一株抗水稻纹枯病菌的解淀粉芽胞杆菌分离与鉴定

【目的】筛选对水稻纹枯病菌(Rhizoctonia solani)具有强拮抗作用的细菌菌株。【方法】用指示菌法筛选拮抗菌株;通过形态观察、生理生化实验、Biolog及16S rDNA序列分析鉴定目标菌株;利用平板双向培养法和滤纸片扩散法测定抑菌谱及拮抗性质。【结果】分离到一株高活力的水稻纹枯病菌拮抗菌株YB-3,该菌株属于解淀粉芽胞杆菌(Bacillus amyloliquefaciens);菌株YB-3对常见的14株病原真菌和7株细菌具有较强的拮抗作用,并发现其对亲缘关系较近的芽孢菌属有较强的拮抗作用;该菌株的抑制活性具有温度稳定、耐酸、但对蛋白酶敏感的特点。【结论】通过指示菌法筛选到一株对水稻纹枯病菌有强拮抗作用的解淀粉芽胞杆菌(B.amyloliquefaciens)YB-3,它具有广谱、高效的植物病原菌拮抗活性。     

天然海鱼共附生放线菌抑菌和抗病毒活性筛选研究

从台湾海峡天然海鱼共附生微生物群落分离得到29株放线菌,通过抗病毒和抑菌活性筛选,分别得到6株对烟草花叶病毒和植物病原真菌具有较强抑制作用的天然菌株.其中,菌株030603对供试植物病原真菌拮抗作用尤为显著,其发酵产物具有广谱抑菌活性,可以作为广谱性植物病原真菌拮抗菌株进行生防菌产业化开发及发酵产物活性成分的化学研究.该菌株形态特征、培养特性、生理生化特性与微白黄链霉菌基本一致,16 S rDNA序列同源性达99.5 %.实验结果表明,利用天然海洋鱼类共附生微生物群落获取植物病原菌拮抗菌株具有取样便捷和活性菌株检出率高的特点.     

大蒜内生细菌的分离及拮抗菌筛选与鉴定

利用常规分离方法对大蒜鳞茎进行内生细菌的分离,采用对峙法和平板涂布法对分离的内生菌进行拮抗试验研究,并对菌株DSP6进行16S rDNA全序列鉴定。结果表明:分离得到19株内生细菌,其中10株菌对2种以上植物病原真菌有不同程度的抑制作用,占分离菌总数的52.6%,DSN7对番茄早疫病的抑菌圈半径最大,为13mm;17株菌对5种病原细菌中至少1种有抑制作用,占分离菌总数的89.5%,其中菌株DSP3对大肠杆菌的抑菌圈半径最大,达到10 mm;菌株DSP6对供试的9种病原菌有较强的抑菌作用,且抑菌圈平均半径最大,为6.88mm;16S rDNA全序列鉴定显示,菌株DSP6与芽孢杆菌属Bacillus axarquiensis相似性为100%,表明菌株DSP6为Bacillus axarquiensis。     

不同品种香蕉内生菌分离及广谱拮抗菌的筛选

旨在探究抗病品种与易感品种香蕉的健康株和病株内生菌与其中广谱拮抗菌的主要分布规律,并对广谱拮抗菌进行拮抗活性的测定。以样品根、球茎、假茎、叶为材料分离培养内生菌,在实验室条件下,筛选对供试的10种香蕉致病菌均有良好拮抗活性的菌株并测定它们的拮抗活性,对活性最强的菌株进行形态学、16S rDNA序列同源性分析。结果显示,分离得到内生菌438株,其中细菌240株,放线菌142株,真菌56株。抗病品种南天黄病株中分离得到的内生菌最多,共计128株。内生菌数量在香蕉植株中的分布呈现规律为:根部球茎部假茎部叶部。内生真菌在各香蕉种病株中的分布最广泛。筛选出具广谱活性的放线菌10株、细菌2株。其中内生放线菌菌株041的广谱拮抗活性最强,最大抑菌带宽为28.13±1.89 mm。对广谱拮抗内生放线菌菌株041、04-1、19-1、03A-1进行的形态学和16S rDNA序列分析表明,它们属于链霉菌属。     

