Role of glucose-6-phosphate dehydrogenase in freezing-induced freezing resistance of Populus suaveolens.

To explore the role of glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) in the enhancement of freezing resistance induced by freezing acclimation, G6PDH was purified from the leaves of 8-week-old Populus suaveolens cuttings. The G6PDH activity in the absence or the presence of reduced dithiothreitol (DTT(red)) were determined, and the changes in superoxide dismutase (SOD), peroxides (POD) and cytosolic G6PDH activities, malondial-dehyde (MDA) content as well as freezing resistance (expressed as LT(50)) of P. suaveolens cuttings during freezing acclimation at -20 degrees C were investigated. The results showed that the purified G6PDH was probably located in the cytosol of P. suaveolens. Freezing acclimation increased the activities of SOD, POD and cytosolic G6PDH, and decreased the MDA content and LT(50) of cuttings, while 2 d of de-acclimation at 25 degrees C resulted in a decrease in SOD, POD and cytosolic G6PDH activities, and caused an increase in MDA content and LT(50). The change in cytosolic G6PDH activity was found to be closely correlated to the levels of SOD, POD and MDA, and to the degree of freezing resistance of cuttings during freezing acclimation. It is suggested that the enhancement of freezing resistance of cuttings induced by freezing acclimation is related to the distinct increase in cytosolic G6PDH activity, which may be involved in the activation of SOD and POD, and the induction of freezing resistance of cuttings.

[Antifreeze proteins and molecular genetic improvement in freezing resistance of plants].

Over the past several years, the proteins and genes associated with freezing resistance and molecular genetic improvement in freezing resistance of plants have been widely studied. The recent progress of research made at home and abroad on low-temperature-induced proteins and antifreeze proteins (AFPs) with thermal hysteresis (THA) and the identification and expression regulation of low-temperature-induced genes are reviewed in this paper. Recent advances in the approaches of gene engineering that have been successfully taken in the molecular improvement of plant freezing resistance are also reviewed. Emphasis is placed on the transformation and expression of the freezing resistance genes cloned from overwintering insects, polar fishes and plant materials. Finally, some unsolved problems and the trend of research on physiological function and mechanism of AFPs, and the application of modern biotechnology to molecular improvement in freezing resistance of plants are discussed.

The role of calcium and calmodulin in freezing-induced freezing resistance of Populus tomentosa cuttings.

To explore the role of calcium-calmodulin messenger system in the transduction of low temperature signal in woody plants, Populus tomentosa cuttings after being treated with CaCl(2) (10 mmol/L), Ca(2+) chelator EGTA (3 mmol/L), Ca(2+) channel inhibitor LaCl(3) (100 mmol/L) or CaM antagonist CPZ (50 mmol/L) were used for freezing acclimation at -3 degrees C. The changes in the calmodulin (CaM) and malonaldehyde (MDA) contents, the activities of superoxide dismutase (SOD), peroxidase (POD) and Ca(2+)-dependent adenosinetriphosphatase (Ca(2+)-ATPase) of mitochondrial membrane as well as freezing resistance (expressed as LT(50)) of cuttings were investigated to elucidate the physiological mechanisms by which trees adapt to freezing. The results showed that freezing acclimation increased the CaM content, the activities of SOD, POD and Ca(2+)-ATPase of mitochondrial membrane as well as freezing resistance of cuttings, and decreased the MDA content as compared with control cuttings. Treatment with CaCl(2) at the time of freezing acclimation enhanced the effect of freezing acclimation on the above-mentioned indexes, but this enhancement was abolished by Ca(2+)chelator EGTA, Ca(2+) channel inhibitor LaCl(3) or CaM antagonist CPZ, indicating that the calcium-calmodulin messenger system was involved in the course of freezing resistance development. The presence of CaCl(2) at the same time of freezing acclimation also reduced the degree of decline in CaM content, and in SOD, POD and Ca(2+)-ATPase activities caused by freezing stress at -14 degrees C, and enhanced the level of increase in CaM content, and in SOD, POD and Ca(2+)-ATPase activity in the recovery periods at 25 degrees C . The change in CaM content was found to be closely correlated to the levels of SOD, POD and Ca(2+)-ATPase, and to the degree of freezing resistance of cuttings during freezing acclimation either with or without CaCl(2) treatment. It was suggested that the increase of CaM content induced by CaCl(2) treatment promote the formation of Ca(2+)-CaM complexes, which effectively activates the activities of SOD, POD and mitochondrial Ca(2+)-ATPase and then further result in the adaptive changes associated with the development and enhancement of freezing resistance. Thus, It could be concluded that Ca(2+)-calmodulin may be involved in the regulation of the increase in SOD, POD and Ca(2+)-ATPase activities, and the induction of freezing resistance of cuttings.