利用組織培養大量繁殖雙鋸齒葉玄參

Abstract

本試驗以台灣產雙鋸齒葉玄參為供試材料，希能利用組織培養大量繁殖，開發成 為台灣原產優質生藥，以提供市場需求。結果發現，將野生雙鋸齒葉玄參及無菌苗之莖項、 莖段，莖節及葉等組織切取為培植體進行不定芽的誘導，其中以無菌苗之莖項和莖節誘導不 定芽的能力較佳。MS基本鹽類添加1~2mg/l BA及0.2 mg/l NAA之培養基，誘導不定芽的 能力最高，每個培植體經兩個月的培養可產生7.5個不定芽，但約有80%的不定芽產生玻璃 質化，且玻璃質化現象隨著cytokinins濃度的提高而增加，因此玻璃質化為大量繁殖中急需 解決的問題。不定芽的生長高度以不含植物生長調節劑之MS基礎培養基最高，隨著cytokinins 濃度的增加而芽高度降低。不定芽之發根，則以全量MS固體培養基添加0.5 mg/l NAA或IBA 最佳。將健康已發根的無菌苗移植到蛭石：泥炭土：田土=2:1:1之介質中其存活率可達80%； 玻璃質化苗則全數死亡。利用HPLC測定市場品、野生品地上部、野生品地下部皮組織培 養苗四種材料之主成分harpagoside含量，結果以野生品地上部最高，野生品地下部及組織 培養苗次之，且兩者間無顯著差異，而市場品最低。雖然台灣雙鋸齒葉玄參非市場品（本經正品），但有效成分較市場品高，因此應可開發為本省優質生藥。 The purpose of this study was to establish the tissue culture techniques for mass propagation of a Chinese medicinal herb Scrophularia yoshimurae Yamazaki which is indigenous to Taiwan. It was found that adventitious buds could be induced from the excised explant directly. The capability of forming adventitious buds from various pant tissue of field grown and in vitro plants was observed. Shoot tip and node of in vitro plant showed the highest ability to form adventitious buds. In average, 7.5 adventitious buds/node explant produced on MS basal medium supplemented with 1-2 mg/l BA and 0.2 mg/l NAA after 2 months in culture. About 80% adventitious buds produced from node were vitrified shoots in this medium. The higher concentration of cytokinin increased the percentage of vitrified plants markedly. All vitrified plants could not survive after being transplanted into soil. Vitrification has then become an urgent problem which needs to be solved in tissue culture propagation of Scrophularia yoshimurae. The adventitious buds showed highest plant height when buds produced from node were cultured on hormone free medium for adventitious buds producing root was basal medium supplemented with 0~0.5 mg/l NAA or IBA. The survival rate was 80% when the health plantlets produced in vitro were transplanted into a mixture compounds of vermiculite: peat moss: soil=2:1:1. Harpagoside was detected from various species and tissue by HPLC. The results indicated that the stem and leaf of wild species had the highest harpagoside content, followed by root of wild species and plantlet, its lowest was market spcies. Although scrophularia yoshimurae wasn't market species, its harpagoside content was higher than that of market species. It is suggested that Scrophularia yoshimurae could be considered as to a high quality Chinese medicinal herb in Taiwan.