Phosphorus and Nitrogen Utilization in Diatoms under Ocean Acidification: Gene Expression and Proteomic Analysis

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Project title

Code/計畫編號

MOST104-2313-B019-003-MY3

Translated Name/計畫中文名

矽藻在海洋酸化狀態下的氮磷吸收基因表現與全蛋白質體分析

Project Coordinator/計畫主持人

Jeng Chang

Funding Organization/主管機關

National Science and Technology Council

Co-Investigator(s)/共同執行人

許邦弘

Department/Unit

Institute of Marine Biology

Website

https://www.grb.gov.tw/search/planDetail?id=11726517

Year

2016

Start date/計畫起

01-08-2016

Expected Completion/計畫迄

01-07-2017

Co-Investigator(s)

Pang-Hung Hsu

Bugetid/研究經費

1552千元

ResearchField/研究領域

生物技術(農)
漁業

"在全球變遷的大環境下，海洋酸化是無可避免的趨勢，而矽藻在遭受酸化衝擊後可能會改變生長效率，然後影響有機碳傳遞至魚類的過程，所以瞭解矽藻對酸化的反應，在評估與預測漁業資源方面有其重要性。有鑑於此，本申請案想要以三年的時間探討環境pH 值降低時如何影響矽藻的基因表現，並特別著重於酸化對吸收磷酸鹽的影響。本項研究首先將根據歐盟的指導原則配製特定pH 值的培養基，再針對在不同酸化條件下培養的模式矽藻建構全轉錄體和全蛋白質體，以觀察所有基因表現的改變。第二部份的研究將選用僅具有第二型磷傳輸基因的窄隙角刺藻和同時含有第二、三型磷傳輸基因的肋紋骨藻，觀察何種遺傳組成比較適合在酸化環境中生長，並使用定量反轉錄聚合脢連鎖反應 (Q-RT-PCR) 偵測磷傳輸相關基因的mRNA 表現量，再使用簡略型的質譜分析偵測蛋白質表現量。全蛋白質體分析部份將邀請臺灣海洋大學許邦弘教授擔任共同主持人補強此項專長。本申請案採用了先進的技術研究具生態意義的議題，預期所產生的結果將有助於建立矽藻在海洋酸化時吸收磷酸鹽和維持生長的分子機制、比較種類之間適應酸化的演化策略、並引申至推估漁業資源的變動。" "Under the overall influences of global changes, ocean acidification is an inevitable trend. For marine diatoms, the impact of ocean acidification may alter their growth efficiency, and subsequently affect the transfer of organic carbon along the grazing food chain. A better understanding of diatom responses to ocean acidification is important to the evaluation and forecasting concerning fishery resources. With these considerations, the present submission proposes a 3-year study plan that aims to investigate how an acidic environment influences gene expressions in diatoms. Special attentions will be placed on responses of the phosphate uptake system. In the first part of the proposed research, culture media with predetermined pH values will be prepared according to guidelines issued by European Commission. These media will be used to culture a model diatom, Phaeodactylum tricornutum, and its transcriptomes as well as proteomes will be constructed to observe expressional variations in all genes. In the second part of this proposal, two diatom spcies, Chaetoceros affinis and Skeletonema costatum, will be used as the experimental subjects. The former species contains a type II Na+/Pi cotransporter gene only while the latter contains both a type II and a type III transporter genes. Growth parameters will be monitored to determine which species adapts better in a low pH environment. Quantitative reversetranscription polymerase chain reaction will be used to measure the expression levels of genes involved in phosphate uptake. In addition, SILAC proteomic analysis specific to transporter proteins will be conducted to quantify gene expressions at the protein level. Professor Pang-Hung Hsu of Taiwan Ocean University is invited to join the project as a co-principal investigator to provide guidance in the construction of proteomes. This proposal employed advanced techniques to study an ecologically important topic. Experimental results generated by this project will shed light on the molecular mechanisms of phosphate uptake and growth maintenance in marine diatoms under ocean acidification. Evolutionary strategies between diatom species will be compared, and implications will be made for variations in fishery resources due to ocean acidification."

Keyword(s)

浮游植物
矽藻
捕食食物鏈
營養鹽
海洋酸化
全轉錄體
全蛋白質體
磷傳輸基因
diatoms
grazing food chain
nutrients
ocean acidification
phosphate transporter gene
phytoplankton
proteome
transcriptome

DSpace CRIS

Phosphorus and Nitrogen Utilization in Diatoms under Ocean Acidification: Gene Expression and Proteomic Analysis

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