Populus tomentosa cuttings were treated with 1mmol·L -1 , 5mmol·L -1 , 10mmol·L -1 or 15mmol·L -1 of CaCl 2 for 1\|7 d, respectively, for studying the effects of different conc...Populus tomentosa cuttings were treated with 1mmol·L -1 , 5mmol·L -1 , 10mmol·L -1 or 15mmol·L -1 of CaCl 2 for 1\|7 d, respectively, for studying the effects of different concentrations of CaCl 2 on freezing resistance. Results indicated that 10?mmol·L -1 of CaCl 2 has greater effect than other concentrations on the enhancement of freezing resistance, and the optimum time of pretreatment was 5?d. In addition, cuttings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 of CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 05?mmol·L -1 of CaM antagonist CPZ or 0 1?mmol·L -1 of Ca 2+ channel inhibitor LaCl 3 The changes in CaM and freezing resistance of all cuttings were investigated. The results showed that cold acclimation at -3℃ increased CaM content and decreased the minimum temperature for 100% survival. The CaCl 2 pretreatment enhanced the effect of cold acclimation and obviously increased CaM content and decreased the minimum temperature for 100% survival, but this effect was strongly inhibited by the EGTA, CPZ or LaCl 3 It is concluded that the effect of CaCl 2 on freezing resistance is associated with its concentration and time of pretreatment, Ca 2+ CaM may be involved in the induction of freezing resistance of the cuttings.展开更多
Populus tomentosa seedlings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 1?mmol·L -1 of Ca 2+ ...Populus tomentosa seedlings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 1?mmol·L -1 of Ca 2+ channel inhibitor LaCl 3,and 0 05?mmol·L -1 of CaM antagonist CPZ. The changes in the contents of total soluble protein and CaM, and freezing resistance in all pretreated seedlings in various periods ( viz: following cold acclimation, chilling stress and recovery) were investigated. Results showed that cold acclimation increased the contents of total soluble protein and CaM, and freezing resistance of seedlings, which could be strongly reduced by the pretreatments of EGTA CPZ and LaCl 3 Cold acclimation combined with CaCl 2 pretreatment enhanced the effect of cold acclimation on freezing resistance, and obviously increased the contents of total soluble protein and CaM, reduced the declining degree of the contents of total soluble protein and CaM caused by chilling stress as compared with cold acclimation, augmented the increase in the level of total soluble protein and CaM during the recovery periods. Further analysis found that an increase in total soluble protein content during cold acclimation with or without CaCl 2 pretreatment mainly resulted from the increase in content of heat stable protein in total soluble protein. It is suggested that Ca 2+ calmodulin may be involved in the synthesis of total soluble protein, and the induction of freezing resistance of seedlings.展开更多
The changes in the contents of total soluble protein and RNA, the activity of RNase in leaves and branches of Populus tomentosa cuttings at various periods (viz: cold acclimation, deacclimation, chilling stress an...The changes in the contents of total soluble protein and RNA, the activity of RNase in leaves and branches of Populus tomentosa cuttings at various periods (viz: cold acclimation, deacclimation, chilling stress and the recovery after chilling stress), and the survival rate and the freezing resistance of cuttings during cold acclimation at -3℃ were investigated. Results showed that cold acclimation not only increased the contents of total soluble protein and RNA, the survival rates and the freezing resistance of cuttings, decreased the activity of RNase, but also reduced the declining degree of total soluble protein and RNA contents, and the increasing level of RNase caused by chilling stress as compared with the controls. In addition, cold acclimation augmented the increase in the level of total soluble protein and RNA, and facilitated the decrease of RNase during the recovery periods. Further analysis found that the DNA content of all treatments kept relative stability at various periods. The changes in total soluble protein, RNA and RNase were closely related to the freezing resistance of cuttings. It appears that the increase of RNA content caused by cold acclimation induced decrease of RNase activity may be involved in the accumulation of total soluble protein and the induction of freezing resistance of cuttings.展开更多
Glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) was purified from the leaves of 8-week-old Populus suaveolens cuttings. The enzyme activity in the absence and presence of reduced dithiothreitol (DTTred) was det...Glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) was purified from the leaves of 8-week-old Populus suaveolens cuttings. The enzyme activity in the absence and presence of reduced dithiothreitol (DTTred) was determined. The results show that the G6PDH activity is not inactivated by pre-incubation with DTTred, indicating that the purified enzyme probably pre- sented in cytosol of P. suaveolens. The catalytic characteristics and kinetic parameters of cytosolic G6PDH purified from P. suaveolens cuttings were also studied. The results show that G6PDH is characterized by Kmvalue of 360 μmol·L–1 for G6P and 16 μmol·L–1 for NADP, a pH range of 7.3–8.9, and the maximum activity around pH 8.2. The enzyme activity is inhibited by various metabolites such as NADPH, NADH, GTP, UTP, ATP, AMP, ADP, CoA, acetyl CoA, fructose-6-phosphate (F6P), eryth- rose-4-phosphate (E4P), ribose-5-phosphate (R5P) and 3-phosphoglycerate (3-PG) (all at 1 mmol·L–1 except for NADPH and NADH) to different extents. NADPH is the most effective inhibitor of enzyme activity, with an inhibition of 72.0%. The addition of metal ions such as MgCl2, CaCl2and KCl (all 1.0 mmol·L–1) to the standard reaction mixture has no remarkable influence on the cytosolic G6PDH activity. However, CdCl2 (1.0 mmol·L–1) causes high inhibitory effect on the enzyme activity. To explore the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation, the changes in the cytosolic G6PDH activity and freezing resistance (expressed as LT50) of P. suaveolens cuttings during freezing acclimation at –20 °C were investigated. The results reveal that freezing acclimation decreases LT50 of cuttings, and increases the activity of cytosolic G6PDH compared with control ones, while 2 d of de-acclimation at 25 °C result in a decrease in cytosolic G6PDH activity, and caused an increase in LT50. Furthermore, the change in cytosolic G6PDH activity is found to be closely correlated to the degree of freezing resistance of cuttings during freez- ing acclimation. It is suggested that cytosolic G6PDH may be involved in the induction of freezing resistance of cuttings.展开更多
The differences of glucose-6P dehydrogenase (G6PDH) activity and freezing resistance induced by freezing acclimation between cuttings of freezing-sensitive P. tomentosa and freezing-resistant P. suaveolens were compar...The differences of glucose-6P dehydrogenase (G6PDH) activity and freezing resistance induced by freezing acclimation between cuttings of freezing-sensitive P. tomentosa and freezing-resistant P. suaveolens were compared for exploring the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation. After 5 d of freezing acclimation at -3 ℃, the LT50 of P. tomentosa has decreased from -6.2 ℃ in control cuttings to -14.3 ℃ in freezing acclimated ones, and the increase of G6PDH activity was observed in freezing acclimated cuttings as compared with control ones. Whereas, when P. suaveolens was freezing acclimated at -20℃ for 5 d, the LT50 has decreased from -27.1℃ in control cuttings to -43.5 ℃ in freezing acclimated ones, and the activity of G6PDH increased considerably. In addition, the increase of LT50 and the decrease of G6PDH activity resulting from 2 d of deacclimation at 25 ℃ were found in two kinds of freezing acclimated cuttings. It is concluded that the increase in the activity of G6PDH may associate with the inherited freezing resistance of species and the enhancement of freezing resistance of cuttings, and may play an important role in the antifreeze process under freezing temperature, which would provide the basis for the study on the molecular mechanism of freezing resistance in P. suaveolens and the cloning of gene associated with freezing resistance.展开更多
We analyzed the relationships linking overwintering death and frost cracking to temperature and sunlight as well as the effects of low temperatures and freeze–thaw cycles on bud-burst rates,relative electrical conduc...We analyzed the relationships linking overwintering death and frost cracking to temperature and sunlight as well as the effects of low temperatures and freeze–thaw cycles on bud-burst rates,relative electrical conductivity,and phloem and cambial ultrastructures of poplar.Overwintering death rates of poplar were not correlated with negative accumulated temperature or winter minimum temperature.Freeze–thaw cycles caused more bud damage than constant exposure to low temperatures.Resistance to freeze–thaw cycles differed among clones,and the budburst rate decreased with increasing exposure to freeze–thaw cycles.Cold-resistant clones had the lowest relative electrical conductivity.Chloroplasts exhibited the fastest and the most obvious reaction to freeze–thaw damage,whereas a single freeze–thaw cycle caused little damage to cambium ultrastructure.Several such cycles resulted in damage to plasma membranes,severe damage to organelles,dehydration of cells and cell death.We conclude that overwintering death of poplar is mainly attributed to the accumulation of effective freeze–thaw damage beyond the limits of freeze–thaw resistance.展开更多
