Zur Kenntnis der OscillatoriaceePseudanabaena galeata (Cyanophyta)

If exsiccated trichomes ofPseudanabaena galeata are immersed in water, striking changes occur as a consequence of lysis and maceration. The locomotion of living trichomes differs from that inOscillatoriaceae. This and other differences make it doubtful, whetherPs. galeata belongs to theOscillatoriaceae; at any rate it occupies an aberrant position.

     

Estimation of Jordanian isolated Pectobacterium cellulase,pectinase concentrations,RAPD polymorphism and their maceration activity

Forty-two Pectobacterium isolates were recovered from contaminated soil and rotted vegetables in Jordan. Twenty of them were belonged to; Pectobacterium carotovorum subsp. Carotovorum (Pbc) (= Erwinia carotovora subsp. carotovora), 11 isolates were belonged to Pectobacterium atrospeticum (= Erwinia carotovora subsp. atroseptica) (Pba) and 11 isolates were not classifiable (Pbs). Maceration activity of the 42 proved their ability to macerate potato, carrot and radish slices. Maceration activity of the isolates either of the same subspecies or in between the isolates of different subspecies isolated from the same host or from different hosts was varied. The measured concentration in μM?ml^?1 of both cellulase and pectinase enzymes was variable too. The Rapid amplified polymorphic DNA-PCR finger printing of total genomic DNA using a pair of 10-mer oligonucleotide primers amplification showed similar DNA bands with some polymorphic variations amongst the isolates.     

Correlative Light and Scanning Electron Microscopy for Observing the Three-Dimensional Ultrastructure of Membranous Cell Organelles in Relation to Their Molecular Components

Although the osmium maceration method has been used to observe three-dimensional (3D) structures of membranous cell organelles with scanning electron microscopy (SEM), the use of osmium tetroxide for membrane fixation and the removal of cytosolic soluble proteins largely impairs the antigenicity of molecules in the specimens. In the present study, we developed a novel method to combine cryosectioning with the maceration method for correlative immunocytochemical analysis. We first immunocytochemically stained a semi-thin cryosection cut from a pituitary tissue block with a cryo-ultramicrotome, according to the Tokuyasu method, before preparing an osmium-macerated specimen from the remaining tissue block. Correlative microscopy was performed by observing the same area between the immunostained section and the adjacent face of the tissue block. Using this correlative method, we could accurately identify the gonadotropes of pituitary glands in various experimental conditions with SEM. At 4 weeks after castration, dilated cisternae of rough endoplasmic reticulum (RER) were distributed throughout the cytoplasm. On the other hand, an extremely dilated cisterna of the RER occupied the large region of the cytoplasm at 12 weeks after castration. This novel method has the potential to analyze the relationship between the distribution of functional molecules and the 3D ultrastructure in different composite tissues.     

次氯酸钠离析法观测甘蓝叶片脉序的处理条件优化筛选

次氯酸钠（NaClO）离析法主要用于植物叶片表皮的观测,在研究过程中发现该法也可用于叶片脉序的观测。以甘蓝（Brassica oleracea var.capitata）为实验材料,采用二次正交设计方法对水煮时间、NaOH浓度、NaOH处理时间、NaClO浓度、NaClO处理温度和处理时间等各种处理条件进行优化筛选,以期得到适合于甘蓝叶片脉序观测的最佳处理条件组合。实验结果表明,新鲜甘蓝叶片水煮3分钟,10%的NaOH溶液60°C水浴处理2.5小时,3%的NaClO溶液40°C水浴离析2小时,叶片脉序的观测效果最佳。     

A Rapid Method for Macerating Phloem

Sieve cells and sieve tube members can be macerated from the phloem of various organs of woody and herbaceous species by au-toclaving the tissue in a mild macerating medium. This treatment does not digest the primary walls or the callose deposits on the sieve areas and sieve plates of the sieve elements. These cells can then be recognized by the fluorescence of their callose after staining with aniline blue. Sometimes adjacent sieve elements fail to separate and one can observe details of their junctures.     

