芽胞气溶胶表面滞留抗力预测模型研究

针对生物威胁的现场处置工作,建立气溶胶芽胞表面滞留抗力的智能预测模型,以准确预测环境表面芽胞污染状况,为大规模的现场洗消任务提供重要依据,有利于实现及时反应、恰当反应和准确防护的目标。以枯草杆菌芽胞为试验菌,在气溶胶实验室进行芽胞的环境因素暴露及活力测定,以模拟环境中芽胞抗力变化规律数据为依据,采用Matlab6.1软件包中的神经网络工具箱进行抗力预测模型研究。根据研究目的、模拟环境条件和数据训练的平滑曲线等特征,设定了5个输入神经元,8个隐层节点和1个输出神经元。‘tansig’、‘purelin’为传递函数,trainlm为训练函数,网络迭代100次。模型回顾预测效率达到100%,前瞻预测效率达到91%。以实验室数据为依据,利用Matlab平台中的BP神经网络建立的芽胞气溶胶表面滞留抗力预测模型能利用环境因素信息有效预测芽胞抗力。     

芽胞表面展示技术研究进展

芽胞表面展示技术作为微生物表面展示技术的一种,因所表达的异源蛋白无需经过跨膜过程及芽胞的抗逆性等独特优势而备受研究者的关注。以下介绍了芽胞的生理结构和形成过程、芽胞表面展示系统构建原则及目前所构建的芽胞表面展示系统种类。芽胞表面展示技术不仅在疫苗生产中应用广泛,在生物催化和细胞工厂研究领域也具有广阔的应用前景。     

需氧芽胞杆菌芽胞核心组分的研究现状

芽胞核心作为芽胞的原生质体,实际上是处于休眠状态的细胞,其化学组化比较复杂,核酸,蛋白质,水,无机离子以及有机小分子共同构成芽胞核心特有的“内环境”各种化学组分的含量及存在的形式均与芽胞工能特性尤其抗性密切相关,对它们的深入研究有助于进一步揭示芽胞抗生的有关机制,本文综述对需氧芽胞杆菌核心组分研究。     

国外对芽胞杆菌属芽胞长久存活机制的研究进展

能够在不利的外界环境下长久存活是一些产芽胞菌的重要特点之一。本文就国际上在芽胞内环境、酶的休眠、芽胞与化学因子、放射线、热的关系等方面的研究状况进行了综述。     

长柄石杉芽胞及芽胞植株形态研究

为研究石杉属植物芽胞形态特征以及芽胞在石杉属植物无性繁殖中的作用,对野外采集和移栽后新生的长柄石杉（Huperzia javanica）芽胞体、芽胞的形态特征、芽胞萌发形成芽胞植株的过程进行了描述,并对芽胞和芽胞残体发生数量进行计数。结果表明,芽胞体由芽胞和芽托部组成,前者是6个小叶围住的小芽,后者为具有6个鳞状叶的柄状体;芽胞脱落后一周开始萌发并进一步发育为芽胞植株;芽胞体发生枝条上平均有5个芽胞发生;芽胞发生具有一定程度的周期性,可以据此估计石杉植株株龄和生长速度。     

芽胞杆菌属芽胞长久存活机制的研究进展

能够在不利的外界环境下长久存活是一些产芽胞菌的重要特点之一,本文就国外对在芽胞内环境,酶休的眠,芽胞与化学因子、放射线、热的关系等方面的研究状况进行了论述。     

3种细菌芽胞遗传同源性及抗力对比研究

比较细菌芽胞的遗传同源性、结构和抗力差异,为炭疽芽胞的替代试验菌的可靠评价提供依据。采用资料检索、生物信息分析、显微镜观察和微生物学技术分析不同芽胞的遗传同源性、超微结构和抗力差异。炭疽芽胞与腊样芽胞的结构和大小相似,生物遗传同源性最近,对热力、UVC和有效氯的抗力相近。腊样芽胞对炭疽芽胞的代表性最好,可以倾向性选用其替代炭疽芽胞进行试验研究。     

