二氢玉米素核苷组细胞分裂素的放射免疫测定法

本文报道二氢玉米素核苷（ＤＨＺＲ）组细胞分裂素放射免疫测定法（ＲＩＡ）的研制结果。以（３Ｈ）二氢玉米素作示踪剂,测得免抗ＤＨＺＲ抗血清的效价为１：１３７０（Ｂ％＝３０％）。该抗血清主要与本组细胞分裂素发生交叉反应。回收率为９６．９％,灵敏度为１２ｆｍｏｌ ＤＨＺＲ／管,检测线性范围为０．１￣１００ｐｍｏｌＤＨＺＲ／管。批内误差ＣＶ＝４．３,批间误差ＣＶ＝２．０％。对基于同一免抗血清的ＤＨＺＲ组细胞     

菊苣薄层培养花芽,营养芽分化中内源激素的动态变化

菊苣（Ｃｉｃｈｏｒｉｕｍ ｉｎｔｙｂｕｓＬ．）花梗薄层细胞培养于ＭＳ附加ＮＡＡ 和ＢＡ 或ＩＡＡ 和ＢＡ 的ＭＳ培养基上有花芽或营养芽分化． 花芽分化中内源ＩＡＡ、ＤＨＺ＋ ＤＨＺＲ、ｉＰＡ 含量明显增加,而Ｚ＋ ＺＲ变化不明显．营养芽分化中内源细胞分裂素含量增加明显,而ＩＡＡ 在培养前７ ｄ 含量下降,随后有所增加,在原基形成时含量达原初水平的２／３． 可见,花芽分化比营养芽分化所需内源ＩＡＡ／ＣＴＫ 比值要高     

用免疫亲和层析和ELISA测定百合花粉萌发过程中细胞分裂素含量的变化

兰州百合（Ｌｉｌｉｕｍ ｄａｖｉｄｉｉＤａｃｈ．）花粉在ＰＥＧ－４００ 中萌发,用高度灵敏和特异的ｔ－ＺＲ－ＩｇＧ和ｉＰＡ－ＩｇＧ 免疫亲和层析分离纯化萌发花粉和萌发液中细胞分裂素,并用酶检疫连锁鉴定法（ＥＬＩＳＡ）测定Ｎ６－异戊烯腺嘌呤核苷（ｔ－ＺＲ）和异戊烯基腺苷（ｉＰＡ）含量。结果表明每克鲜重花粉含有３９ ｎｇ ｔ－ＺＲ和４８ ｎｇ ｉＰＡ。花粉水合后ｔ－ＺＲ 含量略有下降,而ｉＰＡ 含量明显增高,细胞分裂素总量几乎不变。水合过程中有细胞分裂素的消长,这种变化与花粉管的启动有关。在萌发的前３ 小时,花粉管生长速度最快,ｔ－ＺＲ和ｉＰＡ 在花粉管和萌发液中的含量也随着增加,其增长趋势和花粉管生长速度同步     

玉米雌、雄穗与叶片内几种激素含量的比较

以甜玉米为材料,采用扫描电镜观察雌、雄花序发育过程,并用ＥＬＩＳＡ 和ＨＰＬＣ 检测雌、雄花序发育过程中内源激素（ｉＰＡｓ 、ＺＲｓ、ＤＨＺＲｓ、ＺＴ、ＩＡＡ、ＧＡ３） 和内源玉米赤霉烯酮（ｚｅａｒａｌｅｎｏｎｅ, ＺＥＮ） 含量,分析激素含量变化与玉米性别决定的关系。结果表明：玉米花序经过了无性、双性到单性的发育过程,在茎尖伸长和小穗原基突起时期,雄花序中内源ｉＰＡｓ、ＺＲｓ、ＧＡ３ 和ＺＥＮ 含量最高并高于雌花序;在玉米性别决定的关键时期,即花器官原基突起到雌、雄性器官选择退化阶段,雄花序中ｉＰＡｓ、ＺＲｓ、ＺＴ 和ＺＥＮ 含量明显增加并高于雌花序,此时在两种花序中的ＧＡ３ 含量差异不明显。正常果穗上、中、下不同部位果穗内源ｉＰＡｓ、ＺＲｓ、ＺＴ、ＧＡ３ 和ＺＥＮ 含量无明显差异,顶端小穗雄性化的果穗顶部内源ｉＰＡｓ、ＺＲｓ、ＺＴ、ＧＡ３ 和ＺＥＮ 含量急剧增加。这些结果说明高水平的细胞分裂素、ＧＡ３ 和ＺＥＮ 可能有利于玉米雄性器官的发育。     

