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Publication Details
 
Output Category: C1
Strategic Research Area: None
TISI Citations: PlumX StatisticsPlumX Statistics
Scopus Citations:
Journal Impact: 3.45
All Authors: Scafaro, A, Negrini, A, O'Leary, B, Rashid, F, Hayes, L, Fan, Y, Zhang, Y, Chochois, V, Badger, M, Millar, A, Atkin, O Number:
UWA Authors: O'Leary, B., Millar, A. Number: 2
Title: The combination of gas-phase fluorophore technology and automation to enable high-throughput analysis of plant respiration
Journal: Plant Methods   
ISBN/ISSN 1746-4811
Year: 2017
Volume: 13
Issue: 1
Full Reference (Harvard Style): Scafaro, A., Negrini, A., O'Leary, B., Rashid, F., Hayes, L., Fan, Y., Zhang, Y., Chochois, V., Badger, M., Millar, A., Atkin, O. 2017, 'The combination of gas-phase fluorophore technology and automation to enable high-throughput analysis of plant respiration', Plant Methods, 13, 1,
Abstract:

Background: Mitochondrial respiration in the dark (R dark) is a critical plant physiological process, and hence a reliable, efficient and high-throughput method of measuring variation in rates of R dark is essential for agronomic and ecological studies. However, currently methods used to measure R dark in plant tissues are typically low throughput. We assessed a high-throughput automated fluorophore system of detecting multiple O2 consumption rates. The fluorophore technique was compared with O2-electrodes, infrared gas analysers (IRGA), and membrane inlet mass spectrometry, to determine accuracy and speed of detecting respiratory fluxes. Results: The high-throughput fluorophore system provided stable measurements of R dark in detached leaf and root tissues over many hours. High-throughput potential was evident in that the fluorophore system was 10 to 26-fold faster per sample measurement than other conventional methods. The versatility of the technique was evident in its enabling: (1) rapid screening of R dark in 138 genotypes of wheat; and, (2) quantification of rarely-assessed whole-plant R dark through dissection and simultaneous measurements of above- and below-ground organs. Discussion: Variation in absolute R dark was observed between techniques, likely due to variation in sample conditions (i.e. liquid vs. gas-phase, open vs. closed systems), indicating that comparisons between studies using different measuring apparatus may not be feasible. However, the high-throughput protocol we present provided similar values of R dark to the most commonly used IRGA instrument currently employed by plant scientists. Together with the greater than tenfold increase in sample processing speed, we conclude that the high-throughput protocol enables reliable, stable and reproducible measurements of R dark on multiple samples simultaneously, irrespective of plant or tissue type.