Assessing the potential of mRNA-FISH FACS for isolation of functional soil bacterial populations for quantifying biogeochemical cycle interactions
评估 mRNA-FISH FACS 分离功能性土壤细菌群体以量化生物地球化学循环相互作用的潜力
基本信息
- 批准号:NE/J013153/1
- 负责人:
- 金额:$ 6.56万
- 依托单位:
- 依托单位国家:英国
- 项目类别:Research Grant
- 财政年份:2012
- 资助国家:英国
- 起止时间:2012 至 无数据
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The normal growth of all living entities depends on an adequate source of essential elements (e.g. C, N, S, P) and, in this respect, the Earth can be considered a closed system with the supply of essential elements being finite. Therefore, the recycling of these elements through the environment is fundamental to avoid exhaustion and microbes can be viewed as the 'engine room' that drive the component processes responsible for the recycling of these elements in the Earth's biogeochemical cycles. In cycling carbon, soil microbes utilise different organic and inorganic forms of carbon as energy and carbon sources resulting in the transfer carbon between environmental compartments. However, the carbon cycle does not operate on its own but it is closely metabolically linked with that of other essential elements either via the use of these as reductants and oxidants in energy transduction or via their incorporation into biomass (or release from decaying dead biomass) as part of multiple essential element- containing biomolecules (e.g proteins, DNA). Hence, the availability of carbon is a key factor in determining the transformations and cycling of other essential elements whilst the availability of other key elements control the rate at which microbes consume and respire carbon. Such biogeochemical cycle interactions can be illustrated by the soil microbial process of denitrification: the decomposition of organic carbon under low oxygen conditions through the respiration of nitrate resulting in the step-wise reduction of nitrate to dinitrogen gas (N2) with nitrous oxide (N2O) produced as an intermediate. A central goal in microbial ecology is to link biogeochemical processes to specific microbial taxa in the environment so that the role of microbial community structure can be better represented in predictive models. A suite of methods have been developed in the last decade in order achieve this goal without the need for cultivation and characterization of isolates but none of these offer the opportunity to quantify the interactions between biogeochemical cycles in a microbially-oriented way, for example, with respect to the use of a particular carbon source as a reductant to drive denitrification. Gaining the quantitative understanding of the interactions that is required to predict essential element fluxes and feedbacks under perturbed carbon cycle and environmental change scenarios is therefore method- limited.This project will provide proof-of-concept of a new method to quantify use of carbon by bacteria whilst transforming another essential element. The bacterial denitrification pathway will serve as a case study with a focus on the bacteria using carbon to reduce N2O to N2 (the final step in denitrification) due to the crucial role that this group play in regulating the atmospheric concentration of N2O, a potent greenhouse gas. The new method involves: (i) use of C isotopes to trace microbial C consumption; (ii) labelling actively N2O-reducing microbial cells with a fluorescent dye; (iii) sorting the fluorescent cells and quantifying the C isotope content.The proof of concept will be in simple experimental systems involving known N2O-reducing bacteria and soil microcosms incubated under conditions known to promote denitrification. As a case study, we will test a theory concerning the carbon source preference of the N2O-reducing bacteria.The project brings together the complimentary expertise of the investigators (use of C isotopes, fluorescence-labelling and sorting of bacteria, denitrification biogeochemistry) and the project partner (fluorescence labelling of bacteria active in biogeochemical cycling). We will use state-of-the art stable isotope techniques to quantify microbial N2O reduction and exploit advances in instrumentation for cell sorting that enables the accurate detection of bacterial cells extracted from soil.
