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標題: | 水稻過氧化氫相關之微小核醣核酸 miR535 研究 Study of Hydrogen Peroxide Responsive miR535 in Rice (Oryza sativa L.) |
作者: | Chun-Wei Huang 黃俊瑋 |
指導教授: | 鄭石通 |
關鍵字: | 氧化逆境,過氧化氫,微小核糖核酸,定序資料分析,miR535,OsSPL2,OsFLS2, Oxidative stress,H2O2,miRNA,sequencing data analysis,miR535,OsSPL2,OsFLS2, |
出版年 : | 2019 |
學位: | 碩士 |
摘要: | 過氧化氫為活性氧化物質的一種,在濃度適中的狀況下在植物當中扮演得訊息傳遞分子的角色。但當植物面臨傷害、蟲害、乾旱、炎熱等逆境的時候,會產生過量的ROS進而導致氧化逆境而使得植物的生長發育受到影響。微小核糖核酸(microRNA)在植物當中屬於非編碼小核糖核酸中的一員(non-coding small RNA),雖然僅具有21-24個核苷酸,卻在植物基因調控路徑中扮演的相當重要的角色,這些微小核糖核酸會藉由降解核醣核酸或是抑制傳譯的方式來達到基因調控的目的。在近年來許多研究發現微小核糖核酸不只參與在植物生長發育過程當中,更發現它們與植物面對逆境的機制息息相關,但對於氧化逆境中微小核糖核酸的功能的研究仍然不多,因此,我們實驗室利用次世代微小核酸定序以及降解組定序的方式在水稻中預測出了51種已知的微小核糖核酸可能參與在氧化逆境下植物的基因調控之中。本篇研究更近一步驗證次世代定序的結果並且發現了miR535在氧化逆境之下其表現量會受到抑制,同時藉由生物資訊方式預測下游調控之目標基因OsSPL2以及OsFLS2,可能具有調控水稻基因表現量的功能,經由菸草短暫表達實驗驗證,發現所預測之miR535前驅物確實會產生具有功能的成熟miR535,但經由實驗結果驗證後發現,miR535未如預測結果調控OsSPL2,推測其調控反應應為miR156作用,本研究雖未對miR535功能有所定論,未來可利用轉殖方式來去確認其調控功能與機制。 Hydrogen peroxide (H2O2), one of reactive oxygen species (ROS), is a natural by-product of cellular metabolism, and acts as a signal molecule in response to both biotic and abiotic stresses in plants. MicroRNAs (miRNAs), non-coding small RNAs, play critical roles in regulating gene expression through either degradation of miRNA target genes or inhibiting the process of gene translation. To date, only few studies have concentrated on miRNA-mediated gene regulation in response to oxidative stress in plants. In this study, small RNA sequencing has revealed that 51 conserved miRNAs were regulated during the short period of H2O2 treatment in rice seedling roots. The osa-miR535 with novel functions was identified for further study. The expression of osa-miR535 was decreased under short-term oxidative stress. Through the anaylyses of bioinformatics and degradome sequencing two potential target genes of osa-miR535 were OsSPL2 and OsFLS2. Surprisely, the interaction between osa-miR535 and OsSPL2 could not be confirmed by agroinfiltration. However, according to previous reserches and reanalysis of bioinformatics we speculated that the gene regulation of OsSPL2 might be caused not by osa-miR535 but by osa-miR156. Taken together, our results demonstrated that 51 conserved miRNA were regulated under oxidative stress and osa-miR535 play a role in oxidative stress while the function of osa-miR535 still need further investigation. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/21815 |
DOI: | 10.6342/NTU201803876 |
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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