一种简便快速鲜叶表皮制片技术

用透明胶带夹粘住适当大小的新鲜植物叶片,经轻轻挤刮,撕开两胶带,上、下表皮粘附在两胶带上,经NaOCl溶液处理3～5min,以离析粘附在表皮上的叶肉细胞,在流水下用软刷刷洗,凉干后,放在载玻片上盖上盖玻片,即可在显微镜下观察,摄影.该法简便、快速,更适用于叶片较小的植物叶表皮制片.     

易卷曲叶表皮制片技术（NaOCl法）的改进

在利用NaOCl法制备用于光学显微镜观察的叶表皮过程中,一些科/属植物的叶表皮在100%乙醇脱水后遇二甲苯便发生卷曲,增加了叶表皮制片的难度, 甚至无法进行。本文介绍一种简便易行的方法：在系列乙醇脱水后,加盖小块盖玻片,防止叶表皮脱水后卷曲,并使其保持平整,便于显微摄影。这种方法使得叶表皮显微制片技术更加完善。     

易卷曲叶表皮制片技术(NaOCl法)的改进

在利用NaOCl法制备用于光学显微镜观察的叶表皮过程中,一些科/属植物的叶表皮在100％乙醇脱水后遇二甲苯便发生卷曲,增加了叶表皮制片的难度,甚至无法进行。本文介绍一种简便易行的方法：在系列乙醇脱水后,加盖小块盖玻片,防止叶表皮脱水后卷曲,并使其保持平整,便于显微摄影。这种方法使得叶表皮显微制片技术更加完善。     

生物标本石蜡包埋保存法

生物标本石蜡包埋保存法其制作过程基本同显微镜永久玻片标本制片方法相似。新鲜的生物整体材料如植物柔软的果实、花、蕈类、小型哺乳动物、鱼类、两栖动物以及动物的局部器官和组织,经固定、脱水、石蜡渗透等步骤处理,可以制成干制本标。由     

川西高原4种苹果属植物叶片解剖结构与其抗旱性分析

采用常规石蜡制片法和NaOCl法,对川西高原地区4种苹果属植物叶片的解剖结构特征进行了观察,并选取叶片厚度、上下表皮角质层厚度、上下表皮细胞厚度、栅栏组织厚度等14项抗旱性相关指标进行测定,运用方差分析、主成分分析和隶属函数法对4种苹果属植物的抗旱性进行了分析与综合评价。结果显示:(1)4种苹果属植物叶片上下表皮外均附有角质层,表皮细胞排列紧密,上表皮细胞明显大于下表皮细胞,其中变叶海棠的角质层厚度和表皮细胞厚度均较大;气孔均只分布在下表皮,其中花叶海棠的气孔密度最大,花叶海棠和湖北海棠的气孔长、宽最小;栅栏组织由2~3层排列紧密的栅栏细胞组成,海绵组织细胞排列疏松,细胞间隙大,其中山荆子的栅栏组织厚度、栅海比和叶片结构紧密度最大;主脉均为外韧维管束,其中湖北海棠维管束厚度最大,且皮层中有时出现含晶细胞。(2)所选14项指标中除下表皮角质层厚度外,其余13项指标在4个树种间差异显著,经主成分分析筛选出上表皮角质层厚度、下表皮细胞厚度、栅栏组织厚度等8项指标作为4种苹果属植物抗旱性综合评价的代表性指标。(3)隶属函数法分析结果表明,4种苹果属植物抗旱能力大小顺序为:变叶海棠山荆子花叶海棠湖北海棠。     

