武汉几种外附生纤毛虫新种的记述

缘毛类纤毛虫的很多种类借助于柄和鞘广泛地附生在水生动植物和其它基质上,是营自由生活的周丛生物。Nenninger(1948)曾将固着生活的缘毛类纤毛虫分为3个类群:第一类群包括既能在水生植物体上着生、也可在动物体上着生的种类;第二类群是在目(Order)水平上的各类水生动物体上着生的种类;第三类群是从科到种的水平上的各类水生动物体上的种类。我们沿用epizoon一词,是指第二、三类的营自由生活的周丛种类,它们对着生的基质有一定的选择性,反映了这些虫体的生态特性,在分类上是有意义的。王家楫、倪达书(1933)、倪达书、贺云鸞(1943)对我国缘毛类外附生种类有过报道。近来国外有关这方面的报道颇多。       

Ultrastructure of Stylodinium Iittorale (Dinophyceae) with special reference to the stalk and apical stalk complex

The ultrastructure of Stylodinium littorale Horiguchi et Chihara, a marine, sand-dwelling coccoid dinoflagel-late, was investigated with special emphasis on its stalk and the apical stalk complex. The dinoflagellate alternates between non-motile and motile cells in its life cycle. The non-motile cell possesses a long and distinct stalk. The stalk, consisting of a main cylindrical part and a holdfast, is firmly attached to a thecal plate (the apical pore plate). A part of its proximal portion is hollow and V-shaped in section. The V-shaped hollow space is underlain by a projection from the apical pore plate. An apical stalk complex is present in the motile cells and consists of a large apical pore plate and mucilaginous material. The apical pore plate is depressed into the cell, but has a narrow central tubular projection. The mucilaginous stalk-building material is stored between this plate and the outer plate membrane. The tubular projection of the apical pore plate corresponds to the apical pore of other dinoflagellates and its lumen is filled with electron-dense material. The structure of the apical stalk complex is compared with the homologous structure in Bysmatrum arenicola, the only other example of an apical stalk complex that has been investigated. A general ultrastructural survey revealed that S. littorale possesses a typical dinoflagellate cellular structure.     

Bactofilins,a ubiquitous class of cytoskeletal proteins mediating polar localization of a cell wall synthase in Caulobacter crescentus

The cytoskeleton has a key function in the temporal and spatial organization of both prokaryotic and eukaryotic cells. Here, we report the identification of a new class of polymer-forming proteins, termed bactofilins, that are widely conserved among bacteria. In Caulobacter crescentus, two bactofilin paralogues cooperate to form a sheet-like structure lining the cytoplasmic membrane in proximity of the stalked cell pole. These assemblies mediate polar localization of a peptidoglycan synthase involved in stalk morphogenesis, thus complementing the function of the actin-like cytoskeleton and the cell division machinery in the regulation of cell wall biogenesis. In other bacteria, bactofilins can establish rod-shaped filaments or associate with the cell division apparatus, indicating considerable structural and functional flexibility. Bactofilins polymerize spontaneously in the absence of additional cofactors in vitro, forming stable ribbon- or rod-like filament bundles. Our results suggest that these structures have evolved as an alternative to intermediate filaments, serving as versatile molecular scaffolds in a variety of cellular pathways.     

秸秆降解菌的筛选及模拟田间应用效果分析

以透明圈筛选法和滤纸崩解法相结合,建立初筛方法;模拟田间覆土菌种筛选方法,以秸秆腐解率为指标,建立了复筛方法,并进行了模拟田间应用效果分析。Y13、774、HX及J1 4株菌的腐解率较高,比对照分别提高27.04%、29.65%、35.38%和27.86%;其产CO2能力皆高于空白对照,分别提高41.67%、51.85%、30.56%和57.41%;其可溶性糖含量均提高,比对照平均提高15.40%。     

神经氨酸酶茎部氨基酸缺失对H5N1亚型禽流感病毒生物学特性的影响

近年来H5N1亚型禽流感病毒(AIV)神经氨酸酶(NA)茎部15~20个氨基酸的自发缺失时有报道,突变对于AIV生物学特性的影响还没有得到系统研究。应用反向遗传操作技术,拯救获得5株具有不同NA茎部长度的H5N1/PR8重组AIV。重组病毒的内部基因和血凝素(HA)基因来源相同,NA基因来源不同,并在NA茎部进行20个氨基酸的删除或添加突变。通过研究其生物学特性发现,5株重组病毒在SPF鸡胚中繁殖良好,其EID50、MDT和平均病毒滴度相似;NA茎部长短影响病毒的解凝能力,长茎病毒红细胞解脱能力比短茎病毒强;NA茎部15或20个氨基酸删除突变提高了重组病毒在MDCK细胞上的繁殖能力,短茎病毒释放出的病毒粒子数量是长茎病毒的10~100倍,释放时间提前6~10h,短茎病毒在MDCK细胞上形成的空斑也明显比长茎病毒的空斑大。实验结果揭示了AIV NA茎部氨基酸缺失突变的生物学意义,NA茎部15或20个氨基酸删除突变增强了AIV的细胞适应性,可能与现阶段H5N1亚型AIV宿主范围进一步扩大有关。利用反向遗传技术成功拯救了5株H5N1/PR8重组流感病毒,为流感病毒基因功能研究和重组疫苗研究建立了技术平台。通过对AIV NA茎部氨基酸的删除突变提高了病毒在MDCK细胞上的繁殖产量,为流感病毒细胞苗的生产提供了新的思路。     

