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標題: | 缺水對大豆幼苗Glutathione Reductase活性及其基因表現之影響 The Effect of Water Steress on Glutathione Reductase Gene Expression and Enzyme Activity of Soybean |
作者: | 洪美淑 |
出版年 : | 1994 |
學位: | 碩士 |
摘要: | 植物於不同逆境之下會誘導葉片GR(glutathione reductase)活性的增加,因而保護植物免受傷害。本論文主要研究目的在探討大豆幼苗在不同濃度(10%、20%、30%)的PEG處理下GR活性及其基因的表現情形。將14天苗齡的大豆幼苗植株置於不同濃度PEG 3300逆境下,測定不同時間葉片水勢(water potential)變化,結果顯示不同濃度的PEG處理10分鐘後,葉片水勢即明顯的下降,但30分鐘後漸趨穩定,不同的PEG處理濃度及處理時間均會影響GR基因的表現,20% PEG處理2小時可使GR活性達到最高,不同生育苗齡的大豆幼苗對GR活性亦有不同的反應,其中14天的幼苗以PEG處理後,對GR活性增加1.5倍,最?顯著,若以GR酵素抗體檢測GR蛋白質的含量,則可發現30KD與40KD二個polypeptides,在14天的大豆幼苗葉片中GR蛋白質含量,明顯較其他生育階段的幼苗?高。 本試驗也利用GR cDNA 基因之核?酸序列,分別設計二條primer (+1401?+1420與+1731?+1740),並以大豆poly(A+) RNA?範本進行聚合梅連鎖反應(Reverse transcription-Poly merase Chain reaction),分析PCR所得到的產物約340bP,定序後發現此片斷與Tang and Webb (1993, accesion No. L11632) 所發現的大豆SOYGLUTR coding region +1410?+1740間具有99.4%的同源性,因此將此段序列製成探針,以備後續Northern blot 檢定之用。 將不同濃度與不同時間PEG處理的大豆幼苗之RNA與PCR所得到的探針進行Northern hybridization,結果顯示GR基因雜交的條帶約?2.10Kb,而以20% PEG處理2小時可得到最高RNA表現量。由以上的試驗結果可知14天的大豆幼苗,以20% PEG處理後,可明顯增加GR活性及其基因之表現。 Plants (vegetation) under various of stresses induced can bete increase GR (glutathione reductase) activity, therefere, it can protect plants from all sorts of injuries. The very purpose of this study is to discuss the expression of soybean GR gene when its stuated under different conditions of PEG treatment. Given that 14-days old soybean replaced on different PEG concentration stress and different time to measure water potential changes on leaves, after 10 minutes, the result concentration shows decline in water potential, howerer, 30/min later, it gradually becomes stable which means different concentration and time in PEG treatment affect the expression of gene; 20% of PEG treatment with 2/hr duration can make GR activitier to reach its highest. Furthermore, soybean with varied ages identified accordingly different reflection after treatment. Given GR enzyme antibody measurement GR protein content, 30KD and 40KD polypeptides bands are discovered, particularly, 14-days old soybean leaves contain relatively higher amount than other soybean at different growing stages. The very experiment adopted GR cDNA gene amino acids sequence, sepatatelly designed 2 sets of primer (+1401? +1420 and +1731? +1740). and used soybean poly(A+) RNA as template; RT-PCR (reverse transcription polymerase chain reaction) was reflected and its analysis obtained about product of 340bp. The sequence has brought 99.4% homology when its compared to that of Tang and Webb's (1993, L11632) soybean SOYGLUTR coding ragion +1410? +1740. far, the above sequence provider probe and will be used in Northern blotting analysis. RNA and PCR of soybean from different PEG treatment in concent treatment with 2/hr duration implies the highest expression of RNA. Finally, as the experiment has obviously shown, GR activities and gene expression of 14-days old soybean are increased after 20% of PEG treatment. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76057 |
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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