작약(芍藥)의 약(葯)으로부터 분리된 화분소포자(花粉小胞子)를 생장조절제가 첨가되지 않은 MS 배지에 배양하였을때 직접 배(胚)발생 과정을 거쳐서 소포자 유래의 배(胚)가 형성되었고, 1mg/l의 PAA가 첨가된 배지에서는 화분소포자 유래의 캘러스가 형성 되었으며 이들 캘러스를 생장조절제를 함유하지 않은 배지에 이식하였을 때 배양 7주 후에 배(胚)가 형성되었다.

Glucosinolates (GLS) are secondary metabolites commonly occurring in Brassica crops and more than 130 different GLS have been reported in diverse plants. Recent studies have indicated that isothiocyanate (ITC) derived from GLS by hydrolysis had a potential for anticancer activity against several rumor cells on human. In addition, it was found that glucoraphenin (GRE) and glucoraphasatin (GRH) were abundant and differently regulated in radish plant, depending upon organs and developmental stages. Microspores isolated from flower buds of radish were cultured in vitro to obtain doubled haploid (DH; but homozygous) lines in a short time period. The present study was conducted to determine the concentration of GRE and GRH, an immediate precursor of ITC from DH lines of radish plant. Total 41 DH lines were selected based on flow cytometry analysis. The seeds, obtained by bud pollination from the DH lines, were planted and 3-weeks-old young seedlings were used for the major aliphatic GLS analysis. Amounts of GRH were highly variable from the DH lines ranging from 2.3 to 31.5 mg·g-1 dry weight (DW). The donor plant (DP) contained 18.4 mg·g-1 DW. It was noticed that there were 6-fold differences in the amounts of GRE between the highest and lowest DH lines. Among 41 lines tested, 14 DH lines of radish plant were significantly reduced in the amount of sum of GRH and GRE compared those of the donor plant (P<0.05), whereas only three lines increased. The results obtained in the present study will lend to select genotypes with low and high GLS contents of radish plant. In addition, those DH lines will aid to elucidate a biosynthetic pathway of the aliphatic GLS in radish plant, which remain for the most part unsolved.

국내 육성 유채 품종의 소포자 배양을 통한 효율적인 소포자배 생산법의 확립을 위해 '탐미유채'를 대상으로 소포자 배양 시 적당한 꽃봉오리의 크기, 고온처리시간, 배양밀도, 기내 증식조건 등의 배양조건을 구명하고자 실시한 결과, 소포자배 발생은 1핵기말과 2핵기 초기 상태의 소포자가 포함된 3.0~3.5 mm 크기의 꽃봉오리에서 채취한 소포자에서 배발생률이 가장 높았으며, 배지 1 mL 당 약 388개의 소포자 배가 발생하였다. 배양 초기에 32.5℃ 에서 2일간 열처리하였을 때 배발생률이 가장 높았다. 소포자의 배양밀도에 따른 배발생은 5×104 개/mL일 때 가장 높았으며, 배양밀도가 더 이상 높아지면 배발생을 억제하였다. 발생한 소포자배는 0.5mg~cdotL1 NAA와 1mg~cdotL1 BA가 첨가된 MS 고체배지에 치상하여 배양하면 정상적인 신초로 재분화되었고, 발달한 신초를 절취하여 식물성장 조절제가 첨가되지 않은 MS고체배지에서 배양하면 뿌리가 발달하여 정상적인 식물체로 성장하였다.

For genetic mapping studies, biparental segregating populations are often useful, however recombination is limited, giving rise to large genomic regions under QTL, and one can only study alelles present in both parents. In Wageninegn UR, a core collection is being developed representing all Brassica rapa morphotypes and geographic origins. As most B. rapa accessions are heterozygous and heterogeneous, we started a project to fix the collection through microspore culture. The resulting Diversity Fixed Foundation Set will be an interesting resource for association mapping studies, which have as advantage that they present the allelec variation present in the collection, and for mapping studies recombination is increased. Nineteen accessions of eight subspecies of Brassica rapa were used for microspore culture to developdoubled haploid lines. Eight morphotypes were represented: 3 Chinese cabbage, 2 Chinese turnip cabbage, 3 Pak choi, 5 Turnip, 3 Broccolleto, 1 Mizuna, 1 Komatsuna and 1 Turnip greenfrom the 19acessions examined, embryos were obtained for 13, representing six subspecies (Komatsuna and Turnip Green had no response). The embryo yields differed significantly between these 13 accessions. We regenerated normal plants from 10 accessions that survived more than 4 weeks in the soil using microspore culture. Nine accessions flowered after 4 weeks vernalization at 5℃ and seeds were harvested from 5 accessions. From a Mizuna, we obtained 3791 seeds from one plant and total 7318 seeds were harvested from 5 accessions representing 4 subspecies(Chinese cabbage, Chinese turnip cabbage, Pak choi, Mizuna). At present, we carry out experiment for obtain more seeds and induce embryos from the other plant materials.

국내육성 유채품종 간 반수체식물 생산성을 비교하였다. 화아로부터 분리된 소포자를 13%의 sucrose, 0.05mg/ BA 와 0.5mg/ 의 NAA가 첨가된 NLN 배지에서 배양하였다. 반수체 배의 생산에는 유전형이 중요한 요인이였으며, 탐라유채가 가장 높은 반수체 배 생산능을 보여 화아당 176개의 반수체 배가 발생하였으나, 한라유채와 영산유채는 배 생산은 물론 세포분열도 관찰되지 않았다. 발생한 반수체 배를 NLN배지에서 현탁배양하여 Multilobe abnormal embryos를 형성시켰으며, 계속하여 생장조절제가 첨가되지 않은 MS고체 배지에 치상 배양하여 반수체식물체를 유도 하였다. 재생된 반수체 식물체는 성공적으로 순화되었다.

An efficient system of rice microspore culture could contribute to the production of genetically modified rice. The microspores were isolated by mechanical or shed methods. The number of microspores per 100 anthers isolated at uninucleate stage was higher than (or similar to) those at binucleate stage in isolation method with pestle or spatular, but microspore divisions were not easily observed on both stages. On the other hand, pollen division in shed pollen culture was observed more frequently at uninuclear than at binuclear stage. Cold pretreatment at 10~circC for 10 days resulted in the best multicellular division to produce microcalli at 12.5% efficiency in shed microspores. Heat shock at 33~circC for one hour before or after pollen shedding enhanced cell division and callus formation. Out of twelve green regenerants, two were haploids and ten were diploids based on the chromosome analysis of root tips. The size of stoma was 12m m in haploids and 15 ~mu~textrmm in diploids determined by scanning electron microscope (SEM).