拮抗多种植物病原真菌Streptomyces triostinicus C2的分离与鉴定北大核心CSCD

旨在分离筛选获得对植物病原真菌具有抑制作用的拮抗菌。以水稻纹枯病菌为指示菌,采用平板对峙培养法进行拮抗菌的分离筛选;根据菌体形态学观察、生理生化特征以及16S r DNA序列分析,对拮抗菌进行菌种鉴定。结果显示,从采集自云南、广东、安徽、湖北、江西等地的18个土样中分离筛选到一株对水稻纹枯病菌具有良好抑制效果的菌株C2,该菌株初步鉴定为Streptomyces triostinicus,并暂命名为链霉菌C2;进一步测定链霉菌C2的抗菌谱发现,它对水稻纹枯病菌、水稻稻瘟病菌、葡萄炭疽病菌、西瓜枯萎病菌和橘青霉等多种植物病原真菌均具有良好的抑制作用,其抑菌率分别为49.78%、56.62%、80.96%、18.59%和94.10%。链霉菌C2是一株植物病原真菌广谱拮抗菌。     

拮抗多种植物病原真菌Streptomyces triostinicus C2的分离与鉴定

旨在分离筛选获得对植物病原真菌具有抑制作用的拮抗菌。以水稻纹枯病菌为指示菌,采用平板对峙培养法进行拮抗菌的分离筛选;根据菌体形态学观察、生理生化特征以及16S r DNA序列分析,对拮抗菌进行菌种鉴定。结果显示,从采集自云南、广东、安徽、湖北、江西等地的18个土样中分离筛选到一株对水稻纹枯病菌具有良好抑制效果的菌株C2,该菌株初步鉴定为Streptomyces triostinicus,并暂命名为链霉菌C2;进一步测定链霉菌C2的抗菌谱发现,它对水稻纹枯病菌、水稻稻瘟病菌、葡萄炭疽病菌、西瓜枯萎病菌和橘青霉等多种植物病原真菌均具有良好的抑制作用,其抑菌率分别为49.78%、56.62%、80.96%、18.59%和94.10%。链霉菌C2是一株植物病原真菌广谱拮抗菌。     

一株拮抗香蕉枯萎镰刀菌稀有放线菌的分离及鉴定

本研究采用苯酚、干热、SDS和超声波四种不同的预处理方法从五指山原始林区采集的土样中选择性分离得到的702株稀有放线菌,共筛选出4株具有拮抗香蕉枯萎镰刀菌活性的菌株,经过复筛,获得一株高活性的拮抗菌210-1-61.通过对其进行形态鉴定、生理生化特征鉴定和16S rDNA序列分析,结果表明该菌株与M.pattaloongensis JCM12833T进化关系最近,其16S rDNA序列同源性最高,达98.89%;其形态与生理生化特性也与小单孢菌属M.pattaloongensis JCM12833T很接近.此外,我们还构建了与该菌株同源性较高的模式菌株16S rDNA序列的聚类分析图,结果发现该菌株与M.pattaloongensis JCM12833T稳定单独构成一个分支,二者亲缘关系最近,初步鉴定该菌株为Micromonospora. pattaloongensis.本研究为今后探讨小单孢菌属稀有放线菌对香蕉尖孢镰刀菌具有拮抗活性提供研究实验材料.     

一株具有拮抗作用黄精内生菌HJ-3的鉴定及抗菌活性研究CSCD

本研究从安徽道地药材黄精中筛选出一株内生菌HJ-3,采用五点对峙培养法观察该菌株对8种植物病原真菌的拮抗作用,将HJ-3菌株的发酵产物经萃取浓缩配制成不同浓度,通过滤纸片扩散法检测其对4种供试菌的抑菌活性,并用LC-MS检测分析。结果表明:内生菌HJ-3对8种常见植物病原真菌均表现出明显的拮抗活性,其中对立枯丝核菌等7种病原真菌拮抗活性很明显,盘长孢状刺盘孢拮抗活性较弱;另外发现,不同浓度的发酵产物对4种供试菌均有一定抑菌作用,其中对金黄色葡萄球菌抑菌效果较好。经形态学分析及16S rRNA基因测序,将该内生菌HJ-3鉴定命名为贝莱斯芽孢杆菌(Bacillus velezensis)。LC-MS检测分析,获得一种5,5′-methylenebis(2-tert-butyl-4-methylphenol)酚类物质以及三种同系列化合物。研究结果为黄精内生菌药物开发奠定基础,也为植物内生菌药物广泛应用提供依据。     

