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| 1 | One Rapid and Efficient Method for Isolation of Total RNA from Shoots Regenerated in vitro of Populus suaveolens显示文摘Total RNA was isolated from shoots regenerated in vitro of Populus suaveolens by the modified method of CTAB, and two clear bands of rRNA (28S and 18S) were observed in agarose electrophoresis. In addition, the values of OD260/OD280 and OD260/OD230 of extracted RNA were 2.12 and 2.23 respectively. The results show that RNA is little decomposed and the purity of RNA is high. Moreover, RNA isolated by the modified method of CTAB reagent had been successfully used for reverse transcription of P. suaveolens cDNAs and ideal special band was observed. | LinYuanzhen LinShanzhi ZhangZhiyi HeChengzhong GuoHuan ZhangWei | 2004 | Forestry Studies in China2004，6，1： | 5 |
| 2 | Comparative Study on Antioxidative System in Normal and Vitrified Shoots of Populus suaveolens in Tissue Culture显示文摘To explore the physiological and biochemical mechanism of the occurrence of vitrified shoots of Populus suaveolens in tissue culture, the changes in water, chlorphyll, lignin, H2O2, phenylalanine ammonialyase (PAL), malonaldehyde (MDA), protective enzymatic systems, and some key enzymes involved in the ascorbate- glutathione cycle were comparatively studied in both normal and vitrified shoots of P. suaveolens. The results show that the lower activities of peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and PAL, and the less contents of chlorphyll, lignin, ascorbate (ASA) and reduced glutathione (GSH) as well as the lower ratios of ASA / DHA and GSH / GSSG are observed in vitrified shoots than in normal ones during the whole culture period. While in comparison with normal shoots, the higher activity of superoxide dismutase (SOD) and the more concentrations of water, H2O2, MDA, dehydroascorbate (DHA) and oxidized glutathione (GSSG) are found in vitrified shoots. Statistical analysis indicates that the enhanced activity of SOD and the decreased activities of CAT and POD as well as some enzymes involved in the ascorbate-glutathione cycle might be closely correlated to the accumulation of H2O2. The less regeneration of ASA and GSH and the lower capacity of the ascorbate-glutathione cycle observed in vitrified shoots might be due to a significant decrease in APX, MDAR, DHAR and GR activities and a decline in redox status of ASA and GSH. The decreases in chlorphyll content might result in a decline in photosynthesis. The lower activities of POD and PAL could result in the decrease of lignin synthesis and cell wall ligination, which might be the key factor leading to the increase in water content. It is concluded that the deficiency of detoxification capacity caused by the lower capacity of the ascorbate-glutathione pathway and the decreased activity of protective enzymatic system might lead to the large accumulation of H2O2 and the enhancement of membrane lipid peroxidation, which might be the main cause leading to the occurrence of vitrifying shoots of P. suaveolens in tissue culture. | LinShanzhi ZhangZhiyi LinYuanzhen LiuWenfeng GuoHuan ZhangWei ZhangChong | 2004 | Forestry Studies in China2004，6，3： | 4 |
| 3 | Characterization and Role of Glucose-6-phosphate Dehydrogenase of Populus suaveolens in Induction of Freezing Resistance显示文摘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. | LinShanzhi GuoHuan LiuWenfeng LinYuanzhen ZhangQian HuDongmei ZhuBaoqing ZhangZhiyi | 2004 | Forestry Studies in China2004，6，4： | 1 |
| 4 | Identification of CpTI Gene Integration for 2-year-old Transgenic Poplars at DNA Level显示文摘The putative transgenic hybrid triploid poplars [(P. tomentosa P. bolleana) P. tomentosa] with CpTI gene have been outplanted in test field for 2 years. Although the authors previous studies have proved that they are highly resistant to 3 species of poplar-threatening insect pests and contain high content of CpTI protein in foliage, incorporation status of foreign CpTI gene in poplar genome is uncertain. In this present study, the incorporation of foreign CpTI gene in genome of 5 transgenic poplars was confirmed by PCR and Southern blotting analysis. DNA amplification showed that there were clear DNA bands of about 450bp specific to CpTI gene in transgenic lanes, while no corresponding band in non-transgenic lane was observed. Correspondingly, clear DNA hybridization signals and no signal were exhibited on film for DNA Southern blotting analysis in transgenic lanes and non-transgenic lane, respectively, which further confirmed the stable integration of foreign CpTI gene in genome of 2-year-old transgenic poplar. | ZhangQian LinShanzhi LinYuanzhen ZhangZhiyi LiuHaijun ZouYening WangZeliang | 2004 | Forestry Studies in China2004，6，3： | 1 |
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