Frost tolerance and ice formation in Pinus radiata needles: ice management by the endodermis and transfusion tissues
John S. Roden A B D , Martin J. Canny B , Chen X. Huang C and Marilyn C. Ball BA Department of Biology, Southern Oregon University, Ashland, OR 97520, USA.
B Functional Ecology Group, Research School of Biological Sciences, The Australian National University, Canberra, ACT 0200, Australia.
C Electron Microscopy Unit, Research School of Biological Sciences, The Australian National University, Canberra, ACT 0200, Australia.
D Corresponding author. Email: rodenj@sou.edu
Functional Plant Biology 36(2) 180-189 https://doi.org/10.1071/FP08247
Submitted: 24 September 2008 Accepted: 9 December 2008 Published: 5 February 2009
Abstract
Conifers are among the most frost tolerant tree species. Cryo-scanning electron microscopy (cryo-SEM) was used to visualise ice formation in pine needles to better understand how conifer leaves manage extracellular ice. Acclimated and unacclimated needles of Pinus radiata (D.Don) were subjected to freezing treatments (at a rate of 2°C h−1), tested for electrolyte leakage and sampled for cryo-SEM analysis. Half maximal electrolyte leakage occurred at –4 and −12°C for unacclimated and acclimated needles, respectively. Ice nucleation occurred at similar temperatures (−3°C) in both acclimated and unacclimated pine needles, indicating that frost tolerance did not increase supercooling. During freezing and thawing, the tissues outside and inside the endodermis shrank and swelled independently, with little or no transfer of water between the two regions. During freezing, mesophyll cells shrank, exhibiting cytorrhysis, and extracellular ice accumulated in gas spaces of the mesophyll tissue. Mesophyll cells from acclimated needles recovered their structure after thawing, and unacclimated mesophyll showed significant damage. In the vascular cylinder, ice accumulated in transfusion tracheids which expanded to occupy areas made vacant by shrinkage of transfusion parenchyma, Strasburger cells and the endodermis. This behaviour was reversible in acclimated tissue, and may play an important role in the management of ice during freeze/thaw events.
Additional keywords: anatomy, conifer, freezing, leaf, pine, thawing.
Acknowledgements
Jack Egerton and Beth Loveys provided expertise with techniques, and Daniel Harris-Pascal assisted with image analyses. Cheng Huang set up the cryo-SEM system and prepared beautiful samples for imaging. JR thanks Southern Oregon University for providing financial support during his sabbatical year, and MB thanks the Australian Research Council for support provided through Discovery Project grant DP0881009.
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