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Cited by 107Spectroscopy Techniques in Biomedical and Chemical ResearchSpectroscopy and Chemometric AnalysesAlgal biology and biofuel production

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Metabolic responses of eukaryotic microalgae to environmental stress limit the ability of FT-IR spectroscopy for species identification

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Driver, Thomas, Bajhaiya, Amit K., Allwood, J. William, Goodacre, Royston, Pittman, Jon K., Dean, Andrew P.·2015·Algal Research
DOI 10.1016/j.algal.2015.06.009PMC PMC4694091
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Abstract

Fourier Transform Infrared (FT-IR) spectroscopy is a robust method for macromolecular analysis and differentiation of microorganisms. However, most studies are performed in controlled conditions and it is unclear whether this tool is appropriate for the identification of eukaryotic microalgae species from variable environments. In order to address this, nine closely-related species of marine and freshwater microalgae were grown under controlled (non-stressed) and variable (non-stressed and stressed) conditions, including nutrient-stressed and wastewater-stressed conditions. Following optimization of data processing methods, FT-IR spectra from all species and conditions were compared. The substantial metabolic changes that were caused by nutrient starvation restricted the ability of FT-IR spectroscopy to differentiate the microalgal species grown under variable conditions efficiently. Comparison of unsupervised and supervised multivariate data analysis methods found that principal component-discriminant function analysis was able best to differentiate between some species under controlled conditions but still gave poor differentiation under variable environmental conditions.

