A Transcriptome Study of Diatom Nutrient Uptake Mechanisms in Seas Surrounding Taiwan: Identification of Indicator Genes in Representative Species and Influences of Nitrogen and Phosphorus on Gene Expression

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基本資料

Project title

Code/計畫編號

NSC100-2313-B019-006

Translated Name/計畫中文名

以全轉錄體定序研究臺灣海域代表性矽藻之營養鹽吸收機制：指標基因之鑑別與氮磷對基因表現之影響

Project Coordinator/計畫主持人

Jeng Chang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Institute of Marine Biology

Website

https://www.grb.gov.tw/search/planDetail?id=2329261

Year

2011

Start date/計畫起

01-08-2011

Expected Completion/計畫迄

01-07-2012

Bugetid/研究經費

1200千元

ResearchField/研究領域

生物技術(理)
海洋科學
漁業

Description

Abstract

矽藻是台灣附近海域中重要的初級生產者，而監測指標基因表現量是瞭解矽藻如何因應營養鹽濃度與比例變化的有效方法。目前所選用的矽藻缺氮指標基因是硝酸鹽傳輸蛋白，已在東海中完成測試，但是缺磷指標基因面臨在海洋中不易選殖同源基因的困境。解決之道是利用相關角刺藻進行全轉錄體定序，以便快速且大量的搜尋磷酸鹽吸收利用的相關基因。根據上述之研究進度，本申請案計劃針對由全轉錄體定序所顯示的缺磷高表現基因在不同培養條件下進行測試，確認基因表現對磷缺乏的生理狀態具有專一性。然後使用添加與移除磷酸鹽的手段，找出缺磷指標基因的最大與最小表現量，做為日後在海洋中判斷缺磷程度的標準。接下來要在採自東海的樣本中，調查缺磷指標基因的岐異度，做為設計專一性引子的依據。最後為了瞭解基因序列與表現的種間差異，也計劃針對第二種矽藻 (肋紋骨藻) 進行全轉錄體定序。在本申請案執行完畢後，預計可成功得到一個以上的缺磷指標基因並完成相關測試，然後可以與已備妥的缺氮指標基因共同使用於海洋生態的研究上。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能對漁業資源、海洋優養化、以及二氧化碳的吸收提供重要資訊。 Marine diatoms are important primary producers in waters surrounding Taiwan. To understand how diatoms adapt to the ever changing environment in terms of nutrient concentrations and ratios, a good method is to monitor the expression of indicator genes. The diatom nitrate transporter gene has been selected as an indicator for nitrogen deficiency, and field evaluation in the East China Sea is in its final phase. However, genes homologous to laboratory-defined phosphorus-deficiency indicators are difficult to obtain in marine environments. A solution to this problem is to generate a transcriptome data bank for a common diatom, Chaetoceros affinis, so that genes related to phosphorus uptake and utilization can be rapidly and thoroughly searched. Based on progresses achieved to date, this application proposes to perform laboratory tests on genes with high-level expression in the low-phosphorus transcriptome. Various growth conditions will be examined to ensure that the expression levels of candidate indicators exclusively respond to phosphorus deficiency. At the same time, phosphorus addition and deprivation experiments will be used to induce minimal and maximal expression levels of the candidate indicators. A broad and sensitive expression range is necessary for setting a threshold of phosphorus deficiency in the sea. Subsequently, sequence diversity of indicator genes in the East China Sea will be investigated. The result will allow the design of a series of quantitative PCR primers targeting specific diatom genera. Transcriptome analysis of a second diatom species, Skeletonema costatum, is also proposed, and the purpose is to provide information about inter-specific variations in sequence diversity and expression level. After the completion of proposed works, it is expected to obtain more than one well-tested indicator genes for phosphorus deficiency, which can be jointly used with the field-ready nitrogen deficiency indicator to study marine ecology. This proposal employed advanced techniques to study an ecologically important topic. Technology and experimental results generated by this project will provide important information to the management of fishery resources, the control of marine eutrophication, and the absorption and sinking of carbon dioxide.

Keyword(s)

東海
浮游植物
磷鹽缺乏
指標基因
全轉錄體
矽藻
磷酸鹽傳輸基因
鹼性磷酸酶
East China Sea
phytoplankton
nutrient stress
marker genes
transcriptome
diatom
alkaline phosphatase
phosphate transporter