兜兰DEFICIENS(DEF)-和GLOBOSA(GLO)-like基因的克隆及表达分析

以‘同色兜兰’品种为材料,采用RT-PCR和RACE技术获得了DEFICIENS(DEF)-和GLOBOSA(GLO)-like基因的cDNA全长,命名为PcDEF和PcGLO,并用半定量RT-PCR和实时PCR研究了PcDEF和PcGLO在花芽发育过程和不同组织部位的表达特性。结果表明,PcDEF和PcGLO的全长cDNA分别为1 039bp和934bp,分别编码224和210个氨基酸;蛋白比对表明,PcDEF和PcGLO蛋白都具有典型MADS-box蛋白的MADS和K结构域;蛋白同源性分析显示,PcDEF和PcGLO与已登录的其它兰科植物的DEF/AP3和GLO/PI蛋白的相似性分别在75%～96%和87%～98%;系统进化树分析表明,PcDEF和PcGLO分别属于B类MADS-box蛋白家族的AP3和PI亚家族。表达分析显示,PcDEF和PcGLO在花芽发育中均有表达,PcDEF在成熟花、唇瓣和花瓣中的表达量高,在蕊柱、萼片、苞叶和根中次之,在花茎和叶中较低,在子房中几乎不表达;PcGLO在各组织中均有不同丰度的表达。     

土壤含水量对樟子松幼苗非结构性碳水化合物及生长的影响

为明确樟子松幼苗在不同土壤含水量下的生长适应策略,设置不同土壤含水量(80%、60%、40%和20%田间持水量(SFC))和持续时间(15、30、45和60 d),研究不同器官中可溶性糖、淀粉、非结构性碳水化合物及生物量的变化规律。结果表明：随着土壤含水量减少,樟子松幼苗株高和基径及一年生叶、细根和总生物量呈下降趋势。80%SFC条件下,可溶性糖含量在枝干中较高,在当年生叶和细根中较低;淀粉在枝干和粗根中较高。60%和40%SFC条件下,可溶性糖含量在叶片和细根中较高,淀粉含量分别在枝干和一年生叶中较高。20%SFC条件下,可溶性糖含量在当年生叶较高,在一年生叶较低;淀粉含量在细根中较高,在枝干和粗根中较低。因而,樟子松幼苗在80%SFC下具有更高的生长速率及抗逆性,在60%和40%SFC下具有更高的资源利用效率和较优的碳储存策略,在20%SFC下生长停滞。为促进科尔沁沙地樟子松林天然更新,土壤含水量应维持在7%以上。     

北柴胡营养器官中主要化学成分的组织化学定位及其含量比较

应用植物解剖学、组织化学定位和植物化学方法,研究了北柴胡各营养器官中柴胡皂苷和黄酮类化合物的积累分布状态及其含量变化。结果表明,柴胡皂苷在根中分布在中柱鞘和次生韧皮部中;在茎中主要分布在表皮、棱角处的厚角组织以及位于皮层和髓中的分泌道的上皮细胞中;在叶中,则分布在表皮细胞和整个叶肉组织中。而黄酮类化合物在茎中分布在表皮、棱角处的厚角组织、皮层、髓射线和髓鞘细胞中;在叶中,则主要分布在表皮和位于上下表皮内的厚角组织中。同时,北柴胡中柴胡总皂苷在根、茎、叶中的含量的变化规律为根>叶>茎;而总黄酮在根、茎、叶中的含量的变化规律为叶>茎>根;且在叶中含量相当高,从而为北柴胡的综合利用提供依据,对合理利用药材和保护北柴胡资源也有一定意义。     

可持续农业的全球化

在当前西方中心主义的语境下,主流和非主流的话语和被呈现的实践,主要来自西方,难得有处于外围的实践经验得到充分呈现、阐释和尊重。《赛罗庞塔-林区农民的自豪》一文呈现了外围社区的实践,给了我们更多的阅读自由和思想自由。正如资本和压制的全球化在蔓延一样,劳动和抵抗的全球化也在蔓延。在可持续农业和农村发展领域,也是这样。     

