Abstract

Frost-sensitive plant species have a limited ability to tolerate ice formation in their tissues. Most plants can supercool below 0°C and avoid ice formation. Discrepancies exist about the role of intrinsic and extrinsic ice-nucleating agents in initiating ice formation in plants. Previous research has demonstrated the ability of infrared video thermography to directly observe and record the freezing process in plants (Wisniewski et al., 1997. Plant Physiol. 113:4378–4397). In the present study, the ability of droplets of a suspension of the ice-nucleating-active (Ice+) bacterium, Pseudomonas syringae, and droplets of deionized water, to induce ice formation in bean plants was compared. The activity of these agents were also compared to intrinsic ice formation in dry plants. Results indicated that the presence of the Ice+ bacteria in droplets ranging from 0.5–4.0 μL always induced freezing at a warmer temperature than droplets of deionized water alone (no bacteria) or intrinsic nucleators in dry plants. When droplets of Ice+ bacteria were allowed to dry, they were no longer effective but were active again upon rewetting. Droplets of water would often supercool below temperatures at which ice formation was initiated by intrinsic agents. When a silicon grease barrier was placed between the droplets of Ice+ bacteria and the leaf surface, the bacteria were no longer capable of inducing ice formation in the plant, despite the droplets being frozen on the plant surface. This indicates that ice crystals must penetrate the cuticle in order to induce freezing of the plant.

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