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In situ analysis by microspectroscopy reveals triterpenoid compositional patterns within leaf cuticles of Prunus laurocerasus

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Abstract

The cuticular waxes on the leaves of Prunus laurocerasus are arranged in distinct layers differing in triterpenoid concentrations (Jetter et al., Plant Cell Environ 23:619–628, 2000). In addition to this transversal gradient, the lateral distribution of cuticular triterpenoids must be investigated to fully describe the spatial distribution of wax components on the leaf surfaces. In the present investigation, near infrared (NIR) Raman microspectroscopy, coherent anti-Stokes Raman scattering (CARS) microscopy, and third harmonic generation (THG) spectroscopy were employed to map the triterpenoid distribution in isolated cuticles from adaxial and abaxial sides of P. laurocerasus leaves. The relative concentrations of ursolic acid and oleanolic acid were calculated by treating the cuticle spectra as linear combinations of reference spectra from the major compounds found in the wax. Raman maps of the adaxial cuticle showed that the triterpenoids accumulate to relatively high concentrations over the periclinal regions of the pavement cells, while the very long chain aliphatic wax constituents are distributed fairly evenly across the entire adaxial cuticle. In the analysis of the abaxial cuticles, the triterpenoids were found to accumulate in greater amounts over the guard cells relative to the pavement cells. The very long chain aliphatic compounds accumulated in the cuticle above the anticlinal cell walls of the pavement cells, and were found at low concentrations above the periclinals and the guard cells.

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Abbreviations

AFM:

Atomic force microscopy

ATR:

Attenuated total reflectance

BSTFA:

bis-N,N-(trimethylsilyl)trifluoroacetamide

CARS:

Coherent anti-Stokes Raman scattering

CCD:

Charge-coupled device

CM:

Cuticular membrane

FT-IR:

Fourier transform infrared

GC:

Gas chromatography

GC–FID:

Gas chromatography–flame ionization detector

MS:

Mass spectrometry

MX:

Cutin matrix

NIR:

Near infrared

NMR:

Nuclear magnetic resonance

OPA:

Optical parametric amplifier

SEM:

Scanning electron microscopy

TEM:

Transmission electron microscopy

THG:

Third harmonic generation

TIR:

Total internal reflection

Ti:Sapphire:

Titanium:Sapphire

ToF-SIMS:

Time-of-flight secondary ion mass spectrometry

XPS:

X-ray photoelectron spectroscopy

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Acknowledgments

The authors thank the Natural Sciences and Engineering Research Council (NSERC), the Canada Foundation for Innovation (CFI), the British Columbia Knowledge Development Fund (BCKDF), the Canada Research Chairs Program, and the University of British Columbia (UBC) for financial support. Instrumentation and infrastructure for this work was provided by the UBC Laboratory for Advanced Spectroscopy and Imaging Research (LASIR) and Laboratory for Molecular Biophysics (LMB).

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Authors and Affiliations

  1. Department of Chemistry, The University of British Columbia, 2036 Main Mall, Vancouver, BC, Canada, V6T 1Z3

    Marcia M. L. Yu, Stanislav O. Konorov, Michael W. Blades, Robin F. B. Turner & Reinhard Jetter

  2. Michael Smith Laboratories, The University of British Columbia, 2185 East Mall, Vancouver, BC, Canada, V6T 1Z4

    Marcia M. L. Yu, Stanislav O. Konorov, H. Georg Schulze & Robin F. B. Turner

  3. Department of Electrical and Computer Engineering, The University of British Columbia, 2332 Main Mall, Vancouver, BC, Canada, V6T 1Z4

    Robin F. B. Turner

  4. Department of Botany, The University of British Columbia, 6270 University Boulevard, Vancouver, BC, Canada, V6T 1Z4

    Reinhard Jetter

Authors
  1. Marcia M. L. Yu
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  2. Stanislav O. Konorov
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  3. H. Georg Schulze
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  4. Michael W. Blades
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  5. Robin F. B. Turner
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  6. Reinhard Jetter
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Correspondence to Reinhard Jetter.

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Yu, M.M.L., Konorov, S.O., Schulze, H.G. et al. In situ analysis by microspectroscopy reveals triterpenoid compositional patterns within leaf cuticles of Prunus laurocerasus . Planta 227, 823–834 (2008). https://doi.org/10.1007/s00425-007-0659-z

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  • DOI: https://doi.org/10.1007/s00425-007-0659-z

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