Hira基因产物在银鲫和彩鲫卵子发生过程中的动态变化

为进一步研究Hira基因在卵子发生和雌核发育过程中的作用,通过原位杂交和免疫荧光定位的方法检测了Hira mRNA和蛋白质在雌核发育银鲫和两性生殖彩鲫卵子发生过程中的动态变化。结果表明,银鲫和彩鲫卵子发生过程中Hira基因转录产物的变化基本一致,在Ⅰ期卵母细胞的细胞核中大量表达,至Ⅱ期卵母细胞时转至细胞质中均匀分布,在Ⅲ期卵母细胞中,杂交信号逐渐移向细胞的周边,到Ⅳ期时随着卵黄物质大量积累,杂交信号几乎不见。HIRA蛋白在银鲫和彩鲫卵子发生过程中的变化略有差别。HIRA蛋白在银鲫Ⅰ期卵母细胞中没有表达,在Ⅱ期卵母细胞的细胞质中有弱表达,在Ⅲ期早期卵母细胞的周边有强烈表达;而在彩鲫Ⅰ期卵母细胞中就有HIRA蛋白的弱表达,至Ⅱ期时HIRA蛋白在细胞质中大量表达,Ⅲ期早期卵母细胞的细胞质中有弱表达。在银鲫和彩鲫Ⅲ期末和Ⅳ期卵母细胞中都很难观察到荧光信号。Hira mRNA和蛋白质在银鲫和彩鲫早期卵母细胞中有较强表达,且在银鲫和彩鲫卵子发生过程中没有显著差异,说明其可能对于脊椎动物卵子发生和减数分裂没有显著影响,而是在受精和/或胚胎发育过程中起作用。       

昆虫谐波雷达在柑橘大实蝇行为学上的应用研究

本试验分别在橘园中的单株橘树和在整个橘园及它附近森林中,对昆虫谐波雷达的检测率和发现率进行研究,评估昆虫谐波雷达在橘园和森林中对初羽化柑橘大实蝇成虫的扩散行为能否进行有效的跟踪。结果表明,在2013年和2014年,在柑橘园单株橘树试验中,在5 m和3 m条件下,昆虫谐波雷达对带标签的初羽化的柑橘大实蝇成虫的总平均检测率分别为99.1%和100%。按降落位置和释放位置计算,带标签的柑橘大实蝇的总平均发现率分别为87.9%和71.6%。在2014年,在橘园和森林试验中,昆虫谐波雷达对带标签的初羽化的柑橘大实蝇成虫总平均检测率和总平均发现率分别为91.4%和77.8%。结果表明,昆虫谐波雷达对在橘园和森林中初羽化的柑橘大实蝇扩散行为能够进行有效跟踪。     

家蝇3种丝氨酸蛋白酶抑制剂(Serpin)基因在感染白色念珠菌后的表达

研究家蝇丝氨酸蛋白酶抑制剂（ Serpin ）基因 Sp2、Sp13、Sp16 在感染白色 念珠菌后的表达模式。利用白色念珠菌刺激家蝇幼虫,对收集的标本进行逆转录,以 GAPDH 为内参基因,通过qPCR技术检测3条 Serpin 基因在家蝇感染后不同时间点及不同组织中的表达特征,统计分析结果。结果表明,不同时间点感染组和对照组比较, Sp2 mRNA在3 h、6 h和48 h的表达量比对照组高; Sp13 mRNA在6 h的表达量比对照组高,在3 h和24 h时的表达量比对照组低; Sp16 mRNA在6 h、24 h和48 h时的表达量比对照组高,在36 h时的表达量比对照组低。不同组织中感染组和对照组比较, Sp2 mRNA在3 h时,在血淋巴和脂肪体中的表达量比对照组高,在马氏管、气管和肠道的表达量比对照组低;12 h时,在血淋巴中的表达量比对照组高,在马氏管和唾液腺中的表达量比对照组低;24 h时,在脂肪体中的表达量比对照组低;48 h时,在血淋巴中呈高表达,在唾液腺中的表达量比对照组低。 Sp13 mRNA在3 h时,在唾液腺和脂肪体中的表达量比对照组高,在马氏管、体壁、血淋巴和气管中的表达量比对照组低;12 h时,在血淋巴中呈高表达,在体壁和脂肪体中的表达量比对照组低;24 h时,在血淋巴、气管和唾液腺中的表达量比对照组高,在脂肪体中的表达量比对照组低;48 h时,在马氏管、血淋巴、气管和脂肪体中的表达量比对照组高,在体壁和唾液腺中的表达量比对照组低。 Sp16 mRNA在3 h时,在马氏管和气管中的表达量比对照组低;12 h时,在马氏管和脂肪体中其表达量比对照组低;24 h时,其表达量在马氏管中比对照组高,在脂肪体中比对照组低;48 h时,在血淋巴中呈高表达,而在马氏管中的表达量比对照组低。家蝇3种 Serpin基因在白色念珠菌感染后均呈现出差异性表达,推测 Serpin 基因在家蝇免疫调节中具有协同调节作用。     

