意大利蝗痘病毒一些特性研究

电镜观察意大利蝗痘病毒的包涵体大多数为椭圆形,少数为近圆形,大小相差极为悬殊。病毒粒子呈卵圆形,表面桑椹状。病毒粒子髓核呈圆筒状,其纵切面内绳索结构折叠2—3次,横切面可以看到2—3个圆点,侧体也圆筒状,均匀包围在核衣壳外面。DNA经限制性内切酶LoR I酶切产生16个DNA片段,据此求得DNA分子量为135.7×106D。 意大利蝗痘病毒包涵体的裂解释放病毒速度快,在相同条件下比亚洲小车蝗痘病毒包涵体裂解释放病毒粒子快。当温度恒定(37℃),在一定的pH值内(pH 9.0—11.5),随pH值的升高包涵体裂解释放病毒粒子速度加快。当pH值恒定(11.2),在一定的温度范围内(25～40℃),随温度的升高包涵体的裂解释放病毒粒子速度加快。     

意大利蝗痘病毒与西伯利亚蝗痘病毒包涵体蛋白基因序列分析(英文)

本文报道了意大利蝗痘病毒(CiEPV)与西伯利亚蝗痘病毒(GsEPV)包涵体蛋白基因序列分析。CiEPV与GsEPV包涵体蛋白基因分别包含2922bps,2967bps的开放阅读框架,编码109.2kDa,111.1kDa蛋白质。与鳞翅目及鞘翅目昆虫痘病毒包涵体蛋白氨基酸的同源性低于20％,而与其他直翅目昆虫痘病毒包涵体蛋白氨基酸的同源性均高于80％。CiEPV与GsEPV包涵体蛋白分别包含19与21半胱氨酸位点,主要分布在C-末端,半胱氨酸位点的数目与位置均类似于其他直翅目昆虫痘病毒包涵体蛋白。此两种痘病毒包涵体蛋白基因的启动子区域基因序列保守,富含A+T并且具有典型的痘病毒晚期启动子信号TAAATG。同时在此两种痘病毒包涵体基因的下游均克隆了另一个不完整的基因序列,此基因与血黑蝗痘病毒的MSV072基因同源,并且相对于包涵体蛋白基因为反向。     

Maturation and aging of adult fat body and oenocytes in Drosophila as revealed by light microscopic morphometry

A morphological and cytometric analysis of the adult fat body cells and oenocytes was made on sections of abdomens from immature, mature and senescent Drosophila melanogaster of both sexes. There are about 18,000 fat body cells in abdomens of female and mature male flies. Immature and senescent males have about 12,000 and 15,000 cells, respectively. The size of the cells is almost the same for immature flies of both sexes and increases about six-fold to approximately 2600 micron2, so that mature flies of both sexes have equivalent amounts of fat body tissue. The proportions of lipid, glycogen, and background cytoplasm of fat body cells also remain relatively constant throughout adult life, but dense, proteinaceous granules are observed in cells of senescent flies. The amounts of cellular components change dramatically due to change of cell size with age; the amount of lipid shows the greatest sexual difference with about 2x more in the females at all stages studied. The oenocytes number about 6,000 in the abdomens of all but immature male flies, which have approximately 4,000. Although the cells of both sexes triple in size to about 700 micron 2, the oenocytes of males reach maximum size earlier than those of females. The major features of oenocytes appear to be dense background cytoplasm, putative lipid droplets found only in mature flies, and pigmented granules first seen in the cells of mature flies which accumulate with age to 33% of the cytoplasm. The number of cells and their anticipated capacity for protein synthesis is discussed in relation to the production of yolk protein precursors.     

SARS-CoV在Vero细胞培养中的多形性特征

为揭示SARS相关冠状病毒(SARS-associated coronavirus,SARS-CoV)的复制特点,并探讨该病毒的致病机制,利用负染电子显微镜(EM)、超薄切片EM和免疫EM技术研究了SARS-CoV在Vero细胞上的形态学特征。结果显示,成熟病毒颗粒多为圆形或椭圆形,直径80~120nm,包膜上有放射状排列的纤毛样突起,长约20nm,基底窄。此外可见哑铃形、肾形及“钉子样”等多形性成熟病毒颗粒,这些颗粒能与病人恢复期血清和抗S蛋白抗体反应。细胞内病毒颗粒多位于包涵体内,呈显著的多形性,直径为20~400nm,其形状可分为:①圆形、肾形或椭圆形;②管样结构;③不规则形,如三角形、哑铃形等,另外可见一种特殊的分枝样颗粒。颗粒电子密度不一,有实心和空心两种,其中空心颗粒周边的电子密度高。还观察到螺旋形的病毒核衣壳结构。     

