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RAPD Marker를 이용한 Zoysiagrass 수집계통과 종간교배 계통의 유전적 다양성 분석
金斗煥,毛淑淵,林星希 建國大學校 附設 農業資源開發硏究所 1997 農資源開發論集 Vol.19 No.-
ABSTRACT: RAPD markers were analyzed with zoysiagrass accessions and their hybrid lines in order to detect the genetic variation, and the following results were obtained.1. CTAB extraction method produced more and purer DNA than SDS method. RAPD reaction conditions were optimized as follows: 10ng template DNA, 270nM primer, 200μM each of dATP, dGTP and dTTP, 0.5μ unit dynazyme and 10x buffer brought to 15μl final volume with distilled water. The adequate annealing temperature was 36℃ and time was 30sec. Fifty cycles of amplification produced the best RAPD band patterns.2. Electrophoresis of PCR-products produced more polymorphic bands with polyacrylamide gel than with agarose gel. 3.The grouping patterns of zoysiagrass lines at dendrograms by RAPD markers were very different from the expected grouping pattern, and that was probably because of natural interspecific crossings for the long period. Therefore, the classification of the species within Zoysia genus will not be any longer meaningful and zoysiagrasses should be classified by the cultivar names or morphological characteristics such as leaf width.4. The zoysiagrass lines collected from Japan and Korea were belonged to the completely different groups at the dendrograms. The reason is expected to be the genetic drift due to geographical isolation.
캘러스 유기와 식물체 재분화를 이용한 무의 기내 대량증식
김유경,모숙연,최수빈,박한용 한국식물생명공학회 2023 JOURNAL OF PLANT BIOTECHNOLOGY Vol.50 No.3
Radish (Raphanus sativus L.), a root vegetable grown worldwide, is consumed in several ways. In the cross between parental lines to produce F1 seeds of radish, the problem of low purity may arise because of pollen contamination. Therefore, we aimed to establish conditions for callus induction and regeneration so that in vitro cultured plants could be used for the propagation of stock seeds. The most effective hormone combination containing various concentrations of 2,4-D, TDZ, and kinetin was selected for callus induction using radish hypocotyl, and the induced calli were transferred to two types of hormone media to investigate the optimal conditions for shoot regeneration of the callus. The combination of 1 mg/L 2,4-D + 0.05 mg/L kin was the most effective for callus induction of RA2 and RA10, 1 mg/L 2,4-D + 0.1 mg/L kin + 0.025 mg/L TDZ of RA4, and 1 mg/L 2,4-D + 0.2 mg/L kin of RA30. Shoot regeneration of the RA4 callus occurred in both shoot regeneration media, but the frequency was much higher in the 5H+1B medium (1 mg/L NAA + 0.1 mg/L 2,4-D + 1 mg/L IPA + 0.02 mg/L GA3 + 2 mg/L zeatin + 1 mg/L BA). For the in vitro micropropagation of radish, the conditions selected in this study can assist in the propagation and maintenance of stock seeds to produce F1 seeds.
RAPD분석에 의한 잔대와 더덕의 유연관계 비교 및 감별
이미영,모숙연,김두환,오승은,고병섭 韓國藥用作物學會 2001 한국약용작물학회지 Vol.9 No.3
50여개의 primer를 사용하여 잔대 Adenophora triphylla와 층층잔대 A. radiatifolia Nakai, 그리고 더덕 Codonopsis laceolata Trautv의 지역간, 속간의 차이점과 감별여부를 RAPD법으로 시행한 결과, 잔대와 층층잔대의 차이점은 거의 없었으며, 더덕의 지역차이는 0.889의 유전적거리를 나타내었다. 두 종(種)을 구별할 수 있는 특이 band로는 primer 357, 361, 363, 393 이었으며, 건조약재와 비교하였을 때 재현성이 확인되었고, 또한 잔대와 더덕의 건조약재를 각각 혼합시켰을 때 이를 구별할 수 있는 major band가 뚜렷이 나타나 혼용되어있는 건조약재에서의 감별이 가능함을 알 수 있었다.. Dried parts of the two species are difficult to distinguish morphologically, thus Codonopsis radix has been sold instead of Adenophorae radix in herbal medicine market. Therefore, this study was conducted to develop the genetic marker through the examination of the phylogenetic relationships between two Adenophora triphylla(Thunb.) A. DC. var. japonica Hara, two Adenophora radiatifolia Nakai, five Codonopsis lanceolata(Sieb. et Zucc)Trautv. using RAPD analysis. Fifty decarmer oligonucleotide primers were screened for the RAPD analysis, and four primers generated distinct RAPD markers specific to Adenophorae radix and Codonopsis radix. Based on the RAPD patterns, the genetic relationships between three herbal medicine were analyzed by UPGMA method. As a result, Adenophorae radix and Codonopsis radix were classified into two major subgroups on the basis of the genetic similarity coefficient. The specific RAPD patterns generated by the selected primers were reproducible from dried materials. Furthermore, the specific RAPD patterns were produced from the mixture of dried roots of A. triphylla and C. lanceolata. These results prone the usefulness of the RAPD analysis for the discrimination of pure materials from the mixtures of A. triphylla and C. lanceolata.