桔梗科4种植物分离配囊的比较研究

应用酶解技术分离出桔梗科四种植物(桔梗、轮叶沙参、杏叶沙参和羊乳)的胚囊。对胚囊的形态结构和贮藏物质的变化作了观察比较。它们的成熟胚囊结构基本相同,均由卵细胞、一对助细胞和中央细胞组成。反足器退化较早。但形态、大小等有一定差异。桔梗胚囊狭长,细胞质浓密;沙参(包括轮叶沙参与杏叶沙参)的胚囊也较狭长,细胞质不如前者浓密;羊乳胚囊非常宽大,而细胞质相当稀薄,与前三种植物的差异较大。它们均含有贮藏物质,显微化学反应表明主要系多糖物质。多糖颗粒的形状、大小各有特点,在胚囊受精前后的消长变化也有不同。其中羊乳与其它三种植物之间的差别较大。     

Isolation of the lily embryo sac

Summary A 5%–8% yield of isolated embryo sacs of Lilium longiflorum was obtained using an enzymatic isolation procedure. The best results were obtained with a maceration mixture containing mannitol, pectinase, pectolyase, cellulase, hemicellulase, CaCl₂ and NaOH. All developmental stages of the female gametophyte can be isolated in the living condition, although fewer than expected mature stages were observed. Moreover, only some of the more mature embryo sacs showed a positive fluorochromatic reaction (FCR) at the time of liberation. When the embryo sacs were stored in the enzyme or sugar solution after isolation, the positive reaction rapidly diminished for all stages. The isolated embryo sac and its nuclei were similar in size, shape, and position to the in situ embryo sac. Light microscopical observations of sectioned material revealed an intact cellular structure. However, the deleterious effects of the enzyme solution were sometimes observed in the form of lipid-like accumulations inside the isolated embryo sac.In collaboration with: Reconnaissance Cellulaire et Amélioration des Plantes, Université Cl. Bernard-Lyon I; INRA 879, 43, Boulevard du 11 Novembre 1918, F-69622 Villeurbanne Cedex, France     

Enzymatic isolation of living embryo sacs of Petunia

Summary A 20%–25% yield of isolated and living embryo sacs of Petunia hybrida L. was obtained using an enzymatic maceration mixture containing 3% driselase (soluble fraction only), 0.1% MES buffer, pH 5.5, and 8% mannitol. For each maceration ± 450 ovules were incubated in 1 ml enzyme solution for 2 h at 30° C in a shaking waterbath (150 rpm). Subsequently, the enzyme solution was replaced by Brewbaker and Kwack's medium, pH 6.5, supplemented with 10% mannitol (BKM). Gentle agitation of the suspension resulted in the liberation of embryo sacs, which were then collected with a micropipette using a dissecting microscope and transferred to fresh BKM. The embryo sacs isolated are intact and living, and have maintained their original shape and organization When stored in BKM at room temperature the isolated embryo sacs remain alive for 8 h. Storage at 4° C results in a prolongation of viability of up to 80 h. Prolonged incubation of ovules or reincubation of isolated embryo sacs in the maceration mixture results in the liberation of the gametophytic cells as individual, living protoplasts.     

次氯酸钠离析法观测甘蓝叶片脉序的处理条件优化筛选

次氯酸钠(NaClO)离析法主要用于植物叶片表皮的观测, 在研究过程中发现该法也可用于叶片脉序的观测。以甘蓝(Brassica oleracea var. capitata)为实验材料, 采用二次正交设计方法对水煮时间、NaOH浓度、NaOH处理时间、NaClO浓度、NaClO处理温度和处理时间等各种处理条件进行优化筛选, 以期得到适合于甘蓝叶片脉序观测的最佳处理条件组合。实验结果表明, 新鲜甘蓝叶片水煮3分钟, 10%的NaOH溶液60°C水浴处理2.5小时, 3%的NaClO溶液40°C水浴离析2小时,叶片脉序的观测效果最佳。