Diatoms under Interspecific Competition: Expressions of Nitrogen and Phosphorus Utilization Genes and Proteomic Analysis

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基本資料

Project title

Code/計畫編號

NSC102-2313-B019-018-MY2

Translated Name/計畫中文名

矽藻在種間競爭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

Department/Unit

Institute of Marine Biology

Website

https://www.grb.gov.tw/search/planDetail?id=3085177

Year

2013

Start date/計畫起

01-08-2013

Expected Completion/計畫迄

01-07-2014

Co-Investigator(s)

Pang-Hung Hsu

Bugetid/研究經費

1623千元

ResearchField/研究領域

生物技術(農)
漁業

海洋中，矽藻的種類組成與營養鹽充足程度影響橈腳類的攝食和生長效率，並決定有多少有機碳會沿食物鏈傳遞至魚類的食階，所以對評估與預測漁業資源有其重要性。由於傳統的營養鹽限制參數與種間競爭係數在偵測上十分困難，而且會隨著矽藻的生理狀態改變，觀察基因表現可以提供直接證據來建立易於解釋的生態機制。本申請案想要以兩年的時間探討銨鹽和磷酸鹽如何影響矽藻的基因表現，觀察環境中的氮、磷濃度是否獨立調控各種營養鹽傳輸基因。另外還要進一步探討兩種矽藻，Skeletonema 和Chaetoceros，在競爭營養鹽時是否經由降低環境中的營養鹽濃度給予對方壓力。研究的手段是將矽藻單獨或混合培養於各種氮磷組合中，再以定量反轉錄聚合脢連鎖反應偵測硝酸鹽傳輸蛋白、磷酸鹽傳輸蛋白、和鹼性磷酸脢等相關基因的表現量，以及進行競爭狀態下的SILAC 全蛋白質體分析。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營養鹽缺乏和營養鹽競爭的分子機制、解釋矽藻在東海中的分佈、及估計捕食食物鏈和漁業資源的關係。 In marine environment, species composition of diatoms and their levels of nutrient sufficiency influence copepod feeding and growth efficiency, which in turn determine the amount of organic carbon reaching the trophic level of fish via the grazing food chain. Diatom composition and physiological status are thus important to the evaluation of fishery resources. Since conventional parameters of nutrient limitation and interspecific competition are difficult to measure, and these parameters are known to change with physiological status, the observation of gene expression may provide direct evidence to establish a simple model for the explanation of mechanisms. This application proposes a 2-year study plan that aims to study how ammonium and phosphate affect gene expression in 2 species of diatoms, Chaetoceros and Skeletonema. Emphasis will be given to observing whether ammonium and phosphate regulate corresponding transporter genes independently. A second goal is to study nutrient competition between these 2 diatoms and to determine if competition stress solely comes from the lowered ambient nutrient concentrations. Experimental design will include single-species and mixed-species diatom cultures grown at various combinations of ammonium and phosphate concentrations. Quantitative reversetranscription polymerase chain reaction will be used to measure the expression levels of genes involved in nutrient uptake, including nitrate transporter genes, phosphate transporter genes, and alkaline phosphatase genes. In addition, SILAC proteomic analysis will be conducted for diatoms under the status of nutrient competition. 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 reveal molecular mechanisms of nutrient competition in diatoms. These results will also be helpful to explain the distribution of diatoms in the East China Sea and to estimate the relationship between fishery resources and the grazing food chain.

Keyword(s)

浮游植物
矽藻
捕食食物鏈
營養鹽
種間競爭
指標基因
全蛋白質體
diatoms
grazing food chain
indicator genes
interspecific competition
nutrients
phytoplankton
proteome

DSpace CRIS

Diatoms under Interspecific Competition: Expressions of Nitrogen and Phosphorus Utilization Genes and Proteomic Analysis

基本資料

Description