Notes on the Carbowax Method of Making Tissue Sections

In undertaking to employ the water-soluble “Carbo-wax” compounds in making tissues sections, some experimentation was found necessary. These waxes are designated by the average molecular weights of their components except the 1500 grade, which is a mixture of the 1540 variety and the fluid polyethylene glycol 300; no reason is seen for its use in histological work. Different lots of the same designation may vary to some extent in their physical characteristics. The different grades vary greatly in hygroscopicity, the 1000 showing most and the 4000 none, but water taken up is not firmly held. Carbowaxes congeal in crystalline forms which are affected by the way the material is treated; the wax should be melted in the paraffin oven and the blocks cooled in the refrigerator. During infiltration the tissue should be stirred around occasionally, because water is not taken up with avidity. With leprosy skin lesions, infiltrating for 6 hours is preferable to the shorter periods usually specified. Using the 1540 grade (instead of the usual 1500) with the 4000 variety, a 15:85 mixture is satisfactory in our hottest weather, and a 20:80 mixture works well when it is cooler. Blocks which do not ribbon well tend to improve spontaneously, but the trouble may be corrected at once by “doping” the opposite surfaces with 25% beeswax in chloroform. Difficulty in affixing sections is avoided by using dried albuminized slides. Mounting sections on the slide is better done directly than by picking them up from a dish of the flotation material. The effects of a wide range of materials tested as flotants are summarized. The one preferred for ordinary purposes is a very dilute solution of Turgitol 7 with 10% carbowax 1540. Certain aliphatic hydrocarbons are useful for preserving serial relationships, but better is a thin celloidin film on water. Excellent results have been obtained in the staining of leprosy bacilli in carbowax sections.     

Plant Tissue 2

A new five-element electrical model was proposed recently (Zhang,Stout, and Willison, 1990; Zhang and Willison, 1991) to representplant tissues. In previous studies on the relationship betweenelectrical impedance and cold-hardiness, one of us had analyseddata in relation to a simpler three-element electrical model.Here, we have re-analysed these data in relation to the morecomplex model. F-tests showed that the new model always fittedmeasured impedance spectra significantly better (P<0·005)than the earlier model. The previously reported increase inintracellular resistance during cold acclimation was found tobe related to increased resistance of both the cytoplasm andvacuole In the species trial of birdsfood trefoil and alfalfa, coldacclimation was accompanied by an increase in extracellularresistance and a decrease in capacitances of both the plasmamembrane and tonoplast. In the cultivar trial of birdsfoot trefoil,cold acclimation did not affect plasma membrane capacitancein Viking and extracellular resistance in both Leo and Viking.In the species and growth time trial, cold acclimation was accompaniedby a decrease in plasma membrane capacitance in alfalfa butnot in birdsfoot trefoil Key words: Electrical impedance, cold acclimation, equivalent electrical circuit     

Notes on Contagious Distributions in Plant Populations

Past experience has shown that the Poisson series is often inadequateas a model for describing plant populations. Various alternativetwo-parameter models have been suggested in place of the Poissonseries, but they all depend on assumptions which may or maynot hold. In this paper a different approach is put forwardin that attention is concentrated on the mean number of plantsper quadrat and an index of ‘clumping’ or ‘contagiousness’.Examples are given as to the use of these concepts to test fordifferences between the distribution of a plant in two localitiesor between two plants in the same locality.     

植物组织培养的简化

介绍简化植物组织培养中培养基、培养条件、培养器皿以及其他方面的简化所采取的方法和所取得的成果,并提出一套合理的简化植物组织培养、降低培养成本的方案。     

Enzymatic Maceration of Plant Tissue

Tissue cells of Vicia faba were separated from the leaf pieces by the pectinase treatment after which the chlorophyll determination method was employed. The separation proceeded at a constant rate for 3 hours', the maximum rate of separation was at pH 5.3. The optimal pH shifted to 5.5 in the presence of 0.001 M 2Na-ethylenediaminetetraacetic acid (EDTA), which accelerated the separation at pH levels higher than 5.5. The separation was enhanced by NaCl and MgCL₂ and delayed by CaCl₂ and hypertonicity produced by sugars. Indoleacetic acid (IAA) and naphthaleneacetic acid (NAA) accelerated the separation at physiological concentrations. The rate of separation differed markedly by the age of the leaf and the species which offered the substrate material.     

