胡杨液泡膜微囊的纯化及其质子转运活性

 为进一步研究液泡膜及 H+ - ATP酶在胡杨抵御盐胁迫中所起的作用 ,比较了研磨、捣碎和超声破碎三种细胞破碎方法 ,从悬浮培养的胡杨细胞中制备液泡膜微囊的效果 ;并用差速离心和不连续蔗糖密度梯度离心纯化了胡杨液泡膜微囊 .通过测定 H+ - ATP酶对 NO-3 、VO3-4和 Na N3的敏感性 ,以及焦磷酸酶质子转运活性表明 ,液泡膜微囊主要分布在 0 %～ 2 5%的蔗糖界面上 .捣碎法破碎细胞结合差速离心和蔗糖密度梯度离心可获得正向微囊比例高、封闭性好和酶活性高的液泡膜微囊     

玉米根尖细胞液泡膜结合的蛋白激酶的存在及其性质

为了解液泡膜蛋白在植物细胞信号途径中的功能 ,用新型的非放射性同位素方法从玉米根细胞的高纯度液泡膜上鉴定出一种膜内在的蛋白激酶。这种蛋白激酶具有Ca2 依赖、CaM和磷脂酰丝氨酸不依赖等特性 ,与已在多种植物中报道的含有类似钙调素结构域的蛋白激酶CDPK相似。离体实验表明其活性的最适pH值为 6 .5 ,最适Ca2 浓度为 1 0 μmol/L。从最适pH值和去污剂的影响可以推测出其活性位点朝向胞质一侧。Zn2 对其活性没有明显的抑制作用 ,说明该激酶缺少某些哺乳动物的蛋白激酶常含有的锌指结构。当液泡膜蛋白在Ca2 和ATP存在的条件下被预磷酸化后 ,液泡膜H _ATPase的ATP水解和质子转运过程均被激活。激活的活性可以被碱性磷酸酶逆转。以上结果说明玉米根尖细胞的液泡膜中存在一种可能是CDPK的蛋白激酶。由它造成的Ca2 依赖的磷酸化作用激活了液泡膜H _ATPase的活性。这些结果将有助于深入研究CDPK在植物细胞信号转导中的功能。     

大豆液泡膜H~+-ATPase的纯化和重组

通过不连续蔗糖密度梯度离心得到的液泡膜微囊 ,先由胆酸钠和 OG分步破膜抽提、经阴离子交换柱 ( Q- Sepharose)层析分离 .纯化后的酶含 V型 H+ - ATPase的主要亚基 ,与大豆磷脂重组 ,获得了有较高泵活性的脂酶体 .脂酶体的质子泵活性受 Valinomycin激活 ,说明它是致电性的 ,受NO-3 ,DCCD以及特异性的 V型 ATPase抑制剂 Bafilomycin的抑制 .脂酶体的泵活性不受 F型和P型 ATPase抑制剂抑制 ,表明质子转运是由 V型 H+ - ATPase引起的 .     

小麦根H^＋—ATPase与脂质体的重组方法研究

液泡是植物细胞中的大型细胞器,除了维持细胞的渗透压和贮存代谢的中间产物外,其内部还含有多种水解酶、pH值偏酸且具有类似溶酶体的功能。液泡膜ATPase是一种新类型的质子泵。由质子泵作用形成的跨液泡膜质子电化学梯度和质子驱动力为各种溶质(如阳、阴离子、氨基酸和糖类等)分子的主动跨膜转运提供了动力,使液泡成为植物细胞内离子平衡的调节器。液泡膜ATPase和线粒体膜ATPase都具有泵质子的功能,而且都受阴离子激活,其生化性质有许多相似     

NaCl胁迫下大麦根液泡膜H^＋—ATPase活性、离子吸收与钙的关系

NaCl胁迫 2d ,耐盐大麦 (HordeumvulgareL .cv) (“滩引 2号”)根系液泡膜H _ATPase活性增强 ,H _PPase活性下降。以质膜Ca2 通道抑制剂La3 (1mmol/L)或Ca2 螯合剂EGTA (5mmol/L)处理大麦幼苗 ,抑制了NaCl诱导的液泡膜H _ATPase活性的增强 ,但提高了H _PPase活性 ;用CaM拮抗剂三氟拉嗪 (TFP ,2 0 μmol/L)处理 ,也抑制了液泡膜H _ATPase活性的增强。NaCl胁迫下 ,外加La3 ,TFP或La3 TFP处理 ,使Na 吸收增加 ,K 和Ca2 吸收降低。结果表明 ,NaCl胁迫下 ,液泡膜H _ATPase活性提高和离子吸收的变化可能与Ca_CaM系统有关。     

