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JXB Advance Access originally published online on April 8, 2009
Journal of Experimental Botany 2009 60(8):2217-2234; doi:10.1093/jxb/erp081
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© The Author [2009]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

This article appears in the following Journal of Experimental Botany issue: Special Issue: Mesophyll conductance to CO2: mechanisms, modelling, and ecological implications [View the issue table of contents]

REVIEW-ARTICLE

Estimating mesophyll conductance to CO2: methodology, potential errors, and recommendations

Thijs L. Pons1, Jaume Flexas2,*, Susanne von Caemmerer3, John R. Evans3, Bernard Genty4, Miquel Ribas-Carbo2 and Enrico Brugnoli5

1Department of Plant Ecophysiology, Utrecht University, PO Box 80084, 3598 TB Utrecht, The Netherlands
2Research Group on ‘Plant Biology under Mediterranean Conditions’, Department of Biology, Universitat de les Illes Balears, Carretera de Valldemossa Km 7.5, 07122 Palma de Mallorca, Illes Balears, Spain
3Research School of Biological Sciences, The Australian National University, Canberra, Australian Capital Territory 2601, Australia
4CEA, CNRS, Université Aix-Marseille, UMR 6191 Biologie Végétale et Microbiologie Environnementale, Laboratoire d'Ecophysiologie Moléculaire des Plantes, CEA Cadarache, 13108 Saint Paul lez Durance, France
5CNR-Institute of Agro-Environmental Biology and Forestry, Via Marconi 2, I-05010 Porano (TR), Italy

* To whom correspondence should be addressed. E-mail: jaume.flexas{at}uib.es

The three most commonly used methods for estimating mesophyll conductance (gm) are described. They are based on gas exchange measurements either (i) by themselves; (ii) in combination with chlorophyll fluorescence quenching analysis; or (iii) in combination with discrimination against 13CO2. To obtain reliable estimates of gm, the highest possible accuracy of gas exchange is required, particularly when using small leaf chambers. While there may be problems in achieving a high accuracy with leaf chambers that clamp onto a leaf with gaskets, guidelines are provided for making necessary corrections that increase reliability. All methods also rely on models for the calculation of gm and are sensitive to variation in the values of the model parameters. The sensitivity to these factors and to measurement error is analysed and ways to obtain the most reliable gm values are discussed. Small leaf areas can best be measured using one of the fluorescence methods. When larger leaf areas can be measured in larger chambers, the online isotopic methods are preferred. Using the large CO2 draw-down provided by big chambers, and the isotopic method, is particularly important when measuring leaves with high gm that have a small difference in [CO2] between the substomatal cavity and the site of carboxylation in the chloroplast (Ci–Cc gradient). However, equipment for the fluorescence methods is more easily accessible. Carbon isotope discrimination can also be measured in recently synthesized carbohydrates, which has its advantages under field conditions when large number of samples must be processed. The curve-fitting method that uses gas exchange measurements only is not preferred and should only be used when no alternative is available. Since all methods have their weaknesses, the use of two methods for the estimation of gm, which are as independent as possible, is recommended.

Key words: Chlorophyll fluorescence, isotope discrimination, mesophyll conductance, methodology, photosynthesis

Received 19 December 2008; Revised 22 February 2009 Accepted 25 February 2009


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