Photosynthesis Research

, Volume 118, Issue 3, pp 277–295 | Cite as

Anthocyanin contribution to chlorophyll meter readings and its correction

Regular Paper

Abstract

Leaf chlorophyll content is an important physiological parameter which can serve as an indicator of nutritional status, plant stress or senescence. Signals proportional to the chlorophyll content can be measured non-destructively with instruments detecting leaf transmittance (e.g., SPAD-502) or reflectance (e.g., showing normalized differential vegetation index, NDVI) in red and near infrared spectral regions. The measurements are based on the assumption that only chlorophylls absorb in the examined red regions. However, there is a question whether accumulation of other pigments (e.g., anthocyanins) could in some cases affect the chlorophyll meter readings. To answer this question, we cultivated tomato plants (Solanum lycopersicum L.) for a long time under low light conditions and then exposed them for several weeks (4 h a day) to high sunlight containing the UV-A spectral region. The senescent leaves of these plants evolved a high relative content of anthocyanins and visually revealed a distinct blue color. The SPAD and NDVI data were collected and the spectra of diffusive transmittance and reflectance of the leaves were measured using an integration sphere. The content of anthocyanins and chlorophylls was measured analytically. Our results show that SPAD and NDVI measurement can be significantly affected by the accumulated anthocyanins in the leaves with relatively high anthocyanin content. To describe theoretically this effect of anthocyanins, concepts of a specific absorbance and a leaf spectral polarity were developed. Corrective procedures of the chlorophyll meter readings for the anthocyanin contribution are suggested both for the transmittance and reflectance mode.

Keywords

Anthocyanins Chlorophyll Chlorophyll meters Correction NDVI SPAD 

List of symbols

a, b, d

Constant numbers

A535, A640

Absorbance of the sample at 535 and 640 nm, respectively

ACN

Anthocyanins

CACN

Anthocyanin content in leaf disk [nmol cm−2] estimated analytically

CCHL

Chlorophyll a+b content in leaf disk [nmol cm−2] estimated analytically

CRb, CRc

Mean theoretical anthocyanin (b) or chlorophyll (c) molar concentration [M] along the reflection pathway, respectively

CTb, CTc

Mean theoretical anthocyanin (b) or chlorophyll (c) molar concentration [M] along the transmission pathway, respectively

DR(λ)

Overall specific RM absorbance at λ

DRb(λ), DRc(λ)

Specific RM absorbance of anthocyanin (b) or chlorophyll (c) at λ, respectively

DRg(λ)

Mean normalized specific RM absorbance of a leaf without anthocyanins at λ

DRr(λ)

Specific RM absorbance caused by reflection, scatter, refraction, and diffraction at λ

DT(λ)

Overall specific TM absorbance at λ

DTb(λ), DTc)

Specific TM absorbance of anthocyanins (b) or chlorophyll (c) at λ, respectively

DTg(λ)

Mean normalized specific TM absorbance of a leaf without anthocyanins at λ

DTr(λ)

Specific TM absorbance caused by reflection, scatter, refraction, and diffraction at λ

DsR(λ)

Calculated specific RM absorbance at λ

DsT(λ)

Calculated specific TM absorbance at λ

Δb(SPAD)

Relative contribution of anthocyanins (b) to the SPAD signal

ΔRb(λ1R)

Relative contribution of anthocyanins (b) to the specific RM absorbance at λ1R

ΔTb(λ1T)

Relative contribution of anthocyanins (b) to the specific TM absorbance at λ1T

εRb(λ), εRc(λ)

Mean molar absorption coefficients of anthocyanins (b) or chlorophyll (c) along the reflection (R) pathway at λ, respectively

εTb(λ), εTc(λ)

Mean molar absorption coefficients of anthocyanins (b) or chlorophyll (c) along the transverse (T) pathway at λ, respectively

h

Leaf thickness

I0(λ)

Intensity of the incident light at λ

\( I^{\prime}_{ 0} (\lambda ) \)

Intensity of light entering the leaf at λ

IR(λ)

Intensity of light reflected (R) from the leaf at λ

IRe(λ), IRi(λ)

Intensity of light reflected (R) from the leaf surface at λ (external reflection) or from internal leaf structures at λ (internal reflection), respectively

IT(λ)

Intensity of light transmitted (T) through the leaf at λ

K

NDVI correction factor

l

Confidential proportionality coefficient which defines the relative SPAD units

λIR, λR

Wavelength in the infrared (IR) and red (R) region

λ1R, λ2R, λ1T, λ2T

Detection wavelengths for the reflectance (R) and transmittance (T) mode

NDVI

Normalized difference vegetation index

NDVIC

Corrected value of NDVI

Rm(λ)

Measured leaf diffusive reflectance at λ

Re(λ), Ri(λ)

External (surface) or internal reflectance of the leaf at λ, respectively

SPAD

SPAD value

SPADC

Corrected SPAD value

Specific RM absorbance

Specific absorbance in the reflectance mode

Specific TM absorbance

Specific absorbance in the transmittance mode

Tm(λ)

Measured leaf diffusive transmittance at λ

xT, xR

A beam trajectory in the transmittance (T) or reflectance (R) mode, respectively

Upper left index B

Abaxial side

Upper left index D

Adaxial side

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  1. 1.Department of Biophysics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of SciencePalacký University, OlomoucOlomoucCzech Republic

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