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A field-compatible method for measuring alternative respiratory pathway activities in vivo using stable O2 isotopes

Citation
Kornfeld A, Horton TW, Akir D, Searle SY, Griffin KL, Atkin OK, Subke J & Turnbull MH (2012) A field-compatible method for measuring alternative respiratory pathway activities in vivo using stable O2 isotopes, Plant, Cell and Environment, 35 (8), pp. 1518-1532.

Abstract
Plants can alter rates of electron transport through the alternative oxidase (AOX) pathway in response to environmental cues, thus modulating respiratory efficiency, but the 18O discrimination method necessary for measuring electron partitioning in vivo has been restricted to laboratory settings. To overcome this limitation, we developed a field-compatible analytical method. Series of plant tissue subsamples were incubated in 12 mL septum-capped vials for 0.5-4 h before aliquots of incubation air were injected into 3.7 mL evacuated storage vials. Vials were stored for up to 10 months before analysis by mass spectrometry. Measurements were corrected for unavoidable contamination. Additional mathematical tools were developed for detecting and addressing non-linearity (whether intrinsic or due to contamination) in the data used to estimate discrimination values. Initial contamination in the storage vials was 0.03 ± 0.01 atm; storing the gas samples at -17 °C eliminated further contamination effects over 10 months. Discrimination values obtained using our offline incubation and computation method replicated previously reported results over a range of 10-31‰, with precision generally better than ±0.5‰. Our method enables large-scale investigations of plant alternative respiration along natural environmental gradients under field conditions.

Keywords
alternative oxidase; cytochrome c oxidase; oxygen isotope discrimination; respiration; stable isotope ratio mass spectrometry (IRMS)

StatusPublished
AuthorsKornfeld Ari, Horton Travis W, Akir Dan, Searle Stephanie Y, Griffin Kevin L, Atkin Owen K, Subke Jens-Arne, Turnbull Matthew H
Publication date08/2012
Publication date online18/04/2012
Date accepted by journal11/03/2012
PublisherBlackwell
ISSN 0140-7791
LanguageEnglish

Journal
Plant, Cell and Environment: Volume 35, Issue 8

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