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Title: Imaging Nutrient Distribution in the Rhizosphere Using FTIR Imaging

Abstract

Symbiotic associations in the rhizosphere between plants and microorganisms lead to efficient changes in the distribution of nutrients that promote growth and development for each organism involved. Understanding these nutrient fluxes provides insight into the molecular dynamics involved in nutrient transport from one organism to the other. Here, to study such a nutrient flow, a new application of Fourier transform infrared imaging (FTIRI) was developed that entailed growing Populus tremulodes seedlings on a thin, nutrient-enriched Phytagel matrix that allows pixel to pixel measurement of the distribution of nutrients, in particular, nitrate, in the rhizosphere. The FTIR spectra collected from ammonium nitrate in the matrix indicated the greatest changes in the spectra at 1340 cm -1 due to the asymmetric stretching vibrations of nitrate. For quantification of the nitrate concentration in the rhizosphere of experimental plants, a calibration curve was generated that gave the nitrate concentration at each pixel in the chemical image. These images of the poplar rhizosphere showed evidence for symbiotic sharing of nutrients between the plant and the fungi, Laccaria bicolor, where the nitrate concentration was five times higher near mycorrhizal roots than further out into the rhizosphere. This suggested that nitrates are acquired and transported from themore » media toward the plant root by the fungi. Similarly, the sucrose used in the growth media as a carbon source was depleted around the fungi, suggesting its uptake and consumption by the system. In conclusion, this study is the first of its kind to visualize and quantify the nutrient availability associated with mycorrhizal interactions, indicating that FTIRI has the ability to monitor nutrient changes with other microorganisms in the rhizosphere as a key step for understanding nutrient flow processes in more diverse biological systems.« less

Authors:
 [1];  [2];  [3]; ORCiD logo [4];  [3]
  1. Stony Brook Univ., NY (United States). Dept. of Chemistry
  2. Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source II (NSLS-II)
  3. University of Alabama in Huntsville, Huntsville, AL (United States). Department of Biological Science
  4. Stony Brook Univ., NY (United States). Dept. of Chemistry; Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source II (NSLS-II)
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1413940
Report Number(s):
BNL-114710-2017-JA
Journal ID: ISSN 0003-2700; TRN: US1800596
Grant/Contract Number:
SC0012704; SC0006652; AC02-98CH10886
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Analytical Chemistry
Additional Journal Information:
Journal Volume: 89; Journal Issue: 9; Journal ID: ISSN 0003-2700
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; FTIR imaging; mycorrhizal fungi; rhizosphere; Phytagel; nutrient distribution

Citation Formats

Victor, Tiffany, Delpratt, Natalie, Cseke, Sarah Beth, Miller, Lisa M., and Cseke, Leland James. Imaging Nutrient Distribution in the Rhizosphere Using FTIR Imaging. United States: N. p., 2017. Web. doi:10.1021/acs.analchem.6b04376.
Victor, Tiffany, Delpratt, Natalie, Cseke, Sarah Beth, Miller, Lisa M., & Cseke, Leland James. Imaging Nutrient Distribution in the Rhizosphere Using FTIR Imaging. United States. doi:10.1021/acs.analchem.6b04376.
Victor, Tiffany, Delpratt, Natalie, Cseke, Sarah Beth, Miller, Lisa M., and Cseke, Leland James. Mon . "Imaging Nutrient Distribution in the Rhizosphere Using FTIR Imaging". United States. doi:10.1021/acs.analchem.6b04376. https://www.osti.gov/servlets/purl/1413940.
@article{osti_1413940,
title = {Imaging Nutrient Distribution in the Rhizosphere Using FTIR Imaging},
author = {Victor, Tiffany and Delpratt, Natalie and Cseke, Sarah Beth and Miller, Lisa M. and Cseke, Leland James},
abstractNote = {Symbiotic associations in the rhizosphere between plants and microorganisms lead to efficient changes in the distribution of nutrients that promote growth and development for each organism involved. Understanding these nutrient fluxes provides insight into the molecular dynamics involved in nutrient transport from one organism to the other. Here, to study such a nutrient flow, a new application of Fourier transform infrared imaging (FTIRI) was developed that entailed growing Populus tremulodes seedlings on a thin, nutrient-enriched Phytagel matrix that allows pixel to pixel measurement of the distribution of nutrients, in particular, nitrate, in the rhizosphere. The FTIR spectra collected from ammonium nitrate in the matrix indicated the greatest changes in the spectra at 1340 cm-1 due to the asymmetric stretching vibrations of nitrate. For quantification of the nitrate concentration in the rhizosphere of experimental plants, a calibration curve was generated that gave the nitrate concentration at each pixel in the chemical image. These images of the poplar rhizosphere showed evidence for symbiotic sharing of nutrients between the plant and the fungi, Laccaria bicolor, where the nitrate concentration was five times higher near mycorrhizal roots than further out into the rhizosphere. This suggested that nitrates are acquired and transported from the media toward the plant root by the fungi. Similarly, the sucrose used in the growth media as a carbon source was depleted around the fungi, suggesting its uptake and consumption by the system. In conclusion, this study is the first of its kind to visualize and quantify the nutrient availability associated with mycorrhizal interactions, indicating that FTIRI has the ability to monitor nutrient changes with other microorganisms in the rhizosphere as a key step for understanding nutrient flow processes in more diverse biological systems.},
doi = {10.1021/acs.analchem.6b04376},
journal = {Analytical Chemistry},
number = 9,
volume = 89,
place = {United States},
year = {Mon Mar 06 00:00:00 EST 2017},
month = {Mon Mar 06 00:00:00 EST 2017}
}