NaCl胁迫对高粱根、叶鞘和叶片液泡膜ATP酶和焦磷酸酶活性的影响

NaCl胁迫初期 ,Na 主要在根和叶鞘中积累。相应地 ,根和叶鞘液泡膜ATP酶和焦磷酸酶水解活性、依赖ATP和PPi的质子泵活性及Na /H 逆向转运活性均明显增加 ,根和叶鞘的生长没有受到抑制。NaCl胁迫后期 ,Na 开始向地上部分运输并在叶片中积累。此时 ,叶片液泡膜质子泵和Na /H 逆向转运活性开始增加 ,根和叶鞘的Na/K比增加 ,其液泡膜ATP酶和焦磷酸酶水解活性、质子泵活性和Na /H 逆向转运活性下降。相应地 ,根和叶鞘的生长也下降。当保温介质中Na/K比超过 1时 ,液泡膜微囊ATP酶和焦磷酸酶活性均随Na/K比的增加而下降。表明非盐生植物液泡膜质子泵在盐胁迫的初期对Na 在液泡内的积累及其耐盐性起重要作用     

大麦根液泡膜微囊依赖ATP质子泵测定技术的改进

ImprovementofDeterminationofATP－DependentProtonPumpinTonoplastVesiclesfromBarleyRootsZHANGWen－Hua,LIUYou－Liang,YUBing－Jun（DepartmentofAgronomy,NanjingAgriculturalUniversity,Nanjing210095）HEPing－Qing（CentralLaboratory,NanjingAgriculturalUniversity,Nanjing210095）膜微囊是研究质子泵的有效工具,目前一般用不连续葡聚精（蔗糖）密度梯度获取液泡膜微囊「’,‘’。我们在实验中发现,液泡膜微囊提取及其质子泵的测定有一定难度,尤其是跨液泡膜质子梯度（APH）的测定值波动较大,有时甚至…     

胡杨液泡膜H+-ATPase的部分纯化及其耐盐性研究

为了阐明液泡膜H^ -ATPase在盐胁迫下的作用和适应性机制,对悬浮培养的胡杨细胞在50mmol／L盐浓度下处理10d,结果表明液泡膜H^ -ATPase、焦磷酸酶的水解活性、质子泵活性增加。将通过差速离心和不连续蔗糖密度梯度离心富积的液泡膜微囊先由脱氧胆酸钠(DOC)和n-辛基-β-D-葡萄糖(OG)分步破膜抽提,经蔗糖密度梯度离心分离,部分纯化的酶含V型H^ -ATPase的主要亚基。     

玉米根尖细胞液泡膜结合的蛋白激酶的存在及其性质

为了解液泡膜蛋白在植物细胞信号途径中的功能,用新型的非放射性同位素方法从玉米根细胞的高纯度液泡膜上鉴定出一种膜内在的蛋白激酶.这种蛋白激酶具有Ca2+依赖、CaM和磷脂酰丝氨酸不依赖等特性,与已在多种植物中报道的含有类似钙调素结构域的蛋白激酶CDPK相似.离体实验表明其活性的最适pH值为6.5,最适Ca2+浓度为10 μmol/L.从最适pH值和去污剂的影响可以推测出其活性位点朝向胞质一侧.Zn2+对其活性没有明显的抑制作用,说明该激酶缺少某些哺乳动物的蛋白激酶常含有的锌指结构.当液泡膜蛋白在Ca2+和ATP存在的条件下被预磷酸化后,液泡膜H+-ATPase的ATP水解和质子转运过程均被激活.激活的活性可以被碱性磷酸酶逆转.以上结果说明玉米根尖细胞的液泡膜中存在一种可能是CDPK的蛋白激酶.由它造成的Ca2+依赖的磷酸化作用激活了液泡膜H+-ATPase的活性.这些结果将有助于深入研究CDPK在植物细胞信号转导中的功能.     

欧洲山杨液泡膜Na+/H+ 反向运输体的性质鉴定

植物液泡膜Na /H 反向运输体可将细胞质中的Na 转运到液泡内储存,以减少胞内Na 的毒性.但木本植物如杨树是否有同样的机制目前还不清楚.以欧洲山杨的愈伤组织为材料,捣碎破碎愈伤组织细胞,经过差速离心和不连续蔗糖梯度离心得到纯化的欧洲山杨液泡微囊.通过液泡V-ATPase建立质子梯度,该液泡能够利用此梯度调控Na 的转运,表明液泡膜上存在Na /H 反向运输体活性(表观米氏常数Km是11.4mmol/L).Na /H 反向运输体的抑制剂——氨氯吡嗪咪能明显抑制转运体的活性.该Na /H 反向运输体也可以转运K ,但亲和能力比Na 低30%.该结果首次证明木本植物的液泡膜上存在Na /H 反向运输体.初步功能研究表明,愈伤组织在盐胁迫条件下,Na /H 反向运输体活性明显下降,提示该机制可能与山杨不耐盐有关.     

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.     

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.     

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.     

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.     

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。     

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.     

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.