Purification and Characterization of Cytosolic NADP Specific Isocitrate Dehydrogenase from Pisum sativum

Cytosolic NADP-specific isocitrate dehydrogenase was isolated from leaves of Pisum sativum. The purified enzyme was obtained by ammonium sulfate fractionation, ion exchange, affinity, and gel filtration chromatography. The purification procedure yields greater than 50% of the total enzyme activity originally present in the crude extract. The enzyme has a native molecular weight of 90 kilodaltons and is resolved into two catalytically active bands by isoelectric focusing. Purified NADP-isocitrate dehydrogenase exhibited K_m values of 23 micromolar for dl-isocitrate and 10 micromolar for NADP, and displayed optimum activity at pH 8.5 with both Mg²⁺ and Mn²⁺.     

光质对一品红叶离体培养中形态发生及生化变化的影响

本文讨论了将光质应用于一品红(Euphorbia pulcherrima)大规模细胞繁殖及形态发生调控的可行性,并测定了一些形态及生化指标作为这种调控的参数。 以一品红试管苗的叶片作为外植体,置于MS培养基中并分别培养于白(W)、红(R)、黄(Y)、蓝(B)、绿(G)色光及黑暗(D)下,一个月后进行测定。蓝光和黄光明显促进出苗。而愈伤组织形成的情况则正相反,红光利于愈伤组织的形成,黄光效果最差。过氧化物酶、过氧化氢酶活力及蛋白质、RNA、碳水化合物的含量与愈伤组织形成有较为一致的关系,但与分化不直接相关。 实验中还测定了叶绿素a、叶绿素b及类胡萝卜素的含量。白光与蓝光均能促进叶绿素及类胡萝卜素的合成,但蓝光中chl b/chl a的比率较高。此外,本文尚涉及暗处理中的晶质化现象(vitrification)及其成因。     

Measurement of extracellular 5-HT and 5-HIAA in hippocampus of freely moving rats using microdialysis: long-term applications

Extracellular levels of 5-hydroxytryptamine (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) were measured in the ventral hippocampus of the awake rat using microdialysis. The basal level of 5-HT in hippocampal dialysates was very close to the detection limit of our assay. However, addition of a 5-HT re-uptake blocker, citalopram, to the perfusion medium resulted in a 3-fold rise of 5-HT levels in dialysates and provided a stable baseline. Under these conditions, extracellular levels of 5-HT and 5-HIAA could be measured up to 11 days after dialysis probe implantation. 5-HT measured in dialysate was Ca²⁺-dependent both 24 h and 7 days after surgery. In comparison K⁺-induced depolarization caused a 9-fold increase in 5-HT output 24 h after probe implantation and this effect had disappeared 10 days later, although behavioral activation following K⁺ application was similar 24 h and 11 days after surgery. Systematic administration of -trytophan had no significant effect on 5-HT but increased 5-HIAA levels by 90%. These results suggest that in the presence of citalopram, 5-HT in hippocampal dialysates is derived from serotonergic neurones. The microdialysis method appears most useful for measurement of extracellular 5-HT 24–72 h after probe implantation. Finally, our data indicate that extracellular 5-HIAA mainly reflects intraneuronal metabolism of 5-HT prior to release.     

Protoplast isolation and callus production from leaves of tissue-cultured Vitis spp.

Protoplasts were isolated from mesophyll of axenic cultures of grape, Vitis rotundifolia cv. Summit and V. vinifera cv. Cabernet Sauvignon. Enzymes effective for protoplast isolation were Macerozyme R-10 (0.5% and 0.1%) and Cellulase Onozuka R-10 (1.0% and 0.5%) for V. rotundifolia and V. vinifera, respectively. Polyvinylpyrrolidone and 2-(N-morpholino)ethanesulfonic acid were essential in the isolation media. Protoplasts were purified using flotation/centrifugation. The protoplasts of V. rotundifolia cultured in Gamborg's B5 basal medium with 2.2 M 6-benzyladenine, 4.5 M 2,4-dichlorophenoxyacetic acid and 0.4% agarose gave the best plating efficiency of conditions tried in this study. Cell division occurred within 5 to 6 days and visible microcalli developed within one month. After 6 weeks in culture, microcalli transferred to liquid medium exhibited active callus growth. Protoplasts of V. vinifera cultured under these conditions had similar results.Abbreviations MES 2-(N-morpholino)ethanesulfonic acid monohydrate - PVP polyvinylpyrrolidone - PDS potassium dextran sulfate - BA 6-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - FDA fluorescein diacetate     

水生观赏植物的调查和研究

水生绿化观赏植物是美化园林水景的重要植物材料,作者通过几年的调查研究和引种栽培,提出了28种(其中荷花含18个品种,睡莲含20个品种)水生绿化观赏植物,同时介绍了20种(其中荷花含6个品种)观赏价值较高的水生绿化观赏植物。     

猪和牛轮状病毒抗原和抗体的检测

应用ELISA直接双抗体夹心法检查轮状病毒抗原,24份仔猪和29份犊牛的腹泻粪样,分别有12和16份阳性。用病毒RNA电泳分析检查阳性粪样,各出现两种病毒RNA电泳型,用中和试验检查17份成年牛和16份成年猪血清,分别有16和15份病毒抗体阳性。将其与ELISA间接法和结合法进行了比较。     

Ca~(++)促进大麦根K~+吸收的过程分析

吸收溶液中CaCl_2促进了大麦根K~+净吸收,Ca~(++)本身对H~+分泌无影响,Cl~-减少了H~+净分泌量。 含有~(86)Rb的大麦根在1m mol/L KCl溶液中发生~(86)Rb外流,在H_2O、1m mol/L NaCl或0.5 m mol/L CaCl_2中没有明显外流。VO_4~(3-)、NaN_3和4℃低温均可以减少根段在1m mol/L KCl溶液中的~(86)Rb外流量。Ca~(++)抑制K~+(~(86)Rb~+)的外流,EDTA加剧外流,Ca~(++)可以逆转EDTA的效应。 Ca~(++)抑制K~+(~(86)Rb)外流和促进K~+净吸收的趋势相吻合。K~+吸收的通量分析结果表明,Ca~(++)抑制K~+通过质膜的外流,促进K~+由细胞质向液泡中转运,而不影响K~+通过质膜的内流速率。     

高等植物叶片中的硝酸还原酶活性稳定因子

从小麦、油菜、浮萍、番茄、烟草的叶片中分离得到NR-SF。不同植物材料中NR及NR-SF能起交叉反应;不同NR-SF影响NR酶动力学性质相同;不同NR-SF的凝胶电泳谱带显示蛋白和糖蛋白性质。NR-SF广泛存在于植物细胞中。