[Objective] The experiment was aimed to select effective and economical media for container seedling of triploid clones of Populus tomentosa that was carried out. [Method] The sandy loam, peat, perlite, vermiculite, r...[Objective] The experiment was aimed to select effective and economical media for container seedling of triploid clones of Populus tomentosa that was carried out. [Method] The sandy loam, peat, perlite, vermiculite, riversand, sludge were taken as media of hardwood cutting and survival rate, seedling height were taken as indexes to select media for container seedling of triploid clones of Populus tomentosa. [Result] Different mixedmedia had great influence on survival rates of container seedlings. Taking peat and vermiculite with the proportion of 5∶2 (M10) or peat ,vermiculite with the proportion of 7∶2 (M11) or sandy loam (M1) as media would generate higher cutting survival rate that was higher than 90.0%. There were significant differece in height increments of container seedlings. Taking sandy loam, peat and vermiculite with the proportion of 6∶2∶2(M5)or sandy loam (M1), seedling height of 60-days the seedling was over 37.0 cm. [Conclusion] According to cost analysis of nursery medium, the optimum medium for hardwood cuttings container seedling-raising of triploid clones of Populus tomentosa was sandy loam.展开更多
cDNA encoding caffeoyl CoA O-methyltransferase (CCoAOMT) from Chinese white poplar ( Populus tomentosa Carr.) was cloned by RT-PCR and sequenced. Northern analysis displayed that the CCoAOMT was expressed specifically...cDNA encoding caffeoyl CoA O-methyltransferase (CCoAOMT) from Chinese white poplar ( Populus tomentosa Carr.) was cloned by RT-PCR and sequenced. Northern analysis displayed that the CCoAOMT was expressed specifically in the developing secondary xylem and its expression was coincident with lignification. The antisense CCoAOMT cDNA was transformed into P. tremula x P. alba mediated by Agrobacterium tumefaciens ( Smith et Townsend) Conn. Transgenic plants were identified with PCR, PCR-Southern and Southern analysis. Lignin content in 5- to 6-month-old transgenic plants was measured. One of the transgenic lines had significant reduction of 17.9% in Klason lignin content as compared with that of untransformed poplar. The results demonstrate that antisense repression of CCoAOMT is an efficient way to reduce lignin content for improving pulping property in engineered trees.展开更多
The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of pla...The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of plants, such as concentration of plant growth regulators, leaf explant orientation, leaf growth sequence and leaf segment, were studied. The results indicated that the differentiation rate of adventitious shoots was 90% on basal MS medium only supplemented with 1 5?mg·L -1 BA (6 benzyladenine) and reached the highest level(95%) when 1 0?mg·L -1 BA and 0 3?mg·L -1 NAA (naphthaleneacetic acid) were added to MS medium. 90% of differentiation rate of adventitious roots were obtained when 0 3?mg·L -1 NAA was only added to MS medium. It was also found that more adventitious shoots were regenerated from the lower segment of leaf (with petiole) than the other segments, the number of adventitious shoots decreased from top to base of leaf growth sequence and the percentage of adventitious shoot induction with adaxial side downward was higher than that with adaxial side upward.展开更多
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
Acyl-ACP thioesterases （FATs） terminates the fatty acid synthesis and allow the transport of fatty acids out of the plastids, which are the important determinants of cellular metabolism. FATB is a member of FAT enzy...Acyl-ACP thioesterases （FATs） terminates the fatty acid synthesis and allow the transport of fatty acids out of the plastids, which are the important determinants of cellular metabolism. FATB is a member of FAT enzymes that has been described previously in most of the plants. In silico cloning is a new method that utilizes the bioinformatics on the complete genome and available EST database. In this study, a full-length cDNA clone of PtFATB gene was isolated from Populus tomentosa using this approach. It is 1,450 bp in length and the open reading frame encodes a peptide of 421 amino acids. The predicted amino acid sequence shows significant homology with those from other plant species, which contain typical domains owned by FATB proteins. The transcripts of PtFATB were abundant in leaves, and less in roots detected by using semiquantitative RT-PCR. When the shoots were subjected to the stress treatments （cold, dry, NaC1） and ABA （Abscisic acid）, the expression of PtFATB was only slightly reduced under the treatment of low temperature. This suggests that the expression of PtFATB is in a constitutive fashion. This study provides the basis not only for the identification and characterization of this gene but also for the improvement of cold tolerance by controlling the expression of the PtFATB gene in trees in near future.展开更多