胰岛素在小鼠受精卵早期发育中的作用

大量研究已经证实生长因子和激素在胚胎早期发育的细胞增殖与分化过程中起着重要的作用．应用定量ELISA,RT—PCR,免疫印迹和免疫荧光的方法检测胰岛素在小鼠受精卵和卵母细胞中的表达和定位．发现胰岛素均匀分布在卵细胞的胞浆中．同时也检测到mTOR（mammalian target of rapamycin）和p70S6K的表达、活性和定位．在小鼠受精卵中mTOR和p70S6K的表达没有明显不同．二者在G1,G2和M期分布在细胞浆,在S期聚集在原核的周围．在不同时期,mTOR的活性是波动的．利用P13K的特异性抑制剂渥曼青霉素,观察到卵裂率明显减低．当使用mTOR的特异性抑制剂雷帕霉素时,受精卵的第一次有丝分裂延迟．这些结果表明胰岛素存在于小鼠的卵母细胞和受精卵中,并且胰岛素可能通过激活P13K／PKB／mTOR／S6K的信号传导通路在小鼠的早期胚胎发育中发挥功能作用．     

Gas7在大鼠海马和齿状回发育过程中的动态表达

目的研究生长休止蛋白7（Gas7）在大鼠海马和齿状回不同发育阶段的表达。方法采用免疫组织化学方法观察Gas7在SD大鼠胚胎第18d（E18）、新生（P0）、生后第7d（P7）、P14、P21和成年海马和齿状回中的表达和分布。结果在大鼠脑海马和齿状回部位的冠状切片上,Gas7免疫反应阳性产物主要表达在海马的锥体细胞、齿状回的颗粒细胞和门区的多形层细胞。随着发育的进程,在海马,Gas7较早表达在CA3区,其次是CA2和CA1区;在齿状回,Gas7在外臂的表达早于内臂,在颗粒细胞层的表达是按先外层后内层的顺序。在围生期,Gas7在海马和齿状回各区的表达逐渐增强,至P14达到高峰,后逐渐降低,至P21其表达强度和分布趋于恒定至成年水平。结论 Gas7在大鼠海马和齿状回发育过程中的动态表达具有时间和空间上的特异性,提示Gas7可能参与了海马和齿状回形态形成和功能成熟的调控。     

右撒子、左撒子？——人类的利手倾向

在西藏有一个说法：要知道恶魔在你的左手。这一箴言不同程度地存在于各个人种和不同的宗教教义中。在大部分文化群体中,左手被认为是不沽和卑下的。在中东和北美,用右手吃饭的仪式在正统圈子里被严格遵循。在英语中,右（right）意味着正确和礼貌,而左（1eft）从其本意上说意味着懦弱和卑下。     

Scaleable downstream recovery of nematodes used as biopesticides.

