以樟樹及相思樹枝梢材培養牛樟芝之初期生長及三萜類成分分析

The Primary Growth and Triterpene Analysis of Antrodia cinnamomea Cultured on Branch-based Wood Substrates of Cinnamomum camphora (L.) Presl. and Acacia confusa Merr

李世豪(Shih-Hao Lee) ； 謝育玲(Yu-Ling Hsieh) ； 廖宇賡(Yue-Ken Liao)

《作物、環境與生物資訊》 9卷4期 (2012/12) Pp. 220-234

https://doi.org/10.30061/CEB.201212.0002

摘要

本研究以樟樹[Cinnamommum camphora （L.） Presl.; Cc]、相思樹（Acacia confusa Merr.; Ac）及對照組牛樟樹（C. kanehirai Hay.; Ck） 為介質培養牛樟芝（Antrodia cinnamomea T T Chang & W N Chou）。三種樹材皆取自15 y生以上立木側枝，經乾燥、去皮、打碎成木粉後調製成仿木塊（6 cm × 6 cm × 6 cm）作為培養介質。牛樟芝菌絲接種後的初期生長，以Ck仿木塊上菌絲之生長較Cc 與Ac仿木塊上菌絲為快，介質上菌絲生長達4 mon 時有原基（primordia）產生，並以Ac介質上數量最多（p＜0.05）。在平板培養基中繼代之菌絲體（接種源）於培養8 wk後，於菌落外圍採樣可觀察到扣子體（clamp connection）構造，證明接種之菌絲體是已完成質配（plasmogamy） 的雙核菌絲（dikaryotic hyphae），其於繼代培養條件中能持續維持，在菌絲接種9 mon 後生成的原基中仍持續出現。以高效液相層析（HPLC）分析三種樹種之木粉、生長3、6、9 mon 的樟芝以及其原基之萃取物，顯示Ck、Cc 與Ac 三種木質材料中皆無已知牛樟芝之三萜類成分，而所培養之牛樟芝皆可自行合成幾種羊毛甾烷型三萜：dehydroeburicoic acid、15α-acetyl dehydrosulphurenic acid、3β, 15α-dihydroxy-lanosta-7, 9（11）, 24-triene-21-oic acid、dehydrosulphurenic acid以及sulphurenic acid。原基中之成分雖與這些三萜組成分相近，但並無子實體中所含多種麥角甾烷型三萜類存在。本研究證實牛樟芝在樟樹及相思樹介質上培養9 mon之初期生長（扣子體之維持及原基之出現），以及其所含三萜類成分，與培養在牛樟木介質者尚無差異。依據成分分析結果，原基之出現應為菌絲體轉變成子實體之過渡階段。

關鍵字

扣子體 ； 栽培介質 ； 菌絲體 ； 原基 ； 三萜類

並列摘要

This study investigated in vitro growth of Antrodia cinnamomea TT Chang & WN Chou on wood substrates of Cinnamomum camphora (L.) Presl. (Cc), Acacia confuse Merr. (Ac) and C. kanehirae Hay. (Ck; control), which all collected from debarked branches of living trees (15+-y-old). These wood materials were air-dried, processed into fine chips, and then shaped into cubic cakes (6 cm × 6 cm × 6 cm). Preliminary observations showed that the hypha of A. cinnamomea grew slower on Cc and Ac cakes as compared with those on Ck cakes. The primordia of A. cinnamomea was formed on cultures grown for 4 mon, particularly on Ac cakes (p ＜ 0.05). The structure of clamp connection was developed on the mycelial culture of A. cinnamomea (the inoculants) incubated on medium plate for 8 wk, indicating that the mycelial inoculants were dikaryotic hyphae with completion of plasmogamy. Such characteristics were continuously identified during subcultures and were also observed in primordia after cultures being inoculated and grown for 9 mon. By high performance liquid chromatography (HPLC) analysis of wood powders, A. cinnamomea incubating cultures (3-, 6- and 9-mon old) and its primordia, it showed that these powder materials contained no triterpenes. However, the following compounds of lanostane-type triterpenes, dehydroeburicoic acid, 15α-acetyldehydrosulphurenic acid, 3β, 15α-dihydroxylanosta-7, 9(11), 24-triene-21-oic acid, dehydrosulphurenic acid and sulphurenic acid were detected in the cultures of A. cinnamomea and its primordia. Based on triterpene analysis and the monitoring of culture growth, including formation of clamp connection and production of primordia, the 9-mon-old A. cinnamomea cultures grown on Cc and Ac substrates showed no difference as compared with Ck cultures. However, due to the absence of major ergostane-type triterpenes, the development of primordia at this growth phase may just a transition stage prior to the formation of fruiting body.

並列關鍵字

Clamp connection ； Culture substrate ； Mycelia ； Primordia ； Triterpenoids