在植物考据研究中应用进化思想的探讨

探讨了在植物考据研究中如何应用进化思想。我们认为在确定中国古籍文献中记载植物的学名时,对部分植物宜采用集合种（或复合种）的概念。     

药用植物栀子的组织培养

栀子(Gardenia jasminoides)为药用木本植物。以栀子果皮、种子团和种子为外植体,研究不同激素配比及不同培养方式对愈伤组织诱导和芽分化的影响。研究结果表明,培养基成分为MS+0.5 mg·L–12,4-D+0.25 mg·L–16-BA较适宜果皮和种子愈伤组织的诱导,诱导率分别为83.3%和88.5%;培养基成分为MS+1.0 mg·L–12,4-D+1.0 mg·L–16-BA较适宜种子团愈伤组织的诱导,诱导率为78.1%。3种外植体诱导的愈伤组织中,只有种子愈伤组织能通过液体培养分化出芽;TDZ对芽分化有明显的促进作用;最佳的芽分化培养基为MS+0.05 mg·L–1NAA+0.10 mg·L–1TDZ,其愈伤组织分化率为8.75%。该研究以栀子种子为外植体,并获得了再生植株,为药用植物栀子转基因体系的建立奠定了基础。     

薯蓣植物组织培养的研究进展

该文综述了组织培养在薯蓣植物快速繁殖、诱导多倍体及高产无性系筛选等方面的研究状况。褐化是薯蓣植物组培中常见的一种现象,也是阻碍组培发展的一大因素,为此,该文对褐化的原理作了分析并提出了几点解决方法,旨在使组织培养在薯蓣植物上得到更广阔的应用。     

Plant Cell Growth in Tissue

Cell walls are part of the apoplasm pathway that transports water, solutes, and nutrients to cells within plant tissue. Pressures within the apoplasm (cell walls and xylem) are often different from atmospheric pressure during expansive growth of plant cells in tissue. The previously established Augmented Growth Equations are modified to evaluate the turgor pressure, water uptake, and expansive growth of plant cells in tissue when pressures within the apoplasm are lower and higher than atmospheric pressure. Analyses indicate that a step-down and step-up in pressure within the apoplasm will cause an exponential decrease and increase in turgor pressure, respectively, and the rates of water uptake and expansive growth each undergo a rapid decrease and increase, respectively, followed by an exponential return to their initial magnitude. Other analyses indicate that pressure within the apoplasm decreases exponentially to a lower value after a step-down in turgor pressure, which simulates its behavior after an increase in expansive growth rate. Also, analyses indicate that the turgor pressure decays exponentially to a constant value that is the sum of the critical turgor pressure and pressure within the apoplasm during stress relaxation experiments in which pressures within the apoplasm are not atmospheric pressure. Additional analyses indicate that when the turgor pressure is constant (clamped), a decrease in pressure within the apoplasm elicits an increase in elastic expansion followed by an increase in irreversible expansion rate. Some analytical results are supported by prior experimental research, and other analytical results can be verified with existing experimental methods.Cell walls perform many functions for plant, algal, and fungal cells. Physical and chemical protection from the environment and physical support for cells and organs are obvious functions. Cell walls also withstand the stresses imposed by turgor pressure and deform irreversibly and reversible (elastically) during expansive growth. Irreversible wall deformations during expansive growth control cell enlargement, size, and shape. Growing and mature (nongrowing) cell walls undergo elastic deformations after changes in turgor pressure caused by changes in water status and environmental conditions. Elastic wall deformations are fundamental to the water relations of plant, algal, and fungal cells. For plant cells in tissues and organs, cell walls are part of the apoplasm pathway that transports water, solutes, and nutrients to cells.Importantly, pressures within the apoplasm (cell walls and xylem) are frequently different from atmospheric pressure during expansive growth of plant cells in tissues and organs. Lower pressures (tensions) are related to transpiration rates from plant organs and to expansive growth of cells in plant organs, e.g. Boyer (1967, 2001), Molz and Boyer (1978), Nonami and Boyer (1987, 1993), Nonami and Hashimoto (1996), Passioura and Boyer (2003), Boyer and Silk (2004), Koch et al. (2004), Wiegers et al. (2009), and the references within. Higher pressures (root pressures) occur during the spring when the soil is well hydrated (e.g. Kramer, 1932). Bleeding sap from cuts and broken stems is evidence of root pressure. Also, higher pressures may occur diurnally, during the night when transpiration rates are low (e.g. Tang and Boyer, 2008). Guttation drops on leaves in the morning are evidence of these higher pressures.Prior research indicates that a significant amount of chemistry and molecular biology occur within cell walls undergoing irreversible deformation during expansive growth (e.g. Cosgrove, 2005; Boyer, 2009). Two questions arise. First, how do pressures within the wall that are different from atmospheric pressure affect the turgor pressure, water uptake, and growth rate of cells in plant organs such as roots, stems, and leaves? Second, how are relevant chemical reactions affected by lower and higher pressures within the wall? The analyses conducted in this article focus on the first question.Previously, equations derived by Lockhart (1965) for wall deformation and water uptake (Growth Equations) were augmented with terms for elastic wall deformation (Ortega, 1985) and transpiration (Ortega et al., 1988). In this article, the previously established Augmented Growth Equations (Ortega, 1985, 1990, 1994, 2004; Ortega et al., 1988; Geitmann and Ortega, 2009) are modified to evaluate the turgor pressure, water uptake, and expansive growth of plant cells in tissue when pressures within the apoplasm are lower and higher than atmospheric pressure. In addition, the pressure within the apoplasm is evaluated after turgor pressure in cells decrease, thus simulating the condition produced by an increase in expansive growth rate of cells in plant tissues and organs. Also, the modified equations are used to determine how the results of stress relaxation experiments conducted on growing plant organs are affected by pressures within the apoplasm that are not atmospheric pressure. Last, the expansive growth of a plant cell is evaluated when pressure within the apoplasm undergoes a semi-instantaneous change while the turgor pressure remains constant, i.e. clamped. Some analytical results are supported by prior experimental research, and some analytical results can be verified with existing experimental methods.     

