Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/37272
標題: 夏赤箭種子無菌播種與冬赤箭根莖繁殖之研究
Asymbiotic seed germination of Gastrodia flabilabella and in vitro rhizome propagation of G. pubilabiata
作者: 賴漢揚
Lai, Han-Yang
關鍵字: 夏赤箭;Gastrodia flabilabella;冬赤箭;無菌播種;Gastrodia pubilabiata;asymbiotic seed germination
出版社: 農藝學系所
引用: Boufford, D. 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摘要: 
赤箭屬為蘭科植物中最大的真菌異營屬,這一類植物無法行光合作用,需依賴共生菌提供養分來源。台灣分布之赤箭屬植物種類豐富,且其中有幾種為台灣特有種,有人工保育之需求。本研究目的為建立夏赤箭(Gastrodia flabilabella)無菌發芽的最適條件,並且利用組織培養的方式對冬赤箭(G. pubilabiata)根莖進行大量繁殖。結果摘要如下:
觀察野外調查之夏赤箭果莢於授粉後約24~33天達成熟。將野外採集之夏赤箭果莢消毒後進行無菌播種,測試包括播種培養基的基本鹽類組成、氮素組成、植物生長調節劑(BA、NAA與GA)及其濃度、有機添加物(蛋白腖與椰子汁)及其濃度以及種子發芽溫度等因子對種子發芽之影響,結果顯示培養基組成條件中僅NAA、GA及椰子汁具有促進種子發芽之效果,培養溫度則以20℃恆溫或23℃日溫/18℃夜溫較15℃及25℃恆溫培養為佳。以TTC(2,3,5- tetrazolium triphenyl chloride)對播種後32週之種子進行染色處理,結果顯示無論有無合併使用超音波震盪處理,種子皆無明顯染色情形,顯示夏赤箭種子已不具生命力。
以冬赤箭無菌播種養成的根莖為材料,測試根莖生長與繁殖之最佳培養條件,結果顯示根莖適於培養於1/2MS基本鹽類培養基,添加之糖類以蔗糖為最佳,濃度在20 mg/L至40 mg/L之間,添加100 mL/L椰子汁有助於根莖增重與增長,植物生長調節劑的添加明顯有助於根莖的生長與增殖,以1- 2 mg/L BA配合1 mg/L NAA在根莖增重、增長及根莖數增加方面,皆有較佳之表現。液態培養有助於根莖之增重與新生根莖數的增加,為避免頂芽玻璃質化,以2週液態培養後配合4週固態培養可獲得最高的根莖增重與新生根莖數。以4種蜜環菌與冬赤箭根莖進行共培養8週,發現4種蜜環菌均未能與冬赤箭根莖形成共生關係,因此冬赤箭出瓶培養的條件仍待建立。

Gastrodia is the largest mycoheterotrophic genus in Orchidaceae family. Since mycoheterotrophic orchids don’t have chlorophyll for photosynthesis, they require symbiotic fungi serving their nutrition needs. Taiwan has rich Gastrodia resources and among them few endemic species need to be conserved. Objectives of this study were to establish an asymbiotic germination protocoal for G. flabilabella, as well as to establish an in vitro micropropagation method for proliferation rhizomes of G. pubilabiata. Results of this study were summarized as follows:
Maturation of G. flabilabella seed pod was observed 24 to 33 days after pollination in the field. Capsules of G. flabilabella were sterilized before testing various components in the medium including basal salts, nitrogen composition, growth regulators of BA, NAA and GA in various concentrations, organic additives of peptone and coconut milk, and incubation temperature for asymbiotic germination. Results showed that medium containing with NAA, GA and coconut milk were benifical for seed germination. Incubation in consistent temperature of 20℃or 23℃for the day and 18℃for the night were found superior to 15℃ or 25℃ for seed germination. A staining method using 2,3,5- tetrazolium triphenyl chloride (TTC) was applied to the 32-week-old germinating seeds of G. flabilabella however no viability was observed on the seeds with or without additional ultrasonic treatment.
Various components in medium for in vitro proliferation G. pubilabiata rhizomes were tested. Results showed that rhizomes of G. pubilabiata grew well in a half strength MS basal salt medium. Among five tested carbonhydrates, sucrose was found the best carbohydrate source for rhizome growth with suitable concentrations between 20 mg/L to 40 mg/L. In addition, Coconut milk was found increasing rhizome weight and length with concentration of 100 mL/L. Growth regulators of BA and NAA had significant effects on growth and perliferation of G. pubilabiata rhizomes, and the best results were found using 1- 2 mg/L BA in combination with 1 mg/L NAA. Rizomes grown in liquid medium had higher weight and better rhizome induction than that of rhizomes grown on solid medium. A cycle of 2-week liquid culture following with 4-week solid culture was suggested for increasing rhizome growth without vitrification. Rhizomes of G. pubilabiata were inoculated with four strains of Armillaria mellea however no symbiotic hyphae was observed after 8 weeks of coculture. Therefore, ex vitro culture conditions for G. pubilabiata rhizomes need to be studied continuesly in the future.
URI: http://hdl.handle.net/11455/37272
其他識別: U0005-2208201209222200
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