拟南芥菜(Arabidopsis thaliana)子叶中蛋白体形成的超微结构和免疫电镜定位研究

本文对拟南芥菜(Arabidopsis thaliana)种子发育过程中贮藏蛋白的积累和蛋白体的形成进行了超微结构和免疫电镜定位的研究。常规超薄切片的电镜观察表明,在开花后第10天(10 DAF),高电子密度的蛋白质物质开始在子叶细胞的液泡中沉积。这一过程一直延续到种子接近成熟(14 DAF),这时液泡中充满了蛋白质物质,转变成为大的蛋白体。利用了该种植物主要种子贮藏蛋白之一的12 s球蛋白的单克隆抗体作为免疫探针,以蛋白质A-胶体金电镜技术对12 s种子蛋白进行了细胞内定位,证实了在液泡中积累的物质为种子贮藏蛋白。实验结果表明在拟南芥菜中,子叶细胞中的液泡是蛋白体的前体,肯定了蛋白体的发生起源于液泡的观点。本文还对应用胶体金电镜技术进行细胞内定位的某些问题作了初步探讨。     

Techniques for Preparing Seeds with Water-Impermeable Coats for Light and Electron Microscopy

An improved method for fixing and embedding seeds with impermeable coats for microscopic study has been developed. Entry portals are cut into seed coats to permit better penetration of fixative. This makes it possible to obtain semithin sections of whole seeds for light microscopy and thin sections of selected areas for electron microscopy. Seed tissues may thereby be studied relative to their position in the seed and to surrounding tissues. This permits studies of imbibition and developmental morphology of seeds in histological and cytological detail previously possible only with soft or dissected seeds.     

刺楸种子中性与酸性抑制物质的研究

本文研究了刺楸种子抑制物质的提取,分离和鉴定方法,并测定了抑制物质的相对活性及其含量。结果表明,刺揪种子中除含高水平的酸性抑制物外,还存在一种中性抑制物。该中性抑制物同酸性抑制物相似,有较强的生物活性,不但可以抑制白菜种子的萌发与胚根生长,而且对已解除休眠的刺楸种子也有显著作用。利用纸层析,薄层层析,颜色反应和高效液相色谱等手段,证明中性抑制物质为香豆素类物质;酸性抑制物质的主要成分为脱落酸。种子各部位均含这两种抑制物质,但含量有所不同,随种子贮藏时间的延长,抑制物会发生转移。本文还讨论了用不同溶剂提取中性抑制物的效果。     

植物组织石蜡切片的扫描电镜观察方法研究

石蜡切片的扫描电镜观察法有其独到之处:集光镜和扫捕电镜特长于一体,在大量的石蜡切片光镜观察的基础上,挑选具有研究线索的切片,采用此法转移到扫描电镜下作高分辩研究,既可普查切片全貌,又可处得切片中亚微结构的三维图像,这对结构的准确分辩十分有利,且便于作连续切片观察。本文简要介绍这一实验技术。     

古生代有孔虫实体化石研究方法

古生代有孔虫大部分是通过切片进行研究的,但是普通的切片法很难得到非常好的定向标本,这就产生了很多同物异名,限制了有孔虫在古生代生物地层学、古生态学和古海洋学中的应用。另外由于口孔的内部和外部特征在分类学中的地位越来越重要,所以通过实体化石来研究古生代有孔虫理应受到更多的重视。本文论证了在古生代地层中获得实体化石的可能性,对化石提取的实验步骤进行了系统总结,细述了对实体化石进行扫描电镜和切片研究的全过程,并根据笔者的经验,对一些技术细节及注意事项进行了详细介绍。本文最后建议,古生代的有孔虫研究应该尽可能地利用实体化石来进行。     

