Cell Suspension and Protoplast Isolation of Eucalyptus macarthuii and E. radiata

毛皮桉及放射桉之細胞培養與原生質體分離

張淑華(S. H. Chang) ； D. G. M. Donald

《林業試驗所研究報告季刊》 10卷4期 (1995/12) Pp. 501-511

https://doi.org/10.7075/BTFRI.199512.0501

摘要

本研究已成功的建立了毛皮桉(Eucalyptus macarthurii)及放射桉(E. radiata)的懸浮細胞培養，並可經定期的繼代培養維持一年以上。放射桉的懸浮細胞呈現均勻的游離細胞，而毛皮桉則呈細胞團聚集，並隨著培養期間加長而增大經過一年繼代培養後，利用細胞鮮、乾重及pH值測定培養細胞的生長週期，發現此二樹種細胞之生長週期從開始的停滯期(lag phase)到最後的靜止期(stationary phasc)約持續1月。培養基的pH值可用來測定細胞生長週期中的停滯期與指數期(exponential stage)。二樹種由懸浮細胞分離原生質體的產量差異頗大，毛皮桉爲4.7×10^5/mL，放射桉為6.9×10^6/mL。最佳酵素解離液的組成因樹種而異。來自放射桉懸浮細胞之原生質體培養，可觀測到原生質體分裂及細胞壁的形成，唯經培養1個月後，並未再進一步的發展而漸死亡。

關鍵字

細胞培養 ； 原生質體分離 ； 毛皮桉 ； 放射桉

並列摘要

Cell suspension cultures of Eucalyptus macarthurii and E. radiata were established and maintained by regular subculturing for more than one year. Suspension cells of E. radiate appeared as fine cells in the culture medium while those of E. macarthurii formed cell aggregates of increasing size throughout the culture period Cell growth cycles after for subculturing for one year were determined by fresh weight, dry weight, and pH valucs of the culture medium. Cell growth of both species continued for a month between the lag and stationary phases of the growth cycle. The pH value of the culture medium could be used as a method for determining the lag and exponential stages of the growth cycle. The highest yield of the two eucalypts from cell suspension cultures differed: 4.7×10^6/mL for E. macarthurii and 6.9×10^6/mL for E. radiata. The optimal composition of the enzyme mixture depended on the species. Cell wall formation and division of protoplasts derived form E. radiata suspension cells were observed. However, division did not progress beyond this stage and the protoplasts died after one month.

並列關鍵字

cell suspension ； protoplast ； E. macarthurii and E. radiata