利用ERIC-PCR对苏云金芽胞杆菌与蜡状芽胞杆菌进行鉴定的研究

为了探索ERIC-PCR技术在苏云金芽胞杆菌和蜡状芽胞杆菌的鉴定及分型中的应用价值,本研究采用PCR方法初步检测苏云金芽胞杆菌杀虫晶体蛋白基因的组成,并对苏云金芽胞杆菌和蜡状芽胞杆菌的总DNA进行ERIC-PCR扩增,分析ERIC-PCR指纹图谱的特点并采用NTSYS2.10软件对其进行聚类。结果显示,各菌株的ERIC指纹图谱表现出不同程度的多态性,但图谱与菌株所含cry基因的类型存在一定的相关性。聚类分析结果显示,含有相同或相近cry基因类型的Bt菌株在进化树上趋向聚为一类,而不含cry基因的蜡状芽胞杆菌趋向于与不含cry基因的Bt菌株聚为一类或单独聚类。若在多种模式菌株的参考下,该方法可用于苏云金芽胞杆菌的初步鉴定和分型。     

基于芽胞的混凝土微生物原位修复技术研究进展

混凝土是最广泛使用的现代建筑材料,在应力作用下易于开裂,使混凝土结构具有渗透性,影响其耐用性和完整性,进而缩短使用寿命.混凝土微生物原位修复技术是一种廉价、有效、绿色的方式,因其具有良好的生物相容性、延长混凝土服役寿命、减少经济损失与环境污染等特点,已成为研究热点.其中,芽胞杆菌因良好的生物矿化能力且其芽胞具有极强的环...     

苏云金芽胞杆菌G03 spoIVF操纵子敲除对芽胞和晶体形成的影响

spoIVF是一个普遍存在于芽胞杆菌中的操纵子。在枯草芽胞杆菌中,它编码的两个蛋白是芽胞形成所必需的。采用基因重组技术敲除了苏云金芽胞杆菌G03菌株中的spoIVF操纵子,构建了spoIVF缺失株G03(spoIVF-)。研究表明:该突变株丧失了形成芽胞和晶体的能力。lacZ基因与cry1Aa基因的启动子融合表达分析发现:突变株中的cry1Aa基因的活性严重降低。利用载体pSTK携带spoIVF操纵子在突变株中的表达,使突变株部分恢复了产胞和形成杀虫晶体蛋白的能力。这说明spoIVF操纵子是所必需的,同时该操纵子还影响σE因子控制的cry1Aa基因表达。     

藓类植物孢子生活力的快速检测方法初探

通过对温度和光照条件的实验探索,筛选出金发藓（Polytrichum commune）和细叶小羽藓（Haplocladium microphyllum）2种藓类孢子萌发的最适条件。采用碘-碘化钾染色法、TTC染色法、红墨水染色法、亚甲基蓝染色法对6种藓类植物孢子的生活力进行测定,并将测得的生活力结果与孢子萌发率进行了比较分析。结果表明,亚甲基蓝染色法测定的藓类植物孢子平均生活力百分率与孢子平均萌发率最为接近,且染色效果明显,可用于苔藓植物孢子生活力的快速测定。亚甲基蓝染色法测得的孢子生活力（x）与其离体萌发率（y）的相关性达到极显著水平（r=0.976）,其回归方程为y=-8.547+1.069x(P<0.01),可通过孢子生活力方便预测萌发率。     

Analysis of the action of compounds that inhibit the germination of spores of Bacillus species

