黃世佑2006-07-252018-06-282006-07-252018-06-282003-05-29http://ntur.lib.ntu.edu.tw//handle/246246/9341植物體胚發生(somatic embryogenesis)為利用其全潛能性 (totipotency)於體細胞誘導不定胚之技 術, 由於其為人工種子的關鍵之一, 故近年來甚受重視。本研究以芹菜 ( Apium graveolens L .)葉及莖為培植體, 利用SH 培養基誘導癒合組織(含2,4-D 7.5mM)及繼代培養(含2,4-D 1mM, kinetin 1mM), 其中以葉誘導之 癒合組織LC1 於第28 天鮮重平均生長 率為0.39(1/day);以莖誘導之癒合組織 SC1 於第28 天鮮重平均生長率為 0.56(1/day)。懸浮細胞培養於第8 天沉 降體積(SCV)增加一倍。利用回應曲面 法(Response surface method)針對蔗糖 濃度、還原氮濃度、初始pH 值三個因 子, 探討固態培養癒合組織最佳生長 條件。一階RSM 實驗結果顯示蔗糖 2%, 還原氮(NH4 + ) 300 ppm, 初始pH 6.2 的組合有較高的鮮重生長率(0.32 1/day,於第16 天), 而乾溼重比隨蔗糖 濃度增加而上升。未來幾個月內將從 事篩選胚性癒合組織及探討體胚發生 率, 利用部分因子設計, 回應曲面法, 以初始pH, 還原氮濃度, 碳源及培養 液黏度等因子, 尋求最佳體胚誘導條 件, 以作為未來生物反應器培養的依據。Somatic embryogenesis has been receiving much attention due to its inherent tortipotency and favorable in inducing elite plant species and mass production of the plants. Artificial seed production via somatic embryogenesis is the future prospect in breeding. The aim of this work is to investigate the key factors for developing somatic embryos and elaborating the bioreactor configuration suitable for mass production of the embryo. Celery ( Apium graveolens L .) was adopted as a model system. The work began with inducing a callus from leaves with SH medium containing 7.5 mM of 2,4-D, and subcultured with the same medium containing 1 mM of 2,4-D and 1 mM kinetin. The callus obtained from the leaves (LC1) exhibited an Average Growth Rate (AGR) of 0.39 day -1 on day 28. The AGR was the objective function of the optimization. While that obtained from stem (SC1) showed the AGR of 0.56 day -1 on day 28. Embryogenic suspension culture for 8 days resulted in a doubling in settling cell volume (SCV). As a preliminary optimization of culture condition of callus induction, Response Surface Method was employed to search the optimum condition for obtaining the AGR and Dry wt./Fresh wt. Ratio. The result showed that under 2% of sucrose, 300 ppm of NH4 + , and the initial pH of 6.2, the highest AGR(0.32 day -1 ) was obtained (based on 16th day). The other objective function, Dry wt./Fresh wt. increases with increase of sucrose concentration. In coming months, the work will be focused on the screening of embryogenic callus as well as the efficient regeneration rate of embryos. The optimum conditions for inducing somatic embryos will be searched by employing fractional factorial design/ Response Surface Mmethod, taking initial pH, concentrations of NH4 + and sucrose/mannitol and viscosity as the independent variables. The results will throw light on the operation of bioreactor.application/pdf591806 bytesapplication/pdfzh-TW國立臺灣大學化學工程學系暨研究所胚細胞芹菜再生擴大規模生物反應器高濃度細胞胚embryogenic cellsceleryregenerationbioreactorhigh density embryogenic cellreporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/9341/1/912214E002034.pdf