Influence of Tissue Density-specific Mechanical Properties on the Scaling of Plant Height

Tissue density,     

Notes on Technic: A Device for Centering Tissue Blocks on the Porter-Blum Microtome

A method of double embedding fixed tissues in 3% low viscosity nitrocellulose and paraffin is described. Five percent phenol in 80% alcohol during dehydration and 5% glycerin in the nitrocellulose solutions enhance cutting qualities. A modified Ruyter's solution is used to flatten sections. After a section is aflixed to a slide, it is passed through chloroform and acetone to remove the paraftin and celloidin. A 1% celloidin dip insures adherence of the seaion to the slide. Slides are stored in 70% alcohol until they are to be stained. Following staining and dehydration in graded alcohols, clearing should be done in a 1: 3 mixture of terpineol and toluene.     

腰果组织培养与植株再生(简报)

以腰果种子子叶为外植体,进行离体再生体系研究.结果表明,培养基MS 6-BA 5 mg/L 3%蔗糖 8g/L琼脂(暗培养)、b)1/2MS 1mg/L IAA 1mg/L IBA 3mg/L IBA 3mg/L PP333 0.2? 0.2AC 100mg/L VC 3%蔗糖 8g/L琼脂,运用生根法,生根率达到50%.     

Notes on Technic: Mahon's Myelin Stain for Celloidin-Embedded Nervous Tissue

Two methods commonly used to stain myelin sheaths are Kluver and Barrera's luxol fast blue (Kluver and Barrera 1953) and Weil's iron hematoxylin (Weil 1928). Both require differentiation of the stain; in addition, the Kluver-Barrera method specifies 16-24 hour staining. A third method for the selective staining of myelinated axons is that of Mahon (1938), which was introduced for use with paraffin-embedded autopsy tissue. The procedure possesses two distinct advantages since it requires: (1) no differentiation of the stain and (2) only 1 hour staining. Loyez's (1910) myelin stain for celloidin embedded tissue is similar to Mahon's but calls for long staining followed by differentiation. This report describes the application of Mahon's method to celloidin-embedded experimental tissue and emphasizes its utility for staining tissues to be used for reconstructing microelectrode penetrations (fig. 1) and for demonstrating the effect of experimental lesions (fig. 2).     

Deoxyribonucleotide Pools in Plant Tissue Cultures

Two dimensional thin-layer chromatography on anion-exchange cellulose enables the separation of the normally occurring ribo- and deoxyribonucleoside triphosphates. This technique was applied to perchloric acid extracts of callus tissue of sycamore and tobacco and of pine pollen grown in ³²P-orthophosphate labelled media to quantitate the nucleoside triphosphate pools under different growth conditions. The results showed that the ratio of the deoxyribonucleo-side triphosphates to their corresponding ribonucleoside triphosphates is low in plant cells, similar to the ratio previously found for animal cells. During the period of most rapid DNA synthesis in the callus tissue, the deoxyribonucleoside triphosphate pools reach their highest values. This effect is also demonstrated with cells of Escbericbia coli.     

植物组织培养物的超低温保存

对超低温保存技术的研究历史、基本原理、方法、影响因素和国内外植物组织培养物超低温保存的研究进展作了介绍,并对这一问题的前景作了展望.     

一种快速提取植物病毒DNA的方法

介绍在PCR检测体系中一种快速提取病毒DNA的方法。利用高盐缓冲液溶液释放病毒DNA,同时利用葡聚糖凝胶微柱纯化提取液,有效消除样品中PCR抑制物质并直接作为PCR反应模板扩增检测病毒。该法无需任何特殊设备,适合对大量植株进行大通量的检测分析。