SGER: Development of a method to measure feeding by copepods using qPCR

SGER：开发一种使用 qPCR 测量桡足类摄食的方法

• 批准号: 0748963
• 负责人: Edward Durbin
• 金额: --
• 依托单位: University of Rhode Island
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0748963&HistoricalAwards=false
• 关键词: SGER; Development; method; measure; feeding

This project will attempt to development a new method using molecular techniques to quantitatively measure in situ consumption rates of copepods on different phytoplankton and microzooplankton prey species. The approach will be to use the 18S ribosomal RNA gene as a marker to identify different prey species in the guts of copepods and to use quantitative real-time polymerase chain reaction (q-PCR) to measure of the amount of each species of prey in the stomachs of the copepods. By combining this gut content information with DNA digestion rates the investigators will be able to calculate feeding rate on these different prey species. The work will proceed in several phases: (1) develop and test group-specific primers, (2) carry out laboratory experiments to determine DNA digestion rates by copepods, (3) determine the 18S copy number per prey organism for selected prey species, (4) sample Calanus finmarchicus from the Gulf of Maine/Georges Bank region during late spring and generate a clone library from the gut contents using the group-specific primers. Species-specific primers developed from this clone library will be used with qPCR to quantify the prey DNA. Experiments will be carried out with the same group of animals to measure DNA digestion rates for selected prey species in the field. Ingestion rates will be calculated and compared with rates using the gut pigment method. Broader Impacts: While some aspects of the research are high risk, the potential payoff will considerable. Understanding the effect of changing environmental conditions on zooplankton and their role in controlling the fate of phytoplankton requires knowledge of their in situ feeding rates. Because of the strong gradients in the food environment in many regions of the ocean e.g. thin layers, ice edge and under ice regions, we argue traditional bottle incubation techniques for measuring in situ feeding by copepods are not appropriate. The methods being developed will allow direct in situ consumption estimates of different prey species overcoming these previous limitations. This research will result in the development of methods that will have many applications for investigating in situ predator-prey interactions in the marine environment because of its sensitivity and ability to measure ingestion rate of individual prey species present at low concentrations in the environment. Examples include fish larvae, deep-sea fishes, euphausiids, and amphipods. This knowledge would lead to new insights as to how the oceanic food web is functioning. The project will involve participation of a Graduate Student.

该项目将尝试使用分子技术开发一种新方法，以定量测量不同浮游植物和Microzooplankton Prey物种上copepods的原位消耗率。该方法将是使用18S核糖体RNA基因作为标志物，以鉴定copepods肠道中的不同猎物，并使用定量的实时聚合酶链反应（Q-PCR）来测量copepods胃中每种猎物的数量。通过将这些肠道含量信息与DNA消化速率相结合，研究人员将能够计算这些不同猎物物种的饲养率。 The work will proceed in several phases: (1) develop and test group-specific primers, (2) carry out laboratory experiments to determine DNA digestion rates by copepods, (3) determine the 18S copy number per prey organism for selected prey species, (4) sample Calanus finmarchicus from the Gulf of Maine/Georges Bank region during late spring and generate a clone library from the gut contents using the group-specific primers.该克隆文库开发的物种特异性引物将与qPCR一起使用，以量化猎物DNA。实验将与同一组动物进行，以测量该场中选定的猎物物种的DNA消化率。使用肠道色素方法将计算摄入率并将其与速率进行比较。更广泛的影响：虽然研究的某些方面是高风险，但潜在的回报将相当大。了解改变环境条件对浮游动物的影响及其在控制浮游植物命运中的作用需要了解其原位喂养率。由于海洋许多地区的食物环境中的梯度很强，例如我们认为，薄层，冰边缘和冰区域下方，我们认为传统的瓶孵化技术用于测量由Copepods进行原位喂养的方法是不合适的。所开发的方法将允许直接对不同猎物物种的原位消耗估计来克服以前的限制。这项研究将导致发展方法的发展，这些方法将在海洋环境中研究原位捕食者 - 捕食者相互作用，因为其敏感性和测量在环境中低浓度下存在的个别猎物物种的摄入率的能力。例子包括鱼幼虫，深海鱼，euphausiids和两亲脚。这些知识将导致有关海洋食品网的运作方式的新见解。该项目将涉及研究生的参与。