枸杞子 體細肥雜種 合成을 위한 生物工學的 基礎硏究 = A Study on the Basic Information for Somatic Hybrid of Lycium chinense through Biotechnology

다국어 초록 (Multilingual Abstract)

The present study was carried out in order to develop techniques for rapid mass propagation and afford the basic information for somatic hybrid of Lycium chinense, used commonly as landscape purpose and medicinal resource.
Through biotechnology, bud culture, callus culture, protoplast isolation and culture, fusion and gene transformation were examined. Results obtained were summarized as follows ;
1. Plantlets were developed from axillary buds attached to the stem of the current year growth collected in early June and cultured on MS media supplemented with 1.0㎎/l IAA and 2.0㎎/l Kinetin by 56% as the highest yield rate.
2. The highest rate of shoot induction was resulted at 0.5㎎/l kinetin, root induction at MS + 1.0㎎/l IAA + 2.0㎎/l kinetin (56%).
3. The rate of plantlets formation supplemented with 1.0㎎/l IAA and 2.0㎎/l kinetin was the highest and WPM (60%), MS·SH (50%), B_5 (40%) and GD (30%) media gave the result in descending order.
4. Plantlets (10-12 ㎝ of length) survived without fail after being transferred to pots with a mixture of perlite : vermiculite : peat (1:1:1 v/v/v).
5. The leaf collected from in vitro cultured plants cultured on MS media without growth regulators supply gave 60% callus formation and 40% shoot induction.
6. The highest protoplast yield and viability were resulted in the treatment plot of 3.0% macerozyme, 1% cellulase R-10, 0.4M mannitol, pH 6.2 and incubation for 6 hours. And the more effective was at the more juvenile leaf tissue. When protoplasts were cultured at 5x10^4/ml density in MSP_19M medium, the first budding occured at 18 days after culture and 28 days later cell colony was formed.
7. The highest cell fusion yield was 29.6% between leaf cells and 5.1% between leaf and root cell, using 30% (W/V) polyethlene glycol (PEG) concentration, 10 minutes treated and 2x10^5/ml protoplasts density from seedlings in virto.
8. When leaf was inoculated with Agrobacterium tumefaciens ATCC 11157, tumour was formed and rooting was initiated 8 days after inoculation and the growth was continued for 3 weeks. And the rate of tumour formation was 10%. The genetic transformation was confirmed through the opine analysis by the electrophoration.

목차 (Table of Contents)

• Ⅰ. 緖 論
• Ⅱ. 硏 究 史
• Ⅲ. 材料 및 方法
• Ⅳ. 結果 및 考察
• 1. 芽培養, Callus培養
• 2. 原形質體分離 및 培養
• 3. 細胞融合
• 4. 形質轉換
• Ⅴ. 結 論
• · 引用文獻
• · SUMMARY