牡丹冬季到内催花过程中内源激素含量的变化

牡丹品种朱砂垒（Ｐａｅｏｎｉａ ｓｕｆｆｒｕｔｉｃｏｓａ Ａｎｄｒ．ｃｖ．Ｚｈｕｓｈａｌｅｉ）在冬季室内催花过程７种内源激素含量变化不同。玉米素核苷（Ｚ＋ＡＲ）、生长素（ＩＡＡ）和赤霉素（ＧＡ３）的含量在花生长发 处于较高水平;而脱落酸（ＡＢ）、异戊烯基腺苷（ＩＰ＋ＩＰＡ）、二氢玉米素核苷（ＤＨＺ＋ＤＨＺＲ）、赤霉素（ＧＡ４）的含量低于上述３种内源激素。激素平衡方面,ＧＡｓ／ＡＢＡ、ＣＴＫｓ／ＡＢ     

大豆花荚败育期间的植物激素变化

大豆开花结荚期,不同发育阶段的幼蕾与花荚的脱落率不同,其中以花后５ｄ内的幼荚脱落最严重。与败育花荚相比,正常花荚中的干物质积累量均较高。细胞分裂素（ＤＨＺＲｓ,ＺＲｓ,ｉＰＡ）含量也较高,花后３～５ｄ的幼荚中表现更明显。脱落酸（ＡＢＡ）则是以败育幼蕾及花后３～５ｄ的幼荚中含量较高。不同发育阶段的大豆生殖器官中,正常开放花中的玉米赤霉烯酮（ＺＥＮ）含量最高     

外源激素在诱导风信子花被外植体不同部位细胞再生花芽中的作用

外源激素诱导风信子（Ｈｙａｃｉｎｔｈｕｓ ｏｒｉｅｎｔａｌｉｓＬ．）同一发育时期花被外植体不同部位细胞再生花芽的实验表明∶１． 诱导花被外植体细胞再生花芽,外源激素是必需的;２． 仅有细胞分裂素就可以诱导花芽再生,生长素并不是必需的;３． 花被外植体上的不同部位的细胞再生花芽时,需要不同浓度的外源激素． 单独加６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ可以诱导花被下部的细胞再生花芽;６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ和２,４－Ｄ ０．１ ｍ ｇ／Ｌ的组合有利于花被中部的细胞再生花芽;６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ和２,４－Ｄ １．０ ｍ ｇ／Ｌ的组合能促进花被上部的细胞分化花芽     

菜豆下胚轴中的细胞分裂素结合蛋白

用植物体内天然存在并具有很高活性的玉米素（ｔ－Ｚ）和异戊烯基腺苷（ｉＰＡ）作为配基,分别与Ｓｅｐｈａｒｏｓｅ－４Ｂ偶联,制成亲和吸附介质,分离、纯化菜豆（Ｐｈａｓｅｏｌｕｓ ｖｕｌｇａｒｉｓ）黄化幼苗下胚轴中的细胞分裂素结合蛋白。一种分子量为１５．５ ｋＤ（简称ＺＢＰ）,只含一个多肽链;另一种分子量为１６５ ｋＤ（简称ＩＢＰ）,含有两种亚基,分子量分别为４３ ｋＤ和４０ ｋＤ。对ＺＢＰ的结合活性进行了研究,发现ＺＢＰ与ｔ－Ｚ结合时的解离常数（Ｋｄ）为３．２×１０－ ７ ｍ ｏｌ／Ｌ。经计算,每个ＺＢＰ分子只有一个ｔ－Ｚ结合位点     

