Controlled frost sensitivity chamber experiment (Plantae)

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Method synopsis

Plant tissues are subjected to controlled freezing temperatures in a programmable chamber to determine frost damage thresholds. Temperatures are gradually reduced to simulate natural frost events, then damage is assessed visually.

Synthesized from method descriptions across 3 papers using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Frost sensitivity of leaves and flowers of subalpine plants is related to tissue type and phenology. (2016), Journal of Ecology. The protocol was originally introduced by Frost Sensitivity of Subalpine Plants in the Colorado Rocky Mountains: The Effects of Seasonality, Water Content, and Phylogeny (2014). Implementations in other papers (listed below) may differ.

1. Experimental temperature treatments
   Temperature treatments were programmed to model the rate and duration of nighttime frost events at the field site. All frost simulations began at 5°C, the temperature was ramped down from 5°C to one of the five target temperatures (0, -2.5, -5.0, -7.5 or -10°C) over the course of 6 hours, the temperature was held for 2 hours and then brought back up to 5°C in 1 hour. Each temperature treatment was 9 hours total. Plant tissue was placed inside the temperature chamber with liquid CO₂ as cooling agent, and each leaf and flower sample was wrapped individually in its own foil packet to prevent excess water loss and desiccation during the experiment.
   Quantities: 5 temperature treatments per species, approximately 20 samples per temperature treatment for leaves and flowers, 100 samples per species across growing seasonDuration: 9 hours per temperature treatment (6 hours cooling, 2 hours hold, 1 hour warming)Conditions: Controlled cooling rate to match field frost events
   Equipment: Sigma M26-C4 programmable temperature chamber, liquid CO₂, aluminum foil
2. Frost damage assessment
   Once each temperature treatment was complete, individual plant specimens were removed from the temperature chamber and visually examined for frost damage. Frost damage to leaves and flowers was assessed using a binary damage scale where 1 = frost damage, and 0 = no frost damage. For flowers, all floral parts were assessed simultaneously without distinction among specific floral parts. A binary damage scale of continuous damage was used because in the vast majority of cases, if frost damage occurred, it was either complete or extensive throughout the plant tissue.

Typical Equipment

• Sigma M26-C4 programmable temperature chamber
• liquid CO₂
• aluminum foil

Output Measurements

• frost damage binary scores
• LT₁₀, LT₅₀, LT₉₀ temperature thresholds

Papers Using This Protocol (3)

Species Studied With This Protocol