19 and OPM 11. Five plants obtained from the 'Yeonpung were differentiated using the primer OPM 11. Detection of within-line RAPD polymorphisms might be attributed to the fact that cross pollination appear in P. gjnseng and a long period of three to four years required to reach the reproductive stage thereby delay the process to homozygosity.