青脆枝(Nothapodytes foetida)組織培養,植株再生及喜樹鹼含量分析

Tissue Culture, Plant Regeneration and Camptothecin Production of Nothapodytes foetida

賴慧君(Hui-Chun Lai)

指導教授 ： 何錦玟

大同大學/工程學院/生物工程學系所/碩士(2005年)

摘要

取青脆枝(Nothapodytes foetida)無菌實生苗的葉片、下胚軸為培植體，培養在MS basal salts + 0.5 mg/l BA + 0.5 mg/l 2,4-D建立癒合組織。將葉片癒合組織轉移至MS basal salts + 0.1 mg/l picloram再生不定根，並含有0.059% DW CPT。實生苗頂芽再生植株的根段，培養在MS basal salts + 1.0 mg/l BA + 0.1 mg/l 2,4-D誘導得不定根，含有0.0156% DW CPT，每30天的繼代培養可得到鮮重增加1.67倍。
葉片、下胚軸誘導的癒合組織及根段培植體培養在MS basal salts + 0.5 mg/l TDZ可間接再生不定芽，不定芽經BA誘導芽體抽長，再以IBA誘導發根，得到小植株，循序馴化，可出瓶培養。
取實生苗頂芽再生植株的葉片，經MS basal salts + 1 mg/l BA + 2 mg/l 2,4-D培養5個月，可得到胚性癒合組織。胚性癒合組織轉移到MS basal salt medium不添加生長調節劑，培養10週後開始有體胚發生，少數體胚可正常發育成植株，但多數體胚形態不正常。

關鍵字

青脆枝 ； 組織培養 ； 植株再生 ； 體胚發生 ； 喜樹鹼

並列摘要

Callus were induced from leaf and hypocotyl explants of sterile seedlings of Nothapodytes foetida cultured on MS basal salts + 0.5 mg/l BA + 0.5 mg/l 2,4-D. Adventitious roots regenerated from leaf-derived callus on MS basal salts + 0.1 mg/l picloram, and accumulated 0.059% DW CPT. Adventitious roots induced from root segments on MS basal salts + 1.0 mg/l BA + 0.1 mg/l 2,4-D, accumulated 0.0156% DW CPT after 30 days culture, and biomass increased 1.67 fold by subculture routinely.
Adventitious buds formed directly from root explants and indirectly from leaf and hypocotyl derived calli on MS basal salts + 0.5 mg/l TDZ. Adventitious buds elongated on MS basal medium supplemented with BA, and rooting by subculture on medium contained IBA. After acclimation, plantlets survived after transferred to soil.
Embryognenic callus induced from leaf explant of regenerants cultured on MS basal salts + 1 mg/l BA + 2 mg/l 2,4-D after 5 months culture. Then transferred to MS medium without PGRs, after 10 weeks culture, somatic embryogenesis was observed. Only few somatic embryos developed into plantlets, the others were morphologically abnormal.

並列關鍵字

embryogenesis ； Nothapodytes foetida ； plant regeneration ； tissue culture

參考文獻

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