文摘Populus tomentosa cuttings were treated with 1mmol·L -1 , 5mmol·L -1 , 10mmol·L -1 or 15mmol·L -1 of CaCl 2 for 1\|7 d, respectively, for studying the effects of different concentrations of CaCl 2 on freezing resistance. Results indicated that 10?mmol·L -1 of CaCl 2 has greater effect than other concentrations on the enhancement of freezing resistance, and the optimum time of pretreatment was 5?d. In addition, cuttings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 of CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 05?mmol·L -1 of CaM antagonist CPZ or 0 1?mmol·L -1 of Ca 2+ channel inhibitor LaCl 3 The changes in CaM and freezing resistance of all cuttings were investigated. The results showed that cold acclimation at -3℃ increased CaM content and decreased the minimum temperature for 100% survival. The CaCl 2 pretreatment enhanced the effect of cold acclimation and obviously increased CaM content and decreased the minimum temperature for 100% survival, but this effect was strongly inhibited by the EGTA, CPZ or LaCl 3 It is concluded that the effect of CaCl 2 on freezing resistance is associated with its concentration and time of pretreatment, Ca 2+ CaM may be involved in the induction of freezing resistance of the cuttings.
文摘Populus tomentosa seedlings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 1?mmol·L -1 of Ca 2+ channel inhibitor LaCl 3,and 0 05?mmol·L -1 of CaM antagonist CPZ. The changes in the contents of total soluble protein and CaM, and freezing resistance in all pretreated seedlings in various periods ( viz: following cold acclimation, chilling stress and recovery) were investigated. Results showed that cold acclimation increased the contents of total soluble protein and CaM, and freezing resistance of seedlings, which could be strongly reduced by the pretreatments of EGTA CPZ and LaCl 3 Cold acclimation combined with CaCl 2 pretreatment enhanced the effect of cold acclimation on freezing resistance, and obviously increased the contents of total soluble protein and CaM, reduced the declining degree of the contents of total soluble protein and CaM caused by chilling stress as compared with cold acclimation, augmented the increase in the level of total soluble protein and CaM during the recovery periods. Further analysis found that an increase in total soluble protein content during cold acclimation with or without CaCl 2 pretreatment mainly resulted from the increase in content of heat stable protein in total soluble protein. It is suggested that Ca 2+ calmodulin may be involved in the synthesis of total soluble protein, and the induction of freezing resistance of seedlings.
文摘The changes in the contents of total soluble protein and RNA, the activity of RNase in leaves and branches of Populus tomentosa cuttings at various periods (viz: cold acclimation, deacclimation, chilling stress and the recovery after chilling stress), and the survival rate and the freezing resistance of cuttings during cold acclimation at -3℃ were investigated. Results showed that cold acclimation not only increased the contents of total soluble protein and RNA, the survival rates and the freezing resistance of cuttings, decreased the activity of RNase, but also reduced the declining degree of total soluble protein and RNA contents, and the increasing level of RNase caused by chilling stress as compared with the controls. In addition, cold acclimation augmented the increase in the level of total soluble protein and RNA, and facilitated the decrease of RNase during the recovery periods. Further analysis found that the DNA content of all treatments kept relative stability at various periods. The changes in total soluble protein, RNA and RNase were closely related to the freezing resistance of cuttings. It appears that the increase of RNA content caused by cold acclimation induced decrease of RNase activity may be involved in the accumulation of total soluble protein and the induction of freezing resistance of cuttings.