冷蒿和苜蓿对意大利蝗生长及生殖力的影响

以紫花苜蓿 Medicago sativa (L.)、冷蒿 Artemisia frigida Willd.Sp.Pl.以及二者1:1混合饲喂意大利蝗Calliptamus italicus (L.)一龄蝗蝻,冷蒿饲喂不能完成其生活史,两种食料混合饲喂的发育进度要快于紫花苜蓿饲喂的个体。以三种食料分别饲喂意大利蝗成虫,研究其生长及生殖力的变化,结果表明：与取食紫花苜蓿和取食混合食料相比,取食冷蒿抑制了意大利蝗成虫体长、体重、雌雄成虫寿命、交配率和单雌产卵量,导致交配期提前,雌虫死亡率升高,而对卵囊内卵粒数无影响。与取食紫花苜蓿相比,取食混合食料促进了卵囊内卵粒数的增加,其余指标没有显著变化。这些结果表明两种牧草的隔离种植对减轻意大利蝗为害是十分必要的。     

Estimating acridid densities in grassland habitats: a comparison between presence-absence and abundance sampling designs

Sampling methods to estimate acridid density per surface area unit in grassland habitats were compared using presence-absence data and count data. Sampling plans based on 6 yr of surveys were devised to estimate the density of Chorthippus spp., Euchorthippus spp., and Calliptamus italicus L. These acridids represented >90% of species in the study area. Sampling plans based on count data provided a reasonable tool when densities were >1/m(2) and when the level of precision was 0.20-0.30. A binomial sampling plan can be used to estimate C. italicus density with a level of precision >or=0.28. Sampling characteristics, i.e., estimated mean, actual precision, and sample size, were established on validation data sets with bootstrapping analysis. Sampling costs were also calculated according to density-dependent functions. Comparison between binomial sampling and enumerative sampling of C. italicus showed that binomial sampling required less time than enumerative sampling when densities were 0.35. Plot area had no significant effect on sample variances of counts.     

Henneguya adherens N. Sp. (Myxozoa, Myxosporea), Parasite of the Amazonian Fish, Acestrorhynchus falcatus

ABSTRACT. A new species of myxosporean from the gill filaments of the freshwater teleost fish, Acestrorhynchus falcatus collected in the Amazon river is described from light and transmission electron microscope observations. The mature spores (total length 32.3 [30.7–35.1] μ) and all developmental stages were found in the same sporogonic plasmodium. The ellipsoidal spore body consists of 2 unequal shell valves adhering together along the suture lines. Each valve, tapering as a caudal projection, forms a long tail (length 20.5 [18.0–21.7] μm). The tail was surrounded by a homogeneous sheath on its length. The polar capsules measuring 3.1 × 1.2 μm contain 3–4 coils of the polar filament. All surfaces of the immature and mature spores were surrounded by a closely adherent homogenous structural sheath, mainly thicker around the tails. The taxonomic affinities of this parasite to other species are discussed.     

A newly recorded entomopoxvirus B in Anacridium aegyptium (Orthoptera: Acrididae)

An entomopoxvirus infecting fat body cells of Anacridium aegyptium is reported. Spheroid virus occlusion bodies, ranging in size from 3 to 8 μm, contain virions 336–356 nm long and 210–217 nm wide. The virion ultrastructure indicates that the newly discovered virus belongs to the genus Entomopoxvirus B known to infect Lepidoptera and Orthoptera.     

新疆意大利蝗不同地理种群16S rDNA序列差异研究

研究新疆东部和西部不同地理种群意大利蝗Calliptamus italicus(L.)线粒体基因中16SrDNA序列差异。结果表明:(1)意大利蝗A+T的含量(67.7%)明显高于G+C的含量(32.3%);(2)多重序列比对和UPGMA系统发育树的结果表明新疆东、西部不同地理种群的意大利蝗的遗传变异极小。     

STUDIES ON MORPHOLOGY AND DEVELOPMENT OF OEDMEUS ASIATICUS ENTOMOPOXVIRUS PROPAGATED IN OEDALEUS INFERNALIS*

Abstract The morphological characteristics and development of Oedaleus asiaticus entomopoxvirus propagated in Oeddeus infernalis are reported. This virus mainly infected host's fat bodies and hemocytes. Three kinds of spheroids were observed during propagation: big spheroid (30. 41 μm × 25. 40 μm), ellipsoid (6. 58 μm × 4. 78 μm) and small spheroid (3. 35 μm × 2. 60 μm). The virions embeded in them were oval, measuring 230 nm × 176 nm. The typical characteristic of poxviruses as spherical units with the mulberry-like surface could be seen under transmission electron microscope. The lateral body was cylinder-shaped. The rope-like substances in the core folded back only once; therefore two spots in transverse sections were seen. The development of the virions included four stages: the appearance of viro-plasm, the formation of spherical particles, the differentiation of core and capsid. The grasshoppers only in the same genus could be infected by this virus.     