一株白芍内生放线菌的分离、活性及系统发育分析

目的:分离白芍中拮抗农作物致病菌和人类常见病原菌的内生放线菌,并进行系统发育分析。方法:采用3种分离培养基,从白芍根部分离内生放线菌;通过滤纸片法筛选具有拮抗活性的菌株,观察菌丝形态,并进行16S rDNA序列系统发育分析。结果:从白芍中分离得到16株内生放线菌,其中从FYSCA培养基中分离到9株;16株内生放线菌中有6株具有拮抗作用,菌株S-BS033004对5种病原菌有拮抗活性,尤其是对棉花黄萎病菌和小孢拟盘多毛孢菌和耐青霉素类金黄色葡萄球菌的拮抗作用显著,抑菌圈≥20mm。经16S rDNA系统发育分析表明该菌株与Streptomyces anulatus NBRC13369T等6株链霉菌模式菌株亲缘关系较近,相似性均为99.7%。结论:白芍内生放线菌S-BS033004是一株杀菌谱较广的链霉菌,具有很好的开发潜力。     

拮抗枯草芽孢杆菌KC-5的分离鉴定及其发酵优化

为了筛选出对植物病原菌具有拮抗作用的生物防治细菌,采用平板对峙法从蔬菜根际土壤中分离获得一株对多种病原真菌具有抑制作用的枯草芽孢杆菌,并对抑制后的病原菌菌丝进行了观察,结果表明该菌株对尖孢镰刀菌、串珠镰刀菌、层出镰刀菌以及腐皮镰刀菌均具有抑菌活性。通过形态特征、生理生化特征及16S rRNA序列分析,将该菌株鉴定为枯草芽孢杆菌(Bacillus subtilis),并对其发酵培养基进行了优化。     

Increased prostaglandin concentrations in the cervical mucus of pregnant women with bacterial vaginosis.

Microorganisms associated with bacterial vaginosis are commonly recovered from the amniotic fluid and chorion-amnion of patients who deliver prematurely. Bacteria closely related to those causing bacterial vaginosis may play a role in the initiation of uterine contractions, ripening of the cervix and weakening of the fetal membranes by stimulating prostaglandin synthesis. In the present investigation, cervical mucus was collected by brush from early pregnant women with and without bacterial vaginosis. The concentrations of PGE2, PGF2 alpha and 6-keto-PGF1 alpha were determined in the mucus samples by methyl oximation and then radioimmunoassay, utilizing antibodies raised against oximated prostaglandins. It was found that the concentration of PGE2 and PGF2 alpha was significantly higher in the mucus of women with bacterial vaginosis compared with healthy women. The concentration of 6-keto-PGF1 alpha was similar in both study groups. All patients had been instructed to abstain from sexual intercourse for 24 hours before sampling. However, it may be that women with high concentrations in their mucus may have had intercourse anyway. However, it is fairly well possible that the significant differences in the PGE2 and PGF2 alpha values are causally related to the higher rate of preterm labor in women with the commonplace infection of bacterial vaginosis.     

白木香内生真菌抗肿瘤抗菌活性的筛选研究

白本香[Aquilariasinensis（Lour．）Gilg]是我国特有的珍稀濒危药用植物。对已分离得到的白木香内生真菌的发酵液提取物的抗肿瘤抗菌活性进行了筛选。结果表明,对肿瘤细胞SF-268增殖的抑制率在90％以上的菌株有6株,占总菌株数的31．6％。其中有16株对金黄色葡萄球菌具有一定的抑菌活性,占总菌株教的84．2％,有9株表现较强的抑菌活性。另外有1株时白色念珠菌表现较强的抑菌活性;有2株对黑曲霉表现一定的抑菌活性。所有菌株对大肠埃希菌都没有表现明显的抑菌活性。     

