JEM-120EXⅡ电镜测量功能在病毒学研究中的应用

应用ＪＥＭ－１２００ＥＸⅡ透射电子显微镜测量系统观察和测量香石竹斑驳病毒、烟草花叶病毒、马铃薯Ｙ病毒．结果表明：电镜测量的数据准确，测量系统性能可靠．     

鸡后口吸虫的超微结构研究

应用透射电镜对鸡后口吸虫(Postharmostomum gallium)的皮层、盲肠、排泄系及实质细胞的超微结构研究显示：在上述组织细胞核内存在着电子深染的圆形颗粒。这种颗粒具有：①电子密度深，均匀，②数量不一，为1～16个，③形态规则，大多呈球形，少数外周有突起，④大小不定等特性。皮层厚约7．62-13．33μm，属中等厚度皮层的吸虫类，基质有50-80列杆状分泌体，未见体棘。盲肠上皮为合胞体结构，肠腔内容物为鸡血细胞及残骸。排泄系由烙细胞，初、次级毛细管、集合管以及排泄囊所组成，焰细胞的纤毛数为65-76根；集合管和排泄囊的壁可分两部分，靠腔面部分结构致密，均均，另部分结构疏松，富含细胞器。实质细胞形态多样，突起多。本文还对上述组织细胞的结构特点以及胞核内电子深染的圆形颗粒的性质进行了讨论。     

Epidermal Cell Wall Biogenesis with Emphasis on Cuticular Layer Formation during Direct Somatic Embryogenesis in Rice

Abstract

Epidermal cell wall biogenesis with emphasis on cuticular layer formation during direct somatic embryogenesis in rice was examined by transmission electron microscopy. The first phenomenon noticed in the early globular stage of somatic embryos was a high activity of cell wall formation. In this stage, many endoplasmic reticula, Golgi bodies and their vesicles were present and epidermal cells were covered by fibrillar material. Cutin was initially deposited on the epidermal cell wall of late globular or early elongate stage embryos. In the late elongate stage, a lamellated cuticular layer formed, and in the late scutellar stage, a more homogenous continuous cuticular layer formed. The cuticular layer disappeared gradually from the scutellum cell wall as the somatic embryo germinated. The disappearance of the cuticular layer was similar to that of the zygotic embryo. There is no cuticular layer on the scutellar epithelial cell wall of the immature zygotic embryo 5 days after germination. Our observations lead to the conclusion that one of the structural and physiological markers of direct somatic embryogenesis is the transformation of fibrillar material into the cuticular layer.     

Pb、Cd和酸胁迫下枫香萌发种子的生理生化变化及其亚细胞结构

为探讨重金属及酸胁迫下枫香种子萌发过程中营养物质的动态变化规律,本研究采用纸培法,设置不同浓度的Pb、Cd及酸胁迫试验,考察种子萌发过程中可溶性蛋白、可溶性糖、还原糖、淀粉酶活性,并结合傅里叶红外光谱(FTIR)图谱变化及透射电镜(TEM)观察了种子胚乳细胞的超微组织结构变化。结果表明:萌发初中期(4、7 d)高浓度的Pb胁迫显著提高了可溶性蛋白含量及β-淀粉酶活性,α-淀粉酶活性则在Pb-500时活性最高(P0.05)。Cd胁迫对淀粉酶活性影响较大,萌发中后期可溶性蛋白、还原糖和可溶性糖对Cd的响应逐渐加强。FTIR分析表明Pb-250处理刺激膜脂和细胞壁果胶中常见的酯类化合物合成,Pb-500、Cd-25处理利于纤维素、半纤维素、多糖、蛋白质等有机物质的合成,以此来增强对铅镉的抗逆性。酸胁迫降低了可溶性蛋白含量,pH-5.5、pH-4.5提高了可溶性糖、还原糖含量及淀粉酶活性。随酸胁迫的增强特征峰变化值下降,pH-3.5处理组峰值略有提升。大部分Pb~(2+)、Cd~(2+)沉积在胚乳细胞细胞壁及细胞质中,细胞区室化作用显著。枫香可在重金属及酸污染的废弃地正常生长,其种子通过渗透物质相互协调及细胞区室化作用对Pb、Cd和酸胁迫有积极的响应。     

猪带绦虫虫卵及六钩蚴膜超微结构的观察

【目的】观察猪带绦虫虫卵和六钩蚴膜的超微结构。【方法】猪带绦虫病患者用槟榔、南瓜籽法驱虫,收集成熟虫卵。次氯酸钠法破壳后,等渗percoll溶液收集六钩蚴,人工肠液激活六钩蚴。扫描电镜和透射电镜观察虫卵和六钩蚴。【结果】猪带绦虫成熟虫卵呈卵圆形,直径约为50-58μm,由胚膜、内胞质层、六钩蚴膜和六钩蚴等几部分组成;发育期猪带绦虫六钩蚴的膜结构由外向内分别是外胞膜残留层、外胞质层(包括不规则较致密的外层和均质透明的内层)、胚膜(包括胚层和内胞质层)和六钩蚴膜,而成熟六钩蚴的膜结构仅六钩蚴膜一层,厚度约为49-51nm。【结论】猪带绦虫虫卵和六钩蚴膜的超微结构和缩小膜壳绦虫(Hymenolepis diminuta)及肥胖带状绦虫(T.saginata)的超微结构相似,但膜下层的糖原颗粒外周无包膜。     

