Statistical comparison of spectral and biochemical measurements on an example of Norway spruce stands in the Ore Mountains, Czech Republic


  • Markéta Potůčková Department of Applied Geoinformatics and Cartography, Charles University in Prague
  • Lucie Červená Charles University in Prague, Faculty of Science Department of Applied Geoinformatics and Cartography
  • Lucie Kupková Charles University in Prague, Faculty of Science Department of Applied Geoinformatics and Cartography
  • Zuzana Lhotáková Charles University in Prague, Faculty of Science Department of Experimental Plant Biology
  • Jana Albrechtová Charles University in Prague, Faculty of Science Department of Applied Geoinformatics and Cartography



Laboratory spectroscopy, hyperspectral imagery, Ore Mountains, Norway spruce, Chlorophyll, Carotenoids, RWC.


The physiological status of vegetation and changes thereto can be monitored by means of biochemical analysis of collected samples as well as by means of spectroscopic measurements either on the leaf level, using field (or laboratory) spectroradiometers or on the canopy level, applying hyperspectral airborne or spaceborne image data. The presented study focuses on the statistical comparison and ascertainment of relations between three datasets collected from selected Norway spruce forest stands in the Ore Mountains, Czechia. The data sets comprise i) photosynthetic pigments (chlorophylls, carotenoids) and water content of 495 samples collected from 55 trees from three different vertical levels and the first three needle age classes, ii) the spectral reflectance of the same samples measured with an ASD Field Spec 4 Wide-Res spectroradiometer equipped with a plant contact probe, iii) an airborne hyperspecral image acquired with an Apex sensor. The datasets cover two localities in the Ore Mountains that were affected differently by acid deposits in the 1970s and 1980s. A one-way analysis of variance (ANOVA), Tukey’s honest significance test, hot spot analysis and linear regression were applied either on the original measurements (the content of leaf compounds and reflectance spectra) or derived values, i.e., selected spectral indices. The results revealed a generally low correlation between the photosynthetic pigments, water content and spectral measurement. The results of the ANOVA showed significant differences between sites (model areas) only in the case of the leaf compound dataset. Differences between the stands on various levels of significance exist in all three datasets and are explained in detail. The study also proved that the vertical gradient of the biochemical and biophysical parameters in the canopy play a role when the optical properties of the forest stands are modelled.


Jonas Ardö et al. “Satellite Based Estimations of Coniferous Forest Cover Changes:

Krusne Hory, Czech Republic”. In: Ambio 26.3 (1997), pp. 158–166.

ASD Inc. Analytical Spectral Devices. 2013. url:

and-services (visited on 06/03/2016).

ASD Inc. Analytical spectral devices. Ed. by David C Hatchell. Technical Guide 3rd.

Ed. 1999. url: http://www.geo-

New%20Folder%20%282%29/Fieldspec%20fieldguide%20TG_Rev4_web.pdf (visited

on 06/03/2016).

Richard J. Aspinall, W. Andrew Marcus, and Joseph W. Boardman. “Considerations

in collecting, processing, and analysing high spatial resolution hyperspectral data for

environmental investigations”. In: Journal of Geographical Systems 4.1 (2002), pp. 15–

doi: 10.1007/s101090100071.

P. K. Entcheva Campbell et al. “Detection of initial damage in Norway spruce canopies

using hyperspectral airborne data”. In: International Journal of Remote Sensing 25.24

(2004), pp. 5557–5584. doi: 10.1080/01431160410001726058.

Gregory A Carter and Alan K Knapp. “Leaf optical properties in higher plants: linking

spectral characteristics to stress and chlorophyll concentration”. In: American Journal

of Botany 88.4 (2001), pp. 677–684. doi: 10.2307/2657068.

Lucie Červená et al. “Models for estimating leaf pigments and relative water content

in three vertical canopy levels of norway spruce based on laboratory spectroscopy”. In:

EARSeL 34th Symposium Proceedings. Ed. by B. Zagajewski, M. Kycko, and R. Reuter.

EARSeL and University of Warsaw, 16–20 June 2014. doi: 10.12760/03-2014-11. url: symposium- Warsaw/pdf_proceedings/

EARSeL-Symposium-2014_6_1_cervena.pdf (visited on 06/03/2016).

Roshanak Darvishzadeh et al. “Mapping grassland leaf area index with airborne hyper-

spectral imagery: A comparison study of statistical approaches and inversion of radia-

tive transfer models”. In: ISPRS Journal of Photogrammetry and Remote Sensing 66.6

(2011), pp. 894–906. doi: 10.1016/j.isprsjprs.2011.09.013.

Jean-Baptiste Feret et al. “PROSPECT-4 and 5: Advances in the leaf optical properties

model separating photosynthetic pigments”. In: Remote Sensing of Environment 112.6

(2008), pp. 3030–3043. doi: 10.1016/j.rse.2008.02.012.

