一株新粘细菌生物学性质的研究

对粘细菌纤维堆囊菌So ce cpu-1生物学特性进行了研究.结果表明,该粘细菌能利用如滤纸或玉米秸杆等纤维素.So ce cpu-1次级代谢产物的抑茵实验表明,该粘细菌具有广谱抗菌活性:对人白血病细胞K562有较好的抑制效果.     

纤维堆囊菌的供谢产物及其生物学活性分析

纤维堆囊菌不同菌株不但表现细胞和子实体形态的差异,而且供谢产物的组成和生物学活性也存在差异。纤维堆囊菌对革兰氏阴性细菌不表现任何抑制活性,部分菌株可抑制革兰氏阳性细菌;但所有菌株对真菌和肿瘤细胞有广泛和强烈的抑制作用。薄层层析显示,纤维堆囊菌的次级代谢产物组分较多,且大多数组分具有不同程度的抑制真菌和肿细胞的活性。在筛选中发现四株菌的代谢产物能够促进微管蛋白聚合,其中So33-1活性组分的薄层层析Rf值与已知的Epothilone A相似,而So81的则有较大的差异。研究结果表明纤维堆囊菌是很好的筛选抗真核生物活性的天然化合物资源。     

纤维堆囊菌的代谢产物及其生物学活性分析

纤维堆囊菌不同菌株不但表现细胞和子实体形态的差异 ,而且代谢产物的组成和生物学活性也存在差异。纤维堆囊菌对革兰氏阴性细菌不表现任何抑制活性 ,部分菌株可抑制革兰氏阳性细菌 ;但所有菌株对真菌和肿瘤细胞有广泛和强烈的抑制作用。薄层层析显示 ,纤维堆囊菌的次级代谢产物组分较多 ,且大多数组分具有不同程度的抑制真菌和肿瘤细胞的活性。在筛选中发现四株菌的代谢产物能够促进微管蛋白聚合 ,其中So3 3 1活性组分的薄层层析Rf 值与已知的EpothiloneA相似 ,而So81的则有较大的差异。研究结果表明纤维堆囊菌是很好的筛选抗真核生物活性的天然化合物资源。     

纤维堆囊菌生长及限制因素的研究

纤维堆囊菌对可溶性淀粉、木聚糖、纤维素等复杂碳源的利用能力较强,简单碳源中只利用葡萄糖;KNO3、蛋白栋是较好的氮源,大多数氮源都能支持生长,表明该菌营养要求简单,能高效的得用处是里丰富的纤维素资源。堆囊菌的盐耐受能力较低,盐浓度高于1％的培养基中菌体几乎不能生长,Mg^2是生长必须的元素,Ca^2 对生长没有明显的作用,但是子实体的形态发生所必需的。培养基的起始pH为7.0时细胞生长较好,大于8.0或小于5.0不生长。纤维堆囊菌生长具有细胞密度依赖性,低密度的细胞不能生长。固定在滤纸上的细胞淋洗液（纤维素酶降解产物）对细胞生长具有明显促进作用,并能降低细胞生长的密度依赖性。     

不同分离方法对子实体形成和粘细菌分离的影响

【目的】基于模拟原位环境策略、可培养粘细菌的营养策略及细菌互作网络,改良分离培养基,以提高分离粘细菌的多样性。【方法】通过添加土壤浸提液、使用不同种类的诱导菌和改变诱导菌的接种方式设置分离方法,同时以传统的分离方法作对照。【结果】改良的分离方法比对照组诱导出了更多粘细菌子实体种类,采自4个地区的9份样品共分离纯化出40株粘细菌,按形态学和分子生物学,将其归类于原囊菌属(Archangium)、珊瑚菌属(Corallococcus)、软骨霉状菌属(Chondromyces)、粘球菌属(Myxococcus)、侏囊菌属(Nannocystis)、多囊菌属(Polyangium)、匣状球菌属(Pyxidicoccus)。【结论】与传统分离方法相比,添加土壤浸提液,诱导菌点接法能大大提高诱导出的粘细菌子实体种类的数目,革兰氏阳性菌和革兰氏阴性菌作为诱导菌对子实体种类影响较小,但是也发现革兰氏阳性菌特异性诱导出的子实体。虽然本研究通过对分离培养基的改良大大增加了子实体种类,但是纯化出的粘细菌种类远少于观察到的子实体种类,说明除改良分离方法外,还需进一步研究粘细菌的纯化方法,提高分离所得粘细菌的多...     

