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高溫脅迫下菊芋sRNA通路幾個關鍵酶基因的克隆與表達分析

時間:2019-05-25 20:51來源:畢業論文
探究高溫脅迫對菊芋sRNA通路上的幾個關鍵酶基因的影響,嘗試研究高溫脅迫對菊芋的影響過程及其機理,并為篩選或培育耐高溫的菊芋提供一些理論基礎

摘要:本試驗旨在探究高溫脅迫對菊芋sRNA通路上的幾個關鍵酶基因的影響,嘗試研究高溫脅迫對菊芋的影響過程及其機理,并為篩選或培育耐高溫的菊芋提供一些理論基礎。實驗采用電子克隆的方法成功克隆出基因AGO1、AGO5、DCL1、RDR1;采用高溫脅迫(25℃和30℃共2個溫度梯度,分別處理10天、15天、20天)下的菊芋作為實驗材料,提取其RNA,經過反轉錄、RT-PCR,以及轉化、測序等過程來得出結論。熒光定量結果表明經高溫脅迫后的樣品的4種基因的表達量比起未經高溫脅迫處理的樣品的表達量要高,基因AGO5、DCL1、RDR1的表達量隨高溫處理的時間增加有增加的趨勢,而AGO1的表達量則隨處理時間的增加出現了先增加后減少的趨勢,這四種基因可能在對高溫脅迫的應答中起著一定作用,同時這四種基因的表達在應答高溫脅迫的過程中可能會有時間的先后與作用機理的不同。35713
畢業論文關鍵詞:菊芋;高溫脅迫;AGO;DCL;RDR
Expression Analysis of Several Key Enzyme Genes of Jerusalem artichoke sine pathway under high temperature stress
Abstract:The aim of this study was to explore the possible effects of high temperature stress on several key enzyme genes and their expression analysis in sargus sRNA pathway, and to study the effects of high temperature stress on Jerusalem artichoke and its mechanism, and to provide some theoretical basis for screening or cultivating high temperature resistant Jerusalem artichoke. We were successfully cloned genes AGO1, AGO5, DCL1, RDR1 by electronic cloning, the results showed that the cDNA of Jerusalem artichoke was extracted by high temperature stress (25 ℃ and 30 ℃ a total of 2 temperature gradient, respectively, for 10 days, 15 days, 20 days) as the experimental material. The RNA was extracted and analyzed by reverse transcription, RT-PCR, as well as transformation and sequencing. Finally, the gene was quantitatively analyzed by fluorescence quantitative PCR. The gene was then quantitatively analyzed by fluorescence quantitative PCR. The results showed that the length of gene AGO1 in Jerusalem artichoke is 149bp, the length of AGO5 is 153bp, the length of DCL1 is 99bp, the length of RDR1 is 81bp, and the expression of four kinds of genes after high temperature stress was higher than those of samples without high temperature stress. The expression of AGO5, DCL1 and RDR1 increased with the increase of the time of high temperature treatment, while the expression of AGO1 increased first and then decreased with the increase of treatment time. Therefore, the four genes could have some effects on the response of temperature, while the four genes in the high temperature stress response might have different mechanism.

源¥自%六^^維*論-文+網=www.aftnzs.live


Key words: Jerusalem artichoke; high temperature stress; AGO; DCL; RDR
目  錄
摘要1
關鍵詞1
Abstract1
Key words1
引言1
1材料與方法3
1.1實驗材料3
1.2實驗試劑3
1.3儀器與設備3
1.4實驗方法4
1.4.1電子克隆4
1.4.2菊芋的處理4
1.4.3引物合成以及總RNA的提取5
1.4.4 RT-PCR5
1.4.5序列測定與分析 5
1.4.6 實時熒光定量PCR6
1.4.7 數據處理6
2實驗結果6
2.1基因拼接結果6
2.2總RNA的提取6
2.3引物檢測6
2.4實時熒光定量PCR7
3討論8
4結論9
致謝9
參考文獻9
高溫脅迫下菊芋sRNA通路幾個關鍵酶基因的克隆表達
    引言:菊芋,菊科,向日葵屬,多年宿根性草本植物。高1-3米,有塊狀的地下莖及纖維狀根。莖直立,有分枝,被白色短糙毛或剛毛。葉通常對生,有葉柄,但上部葉互生;下部葉卵圓形或卵狀橢圓形。頭狀花序較大,少數或多數,單生于枝端,有1-2個線狀披針形的苞葉,直立,舌狀花通常12-20個,舌片黃色,開展,長橢圓形,管狀花花冠黃色,長6毫米。瘦果小,楔形,上端有2-4個有毛的錐狀扁芒。花期8-9月。具有耐寒耐旱、抗風沙、耐鹽堿等抗性,此外一些品種還具有耐受和富集重金屬的能力[1]。 高溫脅迫下菊芋sRNA通路幾個關鍵酶基因的克隆與表達分析:http://www.aftnzs.live/shengwu/20190525/33791.html
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