Arthur Getis and J Keith Ord. “The analysis of spatial association by use of distance

statistics”. In: Geographical analysis 24.3 (1992), pp. 189–206. doi: 10.1111/j.1538-


Anatoly A Gitelson et al. “Assessing carotenoid content in plant leaves with reflectance

spectroscopy”. In: Photochemistry and Photobiology 75.3 (2002), pp. 272–281. doi: 10.


Driss Haboudane et al. “Integrated narrow-band vegetation indices for prediction of

crop chlorophyll content for application to precision agriculture”. In: Remote sensing of

environment 81.2 (2002), pp. 416–426. doi: 10.1016/s0034-4257(02)00018-4.

E Raymond Hunt and Barrett N Rock. “Detection of changes in leaf water content using

near-and middle-infrared reflectances”. In: Remote sensing of environment 30.1 (1989),

pp. 43–54. doi: 10.1016/0034-4257(89)90046-1.

Stéphane Jacquemoud et al. “PROSPECT+ SAIL models: A review of use for vegetation

characterization”. In: Remote Sensing of Environment 113 (2009), S56–S66. doi: 10.


Raymond F Kokaly and Roger N Clark. “Spectroscopic determination of leaf biochem-

istry using band-depth analysis of absorption features and stepwise multiple linear re-

gression”. In: Remote sensing of environment 67.3 (1999), pp. 267–287. doi: 10.1016/


Raymond F Kokaly et al. “Characterizing canopy biochemistry from imaging spec-

troscopy and its application to ecosystem studies”. In: Remote Sensing of Environment

(2009), S78–S91. doi: 10.1016/j.rse.2008.10.018.

Lucie Kupková et al. “Chlorophyll determination in silver birch and scots pine foliage

from heavy metal polluted regions using spectral reflectance data”. In: EARSeL e-

Proceedings. Vol. 11. 1. 2012, pp. 64–73. url:

vol11_1/11_1_kupkova1.pdf (visited on 06/03/2016).

Zuzana Lhotáková et al. “Detection of multiple stresses in Scots pine growing at post-

mining sites using visible to near-infrared spectroscopy”. In: Environmental Science:

Processes & Impacts 15.11 (2013), pp. 2004–2015. doi: 10.1039/c3em00388d.

Zuzana Lhotáková et al. “Does the azimuth orientation of Norway spruce (Picea abies/L.

/Karst.) branches within sunlit crown part influence the heterogeneity of biochemical,

structural and spectral characteristics of needles?” In: Environmental and experimental

botany 59.3 (2007), pp. 283–292. doi: 10.1016/j.envexpbot.2006.02.003.

Z Malenovský et al. “Applicability of the PROSPECT model for Norway spruce nee-

dles”. In: International Journal of Remote Sensing 27.24 (2006), pp. 5315–5340. doi:


Jan Mišurec et al. “Detection of Spatio-Temporal Changes of Norway Spruce Forest

Stands in Ore Mountains Using Landsat Time Series and Airborne Hyperspectral Im-

agery”. In: Remote Sensing 8.2 (2016), p. 92. doi: 10.3390/rs8020092.

J Peñuelas et al. “Estimation of plant water concentration by the reflectance water

index WI (R900/R970)”. In: International Journal of Remote Sensing 18.13 (1997),

pp. 2869–2875. doi: 10.1080/014311697217396.

RJ Porra, WA Thompson, and PE Kriedemann. “Determination of accurate extinc-

tion coefficients and simultaneous equations for assaying chlorophylls a and b extracted

with four different solvents: verification of the concentration of chlorophyll standards by

atomic absorption spectroscopy”. In: Biochimica et Biophysica Acta (BBA)-Bioenergetics

3 (1989), pp. 384–394. doi: 0.1016/s0005-2728(89)80347-0.

Michael E Schaepman et al. “Advanced radiometry measurements and Earth science

applications with the Airborne Prism Experiment (APEX)”. In: Remote Sensing of

Environment 158 (2015), pp. 207–219. doi: 10.1016/j.rse.2014.11.014.

Daniel A Sims and John A Gamon. “Relationships between leaf pigment content and

spectral reflectance across a wide range of species, leaf structures and developmental

stages”. In: Remote sensing of environment 81.2 (2002), pp. 337–354. doi: 10.1016/


Quan Wang and Pingheng Li. “Canopy vertical heterogeneity plays a critical role in

reflectance simulation”. In: Agricultural and forest meteorology 169 (2013), pp. 111–

doi: 10.1016/j.agrformet.2012.10.004.

Alan R Wellburn. “The spectral determination of chlorophylls a and b, as well as total

carotenoids, using various solvents with spectrophotometers of different resolution”. In:

Journal of plant physiology 144.3 (1994), pp. 307–313. doi: 10.1016/s0176-1617(11)