AIMS: To determine the mechanism of action of inhibitors of the germination of spores of Bacillus species, and where these inhibitors act in the germination process. METHODS AND RESULTS: Spores of various Bacillus species are significant agents of food spoilage and food-borne disease, and inhibition of spore germination is a potential means of reducing such problems. Germination of the following spores was studied: (i) wild-type B. subtilis spores; (ii) B. subtilis spores with a nutrient receptor variant allowing recognition of a novel germinant; (iii) B. subtilis spores with elevated levels of either the variant nutrient receptor or its wild-type allele; (iv) B. subtilis spores lacking all nutrient receptors and (v) wild-type B. megaterium spores. Spores were germinated with a variety of nutrient germinants, Ca2+-dipicolinic acid (DPA) and dodecylamine for B. subtilis spores, and KBr for B. megaterium spores. Compounds tested as inhibitors of germination included alkyl alcohols, a phenol derivative, a fatty acid, ion channel blockers, enzyme inhibitors and several other compounds. Assays used to assess rates of spore germination monitored: (i) the fall in optical density at 600 nm of spore suspensions; (ii) the release of the dormant spore's large depot of DPA; (iii) hydrolysis of the dormant spore's peptidoglycan cortex and (iv) generation of CFU from spores that lacked all nutrient receptors. The results with B. subtilis spores allowed the assignment of inhibitory compounds into two general groups: (i) those that inhibited the action of, or response to, one nutrient receptor and (ii) those that blocked the action of, or response to, several or all of the nutrient receptors. Some of the compounds in groups 1 and 2 also blocked action of at least one cortex lytic enzyme, however, this does not appear to be the primary site of their action in inhibiting spore germination. The inhibitors had rather different effects on germination of B. subtilis spores with nutrients or non-nutrients, consistent with previous work indicating that germination of B. subtilis spores by non-nutrients does not involve the spore's nutrient receptors. In particular, none of the compounds tested inhibited spore germination with dodecylamine, and only three compounds inhibited Ca2+-DPA germination. In contrast, all compounds had very similar effects on the germination of B. megaterium spores with either glucose or KBr. The effects of the inhibitors tested on spores of both Bacillus species were largely reversible. CONCLUSIONS: This work indicates that inhibitors of B. subtilis spore germination fall into two classes: (i) compounds (most alkyl alcohols, N-ethylmaleimide, nifedipine, phenols, potassium sorbate) that inhibit the action of, or response to, primarily one nutrient receptor and (ii) compounds [amiloride, HgCl2, octanoic acid, octanol, phenylmethylsulphonylfluoride (PMSF), quinine, tetracaine, tosyl-l-arginine methyl ester, trifluoperazine] that inhibit the action of, or response to, several nutrient receptors. Action of these inhibitors, is reversible. The similar effects of inhibitors on B. megaterium spore germination by glucose or KBr indicate that inorganic salts likely trigger germination by activating one or more nutrient receptors. The lack of effect of all inhibitors on dodecylamine germination suggests that this compound stimulates germination by creating channels in the spore's inner membrane allowing DPA release. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides new insight into the steps in spore germination that are inhibited by various chemicals, and the mechanism of action of these inhibitors. The work also provides new insights into the process of spore germination itself.     

Effect of mechanical abrasion on the viability, disruption and germination of spores of Bacillus subtilis

AIMS: To elucidate the factors influencing the sensitivity of Bacillus subtilis spores in killing and disrupting by mechanical abrasion, and the mechanism of stimulation of spore germination by abrasion. METHODS AND RESULTS: Spores of B. subtilis strains were abraded by shaking with glass beads in liquid or the dry state, and spore killing, disruption and germination were determined. Dormant spores were more resistant to killing and disruption by abrasion than were growing cells or germinated spores. However, dormant spores of the wild-type strain with or without most coat proteins removed, spores of strains with mutations causing spore coat defects, spores lacking their large depot of dipicolinic acid (DPA) and spores with defects in the germination process exhibited essentially identical rates of killing and disruption by abrasion. When spores lacking all nutrient germinant receptors were enumerated by plating directly on nutrient medium, abrasion increased the plating efficiency of these spores before killing them. Spores lacking all nutrient receptors and either of the two redundant cortex-lytic enzymes behaved similarly in this regard, but the plating efficiency of spores lacking both cortex-lytic enzymes was not stimulated by abrasion. CONCLUSIONS: Dormant spores are more resistant to killing and disruption by abrasion than are growing cells or germinated spores, and neither the complete coats nor DPA are important in spore resistance to such treatments. Germination is not essential for spore killing by abrasion, although abrasion can trigger spore germination by activation of either of the spore's cortex-lytic enzymes. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides new insight into the mechanisms of the killing, disruption and germination of spores by abrasion and makes the surprising finding that at least much of the spore coat is not important in spore resistance to abrasion.     

Fine structure of resting spore formation and germination in Entomophthora virulenta

The fine structure during the formation and germination of resting spores of Entomophthora virulenta is described. There were many microbodies in contact with oil droplets, and the microbodies appeared to participate in spore germination. The mature resting spore had an epispore layer with two regions and an endospore layer with five regions. Dictyosomes, numerous vesicles, and lomasomes were produced during the formation of the endospore layer. Prior to spore germination, the single large oil droplet separated into numerous small oil droplets, and the new cell wall was formed beneath the endospore layer which gradually disintegrated possibly by enzymatic actions. The germ tube perforated the epispore layer mainly by mechanical pressure.     

