台灣金線連之組織培養及其有效成分天麻素之分析

Abstract

台灣金線連（Anoectochilus formosanus Hayata）為我國民間常用的傳統藥材，習用於心血管疾病之治療，開發成保健食品有其必要。利用組織培養繁殖台灣金線連之藥材，以GAP為基礎，進行相關研究刻不容緩。本研究主要探討：一、利用組織培養大量繁殖台灣金線連之培養基中，基本鹽類濃度及馬鈴薯泥及香蕉泥等有機添加物之最適合濃度；及台灣金線連在組織培養系統中最適當之繼代培養次數。二、組織培養瓶苗於溫室栽培後之最適採收時機。 研究結果顯示台灣金線連之芽體培養於含有添加30 g/l 馬鈴薯泥及50 g/l 香蕉泥之1/2 MS培養基三個月後，生長效果最佳。雜交和繼代培養次數試驗中發現，F3和F4世代之繼代培養出現0.8-33%之白化苗；且隨著繼代次數的增加植株於溫室栽培之存活率也有顯著下降。 台灣金線連之組織培養苗定植溫室初期，植株內各元素含量皆高於野生金線連植株，隨著栽培時間增加，各元素含量有遞減的趨勢。於溫室栽培六個月後，植株生長趨減緩，八個月後植株生長幾乎呈現停滯狀態，此時溫室栽培植株內之營養元素含量與野生金線連較為接近。在重金屬檢測方面，溫室栽培植株含量較野生金線連植株低。 在成分分析上，運用薄層色層分析（TLC）與高效液相層析（HPLC）分析天麻素含量，結果顯示溫室栽培一個月後之含量最高，栽培三個月含量最低，栽培八個月後之含量與野生金線連最為接近。

Anoectochilus formosanus Hayata (Orchidaceae) belongs to a group of terrestrial orchids commonly known as “Taiwan Jewel Orchids”, because of their attractive foliage. A. formosanus, one of the nourishing and health drugs, is a popular folk medicine that possesses antioxidant, anti-inflammatory and liver-protective effects and has been used to cure hepatitis, hypertension and cancer in Taiwan. The goal of this study was to test the influence of various strength of MS basal salts and organic additives such as banana pulp and mashed potato on the in vitro shoot growth of A. formosanus. The experimental results indicated that medium containing 1/2 MS basal salts and a mixture of mashed potato (30 g/l) and banana pulp (50 g/l) was found to be the best combination for the growth of lateral buds and plantlets of A. formosanus. The growth of the shoots reduced at each subculture and the generations C3 and C4 resulted in 0.8-33% albino plants. Variation in the survival rate was also observed in these tissue-cultured plants transferred to peat moss and cultured in greenhouse for eight months. Greenhouse-grown plants at earlier growth stage contain higher amounts of micronutrients. After 8 months of transplantation, the contents were found to be close to those of wild type. Heavy metal concentrations in plants grown in greenhouse were also determined and showed lower than those of wild type. Gastrodin, an active phenolic ingredient of A. formosanus have been quantitatively estimated using High Performance Liquid Chromatography (HPLC). Maximum production was observed in one-month-old greenhouse-grown plants. After eight months, the amount of gastrodin in greenhouse grown plants was found to be similar to that of wild type.