黏细菌的分子生物学

黏细菌(myxobacterium)是一类具有复杂多细胞行为,并能产生大量新型活性物质的单细胞原核生物。黏细菌基因组较大,目前黄色黏球菌(Mycococcus xanthus)DK1622及纤维堆囊菌(Sorangium cellulosum)So ce56序列已经测定,这有助于从基因组水平对黏细菌进行深入的研究。本文对黏细菌细胞运动的分子机制、新发现的活性物质等方面进行了综述。     

我国粘细菌(Myxobacteria)资源的分离与鉴定

粘细菌是原核生物中一类具有复杂多细胞行为的革蓝氏阴性细菌[1],能够 在细胞间通过信号的传递和感应,协同摄食、运动和发育形成子实体,具有 显著的社会学特征,被认为是高等的原核生物[2,3].     

基于细菌共现网络的土壤粘细菌分离

粘细菌是一类分布广泛的捕食性细菌,能够产生种类多样、结构新颖、作用机制独特的天然活性物质。但是粘细菌分离纯化困难且耗时,导致其资源匮乏,这已成为粘细菌开发利用的重要瓶颈之一。基于辅助菌的分离方法是当前获得粘细菌资源的重要方法,然而辅助菌多为大肠杆菌（Escherichia coli）等革兰氏阴性菌。为了获得新的辅助菌和粘细菌资源,本研究基于土壤细菌16S rRNA基因高通量测序数据构建细菌共现网络（bacterial co-occurrence network）,发现粘细菌-细菌子网络中有27%的连接节点为放线菌。因此,本研究选择革兰氏阳性菌球形节杆菌（Arthrobacter globiformis） GDMCC 1.1730作为辅助菌,并在捕食培养基上验证其捕食活性。结果表明该辅助菌能够被所有参与测试的粘细菌（12个物种）所捕食。以GDMCC 1.1730作为辅助菌进行粘细菌的分离,共诱导出11种粘细菌子实体,且包括一种仅由GDMCC 1.1730诱导而大肠杆菌未能诱导出的子实体。本研究基于细菌共现网络和捕食活性验证,提供了一株革兰氏阳性菌作为新的粘细菌辅助菌,该辅助菌能够诱导出土壤中的多种粘细菌子实体,为根据细菌共现网络获取未/难培养微生物资源提供了新的证据。     

粘细菌的多细胞形态发生及其分子调控

粘细菌的多细胞形态发生是粘细菌细胞社会性行为的主要表现.包括细胞有序聚集、细胞自溶、子实体发育和粘孢子的分化形成等.粘细菌的形态发生过程涉及复杂的信号系统和调控,与真核生物具有较大的相似性.是研究原核生物细胞分化发育以及生物进化的重要模式材料.     

玉蕈降解木质纤维素的生理生化基础

本文研究了玉蕈在纤维废弃物上生长期间,培养基中主要成分的降解规律及有关的酶学分析。实验结果表明:1.玉蕈分解纤维素和半纤维素的能力较强,分解木素的能力很弱。因此,玉蕈是褐腐型木腐菌。2.纤维素是玉蕈子实体生长阶段的主要碳源。3.玉蕈生长期间可向培养基中释放羧甲基纤维素酶、滤纸纤维素酶、半纤维素酶、淀粉酶和蛋白酶。酶活性在子实体生长阶段显著增加。进一步证明了子实体阶段酶活性增加与培养温度和子实体形成有密切关系。     

Modeling cell proliferation for simulating three-dimensional tissue morphogenesis based on a reversible network reconnection framework

