Growth under elevated [CO 2 ] promoted spring frost damage in field grown seedlings of snow gum (Eucalyptus pauciflora Sieb. ex Spreng.), one of the most frost tolerant of eucalypts. Freezing began in the leaf midvein, consistent with it being a major site of frost damage under field conditions. The average ice nucleation temperature was higher in leaves grown under elevated [CO 2 ] (-5·7°C versus -4·3°C), consistent with the greater incidence of frost damage in these leaves (34% versus 68% of leaves damaged). These results have major implications for agriculture, forestry and vegetation dynamics, as an increase in frost susceptibility may reduce potential gains in productivity from CO 2 fertilization and may affect predictions of vegetation change based on increasing temperature. [CO 2 ]; freeze-induced damage; global climate change; ice nucleation; open-top chambers.
Key-words: Eucalyptus pauciflora; elevated atmospheric
INTRODUCTIONAtmospheric CO 2 concentrations have been increasing since the industrial revolution, and are predicted to reach twice present levels late next century (Boden et al. 1994). It is thought that temperature will increase as [CO 2 ] rises, and that warming may be greater in winter than in summer (Plummer, Lin & Torok 1996). This could lengthen the growing season if evergreen species were able to take advantage of warm conditions earlier in spring when below-ground resources are relatively abundant. However, predictions of plant responses to global climate and atmospheric change are complicated by weather variability (Katz & Brown 1992) and the extent to which elevated [CO 2 ] might affect plant responses to temperature (Long 1991), particularly low temperatures. Some studies have predicted that frost damage may increase for trees that break dormancy too early in spring (Cannell & Smith 1986;Repo, Hänninen & Kellomäki 1996) Here we report the results of a serendipitous experiment. The experiment naturally occurred during a larger fieldbased study aimed at understanding the effects of elevated [CO 2 ] on the interactions between grass and trees during spring. The results were unexpected and showed that one of the most frost-hardy of broadleaved, evergreen species suffered greater frost damage when grown under elevated than ambient [CO 2 ].
MATERIALS AND METHODS
Plant material and growth conditionsSeeds of Eucalyptus pauciflora Sieb. ex Spreng. were collected from three trees growing along the floor of the Orroral Valley at an elevation of 850 m in New South Wales, Australia. The seeds were cold stratified under moist conditions at 3°C for 4 weeks before germinating on sand flats in a mist house. Seedlings of similar size were transferred to individual containers (5 cm diameter, 25 cm deep) and grown out of doors for 6 months before the start of the experiment.In a pasture near Bungendore in southeastern Australia (35°15' S, 149°27' E; elevation 700 m), 10 open-top chambers were installed as five replicate pairs, flushed with air containing either ambient or elevated CO 2 conce...