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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/48762

Title: 多元不飽和醛類對於海洋羽狀矽藻Phaeodactylum tricornutum 死亡特異蛋白基因 (PtDSP) 調控機制之研究
Evaluation of Death Specific Protein gene (PtDSP) expression dynamics in the marine pennate diatom, Phaeodactylum tricornutum, under polyunsaturated aldehydes signaling
Authors: Liu, Tai-Yu
柳泰宇
Contributors: 國立臺灣海洋大學:生命科學暨生物科技學系
Keywords: 矽藻;死亡特異蛋白;不飽和醛類;一氧化氮;過氧化物
diatom;death specific protein;polyunsaturated aldehyde;nitric oxide;reactive oxygen species
Date: 2016
Issue Date: 2018-08-22T06:31:05Z
Abstract: 摘要 有些矽藻遭遇捕食者攻擊或是生長處於緊迫時,會誘發自身產生大量「不飽和醛類」(polyunsaturated aldehyde, PUA) 這類的二次代謝物,以做為防禦外敵或是誘導其族群產生「程式化細胞死亡」(programmed cell death, PCD) 的現象。然而,PUA如何誘導矽藻細胞在緊迫環境下,細胞生理發生反應,這樣的分子調控機制,截至今日仍沒有進行深入的研究。關於矽藻PCD現象,之前有研究指出圓形品系矽藻在生長末期會經由一氧化氮 (nitric oxide, NO) 或過氧化物 (reactive oxygen species, ROS) 做為細胞訊息傳遞媒介,大量誘發「死亡特異蛋白」基因 (death specific protein gene, DSP) 的表現,並推測其可能是促使矽藻細胞發生PCD的重要分子調控途徑。然而在羽狀品系矽藻中沒有DSP的相關研究。因此,ROS、NO、PUAs是否會調控羽狀品系矽藻DSP的表現,是本研究欲探討的主要問題。實驗使用本身不會產生PUA的海洋羽狀品系矽藻Phaeodactylum tricornutum作為模式生物,分別添加heptadienal、octadienal、decadienal等3種不同的PUA,以及H2O2 (提供ROS) 和diethylamine nitric oxide (提供NO) 進行培養,以即時定量PCR與西方墨點法測定不同處理組別,細胞內PtDSP mRNA與蛋白質的表現量。NO 與 ROS 的添加培養,確實會誘發 PtDSP 的大量表現。此外,不同PUA的添加培養實驗結果顯示,只有decadienal會誘發PtDSP mRNA的表現。有趣的是, PtDSP mRNA受到PUA誘發表現情形,並不會發生在以Tempol (ROS清除劑) 或c-PTIO (NO清除劑) 預先添加培養的矽藻細胞中,這意味著羽狀矽藻在緊迫環境下,也能接受外界環境中其他矽藻產生的PUA訊息,經由ROS或NO這類的細胞訊息傳遞媒介,將緊迫訊息傳入細胞中,進而誘發 PtDSP的表現,以調控細胞生理應付外界的環境變化。 關鍵字:矽藻、死亡特異蛋白、不飽和醛類、一氧化氮、過氧化物
Abstract Some diatoms can produce polyunsaturated aldehydes (PUAs), which are a kind of secondary metabolites, to induce self-autolysis via NO or ROS mediated pathway when ambient environment is stressful. However, regulatory mechanism mediated by PUAs in diatoms is still unclear. In addition, previous works indicated that mRNA expression of the death specific protein (DSP), which was suggested to participate in the process of programmed cell death (PCD) in centric diatoms, was regulated by ROS or NO signaling. But, the function of DSP in pennate diatoms is not characterized so far. Therefore, the aim of this study is to clarify whether PUA signaling can regulate DSP expression in pennate diatoms. Phaeodactylum tricornutum was chosen as a model organism because it is not a PUA producer. They were first treated with three different kinds of PUA (heptadienal, octadienal, decadienal), H2O2 (as ROS donor) and diethylamine nitric oxide (as NO donor) sepeartely, and then the levels of PtDSP mRNA and its protein were detected using quantitative reverse transcription PCR (Q-PCR) and weastern blot. The results showed that NO or ROS induced significant mRNA expression of PtDSP. Moreover, only decadienal was able to induce PtDSP mRNA expression. These high expressions were suppressed when the diatoms were pretreated with Tempol (ROS scavenger) or c-PTIO (NO scavenger). These results implied that under stressful environment, P. tricornutum can sense ambient PUA released from other diatoms and then induce PtDSP expression via ROS or NO mediated regulatory pathway. Keywords: diatom, death specific protein, polyunsaturated aldehyde, nitric oxide, reactive oxygen species
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010136011.id
http://ntour.ntou.edu.tw:8080/ir/handle/987654321/48762
Appears in Collections:[生命科學暨生物科技學系] 博碩士論文

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