Tissue morphogenesis in multicellular organisms is accompanied by proliferative cell behaviors: cell division (increase in cell number after each cell cycle) and cell growth (increase in cell volume during each cell cycle). These proliferative cell behaviors can be regulated by multicellular dynamics to achieve proper tissue sizes and shapes in three-dimensional (3D) space. To analyze multicellular dynamics, a reversible network reconnection (RNR) model has been suggested, in which each cell shape is expressed by a single polyhedron. In this study, to apply the RNR model to simulate tissue morphogenesis involving proliferative cell behaviors, we model cell proliferation based on a RNR model framework. In this model, cell division was expressed by dividing a polyhedron at a planar surface for which cell division behaviors were characterized by three quantities: timing, intracellular position, and normal direction of the dividing plane. In addition, cell growth was expressed by volume growth as a function of individual cell times within their respective cell cycles. Numerical simulations using the proposed model showed that tissues grew during successive cell divisions with several cell cycle times. During these processes, the cell number in tissues increased while maintaining individual cell size and shape. Furthermore, tissue morphology dramatically changed based on different regulations of cell division directions. Thus, the proposed model successfully provided a basis for expressing proliferative cell behaviors during morphogenesis based on a RNR model framework.     

Modeling cell apoptosis for simulating three-dimensional multicellular morphogenesis based on a reversible network reconnection framework

Morphogenesis in multicellular organisms is accompanied by apoptotic cell behaviors: cell shrinkage and cell disappearance. The mechanical effects of these behaviors are spatiotemporally regulated within multicellular dynamics to achieve proper tissue sizes and shapes in three-dimensional (3D) space. To analyze 3D multicellular dynamics, 3D vertex models have been suggested, in which a reversible network reconnection (RNR) model has successfully expressed 3D cell rearrangements during large deformations. To analyze the effects of apoptotic cell behaviors on 3D multicellular morphogenesis, we modeled cell apoptosis based on the RNR model framework. Cell shrinkage was modeled by the potential energy as a function of individual cell times during the apoptotic phase. Cell disappearance was modeled by merging neighboring polyhedrons at their boundary surface according to the topological rules of the RNR model. To establish that the apoptotic cell behaviors could be expressed as modeled, we simulated morphogenesis driven by cell apoptosis in two types of tissue topology: 3D monolayer cell sheet and 3D compacted cell aggregate. In both types of tissue topology, the numerical simulations successfully illustrated that cell aggregates gradually shrank because of successive cell apoptosis. During tissue shrinkage, the number of cells in aggregates decreased while maintaining individual cell size and shape. Moreover, in case of localizing apoptotic cells within a part of the 3D monolayer cell aggregate, the cell apoptosis caused the global tissue bending by pulling on surrounding cells. In case of localizing apoptotic cells on the surface of the 3D compacted cell aggregate, the cell apoptosis caused successive, directional cell rearrangements from the inside to the surface. Thus, the proposed model successfully provided a basis for expressing apoptotic cell behaviors during 3D multicellular morphogenesis based on an RNR model framework.     

Mutation in the rel gene of Sorangium cellulosum affects morphological and physiological differentiation

Interruption of the (p)ppGpp synthetase gene ( rel ) of Sorangium cellulosum So ce56 resulted in loss of ppGpp accumulation after norvaline treatment during exponential growth phase. The rel mutant failed to produce wild-type levels of the polyketides chivosazol and etnangien in production media. In wild-type cells expression of the chivosazol biosynthetic operon can be significantly increased by norvaline or α-methylglucoside. This induction does not occur in the rel mutant. The rel mutant also lost the capability to form multicellular fruiting bodies under nutrient starvation.     

Cell migration in multicellular environments

Most experiments observing cell migration use planar plastic or glass surfaces despite these conditions being considerably different from physiological ones. On such planar surfaces, cells take a dorsal-ventral polarity to move two-dimensionally. Cells in tissues, however, interact with surrounding cells and the extracellular matrix such that they transverse three-dimensionally. For this reason, three-dimensional matrices have become more and more popular for cell migration experiments. In addition, recent developments in imaging techniques have enabled high resolution observations of in vivo cell migration. The combination of three-dimensional matrices and such imaging techniques has revealed motile mechanisms in tissues not observable in studies using planar surfaces. Regarding models for such cell migration studies, the cellular slime mould Dictyostelium discoideum is ideal. Single amoeboid cells aggregate into hemispherical mound structures upon starvation to begin a multicellular morphogenesis. These tiny and simple multicellular bodies are suitable for observing the behaviors of individual cells in multicellular structures. Furthermore, the unique life cycle can be exploited to identify which genes are involved in cell migration in multicellular environments. Since mutants lacking such genes are expected to fail to undergo morphogenesis, easy and systematic gene screening is possible by isolating mutants whose developments arrest around the mound stage, which is the case for several mutants lacking specific cytoskeletal proteins. In this article, I discuss the basic elements required for cell migration in multicellular environments and how Dictyostelium can be used to elucidate them.     

