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Nitrogen Fixation by Legumes' Rhizobia

Measuring H2 evolution as a means of determining nitrogenase activity overcome problems of the traditional acetylene reduction assay. The Q-Box Q-NF1LP Nitrogen Fixation Package is completely safe, non-invasive and allows changes in nitrogenase activity to be measured in growing plants.

Nitrogen Fixation Package Q-NF1LP

Q-NF1LP Nitrogen Fixation Package
Q-NF1LP

Q-NF1LP Nitrogen Fixation Package, the only complete experimental package for non-destructively measuring nitrogen fixation in H2-producing legume symbioses. The low range flow-through H2 sensor (Q-S121) measures the rate of H2 production from N2-fixing tissues, allowing in vivo approximation of nitrogenase activity in real time. The package also includes the Q-S151 CO2 analyser (0-2000 ppm), which is used to measure nodulated root respiration. The Q-NF1LP package contains everything needed for successful experiments, including gas exchange pots for growing legumes such as soybeans under nodule-producing conditions, nutrient solution recipes, rhizobia inoculum, and detailed instructions. When combined with the Q-GSS Gas Switching System, nitrogenase activity can be measured in up to 3 legume root systems in parallel (plus a control channel; open flow setting). The N fixation package contains everything needed to carry out a wide range of laboratory studies on the physiology and biochemistry of N2 fixation in an open-flow gas exchange system. The analysers are easy to calibrate, set-up time is minimal and experiments are simple to perform. Applications of the Q-NF1LP range from studies of the effects of temperature/photosynthetic assimilation on nitrogenase activity and root respiration, N fixation efficiency of different genotypes, inhibition of N2 fixation by fertilisation, to the electron allocation coefficient of nitrogenase, etc.

Q-NF1LP Nitrogen Fixation System - Features

  • Non-invasive measurements of nitrogenase activity and respiration from nodulated roots
    • Flow-through gas analysis system for measuring nitrogenase activity in H2-evolving rhizobial symbioses
    • Parallel measurement of CO2 evolution from nodulated roots
  • Easy set-up and calibration
  • Software calculations of nitrogen fixation rates

Options:

  • Multiplexing system, 4 chambers (optional)
  • Custom experimental pots (size, material) available
Components Q-NF1LP Nitrogen Fixation Package OPEN

Product No. Description
Q-NF1LP Complete nitrogen fixation package Q-NF1LP for H2 and CO2 measurements on legumes / rhizobia under laboratory conditions. The system includes the following components:
Q-S121 Hydrogen Analyzer 0-100 ppm, includes a drying column
Q-S151 CO2 Analyzer 0-2000 ppm,  includes CO2 and H2O scrubbers
Q-G131 Plant Growth Pots, 4x, with lids and accessories (fittings, sealant)
Q-G161 Nodule Cuvette
Q-G267 Gas pump and Flow Monitor, 1 L min-1
Q-G268 Gas pump and Flow Monitor, 2 L min-1
Q-S132 Temperature Sensor
Q-C301 Event Switch (Air, Ar)
Q-S122 Large Gas Sample Bags (2x 30L)
Q-C610 LabQuest Mini data interfaces, 3x (9 channels), integrated
Q-C404 Experimental Setup Files "N Fixation", for LoggerPro
Q-C901 Logger Pro Software
Q-NF1LP-Manual Manual
Q-NF1LP-Case Rugged Case, housing analyzers and sensors
Q-NF1LP-Kit Accessory Kit (tubing, connectors, etc.) + N-Fixation Experimentation Starter Kit (Soybean Seeds + Bradyrhizobium japonicum inoculant)
Q-NF1LP-Power Individual power supplies, for stand-alone use of the analyzers, pumps etc.
   
Optional  
Q-GSS Gas Switching System, 4 channels
VSI-Chambers Custom Gas Measurement Chambers; build to user specifications
References Q-NF1LP Package OPEN