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  • Many rhizosphere microorganisms enhance nutrient uptake and plant growth, but their effectiveness can vary with host species and with genotypes within species. This study evaluated the effectiveness of rhizosphere microflora indigenous to the rhizosphere of switchgrass (Panicum virgatum L.) for enhancing seedling yield and nutrient uptake. Switchgrass roots and rhizosphere soil were collected from native prairies and seeded stands in Nebraska, Kansas, Iowa, Missouri, Virginia, and North Carolina. Seedlings of four switchgrass cultivars were inoculated with root fragments and rhizosphere soil from each collection, fertilized with a nutrient solution, and grown in steamed sand for 12 weeks in a greenhouse.more » Seedlings inoculated with rhizosphere microflora produced up to 15-fold greater shoot and root yields, and recovered up to 6-fold more N and 36-fold more P than seedlings inoculated with rhizosphere bacteria only. These responses were consistent for all four switchgrass cultivars and were probably due to arbuscular mycorrhizal fungi. Switchgrass rhizosphere populations were highly variable in their ability to recover N and P and stimulate seedling shoot and root yields. Seedlings inoculated with rhizosphere populations from seeded switchgrass stands averaged 1.5-fold greater shoot and root yields than seedlings inoculated with rhizosphere populations from native prairies. Rhizosphere populations that stimulated the greatest N uptake differed from populations that resulted in the greatest P uptake. Highly effective microbial populations appear to develop in the rhizosphere of seeded switchgrass stands.« less
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  • This short review will discuss and provide perspective into the utilization of mass spectrometry imaging (MSI) in studying the rhizosphere. It also serves to compliment the multi-omic focused review by White et al. in this journal issue, as MSI is capable of elucidating chemical distributions within samples of interest in an in situ fashions, and thus can provide spatial context to MS omics data in complementary experimental endeavors. The majority of reported MSI-based studies of plant-microbe interactions have focused on the phyllosphere and ‘associated rhizosphere’ (e.g., material that is not removed during harvesting), as sample preparation for these in situmore » analyses tends to be a limiting factor. These studies have provided valuable insight into the spatial arrangement of proteins, peptides, lipids, and other metabolites within these systems. We intend for this short review to be a primer about the history of MSI and its role in plant-microbe analysis. Along the way we reference many comprehensive reviews for the interested reader. Lastly, we offer a perspective on the future of MSI and its use in understanding the molecular transformations beyond what we coined as the ‘associated rhizosphere’ to the rest of rhizosphere zone and into the bulk soil.« less
  • In our short review provides perspective regarding the use of mass spectrometry imaging (MSI) to study the rhizosphere. It also serves to complement the multi-omic-focused review by White et al. in this journals’ issue. MSI is capable of elucidating chemical distributions within samples of interest in situ, and thus can provide spatial context to MS omics data in complementary experimental endeavors. Most MSI-based studies of plant-microbe interactions have focused on the phyllosphere and on the “associated rhizosphere” (our term for material that is not removed during harvesting). Sample preparation for these in situ analyses tends to be a limiting factor.more » Our studies, however, have provided valuable insights into the spatial arrangement of proteins, peptides, lipids, and other metabolites within these systems. We intend this short review to be a primer on the fundamentals of MSI and its role in plant-microbe analysis. Finally, we offer a perspective on the future of MSI and its use in understanding the molecular transformations beyond what we call the associated rhizosphere, one which extends to the rest of rhizosphere and into the bulk soil.« less