Antifreeze Evaluation of Two Dehydrin Proteins from <i>Pseudotsuga menziesii</i> and <i>Larix principis-rupprechtii</i> Mayr

Abstract

Dehydrins exist widely in plants and play an important role in abiotic stress resistance. Two low-temperature-induced dehydrin-like genes, PmCAP and LpCAP, from the pine species Pseudotsuga menziesii and Larix principis-rupprechtii Mayr were cloned and found to contain 576 bp and 687 bp, encoding 191 and 228 amino acids, respectively. Both genes were individually assembled into prokaryotic expression vectors and transferred into E. coli cells. When transgenic stains were cultured at −5 °C, the lethal time 50% (LT50) was 50 h and 54 h for PmCAP and LpCAP, respectively, compared with 32 h for the control. When cultured at −20 °C, the LT50 was 38 h, 41 h, and 25 h for PmCAP, LpCAP, and the control. Thermal hysteresis (TH) testing of PmCAP and LpCAP proteins revealed TH values of 0.27 °C and 0.72 °C, respectively, relative to 0.05 °C for the BSA control. These results indicate that the two pine dehydrin proteins have antifreeze characteristics and that their antifreeze levels were well in relation to the environmental conditions of pine growth (Larix principis-rupprechtii Mayr mostly grows in cold and high-altitude zones, while Pseudotsuga menziesii grows in temperate and low-altitude zones). LpCAP, especially, could be a better gene resource for the molecular breeding of plant cold resistance.

Keywords

• dehydrin
• prokaryotic expression
• thermal hysteresis
• pine trees
• antifreeze