Establishment of multiplication protocol on medicinal plant Dysosma pleiantha (Hance) Woodson

本試驗目的在於探討藥用植物八角蓮利用微體繁殖的組織培養技術，建立其增殖系統，方法過程包含建立無菌培植體、癒傷組織誘導、芽體誘導、芽體增殖、發根與健化移植。癒傷組織之誘導以含有BA 0.2 mg.L-1+2,4-D 0.5 mg.L-1之培養基培養誘導效果最佳，誘導率達到88.2%；芽體誘導則以BA 1.0 mg.L-1+TDZ 0.1 mg.L-1之生長調節劑濃度培養基培養，芽體誘導分化率最高，達到33.2%；在芽體增殖方面，運用BA/NAA (0.1-2.0 mg.L-1/0.01-0.2 mg.L-1) 濃度以10:1比值之濃度梯度處理，發現以BA 2.0 mg.L-1+ NAA 0.2 mg.L-1濃度可以獲得最多為8.5個芽體；在發根試驗中，以1/2 MS+ IBA 0.5 mg.L-1 之培養基培養可以於第2週時誘導出根，誘導率約20%，於第4週時根誘導率達到100%；八角蓮在健化移植試驗中，使用發根6週以上之培植體，都可以順利移植，存活率均為100%。The purpose of this test is therefore to examine the use of the tissue culture technology called micropropagation for reproducing the medicinal plant Dysosma and building its multiplication system, which involves methods and processes including preparation of sterile explants, blastemal induction, multiplication, rooting, hardening and transplanting. For the induction of wound healing tissues, placing the petioles of the plant in a culture medium that contained BA 0.2mg.L-1+2,4-D 0.5 mg.L-1 produced the best inductive effect, with an induction rate up to 88.2%. For blastemal induction, incubation using a growth regulator that contained BA 1.0 mg.L-1+TDZ 0.1 mg.L-1 achieved the highest induced blastemal fractionation rate, up to 33.2%. For blastemal multiplication, it was found that treating with BA/NAA in a concentration gradient of 10:1 (0.1-2.0 mg.L-1/0.01-0.2 mg.L-1) produced the largest number of blastemas, with an average of 8.5. In the rooting test, incubation using a culture medium that contained 1/2 MS+ IBA 0.5 mg.L-1 was able to induce rooting in the second week with an induction rate of about 20%, which increased to 100% in the fourth week. In the hardening and transplanting test, explants that had grown roots for at least six weeks were used and all of them, could be successfully transplanted and had a survival rate of 100%.目次 摘要…………………………………………………………………i Abstract……………………………………………………………ii 目次…………………………………………………………………iii 表目次………………………………………………………………v 圖目次………………………………………………………………vi 縮寫表………………………………………………………………vii 前言……………………………………………………………………1 前人研究………………………………………………………………2 材料與方法……………………………………………………………8 (一)材料消毒…………………………………………………………8 (二)培養基配置………………………………………………………8 (三)癒傷組織誘導與增殖試驗………………………………………8 (四)癒傷組織之繼代培養……………………………………………9 (五)芽體誘導分化試驗………………………………………………9 (六)芽體增殖試驗……………………………………………………9 (七)發根………………………………………………………………10 (八)健化移植…………………………………………………………10 (九)培養環境…………………………………………………………10 (十)統計分析方法……………………………………………………10 結果……………………………………………………………………11 一、癒傷組織誘導與增殖試驗………………………………………11 二、不同部位培植體對癒傷組織誘導之影響………………………14 三、不同濃度組合之生長調節劑對芽體分化之影響………………16 四、不同濃度組合生長調節對芽體增殖之影響……………………22 五、不同培養基對發根率之影響……………………………………26 六、健化培養基………………………………………………………29 七、培植體出瓶存活率之觀察………………………………………29 討論……………………………………………………………………31 一、培植體誘導癒傷組織之研究……………………………………31 二、培植體芽體之誘導………………………………………………31 三、植物生長調節劑對培植體芽體增殖之影響……………………32 四、植物生長調節劑對培植體發根之影響…………………………33 五、健化培養基對八角蓮小苗出瓶存活率之影響…………………34 結論……………………………………………………………………35 參考文獻………………………………………………………………36 附錄……………………………………………………………………4