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標題: | 阿拉伯芥澱粉缺失突變種之研究 Studies on Arabidopsis Starch Deficient Mutants |
作者: | Jrhau Lung 龍志浩 |
出版年 : | 1996 |
學位: | 碩士 |
摘要: | 以碘染檢驗澱粉含量的方法,從約一萬株阿拉伯芥的M2植物中篩選得到兩株澱粉含量下降的突變種--ke122與ke478。依遺傳分析結果顯示,ke122與adg1為等位基因(allelic),ke478與pgm為等位基因。adg1、adg2及pgm為前人發表之澱粉含量下降的突變種,adg1不具有ADP-glucose pyrophosphorylase(ADGase)的活性且其兩個次單元體(APL、APS)的蛋白質同時消失,故adg1被認為是調控基因的突變;pgm突變使葉綠體中的Phosphoglucomuatse(PGM)活性消失,故pgm被認為是PGM的基因突變。為瞭解阿拉伯芥ADGase與PGM突變種的基因表現,分析ADGase突變種adg1、ke122和adg2的ADGase基因轉錄的產物。結果顯示ke122只表現APL之mRNA,而無APS之mRNA的存在,顯示其為APS基因突變種,而與ke122不能互補之等位基因突變種adg1卻同時有APL及APS mRNA的表現,顯示adg1和ke122突變在分子層次上的表現仍有差異。利用RFLP分析及基因轉殖互補法確定ke122與ADGase小次單元體基因的關係,結果肯定ke122確實為APS基因的突變種。利用與波菜葉綠體PGM核酸序列具相似性的阿拉伯芥cDNA-120H21T7為探針,檢驗pgm與ke478的RNA表現,結果顯示此兩突變種均能表現120H21T7基因,但是由RFLP的結果卻顯示120H21T7並非與pgm基因相同,故ke478之基因突變特性仍待進一步之探討。 We have isolated two monogenic recessive Arabidopsis mutants, designated ke122 and ke478, which contain less starch. Allelic tests and biochemical study established that ke122 is represented by the mutant previously described as a regulatory gene mutant- adg1 which lost both subunits of ADP glucose pyrophosphorylase (ADGase) and ADGase enzyme activity and ke478 is represented by the mutant previously described as pgm which lost chloroplastic Phosphoglucomutase (PGM) activity. To study the molecular basis of these less starch mutants, adg1 adg2, pgm, ke122 and ke478, the level of mRNAs for ADGase and PGM were examined. The adg1 and adg2 expressed the RNA of both ADGase subunits, however the ke122 only expressed the large subunit RNA. We used RFLP analysis and direct gene complementation to identified the relation between ke122 and ADGase small subunit gene (APS), and the results indicated that ke122 is an APS mutation rather than a previously described regulatory gene mutation. pgm and ke478 expressed the mRNAs of cDNA-120H21T7 which has similarity to the spinach chloroplastic pgm. However, random segregation between 120H21T7 and ke478 in RFLP analysis suggests that ke478 and pgm are not mutations of 120H21T7 gene. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76209 |
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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