所有生物的正常生长都依赖于必需元素(如C、N、S、P)的充足来源,在这方面,地球可以被认为是一个封闭的系统,必需元素的供应是有限的。因此,这些元素通过环境的再循环是避免耗尽的根本,微生物可以被视为“发动机室”,驱动负责地球地球化学循环中这些元素再循环的组成过程。在碳循环中,土壤微生物利用不同的有机和无机形式的碳作为能量和碳源,导致碳在环境分区之间转移。然而,碳循环并不独立运行,而是通过在能量转导中使用这些作为还原剂和氧化剂或通过将其并入生物质(或从腐烂的死亡生物质中释放)作为含有多种必需元素的生物分子(例如蛋白质、DNA)的一部分,与其他必需元素的代谢密切相关。因此,碳的可用性是决定其他必需元素转化和循环的关键因素,而其他关键元素的可用性控制微生物消耗和呼吸碳的速率。土壤微生物的反硝化过程可以说明这种生态地球化学循环的相互作用:在低氧条件下,有机碳通过硝酸盐的呼吸作用分解,导致硝酸盐逐步还原为二氮气体(N2),产生一氧化二氮(N2 O)作为中间产物。微生物生态学的一个中心目标是将生物地球化学过程与环境中特定的微生物类群联系起来,以便在预测模型中更好地代表微生物群落结构的作用。在过去的十年中,已经开发了一套方法,以实现这一目标,而不需要培养和表征分离物,但这些方法都没有提供以微生物为导向的方式量化生物地球化学循环之间的相互作用的机会,例如,关于使用特定的碳源作为还原剂来驱动反硝化。因此,在扰动碳循环和环境变化情景下,获得对预测必需元素通量和反馈所需的相互作用的定量理解是方法有限的。该项目将提供一种新方法的概念验证,以量化细菌对碳的使用,同时转化另一种必需元素。细菌反硝化途径将作为一个案例研究,重点是细菌使用碳将N2 O还原为N2(反硝化的最后一步),因为这一组在调节N2 O(一种有效的温室气体)的大气浓度方面发挥着至关重要的作用。新方法包括:(i)使用碳同位素追踪微生物碳消耗;(ii)用荧光染料主动标记N2 O还原微生物细胞;(iii)对荧光细胞进行分类并量化碳同位素含量。作为一个案例研究,我们将测试一个关于N2 O还原细菌的碳源偏好的理论。该项目汇集了研究人员的互补专业知识(C同位素的使用,细菌的荧光标记和分选,反硝化生物化学)和项目合作伙伴(在生物化学循环中活跃的细菌的荧光标记)。我们将使用最先进的稳定同位素技术来量化微生物N2 O还原,并利用细胞分选仪器的进步,从而能够准确检测从土壤中提取的细菌细胞。
项目成果
期刊论文数量(1)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Elizabeth Shaw其他文献
Cognitive Enhancement and Criminal Behavior
认知增强与犯罪行为
- DOI:
- 发表时间:
2013 - 期刊:
- 影响因子:0
- 作者:
Elizabeth Shaw - 通讯作者:
Elizabeth Shaw
Access to Colposcopy Services for High-risk Canadian Women: Can We Do Better?
为高危加拿大女性提供阴道镜检查服务:我们可以做得更好吗?
- DOI:
10.17269/cjph.95.529 - 发表时间:
2004 - 期刊:
- 影响因子:0
- 作者:
G. Ogilvie;Elizabeth Shaw;Sandra P. Lusk;Joyce Zazulak;J. Kaczorowski - 通讯作者:
J. Kaczorowski
Treatment of a left anterior descending artery chronic total occlusion using a bio-absorbable scaffold, utilising optical coherence tomography
- DOI:
10.1016/j.ijcard.2013.04.009 - 发表时间:
2013-09-01 - 期刊:
- 影响因子:
- 作者:
James Cockburn;Elizabeth Shaw;Ravinay Bhindi;Peter Hansen - 通讯作者:
Peter Hansen
Canadian Task Force on Preventive Health Care: we're back!
加拿大预防保健工作组:我们回来了!
- DOI:
- 发表时间:
2012 - 期刊:
- 影响因子:0
- 作者:
R. Birtwhistle;K. Pottie;Elizabeth Shaw;J. Dickinson;P. Brauer;M. Fortin;N. Bell;Harminder Singh;M. Tonelli;S. Connor Gorber;G. Lewin;M. Joffres;P. Parkin - 通讯作者:
P. Parkin
Free Will Skepticism in Law and Society: An Overview
法律与社会中的自由意志怀疑论:概述
- DOI:
- 发表时间:
2019 - 期刊:
- 影响因子:0
- 作者:
Gregg D. Caruso;Elizabeth Shaw;Derk Pereboom - 通讯作者:
Derk Pereboom
Elizabeth Shaw的其他文献
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{{ truncateString('Elizabeth Shaw', 18)}}的其他基金
Decoding Nitrogen Dynamics in Soil through Novel Integration of in-situ Wireless Soil Sensors with Numerical Modeling
通过原位无线土壤传感器与数值建模的新颖集成来解码土壤中的氮动态
- 批准号:
NE/T010762/1 - 财政年份:2020
- 资助金额:
$ 6.56万 - 项目类别:
Research Grant
Isotope-fluorescence activated cell sorting to allocate C utilization in the soil microbial black box
同位素荧光激活细胞分选分配土壤微生物黑匣子中的碳利用
- 批准号:
BB/F000251/1 - 财政年份:2008
- 资助金额:
$ 6.56万 - 项目类别:
Research Grant
Nanoscale zerovalent iron (nZVI) impact on soil microbial communities
纳米零价铁 (nZVI) 对土壤微生物群落的影响
- 批准号:
NE/F011946/1 - 财政年份:2008
- 资助金额:
$ 6.56万 - 项目类别:
Research Grant
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