胡杨愈伤组织质膜的两相分离法及其H＋-ATPase的特性

以胡杨愈伤组织为材料,用PEG 3350/DextranT 500构成的两相系统提取质膜微囊,研究质膜H+-ATPase的特性.结果显示由6.3% PEG 3350、6.3% Dextran T500、KCl、磷酸缓冲液(pH 7.8)和蔗糖构成的两相系统提取膜微囊的H+-ATPase活性分别被Na3VO4、KNO3、NaN3抑制了约75%、2.6%和1.3%.方向性检测显示原位膜微囊占提取质膜微囊的90%,翻转膜微囊仅占10%.去垢剂对质膜H+-ATPase活性的影响说明0.015%的Triton X-100和0.01%～0.1%的Brij 58适用于测定质膜H+-ATPase活性.Lineweaver-Burk动力学分析该酶的Km值为0.65 mmol*L-1,Vmax为37.59 μmol Pi*mg-1 protein*h-1.研究结果表明两相法提取的质膜微囊主要是正向密闭的膜微囊;胡杨愈伤组织质膜H+-ATPase的最适pH为6.5,最适温度为37℃左右.     

绿豆下胚轴质膜微囊的提取和H+-ATPase 活性研究

以两相法提取纯化绿豆下胚轴质膜微囊,材料与两相体系重量之比为32∶8时,一次洗膜就可以得到纯度较高的质膜微囊。提取缓冲液中牛血清白蛋白的浓度对质膜H+-ATPase的潜在活性有影响。质膜H+-ATPase水解活性依赖于Mg2+,Ca2+对酶活性有明显的促进作用。壳梭孢素（fusicoccin, FC）对酶有明显的刺激作用,活体条件最大刺激达到72％,而离体条件下刺激为30％。     

不同生境下木麻黄脯氨酸含量和质膜ATPase活性的研究

采集生长于恶劣环境和中生环境的普通木麻黄(Casuarina)小枝,超速离心提取粗质膜制剂后,用两相系统法纯化得到质膜微囊,研究不同生境下木麻黄的质膜ATPase活性,并测定木麻黄小枝的游离脯氨酸含量。实验结果表明:同一生境中的木麻黄ATPase活性相对一致,而同一树种木麻黄不同生境下质膜微囊H~+-ATPase、Ca~(2+)-ATPase、K~+-ATPase活性有显著差异,表现出以渗透胁迫为主的恶劣环境下的木麻黄质膜微囊ATPase活性和木麻黄细胞内游离脯氨酸明显高于中生环境下生长的木麻黄。说明普通木麻黄在干旱和盐胁迫下能调整生理生化过程来提高其质膜ATPase活性和增加细胞内脯氨酸含量提高渗透调节能力以保证其在恶劣环境的正常生长。     

灵武长枣果实质膜和液泡膜H~+-ATPase和H~+-PPase活性与果实糖分积累

以不同发育时期灵武长枣(Ziziphus jujuba cv.Lingwuchangzao)的果实为材料,通过测定与分析果肉组织中细胞质膜、液泡膜H+-ATPase和H+-PPase活性、果实糖分含量变化,研究了灵武长枣果实质膜、液泡膜H+-ATPase和H+-PPase活性与糖积累特性的关系。结果表明:(1)果实第二次快速生长期之前主要积累葡萄糖和果糖,之后果实迅速积累蔗糖,葡萄糖和果糖含量则逐渐下降,成熟期果实主要积累蔗糖。(2)在果实发育的缓慢生长期S1,质膜H+-ATPase活性最低;第一次快速生长期,质膜H+-ATPase活性最高;缓慢生长期S2,其活性降低;第二次快速生长期,质膜H+-ATPase活性升至次高;完熟期,质膜H+-ATPase活性下降幅度较大。(3)在果实发育过程中,液泡膜H+-ATPase和H+-PPase活性的变化趋势相似。缓慢生长期S1,液泡膜H+-ATPase和H+-PPase活性较低;从缓慢生长期S1至第一次快速生长期缓慢下降至最低;从第一次快速生长期开始,液泡膜H+-ATPase和H+-PPase活性呈现为逐渐增高的变化趋势;除第二次快速生长期以外,液泡膜H+-PPase活性始终高于H+-ATPase。由此推测,质膜H+-ATPase和液泡膜H+-ATPase、H+-PPase对灵武长枣果实糖分的跨膜次级转运起到重要的调控作用。     

绿豆下胚轴质膜微囊的提取和H+-ATPase活性研究

以两相法提取纯化绿豆下胚轴质膜微囊,材料与两相体系重量之比为32:8时,一次洗膜就可以得到纯度较高的质膜微囊.提取缓冲液中牛血清白蛋白的浓度对质膜H -ATPase的潜在活性有影响.质膜H -ATPase水解活性依赖于Mg2 ,Ca2 对酶活性有明显的促进作用.壳梭孢素(fusicoccin,FC)对酶有明显的刺激作用,活体条件最大刺激达到72%,而离体条件下刺激为30%.     

