頁籤選單縮合
題 名 | 茶樹組織培養的研究 |
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作 者 | 吳振鐸; | 書刊名 | 中華農學會報 |
卷 期 | 93 1976.03[民65.03] |
頁 次 | 頁30-42 |
關鍵詞 | 茶樹; 組織培養; |
語 文 | 中文(Chinese) |
中文摘要 | 植物組織培養法的發展甚為迅速,近年來國內外學者對於蘭花、蕃茄、甘蔗、菸草、香蕉、馬鈴薯以及水稻等草木植物的組織培養已有突出的成就與貢獻。至於木本植物之茶樹的組織培養法,尚鮮有報導。茶為多年生的木本植物,育種所需年限極長,若能利用組織培養,不獨較易增加遺傳值的變異,可望獲得新異的育種材料,且亦有助於優良品種之大量迅速育苗。故作者於二年前(1973年5月)開始以學習的態度從事本研究工作,歷時年餘,至本年(1975)1月始於祁門種子葉之癒合組織中經6~8次的移植,培養出新的植株,至目前為止前後已由祁門種及平水種之子葉癒合組織中培養出28株,這一批小植株是農藝植物界,利用組織培養法獲得成功的第一批小茶樹,目前除衰弱者死亡外,尚有健壯者十二株正在土壤盆中栽培,觀測其生長習性及植株間之外部型態等之變異。茲將茶樹組織培養法研究所得之要點摘列於後,並請參閱表1至表5,圖A1至圖B8,以作今後進一步研究之參考。 (一)材料選擇:初熟時的子葉較易形成癒合組織,幼莖次之,幼芽與幼根較為困難。 (二)誘發癒合組織培養基之調製:無機鹽類參照MURASHIGE-SKOOG 的配方。有機物質的濃度為IAA 2~4 ppm, kinetin 2~4 ppm, 2,4-D 2~8 ppm, myo-inositol 100~200 ppm, glycine 20-40 ppm, nicotinic acid及pyridoxine-HCl 各5~10 ppm, thiamine-HCl 1 ppm coconut milk 10%、sucrose 3%、yeast extract 0.2% agar 1%。 (三)培養基的pH值以調節至5.0最適當。 (四)培養基經1.2kg/cm2之蒸汽壓力,120℃滅菌10分鐘,接種後將培養試管置於28℃之定溫箱內,於黑暗狀態下經10日至二星期的培養,?可誘發癒合組織的形成。 (五)癒合組織的繼代培養:培養基中之無機鹽類仍用MS的配方,有機物質變更為下列配方最有利生長。?IAA 4 ppm, kinetin 2 ppm, 2,4-D 4 ppm, myo-inositol 100 ppm, glycine 20 ppm, nicotinic acid及pyridoxine-HCl 各5 ppm,thiamine-HCl 1 ppm coconut milk 10%、sucrose 3%、yeast extract 0.2%、 agar 1%。 (六)誘導器官分化之培養: 1. 同前培養基中不含2,4-D,在溫度28°~30℃,光強1,700 Lux之生長燈下,每日光照10小時,約經10日?開始有葉綠素的形成。經二個月更換新配之同一培養基繼續培養。 2. 為誘導根及莖葉的分化,培養基中無機鹽類照舊,有機物如下修訂:kinetin 10 ppm, IAA 1 ppm, myo-inositol 100 ppm, nicotinic acid及pyridoxine-HCl 各0.5 ppm, thiamine-HCl 0.1 ppm, glycine 2 ppm, sucrose 3%、yeast extract 0.2%、 agar 1.0%、每經二個月更換新配之同一培養基一次,繼續培養至根及莖葉之誘導分化形成。 (七)分化小植株的培養:將光照增至3,400 Lux,仍在瓶中無菌培養至生長4~5葉時,再移出置於含有至石及水草的盆中栽培,經2~3個月,俟根部健壯再移至土壤盆中栽培,使其漸次適應自然環境,至生長健壯後移於田間栽培。 |
英文摘要 | This experiment was carried out to study the suitable culture media and conditions for the callus fromation, growth and differentiation of tea plant. The author indued callus from segments of the young cotyledon of CAMELLIA sinensis var. sinensis in May 1973. After one and half years and seven times of subculture, the first plantlet was differentiated from the callus on Jan. 18, 1975 (Fig. A5). Untill present, there are 12 plantlets have been derived from the cotyledon calluses of Chyi-men and Pyng-shoei varieties of tea plant, (Figs. A10 and B6). The TTETC No. 3 of the earlist one was ten months old and 31 cm tall (Fig. A9). All of them were transplated from vermiculite to soil on sept. 19, 1975. The morphological characters of the 12 plantlets after measurement are presented in table 5. The results of this experiment are summarized as follows: 1. The young cotyledons without epicotyl (Fig. A1) were easier for the induction of callus than young stem (Fig. A2), shoot tips and root tips. 2. Callus was induced from the yound cotyledons of Chyi-men (Fig. A4) and Pyng-shoei (Fig. B1) varieties of tea plant by using Murashige and Skoog mediom (major and minor elements) containing IAA 2-4 ppm, kinetin 2-4 ppm, 2,4-D 2-8 ppm, myo-inositol 100-200 ppm, glycine 20-40 ppm, 5-10 ppm each of nicotinic acid and pyridoxine-HCl 1 ppm, thiamine-HCl 1 ppm, coconut milk 10%, sucrose 3%, yeast extract 0.2% agar 1%. 3. The optimum pH value for the callus induction and growth was 5.0. 4. Callus was initiated after 10-14 days cultured at 28℃ under darkness. 5. The meduim containing MS. Inorganic salts, IAA 4 ppm, kinetin 2 ppm, 2,4-D 4 ppm, myo-inositol 100 ppm, glycine 20 ppm, 5 ppm each of nicotinic acid and pyridoxine-HCl, thiamine-HCl 1 ppm, coconut milk 10%, sucrose 3%, yeast extract 0.2%, agar 1.0% was appropriate for the growth of callus. After two months, the callus changed from friable to firm one. 6. In the organ differentiation experiment, the medium containing MS. inorganic salts, IAA 1 ppm, kinetin 10 ppm, myo-inositol 100 ppm, 0.5 ppm each of nicotinic acid and pyridoxine-HCl, thiamine-HCl 0.1 ppm, glycine 2 ppm, sucrose 3%, yeast extract 0.2%, agar 1%, without 2,4-D was the best one. Under the conditions of 1700 Lux of light intensity at 28°-30℃ of temperature and 10 hours per day of light period for 10 to 14 days, it could show the chlorophyll formation. The callus must be continuously cultured by transfering into fresh medium every two months. After six to eight times of subculture, it could induce the differentiation of young shoot (Fig. B2) or rootlet (Fig. A5 left). 7. After differentiation, plantlets in vitro were exposed to 3400 lux of light intensity. When they grew 4-5 leaves, the pantlets were transplanted into the pots containing vermiculite (Fig. A8 and B6). After another two months, the young plants were transplanted to the pots containing the soil under higher light intensity, therefore they could adapt to the natural conditions and grow vigorously. |
本系統中英文摘要資訊取自各篇刊載內容。