抗癌胚抗原单链抗体基因在家蚕细胞和幼虫中的表达

将抗癌胚抗原（ＣＥＡ）单链抗体基因插入家蚕杆状病毒转移载体ｐＢａｃＰＡＫＨｉｓ, 与修饰的家蚕核型多角体病毒ＢｍＢａｃＰＡＫＤＮＡ共转染家蚕细胞, 经同源重组得到含有在多角体蛋白基因启动子控制下的抗ＣＥＡＳｃＦｖ 基因的重组病毒ＢｍＢａｃＳｃＦｖ。用重组病毒分别感染家蚕细胞和幼虫, 在两者中均得到了高效表达, 产物分子量为２８ｋＤ, 前者占细胞总蛋白的６ ％ , 后者为０ ．３ ｍｇ／ 蚕。目的基因在家蚕细胞和幼虫中表达产物经Ｎｉ２＋ＩＤＡＳｅｐｈａｒｏｓｅ６Ｂ亲和柱纯化, 前者纯度可达９０％ 以上, 后者纯度较低; 纯化后的融合蛋白具有ＣＥＡ 结合活力, 其亲和常数分别为５ ．４×１０８／ｍｏｌ·Ｌ－ １ 和２．３ ×１０８／ｍｏｌ·Ｌ－１ , 略低于其亲本单抗Ｅ７Ｂ１０ ２．７ ×１０９／ｍｏｌ·Ｌ－ １ 。     

土壤干旱条件下玉米叶片内源激素含量及光合作用的变化

利用ＨＰＬＣ和ＥＬＩＳＡ技术研究了土壤干旱条件下玉米叶片内源ＩＡＡ、ＡＢＡ、ＺＲ、ＤＨＺＲ、ｉＰＡ含量的变化情况,以及叶片光合作用过程中,光合速率（Ｐｎ）,气孔导度（Ｇｓ）,ＰＳⅡ光化学效率（Ｆｖ／Ｆｍ）的变化情况,结果发现：叶片中ＩＡＡ浓度,在整个干旱过程中变化不大,与对照相比下降不明显;ＩＡＡ的浓度对干旱的反应不敏感;叶片中ＡＢＡ浓度在干旱最初３ｄ里急剧升高,直至最大值,之后有所下降;ＡＢＡ的     

Factors affecting dispersion in expanded bed chromatography

Factors affecting the dispersion of solutes in expanded bed chromatography were experimentally investigated to characterize the behavior in small columns. Pulse response curves were measured with a vitamin B¹² tracer, and HETP (height equivalent to a theoretical plate) values were calculated from peak variance and retention time. Approximately 15 min were required to attain a stable steady state expanded bed height with constant HETP values. HETP values ranged from 0.8 to 1.6 cm and did not change appreciably with the degree of expansion (1.5–3.5 fold), column diameter (1.6 and 2.6 cm), column temperature (293–308 K) or settled bed height (ca. 4–11 cm). A very small column (1.6 cm diam. and 4.2 cm-settled bed height) was successfully expanded and axial mixing measured could be useful for conducting scale down experiments.     