This study assesses the suitability of sieving as a scaleable technique for the separation of adult nematodes from infective juveniles, the latter is an effective bioinsecticide whereas the former is waste material resulting from the fermentation process. Batch and semibatch experiments using conventional flow-assisted wet sieving and a novel cross-flow sieving technique were used to study the separation of juveniles from adult nematodes. The experiments were carried out using small-scale devices and the data were analyzed in terms of the screen effectiveness factor. The results were used to identify the sieve size and operating conditions for optimum juvenile recovery. It was found that, for a given species of nematode, optimum recovery was achieved when sieving was carried out in the cross-flow mode, the maximum recovery being a function of the size of the screen. Industrial-scale self-cleaning equipment capable of large-scale continuous screening was used to confirm the capacity of the small-scale operation for scale-up. Experimental results with this unit showed that in continuous operation sieving time is an additional parameter that influences separation performance.     

Replaceable cross-linked polyacrylamide for high performance separation of proteins

An alternative sieving matrix, replaceable cross-linked polyacrylamide (rCPA), was developed for sodium dodecyl sulfate capillary gel electrophoresis (SDS-CGE) separation of proteins. This rCPA could be conveniently pressurized into separation capillaries under a pressure of 80 psi. SDS-CGE separations using this matrix generated high resolutions for a wide range (approximately 4 kD to approximately 300 kD) of proteins. When compared to the most frequently used sieving matrixes, the rCPA permitted the highest resolutions with comparable or increased separation speed for protein separations.     

Separation of DNA fragments and single strand conformation polymorphism analysis in bare capillaries using poly(acrylamide–dimethylacrylamide) as a separation medium

A short chain poly(acrylamide–dimethylacrylamide) (PADMA) was synthesized in aqueous phase using isopropanol as a chain transfer agent, and was characterized according to the chemical composition and molecular mass. This polymer can form a stable dynamic coating on the inner surface of the capillary, thereby suppressing the electroosmotic flow and DNA–capillary wall interaction. The sieving medium has low viscosity and capillary filling with this medium and medium replacement were conveniently carried out by commercial capillary electrophoresis instruments. The effects of components and concentration of copolymers on the separation of DNA fragments were investigated. Highly efficient separation of DNA fragments, successful single strand conformation polymorphism (SSCP) analysis and good reproducibility of the migration time were obtained in bare capillaries using these copolymers as sieving media. Our preliminary results demonstrate that PADMA will become an alternative matrix for DNA separation by capillary electrophoresis.     

无胶筛分毛细管电泳分离DNA片段的现状与展望

毛细管电泳已DNA片段分离分析的重要手段。本简述了毛细管电泳中采用无胶筛分介质分离DNA片段的机理研究,介绍了筛分介质近年的研究发展状况,依据分离介质的化学组成,分单聚物、共聚物和混聚物等3个部分进行了评述,并对其发展前景进行了展望。     

Analysis of morphological characteristics of Solanum chrysotrichum cell suspension cultures

Cell size distribution of Solanum chrysotrichum cell suspension cultures was determined using mechanical sieving and an image analysis system. The results were compared using the sieve size (<0.25, 0.25–0.50, 0.5–1.0 and >1.00 mm) as the criterion. Mechanical sieving as well as image analysis showed that S. chrysotrichum cultures developed in shake flasks present a similar tendency to increase in aggregate size as growth persists. However, there are considerable differences in the values of each fraction. Fractions obtained by mechanical sieving were characterized by image analysis demonstrating that an inefficient separation of the cell population exists. The results demonstrate that digital image analysis was more precise than mechanical sieving to describe the cell size distribution changes occurring during cell growth. It was also possible to achieve a total characterization of S. chrysotrichum morphology.     

Live sieving of freshwater zooplankton: a technique for monitoring community size structure

A simple method is described for a quick determination of thesize structure of zooplankton communities. This is based onsieving of a live sample, using one or two sieves of known meshsize, and determining the dry weight of the size fractions.Size separation by live sieving is validated statistically onDaphnia galeata, a common planktonic cladoceran, by comparingsize-sieved fractions against direct microscopic measurementsof individuals. A non-linear model was used for assessment ofsieving statistics. The sieving technique facilitates the gatheringof size-structured biomass data for zooplankton for routinemonitoring or long-term studies.     

