喜樹癒合組織之培養與喜樹鹼含量分析

Callus Culture and Camptothecin Production of Camptotheca acuminata

張淑華(Shu-Hwa Chang) ； 蔡錦瑩(Jin-Yin Tsay) ； 何政坤(Cheng-Kuen Ho) ； 黃芷雲(Chih-Yun Huang)

《台灣林業科學》 21卷4期 (2006/12) Pp. 513-521

https://doi.org/10.7075/TJFS.200612.0513

摘要

以喜樹下胚軸與2個月大試管種子苗之葉片，建立癒合組織誘導與培養方法，結果兩者癒合組織生長最佳的培養基均為WPM基本培養基添加1~3mg L^(-1)NAA或2,4-D。唯NAA所誘導的癒合組織較硬，而2,4-D誘導的癒合組織較為鬆軟。分析19種來自下胚軸、葉片、莖段之3個月大癒合組織的喜樹鹼(CPT)與10-烴基喜樹鹼(HCPT)，發現CPT含量與材料來源無關，而HCPT只存在於莖段癒合組織，兩個化合物在不同細胞系間差異很大，CPT含量最高為0.0307%（細胞乾重），HCPT最高為0.0021%，最低則都趨近於0。調查6個經3年長期培養於NAA與2,4-D的癒合組織之喜樹鹼含量，發現只有1個培養於NAA的癒合組織，在培養的3年裡，可穩定維持生產喜樹鹼，另外有4個癒合組織的喜樹鹼含量會逐漸下降，尤其是培養於2,4-D之癒合組織幾乎完全喪失了生產喜樹鹼的能力，本研究結果顯示植物生長激素的選擇與細胞株的篩選，對利用細胞培養生產喜樹鹼而言，非常重要。

關鍵字

喜樹 ； 癒合組織 ； 喜樹鹼

並列摘要

Callus cultures of Camptotheca acuminata Decne. were initiated from hypocotyl and leaf explants excised from 2-mo-old plants in vitro. The optimal culture medium for callus growth for both explants was woody plant medium (WPM) containing 1~3mg L-1 NAA or 2,4-D. The calli appeared compact in NAA medium, but friable in 2,4-D medium. Comparing HCPT (10-hydroxy camptothecin) and CPT (camptothecin) contents of calli derived from 19 sources including the hypocotyls, leaves, and stems of different donor plants, we found that CPT existed in all 3 organ-derived calli but HCPT was present only in stem-derived calli. Both CPT and HCPT contents in different callus lines varied greatly. CPT and HCPT contents in a superior line were 0.0307% and 0.0021%, respectively, while those of the poor lines were close to 0. Investigating the stability of CPT content in long-term cultures, 6 calli subcultured monthly in NAA and 2,4-D media for over 3 yr were analyzed once every 6 mo. Only 1 callus line cultured in NAA medium maintained a stable CPT content of between 0.01% and 0.014%. Four lines gradually lost the ability to produce CPT, especially those cultured in 2,4-D medium. This indicates that the type of auxin used and the callus line are very critical for CPT production via cell culture.

並列關鍵字

Camptotheca acuminata ； callus culture ； camptothecin