禽副粘病毒-2 膜融合相关多肽基因的构建与表达

囊膜病毒与宿主细胞的膜融合是病毒入侵宿主细胞的第一步 . 禽副粘病毒 -2 (APMV-2) 囊膜表面糖蛋白有 2 种,与宿主受体结合的血凝素神经氨酸酶 (HN) 及介导膜融合的融合糖蛋白 (F). HN 蛋白的茎部区域 (stalk region) 与球状头部区域 (globular head region) ,以及 F 蛋白的 3 段七肽重复区域 (heptad repeat , HR) 都可能与膜融合直接相关,将 5 段多肽进行基因的构建与表达研究 . 根据已发表的禽副粘病毒 -1 (APMV-1) 氨基酸序列,应用 BLAST 程序进行同源性分析,以确定 APMV-2 相应区域,使用搭桥 PCR 或普通 PCR 方法构建基因,分别克隆入表达载体 pGEX-6p- Ⅰ获得重组质粒,阳性重组质粒转化入大肠杆菌 BL21 (DE₃) ,表达后获得可溶性融合蛋白, 3C 蛋白酶酶切后的蛋白质混合物经 Glutathione-Sepharose 4B 亲和层析纯化,最终获得可溶性、高纯度的 5 段多肽 . 应用 LearnCoil-VMF 软件与 ExPASy 系列软件对蛋白质结构与功能进行预测与分析 . 分子筛实验结果表明, F 蛋白的 HR1 与 HR2 可形成六聚体结构,圆二色谱实验结果则表明,六聚体蛋白富含 α 琢螺旋结构 .     

Effect of genotype and environment on branching in weedy green millet (Setaria viridis) and domesticated foxtail millet (Setaria italica) (Poaceae)

Many domesticated crops are derived from species whose life history includes weedy characteristics, such as the ability to vary branching patterns in response to environmental conditions. However, domesticated crop plants are characterized by less variable plant architecture, as well as by a general reduction in vegetative branching compared to their progenitor species. Here we examine weedy green millet and its domesticate foxtail millet that differ in the number of tillers (basal branches) and axillary branches along each tiller. Branch number in F(2:3) progeny of a cross between the two species varies with genotype, planting density, and other environmental variables, with significant genotype-environment interactions (GEI). This is shown by a complex pattern of reaction norms and by variation in the pattern of significant quantitative trait loci (QTL) amongst trials. Individual and joint analyses of high and low density trials indicate that most QTL have significant GEI. Dominance and epistasis also explain some variation in branching. Likely candidate genes underlying the QTL (based on map position and phenotypic effect) include teosinte branched1 and barren stalk1. Phytochrome B, which has been found to affect response to shading in other plants, explains little or no variation. Much variation in branching is explained by QTL that do not have obvious candidate genes from maize or rice.     

The ntp operon encoding the Na+ V-ATPase of the thermophile Caloramator fervidus

The V-type ATPase of the thermophile Caloramator fervidus is an ATP-driven Na⁺ pump. The nucleotide sequence of the ntpFIKECGABD operon containing the structural genes coding for the nine subunits of the enzyme complex was determined. The identity of the proteins in two pairs of subunits (D, E and F, G) that have very similar mobilities on SDS-PAGE of the purified complex (24.3 and 22.7 kDa, and 12.3 and 11.6 kDa) was established by tryptic digestion of the protein bands followed by mass spectrometric analysis of the peptides.     

Conformational changes in the Escherichia coli ATP synthase b-dimer upon binding to F1-ATPase

Conformational changes within the subunit b-dimer of the E. coli ATP synthase occur upon binding to the F₁ sector. ESR spectra of spin-labeled b at room temperature indicated a pivotal point in the b-structure at residue 62. Spectra of frozen b ± F₁ and calculated interspin distances suggested that where contact between b ₂ and F₁ occurs (above about residue 80), the structure of the dimer changes minimally. Between b-residues 33 and 64 inter-subunit distances in the F₁-bound b-dimer were found to be too large to accommodate tightly coiled coil packing and therefore suggest a dissociation and disengagement of the dimer upon F₁-binding. Mechanistic implications of this “bubble” formation in the tether domain of ATP synthase b ₂ are discussed. This work was supported by grant from the National Science Foundation (MCB 0415713) to PDV     

Structural Relationships Among the Ribosomal Stalk Proteins from the Three Domains of Life

The GTPase center of the large ribosomal subunit, being a landing platform for translation factors, and regarded as one of the oldest structures in the ribosome, is a universally conserved structure in all domains of life. It is thought that this structure could be responsible for the major breakthrough on the way to the RNA/protein world, because its appearance would have dramatically increased the rate and accuracy of protein synthesis. The major part of this center is recognized as a distinct structural entity, called the stalk. The main functional part of the stalk in all domains of life is composed of small L12/P proteins, which are believed to form an evolutionarily conserved group. However, some data indicate that the bacterial and archaeo/eukaryal proteins are not related to each other structurally, and only a functional relationship may be clearly recognized. To clarify this point, we performed a comprehensive comparative analysis of the L12/P proteins from the three domains of life. The results show that bacterial and archaeo/eukaryal L12/P-proteins are not structurally related and, therefore, might not be linked evolutionarily either. Consequently, these proteins should be regarded as analogous rather than homologous systems and probably appeared on the ribosomal particle in two independent events in the course of evolution.