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Fig. 1 FT-IR spectra from C. reinhardtii. Typical FT-IR spectra (wavenumbers 4000–600 cm− 1) C. reinhardtii grown in either standard TAP or Low N TAP media (a). Both spectra are normalized to the amide I peak at 1655 cm− 1. Major band assignments, including bands associated with lipids, proteins and carbohydrates, are indicated by arrows and defined in the list below the graph. Comparison of averaged spectra, following EMSC2 normalisation, generated from C. reinhardtii grown in TAP (from 18 replicates), Low P TAP (from 18 replicates), or Low N TAP media (from 9 replicates) (b). The spectral region referred to as cut-down spectra (wavenumbers 1800–950 cm− 1) in subsequent experiments is indicated by dotted lines.
Fig. 1 FT-IR spectra from C. reinhardtii. Typical FT-IR spectra (wavenumbers 4000–600 cm− 1) C. reinhardtii grown in either standard TAP or Low N TAP media (a). Both spectra are normalized to the amide I peak at 1655 cm− 1. Major band assignments, including bands associated with lipids, proteins and carbohydrates, are indicated by arrows and defined in the list below the graph. Comparison of averaged spectra, following EMSC2 normalisation, generated from C. reinhardtii grown in TAP (from 18 replicates), Low P TAP (from 18 replicates), or Low N TAP media (from 9 replicates) (b). The spectral region referred to as cut-down spectra (wavenumbers 1800–950 cm− 1) in subsequent experiments is indicated by dotted lines.CC BY · PubMed Central
Fig. 2 Comparison of the effect of full or cut-down FT-IR spectral ranges on sample clustering. PCA scores plots (a) and PC1 and PC2 loading plots (b) for full spectra (wavenumbers 4000–600 cm− 1) and scores (c) and loading plots (d) for cut-down spectra (wavenumbers 1800–950 cm− 1) generated from C. reinhardtii grown in either standard TAP (18 replicates), Low P TAP (18 replicates), or Low N TAP media (9 replicates).
Fig. 2 Comparison of the effect of full or cut-down FT-IR spectral ranges on sample clustering. PCA scores plots (a) and PC1 and PC2 loading plots (b) for full spectra (wavenumbers 4000–600 cm− 1) and scores (c) and loading plots (d) for cut-down spectra (wavenumbers 1800–950 cm− 1) generated from C. reinhardtii grown in either standard TAP (18 replicates), Low P TAP (18 replicates), or Low N TAP media (9 replicates).CC BY · PubMed Central
Fig. 3 An example of nutrient stress-induced metabolic response determined by FT-IR spectroscopy in a marine microalga. Mean EMSC2 normalised FT-IR spectra (wavenumbers 1800–950 cm− 1) generated from D. tertiolecta grown in artificial seawater (ASP2) media or fertilised seawater media (a). PCA score plot (b) and PC1 loading plot (c) derived from the replicate spectra. Carbohydrate:amide I and lipid:amide I peak height ratio values generated from replicate spectra (d). Data are mean values ± SE of three replicates. Significant differences (p < 0.05) are indicated with an asterisk.
Fig. 3 An example of nutrient stress-induced metabolic response determined by FT-IR spectroscopy in a marine microalga. Mean EMSC2 normalised FT-IR spectra (wavenumbers 1800–950 cm− 1) generated from D. tertiolecta grown in artificial seawater (ASP2) media or fertilised seawater media (a). PCA score plot (b) and PC1 loading plot (c) derived from the replicate spectra. Carbohydrate:amide I and lipid:amide I peak height ratio values generated from replicate spectra (d). Data are mean values ± SE of three replicates. Significant differences (p < 0.05) are indicated with an asterisk.CC BY · PubMed Central
Fig. 4 PCA score plots of EMSC2 normalised cut-down FT-IR spectra of nine algal species grown for 7 days in non-stressed conditions (a) and a combined plot of the species grown in non-stressed and stressed conditions (b). Different species are highlighted using different symbols. C.r. = C. reinhardtii, C.c. = C. concordia, D.t. = D. tertiolecta, P.k. = P. kessleri, C.l. = C. luteoviridis, H.t. = H. tetrachotoma, D.i. = D. intermedius, C.d. = C. debaryana, C.v. = C. vulgaris. Mean spectra for each species is shown in Fig. S3.
Fig. 4 PCA score plots of EMSC2 normalised cut-down FT-IR spectra of nine algal species grown for 7 days in non-stressed conditions (a) and a combined plot of the species grown in non-stressed and stressed conditions (b). Different species are highlighted using different symbols. C.r. = C. reinhardtii, C.c. = C. concordia, D.t. = D. tertiolecta, P.k. = P. kessleri, C.l. = C. luteoviridis, H.t. = H. tetrachotoma, D.i. = D. intermedius, C.d. = C. debaryana, C.v. = C. vulgaris. Mean spectra for each species is shown in Fig. S3.CC BY · PubMed Central
Fig. 5 PC-DFA scores plots of EMSC2 normalised cut-down FT-IR spectra of nine algal species grown for 7 days in non-stressed conditions (a) and a combined plot of the species grown in non-stressed and stressed conditions (b). Different species are highlighted using different symbols as described in Fig. 4.
Fig. 5 PC-DFA scores plots of EMSC2 normalised cut-down FT-IR spectra of nine algal species grown for 7 days in non-stressed conditions (a) and a combined plot of the species grown in non-stressed and stressed conditions (b). Different species are highlighted using different symbols as described in Fig. 4.CC BY · PubMed Central

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Metadata

Authors
Driver, T., Bajhaiya, A. K., Allwood, J. W., Goodacre, R., Pittman, J. K., & Dean, A. P.
Affiliations
  • University of Manchester (GB)
  • James Hutton Institute (GB)
Corresponding author
  • Pittman, Jon K.
  • Dean, Andrew P.
ORCID
Venue
Algal Research
Publisher
Elsevier BV
Published
2015-09
Volume
11
Pages
148-155
ISSN
2211-9264 (ISSN-L)
DOI
10.1016/j.algal.2015.06.009
PMC
PMC4694091
OpenAlex
W1766987757
Cited by
107
References
31
Citations by year
2026: 10 · 2025: 3 · 2024: 14 · 2023: 6 · 2022: 9 · 2021: 13 · 2020: 23 · 2019: 9 · 2018: 10 · 2017: 3 · 2016: 6 · 2015: 1
FWCI
4.03
Type
journal article
Language
en
Open access
Licence
Locations
  • Algal Research (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Research Explorer (The University of Manchester) (publishedVersion)
Funders
Carbon Trust (082)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
  • Physical Sciences › Chemistry › Analytical Chemistry › Spectroscopy and Chemometric Analyses
  • Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
Keywords
eukaryotic microalgae, species identification, FTIR spectroscopy, environmental stress, nutrient deprivation, metabolic response, multivariate analysis, marine microalgae, freshwater microalgae

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