  • Bulgarelli RG, et al. (2017) Mycorrhizae enhance nitrogen fixation and photosynthesis in phosphorus-starved soybean (Glycine max L. Merrill). Environmental and Experimental Botany 140: 26-33  doi: 10.1016/j.envexpbot.2017.05.015
  • Cabeza R et al. (2014) An RNA sequencing transcriptome analysis reveals novel insights into molecular aspects of the nitrate impact on nodule activity of Medicago truncatula.  Plant Physiology 164:400-411 doi: 10.1104/pp.113.228312
  • Ferreira C, et al. (2016) pH effects on nodulation and biological nitrogen fixation in Calopogonium mucunoides. Brazilian Journal of Botany 39, 1015–1020 doi: 10.1007/s40415-016-0300-0
  • Justino GC, Sodek L. (2013) Recovery of nitrogen fixation after short-term flooding of the nodulated root system of soybean. Journal of Plant Physiology 170: 235-241 10.1016/j.jplph.2012.10.006
  • Kassaw T, Bridges W, Frugoli J. (2015) Multiple Autoregulation of Nodulation (AON) Signals Identified through Split Root Analysis of Medicago truncatula sunn and rdn1 Mutants. Plants 4(2), 209-224; doi: 10.3390/plants4020209
  • Wang X, et al. (2018) Coupling between nitrogen fixation and tetrachlorobiphenyl dechlorination in a rhizobium–legume symbiosis. Environ Sci. Technol. 52: 22178-2224 doi: 10.1021/acs.est.7b05667
Q-NF1LP Nitroxen Fixation Analyzer, with Pot
Q-NF1LP, software live data view
Q-NF1LP, software calculation page
Get Quote

Contact us to receive the Q-NF1LP Nitrogen Fixation Package Manual.



Measuring Principle: Why is H2 evolution related to Nitrogen Fixation by Legumes?

Biological nitrogen fixation (BNF) in economically important food and forage crops has attracted attention worldwide to achieve sustainable agricultural goals. Continue reading

Diazotrophic microorganisms from the bacterial or archaeal domains are responsible for BNF, and only some prokaryotes are able to use atmospheric nitrogen through BNF by encoding nitrogenase, an enzyme that catalyses the conversion of N2 gas to ammonia (NH3). The symbiotic relationship between soil bacteria, collectively known as rhizobia (including the genera Rhizobium, Bradyrhizobium, etc.), and legume roots produces nodules that fix atmospheric nitrogen through the action of the enzyme nitrogenase, releasing approximately one molecule of H2 per molecule of N2 fixed. Hydrogen emission plays a crucial role in the symbiotic relationship between legumes and nitrogen-fixing microorganisms, rhizobia, within their root nodules. In this process, legumes release hydrogen ions (H+) into the surrounding soil. These hydrogen ions help creating an oxygen-deficient environment that favours the activity of nitrogenase enzymes produced by the rhizobia. The emitted hydrogen serves a dual purpose: first, it acts as a signalling molecule that attracts rhizobia to the root hairs of the legume. Secondly, the hydrogen creates an oxygen-deficient environment within the nodule, protecting the often highly O2-sensitive nitrogenase enzymes. This allows efficient N2 fixation to take place, providing the legume with a vital source of nitrogen essential for its growth and development. Gaseous H2 emissions can therefore be directly related to nitrogen fixation using measurements with the Q-NF1LP system.

Hydrogen Production by Free-Living Diazotrophs

Free-living diazotrophs correspond to a small fraction of the plant rhizospheres ecosystem, and they belong to alphaproteobacteria (Rhizobia, Bradyrhizobia, Rhodobacteria), betaproteobacteria (Burkholderia, Nitrosospira), gammaproteobacteria (Pseudomonas, Xanthomonus), firmicutes, and cyanobacteria. These microorganisms can produce molecular hydrogen. Show more

Nitrogenases and hydrogenases are involved in H2 production as described above and this can be exploited to increase hydrogen production on an industrial scale. Understanding the underlying mechanisms of hydrogen production in (cyano-)bacteria and improving production methods will enable the commercialisation of molecular hydrogen as an efficient and environmentally friendly energy source. Although the Q-NF1LP Nitrogen Fixation Package was originally designed for plant-focused research, it can also be used to study H2 emission from bacterial and algal suspensions with minor modifications (i.e. cuvette with headspace) similar to the Q-FL23 Algae Photosynthesis Package. Contact us for a customised system to suit your research needs.


CO2 and O2 Combi Gas Analyser

For parallel measurement of CO2 and O2 in the gas phase with a single analyser, consider the Q-S159 CO2 + O2 analyser. Various other Gas Analysers available for CO2, O2, Methane. 


Gas pumps, flow meters

See our range of multiplexers, gas pumps and flow meters, and Gas Blending and Gas Control Systems to set up custom gas analysis systems. Get in contact for advice.



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