FROZEN THIN SECTIONS OF FRESH TISSUE FOR ELECTRON MICROSCOPY, WITH A DESCRIPTION OF PANCREAS AND LIVER

A simple method has been developed that allows frozen thin sections of fresh-frozen tissue to be cut on a virtually unmodified ultramicrotome kept at room temperature. A bowl-shaped Dewar flask with a knifeholder in its depths replaces the stage of the microtome; a bar extends down into the bowl from the microtome's cutting arm and bears the frozen tissue near its lower end. When the microtome is operated, the tissue passes a glass or diamond knife in the depths of the bowl as in normal cutting. The cutting temperature is maintained by flushing the bowl with cold nitrogen gas, and can be set anywhere from about -160°C up to about -30°C. The microtome is set for a cutting thickness of 540–1000 A. Sections are picked up from the dry knife edge, and are placed on membrane-coated grids, flattened with the polished end of a copper rod, and either dried in nitrogen gas or freeze-dried. Throughout the entire process the tissue is kept cold and does not come in contact with any solvent. The morphology seen in frozen thin sections of rat pancreas and liver generally resembles that in conventional preparations, although freezing damage and low contrast limit the detail that can be discerned. Among unusual findings is a frequent abundance of mitochondrial granules in material prepared by this method.     

Ultrastructural localization of soybean agglutinin on thin sections of Glycine max (soybean) var. Altona by the gold method

Summary Soybean agglutinin (SBA) has been localized in Glycine max (soybean) var. Altona at the ultrastructural level by the gold method. SBA was detected by marking thin sections of different part of the seed with gold granules (12 nm in size) labelled with anti-SBA antiserum. Upon examination by transmission electron microscopy, the lectin was found uniformly distributed in most of the protein bodies of the cotyledon. SBA was also present in the embryo axis.     

Measurement of light within thin plant tissues with fiber optic microprobes

Vogelmann, T. C., Knapp, A. K., McClean, T. M. and Smith, W. K. 1988. Measurement of light within thin plant tissues with fiber optic microprobes. - Physiol. Plant. 72: 623–630.
The measurement of light with fiber optic microprobes has been extended to thin (200–300 μm) plant tissue samples. To test the method, light measurements were made in thin aqueous films and paradermal sections from 10-day-old etiolated Cucurbita pepo L. cv. Fordhook cotyledons. The measurements obtained were highly reproducible. Paradermal sections of spongy mesophyll that were irradiated with collimated light scattered light more effectively than the palisade layer of intact cotyledons. These results demonstrate that different plant tissues have different light scattering characteristics. The successful extension of the fiber optic microprobe technique to thin systems makes it possible to examine the optical properties of different cell layers within leaves and other plant organs.     

鸡冠花种子营养成分的研究

鸡冠花种子可供食用和药用。作者采用常规方法系统地分析测定了其营养成分,并和一些谷类,豆类食物进行了比较。结果显示,鸡冠花种子含各种营养成分:蛋白质、脂肪、碳水化合物、膳食纤维、氨基酸、维生素、无机元素等,不仅含量丰富且高于谷类。这为开发利用鸡冠花种子提供了科学依据。     

STRUCTURAL BIOLOGY OF THE FIBRES OF THE COLLAGENOUS AND ELASTIC SYSTEMS

The different types of fibres of the collagenous and elastic systems can be demonstrated specifically in tissue sections by comparing the typical ultrastructural picture of each of the fibre types with studies using selective staining techniques for light microscopy. A practicalmodus operandi, which includes the recommended staining procedures and interpretation of the results, is presented. Micrographs and tables are provided to summarize the differential procedures. Reticulin fibres display a distinct argyrophilia when studied by means of silver impregnation techniques, and show up as a thin meshwork of weakly birefringent, greenish fibres when examined with the aid of the Picrosirius-polarization method. In addition, electron-microscopic studies showed that reticulin fibres are composed of a small number of thin collagen fibrils, contrasting with the very many thicker fibrils that could be localized ultrastructurally to the sites where non-argyrophilic, coarse collagen fibres had been characterized by the histochemical methods used. The three different fibre types of the elastic system belong to a continuous series: oxytalan—elaunin—elastic (all of the fibre types comprising collections of microfibrils with, in the given sequence, increasing amounts of elastin). The three distinct types of elastic system fibres have different staining characteristics and ultrastructural patterns. Ultrastructurally, a characteristic elastic fibre consists of two morphologically different components: a centrally located solid cylinder of amorphous and homogeneous elastin surrounded by tubular microfibrils. An oxytalan fibre is composed of a bundle of microfibrils, identical to the elastic fibre microfibrils, without amorphous material. In elaunin fibres, dispersed amorphous material (elastin) is intermingled among the microfibrils.