基金Supported by National Key Technology R&D Program during the 11th Five-year Plan Period(2006BAD32B01)~~
文摘[Objective] The experiment was aimed to select effective and economical media for container seedling of triploid clones of Populus tomentosa that was carried out. [Method] The sandy loam, peat, perlite, vermiculite, riversand, sludge were taken as media of hardwood cutting and survival rate, seedling height were taken as indexes to select media for container seedling of triploid clones of Populus tomentosa. [Result] Different mixedmedia had great influence on survival rates of container seedlings. Taking peat and vermiculite with the proportion of 5∶2 (M10) or peat ,vermiculite with the proportion of 7∶2 (M11) or sandy loam (M1) as media would generate higher cutting survival rate that was higher than 90.0%. There were significant differece in height increments of container seedlings. Taking sandy loam, peat and vermiculite with the proportion of 6∶2∶2(M5)or sandy loam (M1), seedling height of 60-days the seedling was over 37.0 cm. [Conclusion] According to cost analysis of nursery medium, the optimum medium for hardwood cuttings container seedling-raising of triploid clones of Populus tomentosa was sandy loam.
文摘cDNA encoding caffeoyl CoA O-methyltransferase (CCoAOMT) from Chinese white poplar ( Populus tomentosa Carr.) was cloned by RT-PCR and sequenced. Northern analysis displayed that the CCoAOMT was expressed specifically in the developing secondary xylem and its expression was coincident with lignification. The antisense CCoAOMT cDNA was transformed into P. tremula x P. alba mediated by Agrobacterium tumefaciens ( Smith et Townsend) Conn. Transgenic plants were identified with PCR, PCR-Southern and Southern analysis. Lignin content in 5- to 6-month-old transgenic plants was measured. One of the transgenic lines had significant reduction of 17.9% in Klason lignin content as compared with that of untransformed poplar. The results demonstrate that antisense repression of CCoAOMT is an efficient way to reduce lignin content for improving pulping property in engineered trees.
文摘The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of plants, such as concentration of plant growth regulators, leaf explant orientation, leaf growth sequence and leaf segment, were studied. The results indicated that the differentiation rate of adventitious shoots was 90% on basal MS medium only supplemented with 1 5?mg·L -1 BA (6 benzyladenine) and reached the highest level(95%) when 1 0?mg·L -1 BA and 0 3?mg·L -1 NAA (naphthaleneacetic acid) were added to MS medium. 90% of differentiation rate of adventitious roots were obtained when 0 3?mg·L -1 NAA was only added to MS medium. It was also found that more adventitious shoots were regenerated from the lower segment of leaf (with petiole) than the other segments, the number of adventitious shoots decreased from top to base of leaf growth sequence and the percentage of adventitious shoot induction with adaxial side downward was higher than that with adaxial side upward.
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This work was supported by project "Regulation of Composition and Saturation of Fatty Acid in Trees by Genetic Engineering", Introduction of Foreign Advanced Agricultural Science and Technology into China (No. 2005-4-52).
文摘Acyl-ACP thioesterases （FATs） terminates the fatty acid synthesis and allow the transport of fatty acids out of the plastids, which are the important determinants of cellular metabolism. FATB is a member of FAT enzymes that has been described previously in most of the plants. In silico cloning is a new method that utilizes the bioinformatics on the complete genome and available EST database. In this study, a full-length cDNA clone of PtFATB gene was isolated from Populus tomentosa using this approach. It is 1,450 bp in length and the open reading frame encodes a peptide of 421 amino acids. The predicted amino acid sequence shows significant homology with those from other plant species, which contain typical domains owned by FATB proteins. The transcripts of PtFATB were abundant in leaves, and less in roots detected by using semiquantitative RT-PCR. When the shoots were subjected to the stress treatments （cold, dry, NaC1） and ABA （Abscisic acid）, the expression of PtFATB was only slightly reduced under the treatment of low temperature. This suggests that the expression of PtFATB is in a constitutive fashion. This study provides the basis not only for the identification and characterization of this gene but also for the improvement of cold tolerance by controlling the expression of the PtFATB gene in trees in near future.