3D-SIM结构照明超分辨率显微镜实现蛋白质在植物亚细胞器内的定位

基因表达产物蛋白质的亚细胞定位是解析基因生物学功能的重要证据之一。近年来出现的超分辨率光学成像技术已成功应用于人类和动物细胞中,预示着显微成像技术继激光共聚焦技术后的又一重要进步。由于植物细胞的特殊性和成像技术的研发取向,超分辨率光学成像技术在植物细胞蛋白质亚细胞定位的应用尚未见报道。该研究利用Delta Vision OMX显微镜技术,克服了叶绿体基粒中叶绿素自发荧光与融合蛋白荧光不易区分的缺陷,解决了受分辨率局限无法将植物细胞中蛋白质在亚细胞器内可视化精确定位的技术难题,成功地将植物蔗糖合成酶Zm SUS-SH1定位在烟草表皮细胞叶绿体基粒周围。该研究同时建立了一套基于撕片制片法的简便OMX显微镜制片方法,并针对OMX显微成像技术在植物细胞中蛋白质亚细胞定位的应用进行了讨论。     

福建荸荠属果皮微形态学研究

本文报道了在立体显微镜和扫描电子显微镜下对福建荸荠属8种、1变种植物果实表皮微形态特征的观察结果。根据果皮纹饰的不同,可分为两种类型:网状纹饰、拟网状纹饰;荸荠属植物果实表皮微形态特征具有丰富的多样性,表型可塑性小,不同类群之间具有明显的种间差异。观察结果表明:荸荠属果实表皮微形态特征是该属分类与系统研究中不可忽视的性状。     

一种禾本科植物鲜叶下表皮制片方法

介绍一种禾本科植物鲜叶下表皮制片的快速方法,该方法也能适用于多数单子叶植物鲜叶和经FAA固定液固定的单子叶植物叶片的气孔观察。该方法制片能有效防止叶表皮脱水、透明后发生卷曲,使保持完整的结构,便于准确的显微观察。具操作简单、材料易得、可靠性和稳定性高等优点。     

禾本科植物叶片表皮气孔观察的样品制备方法改良

对现有的禾本科植物叶片气孔观察的样品制作方法中的不足作了一些改良。改良后的方法操作简便、耗时少、样品制备成功率高,且放大后的效果好,不会造成气孔形态的改变。改良方法适用于禾本科植物和其他叶肉紧实不易剥离的植物叶片。50%NaClO处理3min最适用于小麦旗叶表皮样品的制备。     

景深合成技术在植物光学微形态研究上的应用

介绍了利用电脑景深合成技术在光学显微镜上获得植物标本大景深图片的摄影方法。概述了此方法获取植物光学微形态特征清晰图片的基本步骤和优缺点。该方法制片简单,可利用最普通的生物显微镜、电子目镜结合计算机进行摄影,成本低,观察特征明显,获得的图片具有真色彩,并可拍出植物最鲜活的状态。此方法有可能成为未来植物微形态研究的一个重要手段。     

A simple and sensitive method for detection of Bacillus anthracis by loop‐mediated isothermal amplification

Aims: To develop a rapid and simple system for detection of Bacillus anthracis using a loop‐mediated isothermal amplification (LAMP) method and determine the suitability of LAMP for rapid identification of B. anthracis infection. Methods and Results: A specific LAMP assay targeting unique gene sequences in the bacterial chromosome and two virulence plasmids, pXO1 and pXO2, was designed. With this assay, it was possible to detect more than 10 fg of bacterial DNA per reaction and obtain results within 30–40 min under isothermal conditions at 63°C. No cross‐reactivity was observed among Bacillus cereus group and other Bacillus species. Furthermore, in tests using blood specimens from mice inoculated intranasally with B. anthracis spores, the sensitivity of the LAMP assay following DNA extraction methods using a Qiagen DNeasy kit or boiling protocol was examined. Samples prepared by both methods showed almost equivalent sensitivities in LAMP assay. The detection limit was 3·6 CFU per test. Conclusions: The LAMP assay is a simple, rapid and sensitive method for detecting B. anthracis. Significance and Impact of the Study: The LAMP assay combined with boiling extraction could be used as a simple diagnostic method for identification of B. anthracis infection.     