Chromatography of human prothrombin from Nitschmann fraction III on Q Sepharose Fast Flow using axial and radial flow column

An axial column (Hitrap Q 5 ml, 2.5 2 1.6 cm) and a radial flow column (3.5 2 5 cm) packed with Q Sepharose Fast Flow media had been evaluated for the separation of human prothrombin. Nitschmann fraction III dissolved in buffered saline (0.10 M sodium chloride buffered with 0.06 M Tris/HCl to pH 7.5) was the starting material. Effects of sample flow rate of the two columns were screened. Under radial flow conditions using the radial column, sample flow rate up to 15 ml/min (i.e. 18 bed volumes/h) was achieved and the operating pressure was below 0.2 MPa eventhough the elution velocity was 30 ml/min. Breakthrough capacity was determined by analyzing the total protein and prothrombin activity of the target protein-containing fraction under subsaturating conditions and both columns had almost the same breakthrough capacity per ml media, indicating that the sample loading was independent of radial column geometry. It was concluded that the radial column is an attractive alternate to traditional axial packed bed column, exhibiting very good potential for use in the separation of human prothrombin.     

Evaluation of radial chromatography versus axial chromatography, practical approach

Developments in packing and packing port design of radial columns in recent years have resulted in a claimed significant increase in performance of this process chromatography technology. In this first study, the main chromatographic parameters as efficiency, capacity factor, asymmetry and resolution were evaluated in a unique one-to-one comparison between a 120 ml bed-volume and 6 cm bed length radial chromatography mini-process column against a 50 mm diameter, 6 cm bed height and 120 ml bed-volume axial chromatography column. Radial chromatography showed an increase in efficiency by 31% in the number of plates per meter while the equilibration could be reduced by 0.4-0.5 column volumes. The asymmetry factor for bovine serum albumin in radial chromatography showed a reduction of 20% while the reduction of the asymmetry factor of the smaller protein ovotransferrin decreased even by 46% in comparison to the performance of the comparative axial chromatography column. Therefore in radial chromatography resolution improved up to 20%. The retention volume was similar in both cases. For radial chromatography, the decrease in "width at half height" at Height Equivalent of Theoretical Plates (HETP) measurements was 40% while the decrease of the over-all width of the peak was 27%. For adsorbed/desorbed proteins, the elution peak showed similar results: "width at half height" decreased to 45% while the over-all width of the peak decreased by 28%. The concentration of the non-retained protein in the flow-through (lysozyme), increased by 35% while the concentration of the eluted fraction (serum albumin bovine), increased with 40% in the radial chromatography columns. The better results obtained with the radial column were probably the consequence of the geometrical design of this device (larger inlet surface area and small outlet surface area which concentrate the eluted fraction).     

Chromatography of hen egg-white proteins on anion-exchange cellulose at high flow rates using a radial flow column.

The influence of column configuration on the separation of hen egg-white proteins using Whatman DE52 and QA52 anion-exchange cellulose has been investigated. Using a 100 ml volume axial flow column (6.6 cm x 4.4 cm i.d.) we achieved flow rates of up to 25 ml/min i.e. 15 bed volumes/h after which higher flow was restricted due to pressure constraints within the system. Under radial flow conditions using a 100 ml column flow rates of up to 150 ml/min i.e. 90 bed volumes/h were achieved using DE52 and QA52. While chromatographic resolution was superior under axial flow at the lower flow rates excellent resolution was maintained at up to 150 ml/min using the radial flow column. This is a consequence of the fast kinetics of adsorption/desorption exhibited by DE52 and QA52. The data indicate that it is the column configuration and not the cellulose matrix which influences flow performance.     

High Performance Liquid Chromatographic Analysis of Cytokinins in Sorghum bicolor Leaves

High performance liquid chromatography with octadecylsilica (Bondapak C₁₈/Poracil B) column packing was used to purify and separate cytokinins in Sorghum leaf extracts. The column size was 56 × 0.21 cm i.d. By gradient elution, using acidified water with increasing amounts of methanol, the major peaks of cytokinin activity, as determined by the callus tissue bioassay. were effectively separated from large amounts of extraneous impurities. These cytokinins were further separated on a microoctadecylsilica column (μBondapak C₁₈, 30 × 0.4 cm i.d.) with a gradient of acidified water-acetonitrile. Zeatin and zeatin riboside gave distinct ultra violet absorption peaks which could be used for quantitative estimation. Biological activity corresponded to the elution of these peaks. These two cytokinins are the major cytokinins in Sorghum leaves.     