The fungal type II myosin in Penicillium marneffei, MyoB, is essential for chitin deposition at nascent septation sites but not actin localization

Cytokinesis is essential for proliferative growth but also plays equally important roles during morphogenesis and development. The human pathogen Penicillium marneffei is capable of dimorphic switching in response to temperature, growing in a multicellular filamentous hyphal form at 25°C and in a unicellular yeast form at 37°C. P. marneffei also undergoes asexual development at 25°C to produce multicellular differentiated conidiophores. Thus, P. marneffei exhibits cell division with and without cytokinesis and division by budding and fission, depending on the cell type. The type II myosin gene, myoB, from P. marneffei plays important roles in the morphogenesis of these cell types. Deletion of myoB leads to chitin deposition defects at sites of cell division without perturbing actin localization. In addition to aberrant hyphal cells, distinct conidiophore cell types are lacking due to malformed septa and nuclear division defects. At 37°C, deletion of myoB prevents uninucleate yeast cell formation, instead producing long filaments resembling hyphae at 25°C. The ΔmyoB cells also often lyse due to defects in cell wall biogenesis. Thus, MyoB is essential for correct morphogenesis of all cell types regardless of division mode (budding or fission) and defines differences between the different types of growth.     

Nutrient regulation of epothilone biosynthesis in heterologous and native production strains

Fermentation media with different initial concentrations of ammonium and phosphate salts were used to study the inhibitory effects of those ions on growth and production of epothilone in Sorangium cellulosum and Myxococcus xanthus. The native epothilone producer, S. cellulosum was more sensitive to ammonium and phosphate than the heterologous producer, M. xanthus. An ammonium concentration of 12 mM reduced epothilone titers by 90% in S. cellulosum but by only 40% in M. xanthus. When 5 mM phosphate was added to the medium, production in both strains was 60% lower. Higher phosphate concentrations had little additional effect on M. xanthus titers, but epothilone production with 17 mM extra-cellular phosphate in S. cellulosum was 95% lower than in the control condition. The effect of iron supplementation to the fermentation medium was also investigated. Both strains showed best production with 20 microM iron added to the medium.     

Self-similar colony morphogenesis by gram-negative rods as the experimental model of fractal growth by a cell population.

The ability to form a fractal colony was shown to be common among several species of the family Enterobacteriaceae. Bacterial spreading growth in a two-dimensional field of nutrient concentration was indicated to be important for this experimental self-similar morphogenesis. As a basic analogy, the diffusion-limited aggregation model was suggested. Fractal dimensions of colonies were mostly in the range of values from 1.7 to 1.8, similar to those of the two-dimensional diffusion-limited aggregation model. Bacterial characteristics and culture conditions inducing changes in fractal patterns and growth rates were identified. The contribution of the bacterial multicellular nature to fractal morphogenesis is discussed.     

Self-similar colony morphogenesis by gram-negative rods as the experimental model of fractal growth by a cell population.

The ability to form a fractal colony was shown to be common among several species of the family Enterobacteriaceae. Bacterial spreading growth in a two-dimensional field of nutrient concentration was indicated to be important for this experimental self-similar morphogenesis. As a basic analogy, the diffusion-limited aggregation model was suggested. Fractal dimensions of colonies were mostly in the range of values from 1.7 to 1.8, similar to those of the two-dimensional diffusion-limited aggregation model. Bacterial characteristics and culture conditions inducing changes in fractal patterns and growth rates were identified. The contribution of the bacterial multicellular nature to fractal morphogenesis is discussed.