新疆草原七种蝗虫口器结构差异研究

7种新疆草原蝗虫口器有明显差异,主要体现在上颚切齿和臼齿的形状、长度及排列方式等。根据上颚特征及差异,将意大利蝗Calliptamus italicus italicus和朱腿痂蝗Bryodema gebleri gebleri的上颚归属于齿型,西伯利亚蝗Gomphocerus sibiricus和黑条小车蝗Oedaleus decorus划分为磨型,而蓝斑翅蝗Oedipoda coerulescens、红斑翅蝗Obdipoda miniata miniata和黑腿星翅蝗Calliptamus barbarus cephalotes则属于中间型。蝗虫上颚齿型不同与其不同寄主植物选择有密切关系,齿型蝗虫嗜食纤维素含量少的菊科、十字花科和紫草科植物;磨型蝗虫则嗜食纤维素含量较多的禾本科、莎草科植物。     

Temperature-sensitive mutants with lesions in the vaccinia virus F10 kinase undergo arrest at the earliest stage of virion morphogenesis.

Vaccinia virus encodes two protein kinases; the B1 kinase is expressed early and appears to play a role during DNA replication, whereas the F10 kinase is expressed late and is encapsidated in virions. Here we report that the F10 kinase gene is the locus affected in a complementation group of temperature-sensitive mutants composed of ts15, ts28, ts54, and ts61. Although these mutants have a biochemically normal phenotype at the nonpermissive temperature, directing the full program of viral gene expression, they fail to form mature virions. Electron microscopic analysis indicates that morphogenesis undergoes arrest at a very early stage, prior to the formation of membrane crescents or immature virions. An essential role for the F10 protein kinase in orchestrating the onset of virion assembly is implied.     

Golgi vesicles of uncommon morphology and wall formation in the red alga,Polysiphonia

Summary Golgi bodies of immature carposporangia ofPolysiphonia sp. are composed of a polarized stack of six to ten curved cisternae. The cisternae are surrounded by 50–200 nm diameter slightly granular vesicles.Hypertrophied, fibrillar Golgi cisternae occur in mature carposporangia. Secretory vesicles originate from ends of cisternae and by complete vesiculation of terminal cisternae; 0.6–1.2 m diameter, fibrous vesicles, many with electron dense nucleoids are abundant throughout the cytoplasm of mature sporangia. Vesicles expand, fuse with each other and cluster around starch granules. Some vesicles secrete their content into the spore wall. Morphological analyses of starch granules as well as topographical relations between vesicles, starch granules and the adjacent cytoplasm suggest that these Golgi vesicles function like lysosomes. The significance of these observations is discussed in relation to the composition of plant cell walls and cellular expansion.     

草鱼呼肠孤病毒在CIK细胞中复制及形态发生的研究

以草鱼呼肠孤病毒（ＧＣＲＶ）感染的草鱼肾细胞系（ＣＩＫ）为模型,进行了草鱼呼肠孤病毒在细胞内的形态发生的研究。当病毒以感染复数为５￣１０ＰＦＵ／ＣＥＬＬ感染ＣＩＫ细胞时,在病毒感染细胞４ｈ以内的切片中,可观察到脱去部分外层衣壳的不完整病毒颗粒。感染细胞８ｈ,可观察到浆胞内病毒发生基质,其内含有大量的直径约５０ｎｍ的亚病毒颗粒,无外层蛋白结构。感染１２￣１６ｈ后,这些亚病毒颗粒装配上外层蛋白结构,形     

