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標題: | 水稻β-澱粉水解?表現及其缺失品種之研究 Expression of β-Amylase in deficient and normal rice cultivars |
作者: | Fu-Mee Yang 楊富米 |
出版年 : | 2001 |
學位: | 碩士 |
摘要: | 澱粉是水稻穀粒中含量最多的儲存性養分,種子萌芽後可經由水解酵素分解澱粉的作用,而提供小苗初期生長所需的碳源和能源。本論文旨在分析水稻個體中β-澱粉水解?的活性表現,並比較兩個水稻品種台農67號(TNG)和日本晴(NPB)之β-澱粉水解?表現的差異性。以電泳分析由內子葉、地上部及根誘導出來的TNG和NPB癒傷組織,顯示它們都表現相同的澱粉水解?圖譜,但都沒有β-澱粉水解?的出現。依南方墨點分析,得知此兩個品種至少有一個β-澱粉水解?基因,但由不同萌芽時間澱粉水解?活性的變化和組織特異性的分析結果,都顯示NPB確實是一個β-澱粉水解?缺失的水稻品種。本研究試圖對NPB不表現β-澱粉水解?基因做初步的瞭解,分析雜交種子中β-澱粉水解?活性與TNG之基因組中的重複次數之關係,結果發現NPB中β-澱粉水解?不表現並非其轉錄因數的問題,而是基因本身有缺陷。將TNG之β-澱粉水解?基因啟動子接上GUS報導基因,利用農桿菌感染法轉殖於菸草,分析轉殖菸草,在種子萌芽2-4天時,在其子葉中可以看到GUS活性的表現,之後很快便消失了;而在菸草轉殖株的子房壁及嫩葉中卻觀察不到GUS活性的表現,顯示水稻β-澱粉水解?基因的啟動子在單、雙子葉植物個體中有特殊的專一性亦有其歧異性。 Starch is the major component of storage reserves accumulated in rice kernels during seed development. Upon germination, amylases secreted by aleurone layers and scutella catalyze the degradation of starch and generate soluble sugar, which serves as energy and carbon source for the growth of seedlings. By using β-amylase-specific assay or amylolytic staining on gel, expression of β-amylase in various tissues of two rice cultivars, TNG67 and Nipponbare, was examined. No β-amylase activity was detected in the calli derived from either scutella or leaves of both cultivars. Beta-amylase activity did express in germinating kernels and leaves of TNG67 but not Nipponbare. Results from Southern analysis suggested that there might be two β-amylase genes in the genome of both rice cultivars. All results of amylolytic examination indicated that Nipponbare is a β-amylase deficient mutant. To analyze the property of the mutation, β-amylase activity in germinating hybrid kernels was examined. It is suggested that Nipponbare is defected in β-amylase gene itself and there is no trans-acting elements involved. A Ti-vector having GUS gene directed by OsBA promoter was constructed and was used to transform into tobacco. In transgenic plants, GUS activity is expressed in cotyledons only in the early stage of germination. Tissue specificity of β-amylase expression in monocots and dicots was discussed. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75228 |
全文授權: | 未授權 |
顯示於系所單位: | 植物科學研究所 |
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