Non-destructive measurement of chlorophyll b : a ratios and identification of photosynthetic pathways in grasses by reflectance spectroscopy
Katharina Siebke A B and Marilyn C. Ball A CA Division of Plant Sciences, Research School of Biology, Australian National University, Canberra, ACT 0200, Australia.
B Present address: Heinz Walz GmbH, Eichenring 6, 91090 Effeltrich, Germany.
C Corresponding author. Email: marilyn.ball@anu.edu.au
This paper originates from a presentation at the 1st International Plant Phenomics Symposium, Canberra, Australia, April 2009.
Functional Plant Biology 36(11) 857-866 https://doi.org/10.1071/FP09201
Submitted: 29 July 2009 Accepted: 28 September 2009 Published: 5 November 2009
Abstract
Equations for non-destructive determination of chlorophyll b : a ratios in grasses were developed from reflectance spectra of intact leaves of barley (Hordeum vulgare L.) and two barley mutants: clorina f2, which lacks chlorophyll b and clorina f104, which has a low chlorophyll b content. These plants enabled separation of effects of chlorophyll composition on reflectance spectra due to differential light absorption by chlorophylls a and b and to measure the effects of chlorophyll b on the contribution of fluorescence emitted by chlorophyll a to the reflectance spectra. Indices developed from these data were then tested on growth chamber-grown leaves from six C3 and 17 C4 grass species (7 NAD-ME and 10 NADP-ME subtypes). We used the chlorophyll b : a ratio because the data were less skewed than the chlorophyll a : b ratio. The best index for determination of the chlorophyll b : a ratio utilised wavelengths affected by chlorophyll absorbance: [R626 – 0.5 (R603 + R647)]/[R552– R626]. The chlorophyll b : a ratio was significantly lower in the C4 than C3 grasses, but was not sufficient in itself to separate these two functional groups. However, because of differences in fluorescence characteristics, C3 and C4 species could be distinguished by an index based on wavelengths affected by chlorophyll fluorescence: [R696 to 709/R545 to 567].
Additional keywords: C3-photosynthesis, C4-photosynthesis, leaf pigments, remote sensing.
Acknowledgements
We thank Professors Terry Caelli, Paul Kriedemann and Jochen Zeil and Drs Adam Gilmore and Adrienne Nicotra for critical discussion of previous versions of the manuscript, Dr Oula Ghannoum for providing C4 grass seeds, Dr David Barker for pigment determinations, Drs Luigi Ranzullo, Glenn Newnham and Susanne Thulin for inspiring discussions.
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