不同生境下木麻黄脯氨酸含量和质膜ATPase活性的研究

采集生长于恶劣环境和中生环境的普通木麻黄(Casuarina)小枝,超速离心提取粗质膜制剂后,用两相系统法纯化得到质膜微囊,研究不同生境下木麻黄的质膜ATPase活性,并测定木麻黄小枝的游离脯氨酸含量。实验结果表明：同一生境中的木麻黄ATPase活性相对一致,而同一树种木麻黄不同生境下质膜微囊H^ -ATPase、Ca^2 ATPase、K^ -ATPase活性有显著差异,表现出以渗透胁迫为主的恶劣环境下的木麻黄质膜微囊ATPase活性和木麻黄细胞内游离脯氨酸明显高于中生环境下生长的木麻黄。说明普通木麻黄在干旱和盐胁迫下能调整生理生化过程来提高其质膜ATPase活性和增加细胞内脯氨酸含量提高渗透调节能力以保证其在恶劣环境的正常生长。     

NaCl胁迫对盐芥质膜和液泡膜ATPase活性的影响

以盐生植物盐芥和中生植物拟南芥幼苗为材料,研究了盐胁迫对它们叶片和根质膜、液泡膜H+-ATPase、Ca2+-ATPases和K+-ATPase活性以及H+-ATPase、Na+/H+ 逆向转运蛋白表达的影响.结果显示:在NaCl胁迫下,盐芥叶片和根质膜的H+-ATPase活性分别比对照显著升高41%～212%和35%～53%,液泡膜的H+-ATPase分别显著升高281%～373%和4%～38%,而拟南芥却比相应对照都显著降低;相同盐浓度胁迫下,盐芥叶片的H+-ATPase活性比根部高4～8倍,盐芥根也远高于拟南芥.在NaCl胁迫下,盐芥叶片和根的液泡膜H+-ATPase蛋白质β亚基含量变化与其酶活性变化趋势一致,质膜Na+/H+ 逆向转运蛋白的表达量与Na+含量变化趋势一致.盐胁迫下盐芥根中Ca2+-ATPases和K+-ATPase活性的增加与根中Ca2+和K+含量呈显著正相关.研究发现,在盐胁迫条件下,盐芥能有效增强H+-ATPase蛋白和Na+/H+逆向转运蛋白表达,显著提高其根系与叶片质膜和液泡膜的H+-ATPase、Ca2+-ATPase和K+-ATPase活性,维持细胞质中较高的Ca2+和K+水平,从而缓解盐胁迫的伤害,增强耐盐性.     

茉莉酸甲酯和脱落酸对绿豆下胚轴质膜H+-ATPase水解活性的影响

茉莉酸甲酯(MeJA)促进绿豆下胚轴质膜H+-ATPase水解活性.活体条件下,50μmol·L-1 MeJA处理7 h的酶活性提高30%;离体条件下,10 μmol·L-1 MeJA处理2 h的酶活性最大,即提高30%.壳梭孢素(FC)和MeJA在离体条件下对H+-ATPase活性的促进效应相同,均提高30%左右,无协同效应;活体条件下,FC促进质膜H+-ATPase水解活性可达70%,而MeJA仅为30%.离体条件下,脱落酸(ABA)对H+-ATPase水解活性无明显促进;而活体条件下则有一定的抑制.     

氯化镁预处理对美人蕉根部质膜H+-ATPase活性与农田废水硝态氮吸收效率的影响

以湿地挺水植物七叶红色大花美人蕉(Cana generalis Bailey)幼苗为实验材料,在温室中用150μmol/L氯化镁溶液预处理12h,然后置于硝态氮(NO3--N)浓度为10mg/L的农田废水中处理,考察氯化镁预处理对美人蕉吸收农田废水中硝态氮效率的影响,以及处理期间根部质膜H+-ATPase活性、H+-泵活性、14-3-3蛋白与质膜H+-ATPase相互作用。结果显示:经过150μmol/L氯化镁溶液预处理12h后,美人蕉吸收农田废水中硝态氮的效率比对照增加11%;与没有预处理的植株相比,氯化镁预处理美人蕉根中质膜H+-ATPase和H+-泵的活性均显著提高,质膜H+-ATPase和14-3-3蛋白的相互作用显著增强。研究表明,氯化镁能够通过增强质膜H+-ATPase与14-3-3蛋白的相互作用来提高质膜H+-ATPase的活性,从而增加美人蕉对硝态氮的吸收效率。     

NaCl胁迫对宁夏枸杞幼苗根系质膜和液泡膜H+-ATPase活性的影响

以宁夏枸杞为材料,研究NaCl胁迫下枸杞幼苗组织含水量、干物质重量、无机离子分布、质膜透性、丙二醛 (MDA)含量及幼根质膜、液泡膜H -ATPase活性和耐盐性之间的关系。结果表明,在0．3％NaCl胁迫下,干物质重量、质膜透性、MDA含量略有增加,组织含水量略有下降;随着NaCl胁迫强度的增加,Na 、Cl-含量逐渐增加, K 含量则呈现下降的趋势;质膜透性增大与MDA含量升高呈显著正相关;质膜和液泡膜H -ATPase活性逐渐增加,且液泡膜H -ATPase活性显著高于质膜H -ATPase活性,表明枸杞具有较强的抗盐性。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.