On-line combination of capillary isoelectric focusing and capillary non-gel sieving electrophoresis using a hollow-fiber membrane interface: a novel two-dimensional separation system for proteins

A novel two-dimensional (2D) separation system for proteins was reported. In the system, a piece of dialysis hollow-fiber membrane was employed as the interface for on-line combination of capillary isoelectric focusing (CIEF) and capillary non-gel sieving electrophoresis (CNGSE). The system is similar equivalent to two-dimensional polyacrylamide gel electrophoresis (2D PAGE), by transferring the principal of 2D PAGE separation to the capillary format. Proteins were focused and separated in first dimension CIEF based on their differences in isoelectric points (pIs). Focused protein zones was transferred to the dialysis hollow-fiber interface, where proteins hydrophobically complexed with sodium dodecyl sulfate (SDS). The negatively charged proteins were electromigrated and further resolved by their differences in size in the second dimension CNGSE, in which dextran solution, a replaceable sieving matrix instead of cross-linked polyacrylamide gel was employed for size-dependent separation of proteins. The combination of the two techniques was attributed to high efficiency of the dialysis membrane interface. The feasibility and the orthogonality of the combined CIEF-CNGSE separation technique, an important factor for maximizing peak capacity or resolution elements, were demonstrated by examining each technique independently for the separation of hemoglobin and protein mixtures excreting from lung cancer cells of rat. The 2D separation strategy was found to greatly increase the resolving power and overall peak capacity over those obtained for either dimension alone.     

Collecting isolated microvertebrate fossils

A review is presented of the techniques currently used in the collection and separation of isolated teeth and bones of fossil vertebrates. These involve the collection and disaggregation of the sediment, its sieving, concentration and sorting of the residue, and curation of the fossils obtained.     

Correlation of diafiltration sieving behavior of lysozyme-BSA mixtures with osmotic second virial cross-coefficients

The role of protein-protein interactions in membrane separations of protein mixtures remains incompletely understood, largely due to the difficulty of characterizing protein self- and, especially, cross-association. Recently, a novel technique, cross-interaction chromatography, has been developed to measure weak protein cross-association in terms of the osmotic second virial cross-coefficient. In this work the relationship between protein cross-association and the sieving behavior of lysozyme in the presence of BSA has been investigated. Sieving coefficients were measured using a stirred diafiltration cell over a range of pH and ionic strength, and a striking correlation between the lysozyme sieving and second virial cross-coefficients for BSA/lysozyme mixtures has been found: when the protein cross-interactions are most attractive (negative second virial cross-coefficient), the lysozyme sieving coefficients are lowest, and vice versa. The correlation between the sieving and second virial cross-coefficients may be due to the physically similar environments in the chromatography and filtration experiments since one protein is passed through a concentrated region of the second protein either immobilized on the column or accumulated at the membrane surface, and the migration rate of the mobile protein in both cases is influenced by protein cross-association. This study represents the first time that molecular interactions in binary mixtures have been related directly to filtration behavior, and may provide a useful approach to optimize the separation of other binary protein mixtures.     

Automated egg-collecting and pupa-separator system for medfly mass-rearing facilities

The new automated egg-collecting system and pupa-separator device described here substantially reduces labour during medfly [ Ceratitis capitata (Wiedemann) (Diptera: Tephritidae)] mass production. The egg-collecting system gathers eggs dropping from the cage's oviposition net into a continuous stream of water, after which they accumulate in an egg-collecting sieve. The system provides an optimal environment for eggs, keeping them in a slow stream of well-oxygenated water. A pupa separator was designed to enable the separation of pupae from the pupation medium by sucking off the medium through a slowly moving sieving belt. The smaller particles of pupation medium are removed by a vacuum cleaner located under the circular sieving belt, and the pupae are collected in a collecting box. Quality control tests of the eggs collected and pupae separated have shown that neither system has a negative effect on the quality of the eggs or the pupae.