宫川棘口吸虫和似锥低颈吸虫的卵黄细胞发育（复殖目：棘口科）

用透射电镜观察了棘口科（Ｅｃｈｉｎｏｓｔｏｍａｔｉｄａｅ）、棘口属（Ｅｃｈｉｎｏｓｔｏｍａ）的宫川棘口吸虫（Ｅ．ｍｉｙａｇａｗａｉ），低颈属（Ｈｙｐｏｄｅｒａｅｕｍ）的似锥低颈吸虫（Ｈ．ｃｏｎｏｉｄｅｕｍ）的卵黄细胞发育超微结构变化。两种吸虫卵黄细胞发育过程比较相似。卵黄小叶外被覆一层基膜。未发育卵黄细胞含有较少细胞质，大量游离核糖体。发育期卵黄细胞含有较多细胞质，丰富的粗面内质网，高尔基复合体和卵黄球。成熟期卵黄细胞的胞质丰富，粗面内质网减少，位于核周围和细胞边缘，胞质内还出现大量糖原和包涵体。在似锥低颈吸虫的发育期和成熟期卵黄细胞内，还出现脂滴。     

Ultrastructural and biomolecular detection of Rickettsiales‐like organisms in tissues of rainbow trout with Red Mark Syndrome

Red mark syndrome (RMS) and US strawberry disease (US SD) are skin disorders affecting rainbow trout farmed in Europe and USA. The disease etiology has not yet been established. In spite of specific investigations, identifying Rickettsia‐like organism (RLO)‐ and Midichloria‐like organism (MLO)‐related DNA in affected individuals, these pathogens have never been observed. We performed histological, ultrastructural and biomolecular analysis on skin and spleen samples of trout with RMS. Examination by TEM revealed the presence of intracytoplasmic microorganisms resembling Rickettsiales within macrophages, fibroblasts and erythrocytes. The microorganisms were oval or short rod shaped (400–800 nm in length and 100–200 nm in width) and often showed a cell wall similar to Gram‐negative bacteria. PCR analysis for Rickettsiales supported these findings: 53% of affected trout were positive by both PCR and TEM The primers RiFCfw‐RiFCrev were used to anneal both the RLO 16S DNA sequence and the MLO 16S DNA sequence. For this reason, and in agreement with previous studies confirming the presence of Rickettsiales‐related DNA in trout with RMS, we assume that TEM detected microorganisms morphologically consistent with bacteria belonging to Rickettsiales order and could be considered as possible causative agents of RMS.     

日本柑橘黄龙病病原体电镜观察及PCR检测

对日本柑橘黄龙病病原体的形态结构进行了电镜观察,结果发现柑橘黄龙病病菌日本株系绝大多数是球形和短椭圆形,极少观察到棒状形态,直径为100～500nm,外被一层胶状物质包围,厚约10～50nm,这与其他地区发现的柑橘黄龙病病原体形态不同．PCR检测表明,以硅藻土法提取的DNA为模板,利用Ready-To-Go^TM PCR bead法可以得到理想结果。     

The effect of dietary sodium chloride on some osmoregulatory parameters of the teleost, Oreochromis niloticus, after transfer from freshwater to seawater

The aim of this work was to determine the effects of supplemental dietary sodium chloride on salt water acclimation of tilapia Oreochromis niloticus. Fish were fed a basal diet supplemented with NaCl (8%) during three weeks in fresh water (FW) and then transferred to salt water (SW) at 15 and 20. Changes in plasma osmolality, chloride ion concentration (Cl^–), plasma level of cortisol and gill Na⁺, K⁺-ATPase activity were measured at 6, 12, 24, 48, 72 and 168 h after transfer to 15SW, while the higher strength SW group (20) was only monitored up to 24 h. Morphological changes in the gill mitochondria-rich (MR) cells were examined in relation to environmental salinity. The changes associated with dietary NaCl were sporadic and of small magnitude. The plasma osmolality and Cl^– increased immediately after transfer up to 12–24 h, but fish fed dietary salt (S) showed lower values than the control group (C). The S group showed higher plasma levels of cortisol than the control, which maintained its initial levels during the experiment. Gill Na⁺, K⁺-ATPase activity of the S group began to increase in the first hours after transfer, reaching maximum at 12 h and returned to basal level at 24 h, while the control group maintained basal levels. The differences between gill Na⁺, K⁺-ATPase activity of S and C fish were significant (p < 0.05) at 12 h. Transmission electron microscopy (TEM) revealed that MR cells in SW show more mitochondria and a more developed tubular system arising from the basolateral membrane. The MR cells of both groups frequently formed a multicellular complex in SW, consisting of a main MR and one or more accessory cells. Such complexes are rarely observed in FW. Some MR cells of fish fed supplemented dietary salt displayed convex apical membrane in FW.     

枝管藻细胞壁内、外多糖的细胞化学定位研究

为进行经济海藻枝管藻细胞壁多糖和胞外多糖的超微定位,本文以钌红电镜细胞化学方法制备枝管藻透射电镜样品。结果表明,钌红染色方法不仅比常规电镜制样方法能够更完整地显示海藻细胞壁的超微结构,而且使枝管藻多糖在细胞壁内、外的分布在透射电镜下直观可见:细胞壁内的多糖物质主要呈多层紧密排列的微纤维状,细胞表面的多糖物质则呈松散的网状。本结果为研究海藻细胞壁的超微结构和海藻吸附重金属的作用部位提供了技术支持。