Lin et al. (2011) used the following protocol to determine the temperature at which freezing was initiated in the plants. Stems were cut at the base, sealed with vacuum grease, placed in Petri dishes in an environmental chamber, cooled, and a high-resolution infrared camera was used to detect freezing events. The mean freezing temperature of the transgenic lines expressing both DAFPs-1 and -4 was significantly lower than in wild-type plants (2.9 ◦ Clower), and lines producing only DAFP-1 or DAFP-4. The freezing
temperature of the lines producing only DAFP-1 was also significantly lower than in the wild-type (1.8 ◦ C lower), but was not in those lines producing only DAFP-4. Tests with individual leaves excised from these plants showed similar trends. Although these results were again encouraging, a greater level of freeze avoidance was expected in the lines producing both DAFP-1 and -4 due to expected synergy between the two DAFPs. A greater level of thermal hystereis in the apoplastic fluid of these lines was also expected.
Lin et al. (2011) used the following protocol to determine the temperature at which freezing was initiated in the plants. Stems were cut at the base, sealed with vacuum grease, placed in Petri dishes in an environmental chamber, cooled, and a high-resolution infrared camera was used to detect freezing events. The mean freezing temperature of the transgenic lines expressing both DAFPs-1 and -4 was significantly lower than in wild-type plants (2.9 ◦ Clower), and lines producing only DAFP-1 or DAFP-4. The freezing
temperature of the lines producing only DAFP-1 was also significantly lower than in the wild-type (1.8 ◦ C lower), but was not in those lines producing only DAFP-4. Tests with individual leaves excised from these plants showed similar trends. Although these results were again encouraging, a greater level of freeze avoidance was expected in the lines producing both DAFP-1 and -4 due to expected synergy between the two DAFPs. A greater level of thermal hystereis in the apoplastic fluid of these lines was also expected.
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