The relationship between bark peeling rate and the distribution and mortality of two epiphyte species

Tree bark characteristics influence epiphyte establishment and survival and consequently the way in which epiphytes are distributed on trees. Tree species with peeling bark have been reported as poor epiphyte hosts. We analyzed the distribution and seedling mortality of two Tillandsia species (Bromeliaceae) in relation to rate of bark peeling of Bursera fagaroides (Burseraceae). The highest peeling rate (0.12% per day) took place on the trunk and the lowest rate on twigs (0.04% per day; branches ≤2 cm in diameter). The highest proportion of Tillandsia plants appeared on twigs. The distributions of juvenile and adult plants on twigs were higher than those expected based on the distribution of first-year seedlings, suggesting that on twigs, survival could be greater than on trunks and branches, canopy areas where peeling is faster. On the trunk and branches, in contrast, the proportion of juveniles and adults were similar to or less than that expected for first-year seedlings. The main cause of mortality was peeling and the area of minor overall mortality was the trunk, suggesting that this area should be favored as the main distribution area for the Tillandsia species but is not. Our results show that the peeling rate of B. fagaroides depends on branch size and suggest that the Tillandsia distribution depends not only on peeling rate but also on seed dispersion. We suggest that to colonize B. fagaroides epiphytes would either have adaptations to counteract the peeling rate or should occur in the areas of lowest peeling rate located in the exterior crown of trees.     

The isolation of RNA from raspberry (Rubus idaeus) fruit

Previous attempts to extract high-quality, total RNA from raspberry (Rubus idaeus) fruits using published protocols have proven to be unsuccessful. Even the use of protocols developed for the extraction of RNA from other fruit tissue has resulted in low yields (1) or the isolation of degraded RNA (2). Here, we report on the development of a quick and simple method of extracting total RNA from raspberry fruit. Using this method, high yields of good quality, undegraded RNA were obtained from fruit at all stages of ripening. The RNA is of sufficient quality for northern analysis and cDNA library construction.     

Lysis efficiency of standard DNA extraction methods for Halococcus spp. in an organic rich environment

The extraction of nucleic acids from a given environment marks a crucial and essential starting point in any molecular investigation. Members of Halococcus spp. are known for their rigid cell walls, and are thus difficult to lyse and could potentially be overlooked in an environment. Furthermore, the lack of a suitable lysis method hinders subsequent molecular analysis. The effects of six different DNA extraction methods were tested on Halococcus hamelinensis, Halococcus saccharolyticus and Halobacterium salinarum NRC-1 as well as on an organic rich, highly carbonated sediment from stromatolites spiked with Halococcus hamelinensis. The methods tested were based on physical disruption (boiling and freeze/thawing), chemical lysis (Triton X-100, potassium ethyl xanthogenate (XS) buffer and CTAB) and on enzymatic lysis (lysozyme). Results showed that boiling and freeze/thawing had little effect on the lysis of both Halococcus strains. Methods based on chemical lysis (Triton X-100, XS-buffer, and CTAB) showed the best results, however, Triton X-100 treatment failed to produce visible DNA fragments. Using a combination of bead beating, chemical lysis with lysozyme, and thermal shock, lysis of cells was achieved however DNA was badly sheared. Lysis of cells and DNA extraction of samples from spiked sediment proved to be difficult, with the XS-buffer method indicating the best results. This study provides an evaluation of six commonly used methods of cell lysis and DNA extraction of Halococcus spp., and the suitability of the resulting DNA for molecular analysis.     

A simple and rapid method for preparing genomic DNA from gram-positive bacteria

Molecular epidemiologic and other studies may require preparation of genomic DNA from large numbers of bacteria in sufficiently pure form for restriction endonuclease digestion, cloning, RAPD-PCR, Southern hybridization, and so on.Staphylococcus and other Gram-positive bacteria have a rigid cell wall and can be difficult to lyse. Here, a simple and rapid method for the preparation of genomic DNA from multiple samples is reported. This method produces clean DNA for use in most molecular biology methods in <90 min.     