基金the Foundation ofState-designated Base for Biology Researching and Teaching in Beijing Forestry University
文摘Glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) was purified from the leaves of 8-week-old Populus suaveolens cuttings. The enzyme activity in the absence and presence of reduced dithiothreitol (DTTred) was determined. The results show that the G6PDH activity is not inactivated by pre-incubation with DTTred, indicating that the purified enzyme probably pre- sented in cytosol of P. suaveolens. The catalytic characteristics and kinetic parameters of cytosolic G6PDH purified from P. suaveolens cuttings were also studied. The results show that G6PDH is characterized by Kmvalue of 360 μmol·L–1 for G6P and 16 μmol·L–1 for NADP, a pH range of 7.3–8.9, and the maximum activity around pH 8.2. The enzyme activity is inhibited by various metabolites such as NADPH, NADH, GTP, UTP, ATP, AMP, ADP, CoA, acetyl CoA, fructose-6-phosphate (F6P), eryth- rose-4-phosphate (E4P), ribose-5-phosphate (R5P) and 3-phosphoglycerate (3-PG) (all at 1 mmol·L–1 except for NADPH and NADH) to different extents. NADPH is the most effective inhibitor of enzyme activity, with an inhibition of 72.0%. The addition of metal ions such as MgCl2, CaCl2and KCl (all 1.0 mmol·L–1) to the standard reaction mixture has no remarkable influence on the cytosolic G6PDH activity. However, CdCl2 (1.0 mmol·L–1) causes high inhibitory effect on the enzyme activity. To explore the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation, the changes in the cytosolic G6PDH activity and freezing resistance (expressed as LT50) of P. suaveolens cuttings during freezing acclimation at –20 °C were investigated. The results reveal that freezing acclimation decreases LT50 of cuttings, and increases the activity of cytosolic G6PDH compared with control ones, while 2 d of de-acclimation at 25 °C result in a decrease in cytosolic G6PDH activity, and caused an increase in LT50. Furthermore, the change in cytosolic G6PDH activity is found to be closely correlated to the degree of freezing resistance of cuttings during freez- ing acclimation. It is suggested that cytosolic G6PDH may be involved in the induction of freezing resistance of cuttings.
基金National Natural Science Foundation of China(Grant No.30271093)
文摘The differences of glucose-6P dehydrogenase (G6PDH) activity and freezing resistance induced by freezing acclimation between cuttings of freezing-sensitive P. tomentosa and freezing-resistant P. suaveolens were compared for exploring the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation. After 5 d of freezing acclimation at -3 ℃, the LT50 of P. tomentosa has decreased from -6.2 ℃ in control cuttings to -14.3 ℃ in freezing acclimated ones, and the increase of G6PDH activity was observed in freezing acclimated cuttings as compared with control ones. Whereas, when P. suaveolens was freezing acclimated at -20℃ for 5 d, the LT50 has decreased from -27.1℃ in control cuttings to -43.5 ℃ in freezing acclimated ones, and the activity of G6PDH increased considerably. In addition, the increase of LT50 and the decrease of G6PDH activity resulting from 2 d of deacclimation at 25 ℃ were found in two kinds of freezing acclimated cuttings. It is concluded that the increase in the activity of G6PDH may associate with the inherited freezing resistance of species and the enhancement of freezing resistance of cuttings, and may play an important role in the antifreeze process under freezing temperature, which would provide the basis for the study on the molecular mechanism of freezing resistance in P. suaveolens and the cloning of gene associated with freezing resistance.
基金supported by grants from the National Key Research and Development Program of China(Grant No.2016YFD0600401)the Liaoning Provincial Key Research Project for Agriculture(Grant No.2015103002)
文摘We analyzed the relationships linking overwintering death and frost cracking to temperature and sunlight as well as the effects of low temperatures and freeze–thaw cycles on bud-burst rates,relative electrical conductivity,and phloem and cambial ultrastructures of poplar.Overwintering death rates of poplar were not correlated with negative accumulated temperature or winter minimum temperature.Freeze–thaw cycles caused more bud damage than constant exposure to low temperatures.Resistance to freeze–thaw cycles differed among clones,and the budburst rate decreased with increasing exposure to freeze–thaw cycles.Cold-resistant clones had the lowest relative electrical conductivity.Chloroplasts exhibited the fastest and the most obvious reaction to freeze–thaw damage,whereas a single freeze–thaw cycle caused little damage to cambium ultrastructure.Several such cycles resulted in damage to plasma membranes,severe damage to organelles,dehydration of cells and cell death.We conclude that overwintering death of poplar is mainly attributed to the accumulation of effective freeze–thaw damage beyond the limits of freeze–thaw resistance.
