Effects of Global Change on Phytoplankton Ecology in the Seas Surrounding Taiwan in the Northwest Pacific: Application of Fluorescence in situ Hybridization for Marker Gene Quantification

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Project title

Code/計畫編號

MOST104-2611-M019-008

Translated Name/計畫中文名

全球變遷對西北太平洋臺灣海域海洋生物地球化學與生態系統影響之長期觀測與研究(Ⅱ)---子計畫：全球變遷因子對東海及西北太平洋黑潮海域浮游植物生態之影響：以原位雜合顯示細胞在分子層次之反應

Project Coordinator/計畫主持人

Lee-Kuo Kang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Institute of Marine Environment and Ecology

Website

https://www.grb.gov.tw/search/planDetail?id=11575048

Year

2015

Start date/計畫起

01-08-2015

Expected Completion/計畫迄

31-07-2016

Bugetid/研究經費

1040千元

ResearchField/研究領域

海洋科學
大氣科學
生物技術(理)

本申請案為全球變遷對西北太平洋臺灣海域海洋生物地球化學與生態系統影響之長期觀測與研究（二）整合型計劃中的子計劃之一。浮游植物在海洋中扮演重要的基礎生產者角色，其亦可行光合作用吸收二氧化碳，是探討全球環境變遷的議題上不可或缺的重要組成。然而浮游植物是一群種類歧異度相當大的微生物，不同類群在海洋生態系中各扮演不同的功能角色，因此以高靈敏度、高專一性的分子指標來評估個別類群之生理反應，將可提供不同浮游植物受環境變遷的影響。在本計晝中主要希望發展螢光原位雜合技術用來偵測野外浮游植物之指標基因表現量，也就是以螢光直接標定方式顯示指標基因在浮游植物細胞内的表現情形，以單個細胞的數量級進行獨立的檢測，讓我們可以直接”看”到海洋浮游植物細胞的反應。所以申請人擬先申請一年經費首度嘗試將螢光原位雜合技術拓展至偵測浮游植物細胞之基因表現量，並透過參與東海南部侧線之採樣進行海上實際測試。預期此一新技術的拓展，不僅提供團隊在測量野外指標基因表現的另一種選擇外，這樣的技術也可延伸應用至其他子計劃中締結更緊密的交流合作。This proposal is the branch project in the integrated research project "Effects of Global Change on Ocean Bio geo chemistry and Ecosystems in the Seas surrounding Taiwan in the Northwest Pacific-II" (ECOBEST-II). Marine phytoplankton are responsible for more than 45% of the global primary productivity and play a vital role in global carbon cycling. Consequently it is extremely important to understand the impact that global climate change will have on the ecological performance of these organisms. However, marine phytoplankton are a diverse group of organisms. Each species has different requirements for growth and each contributes differently to carbon flows in marine food webs. The ability to evaluate the status of phytoplankton health in natural environments is thus important to understanding how phytoplankton tolerates environmental stresses and copes with interspecific competition. With advances in molecular biological techniques, molecular markers were proposed as powerful indicators to detect the physiological status of phytoplankton. In this proposal, I plan to use Fluorescence in situ hybridization (FISH) based on padlock probes and in situ target-primed rolling-circle amplification to detect single mRNA molecules in marine phytoplankton. During the first year, this method will be used to detect the transcript levels of nitrate transporter genes in natural assemblages of phytoplankton for evaluating their nitrogen status in the East China Sea. This project will not only bring an alternative method to detect marker gene expression in marine phytoplankton, but also provide a powerful method to collaborate with other projects under the ECOBEST.

Keyword(s)

浮游植物
螢光原位雜合技術
單細胞基因定量
指標基因表現
分子指標
phytoplankton
FISH (fluorescent in situ hybridization)
individual mRNA molecules
marker gene expression
molecular probe

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Effects of Global Change on Phytoplankton Ecology in the Seas Surrounding Taiwan in the Northwest Pacific: Application of Fluorescence in situ Hybridization for Marker Gene Quantification

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