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研究生:張肇麟
研究生(外文):Chang, Chao-Lin
論文名稱:臺灣雲杉體胚之誘導
論文名稱(外文):The Induction of Somatic Embryo of Picea morrisonicola Hayata
指導教授:王亞男王亞男引用關係姜家華姜家華引用關係---
指導教授(外文):Wang Ya-NanChiang Chia-Hua
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:森林學系
學門:農業科學學門
學類:林業學類
論文種類:學術論文
論文出版年:1998
畢業學年度:86
語文別:中文
論文頁數:49
中文關鍵詞:體胚臺灣雲杉胚原性胚柄細胞團
外文關鍵詞:Somatic EmbryoPicea morrisonicolaembronal suspension mass
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以台灣雲杉(Picea morrisonicola Hayata)成熟胚為培植體進行培
養。將成熟種子經48小時流水處理,以70%酒精浸泡2分鐘及浸
在4% NaOCl水溶液(含約1%(v/v) Tween 20展著劑)中超音波震
盪10分鐘,污染率為零。取成熟胚進行胚培養,於每一個50 ml
三角錐形瓶中的20 ml固體培養基上放置10個胚。在全光照及黑
暗的環境中培養,產生癒合組織的不同:全光照下誘導出綠色、
緊實之癒合組織,在黑暗下則為黃色、鬆軟之癒合組織。將黃色、
鬆軟癒合組織培養於添加2ppm 2,4-D、1 ppm BA的MS培養基
中,在黑暗環境下經過四個月,可誘導出白色黏質的胚原性癒合
組織,其含有具胚性端及胚柄細胞的前子葉期體胚;最後將胚原
性癒合組織移至添加0.1ppm2,4-D、1ppm BA與2 ppm ABA的MS
固態培養基中,經過兩個月全光照的培養,則可誘導出綠色短軸
之子葉期體胚。
Results of mature embryo culture and somatic embryo induction of Picea
morrisonicola Hayata were as follows: mature seeds were first treated with
running water for 48 hours, then soaked in 70% ethanol solution for 2
minutes. Soaked in 4% NaOCl (supplemented 1% (v/v) Tween 20) and
treated with ultrasonic shaker for 10 minutes. Mature embryos were them
isolated from the seed and 10 mature embryos were transferred into 50 ml
conical flask containing 20 ml MS solid culture medium. Callus with
different phenotype were induced after they were culture under light and
dark conditions respectively. Under light conditions, green and compact
callus were formed, but yellow and soft callus were formed under dark.
After yellow and soft callus were transferred to medium containing 2 ppm
2,4-D and 1 ppm BA, and cultured under dark conditions for 4 weeks,
white and sticky embrygenic callus were induced. These callus contained
precotyledonary phase somatic embryos with embryonal head and
suspensor . Cotyledonary phase somatic embryos with green and short axis
were induced after these callus were transferred onto MS solid medium
supplemented with 1 ppm 2,4-D,1 ppm NAA and 2 ppm ABA and cultured
at light conditions for 2 months.
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