The role of capsid maturation on adenovirus priming for sequential uncoating

Adenovirus assembly concludes with proteolytic processing of several capsid and core proteins. Immature virions containing precursor proteins lack infectivity because they cannot properly uncoat, becoming trapped in early endosomes. Structural studies have shown that precursors increase the network of interactions maintaining virion integrity. Using different biophysical techniques to analyze capsid disruption in vitro, we show that immature virions are more stable than the mature ones under a variety of stress conditions and that maturation primes adenovirus for highly cooperative DNA release. Cryoelectron tomography reveals that under mildly acidic conditions mimicking the early endosome, mature virions release pentons and peripheral core contents. At higher stress levels, both mature and immature capsids crack open. The virus core is completely released from cracked capsids in mature virions, but it remains connected to shell fragments in the immature particle. The extra stability of immature adenovirus does not equate with greater rigidity, because in nanoindentation assays immature virions exhibit greater elasticity than the mature particles. Our results have implications for the role of proteolytic maturation in adenovirus assembly and uncoating. Precursor proteins favor assembly by establishing stable interactions with the appropriate curvature and preventing premature ejection of contents by tightly sealing the capsid vertices. Upon maturation, core organization is looser, particularly at the periphery, and interactions preserving capsid curvature are weakened. The capsid becomes brittle, and pentons are more easily released. Based on these results, we hypothesize that changes in core compaction during maturation may increase capsid internal pressure to trigger proper uncoating of adenovirus.     

Sphaerospora epinepheli N. Sp. (Myxosporea: Sphaerosporidae) Observed in Grouper (Epinephelus malabaricus)

Sphaerospora epinepheli n. sp. is described from grouper, Epinephelus malabaricus , in cage-cultured and wild fish collected from both coastal lines of southern Thailand. Subspherical to spherical spores and mono- or disporous pseudoplasmodia were observed in the lumen of kidney tubules. Pseudoplasmodia were round to elongate, size range 15.6–22.9 μm (length) × 8.4–21.6 μm (width). Spores were 7.8–10.0 μm (length) × 12.3–14.5 μm (thickness), and 7.0–9.5 μm (width) with two spherical polar capsules of equal size measuring 2.9–4.4 μm in diameter and containing polar filaments with six or seven windings. Two uninucleate sporoplasms showed iodine vacuoles. Blood stages, similar to C-blood protozoans observed from freshwater fish in Europe, were found from peripheral blood smears of grouper. Ultrastructural studies of blood stages showed a similar structure to unidentified mobile protozoans from the blood of carp. Electron dense bodies were observed in the cytoplasm of the primary cell blood stages. Infected proximal-tubular epithelial cells showed highly vacuolated cytoplasm and pycnotic nuclei.     

Process of infection with bacteriophage phi X174 XLI. Synthesis of defective phi X particles at 15 degrees C.

At 15 degrees C, phi X174-infected cells make single-stranded viral DNA fragments, varying in size from 0.2 to 0.9 times that of phi X DNA. In non-deproteinized lysates, this single-stranded DNA is found associated with proteins in particles sedimenting heterogeneously with an S20, w average of 80 to 90S. These particles do not differ appreciably from mature virus in polypeptide composition. Chase experiments, at 37 degrees C, of the label incorporated into this DNA at 15 degrees C suggest that both the single-stranded DNA fragments and the 80 to 90S particles are not precursors of virions but are defective end products.     

红胫戟纹蝗痘病毒形态及理化性质研究

红胫戟纹蝗Dociostaurus kraussi是新疆草原优势种蝗虫。1989年首次从新疆玛纳斯红胫戟纹蝗上分离到痘病毒Dociostaurus kraussi ntomopoxvirus(DkEPV),1992年又在新疆巴里坤发现, 自然流行率达23.3%。显微镜观察表明该病毒主要感染脂肪体。病毒球状体为圆球状,直径为2—7μm,大小差异悬殊,病毒粒子砖形或椭圆形,表面呈桑椹结构, 大小平均为144nlnx269nn。病毒DNA具有典型的核酸紫外吸收光谱。根据热变性曲线测得DkEPV—DNA的T_m,值为79.0,(G+C)%为23.7%。病毒DNA经限制性内切酶EcoRI、Bgl IIH和Hind III酶切后,分别得到29、21和18个片段。以λDNA Hind III酶切片段为标准分子量,计算出各酶切片段的分子量为155.45x10⁶、155.69x10⁶和155.40x10⁶D, 由此得出DkEPV—DNA总分子量为55.5x10⁶D。     

Filamentous structures associated with Epstein-Barr virus-infected cells.

After the onset of Epstein-Barr virus DNA and protein synthesis 10 h after superinfection of Raji cells (a cell line containing Epstein-Barr virus DNA but not producing virus), filamentous structures 25 nm in diameter and 0.2 to 1.4 micrometers in length could be detected in the cell cytoplasm by electron microscopy. These structures banded in metrizamide gradients with viral DNA and proteins, but at a density different from that of virions or nucleocapsids. These filaments, enriched in a 155,000-dalton protein similar in size to a major nucleocapsid protein of Epstein-Barr virus, may represent intermediates in viral nucleocapsid assembly.