Studies on the mechanism of the osmoresistance of spores of Bacillus subtilis

AIMS: To determine the reason that spores of Bacillus species, in particular Bacillus subtilis, are able to form colonies with high efficiency on media with very high salt concentrations. METHODS AND RESULTS: Spores of various Bacillus species have a significantly higher plating efficiency on media with high salt concentration (termed osmoresistance) than do log or stationary phase cells. This spore osmoresistance is higher on richer media. Bacillus subtilis spores lacking various small, acid-soluble spore proteins (SASP) were generally significantly less osmoresistant than were wild-type spores, as shown previously (Ruzal et al. 1994). Other results included: (a) spore osmoresistance varied significantly between species; (b) the osmoresistance of spores lacking SASP was not restored well by amino acid osmolytes added to plating media, but was completely restored by glucose; (c) the osmoresistance of spores lacking SASP was restored upon brief germination in the absence of salt in a process that did not require protein synthesis; (d) significant amounts of amino acids generated by SASP degradation were retained within spores upon germination in a medium with high but not low salt; (e) slowing but not abolishing SASP degradation by loss of the SASP-specific germination protease (GPR) did not affect spore osmoresistance; (f) sporulation at higher temperatures produced less osmoresistant spores; and (g) spore osmoresistance was not decreased markedly by the absence of the stress sigma factor for RNA polymerase, sigmaB. CONCLUSIONS: Spore osmoresistance appears as a result of three major factors: (1) specific characteristics of spores and cells of individual species; (2) the precise sporulation conditions that produce the spores; and (3) sufficient energy generation by the germinating and outgrowing spore to allow the spore to adapt to conditions of high osmotic strength; the substrates for this energy generation can come from either the endogenous generation of amino acids by SASP degradation or from the spore's environment, in the form of a readily taken up and metabolized energy source such as glucose. SIGNFICANCE AND IMPACT OF STUDY: These results provide information on the mechanisms of spore osmoresistance, a spore property that can be of major applied significance given the use of high osmotic strength with or without high salt as a means of food preservation.     

Spore coat is altered in modB glycosylation mutants of Dictyostelium discoideum.

The modB mutation eliminates specific carbohydrate epitopes from glycoproteins which are expressed primarily in prespore and spore cells of differentiating Dictyostelium discoideum. Spores formed by the mutant show several phenotypes. Whereas mutant spores germinate efficiently after heat activation, they germinate poorly after urea activation. Following germination, at least one glycosylation-defective glycoprotein is cleaved, and the larger fragment is released in soluble form from the spore coat. However, an earlier difference in the spore coat can be traced back to the nongerminated spore coat, as detected by the elutability of protein from intact spores by chemical extraction. An altered character of the pregermination spore coat is also suggested by increased labeling by a fluorescent lectin which binds to its interior. The findings are consistent with a change in the character of certain molecular contacts leading to altered characteristics of the mutant spore coat, which are specific because they are distinctive from changes observed in another glycosylation mutant which affects a different epitope.     

Longevity and germination of Edhazardia aedis (Microspora: Amblyosporidae) spores

Edhazardia aedis is a polymorphic microsporidium of mosquitoes that is both horizontally and vertically transmitted to its host. Because it is being developed for biological control of mosquitoes, detailed knowledge is needed regarding the longevity and germation of its fragile, mosquito‐infectious spore. Spores responsible for horizontal transmission were extracted from 7–10‐day‐old larvae (reared from infected Aedes aegypti eggs) and purified by Ludox density gradient centrifugation. Mature spores were variable in specific gravity, being found throughout the 20 and 60% zone in Ludox gradients. The optimal environment for spore germination was dilute KCl at pH 10.0–11.0; ammonia inhibited germination. Osmotic inhibition was almost complete in both sucrose and polyethylene glycol at concentrations equivalent to 40 atm. The spores retained their viability for a maximum of 21 days at 23±2°C, whereas when held at 5±2°C, their viability was completely lost within two days post‐harvest. Potential for germination decreased along with infectivity, providing a simple assay for spore viability.     

Xanthine oxidase induced depolymerization of hyaluronic acid in the presence of ferritin

Oxygen free radicals generated by xanthine oxidase are able to depolymerize hyaluronic acid in the presence of ferritin-bound iron. This suggests that ferritin can catalyse the Haber-Weiss reaction, leading to the formation of highly damaging hydroxyl radicals.     

甘蓝根肿病菌的生物学特性研究*

对甘蓝根肿病菌生物学特性研究表明,该菌休眠孢子萌发的最适温度24℃,最适pH值6.0～6.7,致死温度45℃,肿根腐烂处理可以显著提高萌发率,光对休眠孢子萌发有明显抑制作用。该菌休眠孢子在感病寄主的根分泌物溶液中萌发率最高,达75%。耐病甘蓝品种及番茄的根分泌物均能刺激休眠孢子萌发。通过电镜观察,根肿病菌休眠孢子为近球形,孢壁有乳状突起,直径2.1～3.1mm(平均直径2.5mm)。游动孢子为近球形或椭圆形,大小为1.6～3.6mm,同侧着生不等长尾鞭式双鞭毛。