臺灣龍膽細胞懸浮培養之建立及培養條件之探討   全文下載

Studies on Factors Affecting the Establishment of Gentiana davidii var. formosana (Hayata) T. N. Ho Cell Suspension Cultures

龍膽（Gentianae Radix）首載於神農本草經，列為上品，為常用之苦味健胃消炎藥。近年來由於龍膽藥材市場需求量遠超出其自然繁殖量，使得野生資源瀕臨枯竭。省產優質生藥的開發，藉由細胞懸浮培養技術，進行藥用植物二次代謝物之生產，除具保育作用外，亦有助於臺灣製藥工業競爭力之提升，本研究乃進行臺灣龍膽[Gentiana davidii var. formosana （Hayata） T. N. Ho] 細胞懸浮培養之建立。試驗結果顯示利用含有0.2 mg／L kinetin 及1.0 mg／L NAA之MS （Murashige & Skoog, 1962）基本鹽類培養基培養臺灣龍膽莖段所誘導之癒合組織，經繼代培養在MS基本鹽類添加0.2 mg／L kinetin 、3% sucrose 及pH 4.2~5.2 之液體培養基，於2.33 μE‧m-2‧s-1光照、25 ± 1°C恆溫及80~100 rev／min 轉速下培養，可得一分散均勻，生長良好之懸浮細胞。本研究所建立之臺灣龍膽懸浮培養細胞，將有助於其二次代謝物龍膽皂?（gentiopicroside）及當藥皂?（swertiamarin）之生產及高產細胞系之篩選。"

Gentiana davidii var. formosana (Hayata) T. N. Ho (Gentianaceae), commonly known as long-dan in Chinese, is a perennial herb indigenous to Taiwan. It is distributed throughout the island, ranging from low to high elevations. The roots, which contain bitter-tasting secoiridoid glucosides, are used in traditional Chinese medicine. It is mainly used in the treatment of gastrointestinal tract diseases. Continuous collection of plant material from natural habitat has led to the depletion of Gentiana population. The purpose of this study was to establish the cell suspension cultures of Gentiana, which could be used for large-scale production of active principles such as gentiopicroside and swertiamarin. Callus was initiated by culturing stem explants of G. davidii var. formosana on Murashige and Skoog’s (1962) basal medium supplemented with 0.2 mg/L 6-furfurylaminopurine (kinetin) and 1.0 mg/L α-naphthaleneacetic acid (NAA). Fast-growing suspension cell cultures were established by subculturing the callus in MS basal medium (pH 4.2-5.2) supplemented with 0.2 mg/L kinetin and 3% sucrose. The cultures were incubated on an orbital shaker (80-100 rev/min) at 25 ± 1°C and low light intensity (2.33 μE•m-2•s-1). 龍膽（Gentianae Radix）首載於神農本草經，列為上品，為常用之苦味健胃消炎藥。近年來由於龍膽藥材市場需求量遠超出其自然繁殖量，使得野生資源瀕臨枯竭。省產優質生藥的開發，藉由細胞懸浮培養技術，進行藥用植物二次代謝物之生產，除具保育作用外，亦有助於臺灣製藥工業競爭力之提升，本研究乃進行臺灣龍膽[Gentiana davidii var. formosana （Hayata） T. N. Ho] 細胞懸浮培養之建立。試驗結果顯示利用含有0.2 mg／L kinetin 及1.0 mg／L NAA之MS （Murashige & Skoog, 1962）基本鹽類培養基培養臺灣龍膽莖段所誘導之癒合組織，經繼代培養在MS基本鹽類添加0.2 mg／L kinetin 、3% sucrose 及pH 4.2～5.2 之液體培養基，於2.33 μE•m-2•s-1光照、25 ± 1°C恆溫及80～100 rev／min 轉速下培養，可得一分散均勻，生長良好之懸浮細胞。本研究所建立之臺灣龍膽懸浮培養細胞，將有助於其二次代謝物龍膽皂苷（gentiopicroside）及當藥皂苷（swertiamarin）之生產及高產細胞系之篩選。