Isolation of human fibrinogen by axial and radial flow chromatography from Nitschmann fraction I

An axial column (3×2.6 cm) and a radial flow column (3.5×5 cm) packed with DEAE Sepharose Fast Flow media had been evaluated for the separation of human fibrinogen. Nitschmann fraction I dissolved in buffered saline (0.015 M NaCl buffered with 0.06 M Tris/HCl to pH 7.5) was the starting material. Under radial flow conditions, sample flow up to 15 ml min^–1 (i.e., 18 bed volumes h^–1) was achieved. The operating pressures were below 0.2 MPa, even though the elution velocity was 30 ml min^–1 (i.e., 36 bed volumes h^–1).     

Fixed-Bed Studies on Removal of Arsenic from Simulated Aqueous Solutions Using Chitosan Nanoparticles

Arsenic is a toxic element and may be found in natural as well as in industrial water; therefore, before using water for drinking purpose, its proper treatment is required. Thus, the aim of this work was to evaluate the potential of chitosan nanoparticles, in a continuous-flow method, for the removal of arsenic (III) and (V) from aqueous solutions. All experiments were conducted in fixed-bed columns. Experiments were carried out as a function of varying liquid flow rate (0.3–1.0 ml/min), initial metal concentration (0.5–1.5 mg/L), and bed height (3–9 cm) of adsorbent. The total adsorbed quantity, equilibrium uptake, and total percentage removal of arsenic ions were determined by evaluating the breakthrough curves obtained at different flow rates, initial concentrations, and bed heights. The results showed that the column performed well at the lowest flow rate. Also, column bed capacity and exhaustion time were found to increase with increasing bed height. When initial metal ion concentration was increased from 0.5 to 1.5 mg/L, the corresponding adsorption bed capacity decreased from 0.076 to 0.028 mg/g. The bed depth service time model (BDST) model was used to analyze the experimental data and the model parameters were evaluated. The calculated values of N _o and K _a were found to be 19.28 × 10^?2 mg/L and 0.662 L/mg·min, respectively. Good agreement was found between the experimental breakthrough curves and the model predictions.     

玉米素核苷的酶标免疫测定法

牛血清白蛋白-玉米素核苷(BSA-ZR)的兔抗血清对玉米素核苷具有很高的亲和性,而且专一性强,除了玉米素外,对其它一些细胞分裂素如激动素(KT)的交叉反应甚微。用辣根过氧化物酶(HRP)作为标记物的酶标免疫法,由于它的灵敏度高,相当于几十毫克量的样品就可以测出细胞分裂素的含量。测定范围在0.25—50pmol之间,测定范围较广。由于该方法专一性高,植物组织的粗提取物可以直接用于测定。避免了提取分离的繁琐程序,使得测定方法较简便、快速,可成批进行,适用于一般实验室。用该方法测得风信子(Hyacinthus orientalis L.)各部分的细胞分裂含量(以玉米素核苷计)在10—60×10~(-9)克/克鲜重,即10—60ng/g F.W.。     

Isomers of zeatin and zeatin riboside in clubroot tissue: evidence for trans-zeatin biosynthesis by Plasmodiophora brassicae

Cytokinin-like substances in both healthy and infected ( Plasmodiophora brassicae ) Wor. strain S) roots of Brassica campestris L. ssp. pekinensis cv. Granat have been tentatively identified and quantified by HPLC. The isomers of the cytokinins could be seperated on a reversed phase column using a gradient elution with increasing amounts of methanol. Secondary plasmodia were isolated mechanically from Plasmodiophora brassicae infected roots. The time course of adenine uptake and its conversion to cytokinins were investigated. Evidence is presented for the incorporation of [U-¹⁴C]-adenine into trans -zeatin by secondary plasmodia.