ISHS Acta Horticulturae 319: International Symposium on Transplant Production Systems
MEASUREMENT OF CO2 CONCENTRATION IN TISSUE CULTURE VESSELS FOR SUGAR-FREE MICROPROPAGATION AND CLIMATE CHANGE STUDIES |
| Authors: | H. Davies, J. Hobbs, H. Kroese, J. Fiedler, J. Aitken-Christie |
| Keywords: | Pinus radiata (radiata pine), carbon dioxide analysis |
| Abstract:
The design of a system to investigate the response of Pinus radiata D. Don shoots grown in vitro for sugar-free micropropagation and climate change studies at ambient (≈ 350 μmol mol-1) and enriched (≈ 700 μmol mol-1) concentrations of carbon dioxide (CO2) is described. The CO2 concentration in two growth cabinets was controlled at ambient and enriched CO2 concentrations. The doors on the cabinets were replaced by 5 mm acrylic sheet with two ports for a person's arms. This enabled the CO2 concentration in the vessels containing the tissue culture material to be sampled without contamination and with minimal disturbance. A semi-permeable membrane consisting of a hydrophobic laminate of microporous polypropylene (Celgard ® 4410) was incorporated into the lid of each tissue culture vessel to allow exchange of air between the tissue culture material and the air in the growth cabinet. In addition, two valves were incorporated into the lids of some vessels to allow sterile sampling of the CO2 concentration within the vessel using a syringe. The concentration of CO2 in a sample was measured using a gas chromatograph, employing a post-column methaniser to reduce the CO2 fraction to methane (CH4) before being analysed in a flame-ionisation detector (FID). Measurements from the vessels show that this was a successful method to control the CO2 concentration in the vessels and that the sampling method was accurate and reliable. | |
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