一种简便制作小型鱼类形态标本的方法

本文介绍一种简便制作小型鱼类形态标本的方法.包括以下步骤:1.制作前准备;2.剥离皮肤;3.支撑架制作与放置;4.缝合;5.配制填充材料;6.填充;7.装义眼与整理姿态;8.干燥,上色,生境制作与固定.与其他制作小型鱼类形态标本的方法相比,新方法中先缝合再填充的步骤,难度小,容易控制填充物的量,做出的标本形态更逼真.     

无患子果实生长发育及其内含物的变化特征北大核心CSCD

以福建建宁县无患子为材料,观测果实生长动态,并采用Logistic方程对生长指标进行曲线拟合,观察生长过程中果皮显微结构变化,确定无患子果实生长发育的重要时期,为无患子的高效栽培管理和果实采收策略的制定提供科学依据。结果表明:(1)无患子果实生长、果皮总皂苷和种子油脂的积累规律相似,总体上呈Logistic增长模型的单“S”型曲线;果皮总皂苷和种子油脂的主要积累时期分别为45~90 DAP(花授粉后天数)和75~105 DAP。(2)无患子果皮除含有角质层、表皮细胞、厚角细胞、薄壁细胞、维管束、石细胞等基本结构外,还含有溶生式分泌腔和草酸钙簇晶等特征性结构;随着果实生长,分泌腔体积逐渐变大。(3)根据果实生长变化规律和Logistic方程拟合结果,可将无患子果实生长发育进程划分为生长初期(0~30 DAP)、速生期(30~90 DAP)、生长后期(90~120 DAP)和果实成熟期(120~150 DAP)4个阶段。在生产实践中果实成熟期内均适合果实采收,其中135和150 DAP分别为果实皂用和油用采收的最佳时期,此外还应根据每年天气状况对果实采收时间进行适当调整。     

In situ feeding tactics of short-nosed fruit bat (Cynopterus sphinx) on mango fruits: evidence of extractive foraging in a flying mammal

We report a sequence of behaviors exhibited by the short-nosed fruit bat Cynopterus sphinx while feeding on fruits of Mangifera indica. They peel off the outer skin to form a feeding area of about 3–6 cm diameter. Such food preparatory behaviors were more pronounced on larger mangoes. Bats competed among themselves to feed on the mangoes that had such feeding areas exposed. Individuals that spent a considerable amount of time on food preparatory behaviors actively secured the fruits. Altogether, these behaviors indicate that Cynopterus bats might have learnt, over evolutionary time, and developed behaviors that facilitate efficient processing and feeding of fruits such as mangoes. It appears that actions exhibited by C. sphinx in peeling off the outer skin of mangoes exemplify “extractive foraging”, a behavior that is prominently known in large-brained mammals. Thus, our findings will have implications on the distribution and evolution of extractive foraging and “technical intelligence” among mammalian lineages.     

An immunocytochemical method for histamine: application to the planarians

Histamine-immunoreactivity was investigated in the planarians Dugesia tigrina and Polycelis nigra. Specific antisera against a histamine-protein conjugate were used, and 1-ethyl—3 (3-dimethyl-aminopropyl) carbodiimide was used both as coupling agent to prepare the antigen and as a tissue fixative. In D. tigrina, histamine-immunoreactivity was restricted to photoreceptor cells in the cerebral eye. In P. nigra, nerve fibers were found in the ventral nerve cord and nerves running laterally from these. The epidermal eyes did not display histamine-immunoreactivity. The results suggest that histamine may be a transmitter in some of the most primitive animals. They also suggest that the distribution of histamine may differ in planarians.     

Traditional preparation and uses of cassava in Nigeria

Various methods of cassava preparation are practised by different ethnic groups in Nigeria. These methods involve peeling cassava roots, soaking roots in streams, grating cassava, and pressing grated cassava. Other methods include heating sieved, grated cassava, boiling peeled cassava roots, and pounding boiled or dried cassava roots. The traditional, cassava-based products aregari, fufu, akpu, cassava flour, edible starch, and tapioca. Detoxification of fresh cassava roots is partly achieved through cell rupture during cutting and grating, soaking in running or standing water in earthen pots for 3–5 days, heating, drying, and boiling.