1株拮抗水稻纹枯病菌鼠乳杆菌的筛选与鉴定

采用平板对峙法从浙江省金华市水稻田土壤中筛选获得1株拮抗水稻纹枯病菌的细菌QT-0304菌株,最大拮抗带宽达到20 mm。根据形态特征、生理生化特征、16S rRNA基因比对及进化树构树分析,鉴定QT-0304菌株为鼠乳杆菌（Lactobacillus murinus）。抗菌谱实验结果表明：QT-0304菌株对苹果腐烂病菌（Valsa mali）和杨树溃疡病病菌（ Botryosphaeria dothidea）均有较好的抑制作用,拮抗带宽分别达到34.5 mm和23.0 mm。     

Phage pilH alpha: a phage which adsorbs to IncHI and IncHII plasmid-coded pili

Phage pilH alpha was specific for bacterial strains, of various genera, harbouring plasmids of the HI and HII incompatibility groups. Plaque formation was temperature sensitive in that plaques formed at 26 degrees C but not at 37 degrees C. Plaques were fairly clear, irregular in outline and varied from pin point to about 2 mm in diameter on all hosts where plaques were detected. The phage had an isometric hexagonal outline with a diameter of 25 nm. It contained RNA but differed from all but one other plasmid-dependent RNA phage by being sensitive to chloroform. It adsorbed along the length of the shafts of IncHI and HII plasmid-coded pili.     

烫伤创面绿脓杆菌定植动态的实验研究

本文从细菌以多细胞生理活动观点出发,以认识定植稳定过程为目的。进行了实验大白鼠烫伤创面绿脓杆菌定植与抗定植动态观察。通过用铁浸染色法对细菌群体结构定量化研究,用糖包被负染法对群体结构内部结构观察,对粘附在组织表面细菌数量的测定,反映结构与粘附力的关系。进一步结合电镜观察及细菌生长状态的分析,证明了烫伤创面上绿脓杆菌群体结构的形成是细菌分裂繁殖所致。通过群体结构,糖包被,粘附力及生长状态的动态观察,表现出与定植的稳定程度呈平行关系,显示其重要性。联系抗定植力研究,表明定植与抗定植的一致性。最后分析了定植三个主要条件,和稳定性定植的三要素。     

Human Hsp40 proteins, DNAJA1 and DNAJA2, as potential targets of the immune response triggered by bacterial DnaJ in rheumatoid arthritis

Hsp40 proteins of bacterial and human origin are suspected to be involved in the pathogenesis of rheumatoid arthritis (RA). It has been shown that sera of RA patients contain increased levels of antibodies directed to bacterial and human Hsp40s. The aim of this work was to explore immunological similarities between the bacterial (DnaJ) and human (DNAJA1 and DNAJA2) Hsp40 proteins in relation to their possible involvement in the RA. Using polyclonal antibodies directed against a full-length DnaJ or its domains, against DNAJA1 and DNAJA2, as well as monoclonal anti-DnaJ antibodies, we found immunological similarities between the bacterial and human Hsp40s. Both ELISA and Western blotting showed that these similarities were not restricted to the conserved J domains but were also present in the C-terminal variable regions. We also found a positive correlation between the levels of the anti-DnaJ and anti-DNAJA1 antibodies in the sera of RA patients. This finding supports the molecular mimicry hypothesis that human Hsp40 could be the targets of antibodies originally directed against bacterial DnaJ in RA.     

Trichoplaxin — A new membrane-active antimicrobial peptide from placozoan cDNA

A method based on the use of signal peptide sequences from antimicrobial peptide (AMP) precursors was used to mine a placozoa expressed sequence tag database and identified a potential antimicrobial peptide from Trichoplax adhaerens. This peptide, with predicted sequence FFGRLKSVWSAVKHGWKAAKSR is the first AMP from a placozoan species, and was named trichoplaxin. It was chemically synthesized and its structural properties, biological activities and membrane selectivity were investigated. It adopts an α-helical structure in contact with membrane-like environments and is active against both Gram-negative and Gram-positive bacterial species (including MRSA), as well as yeasts from the Candida genus. The cytotoxic activity, as assessed by the haemolytic activity against rat erythrocytes, U937 cell permeabilization to propidium iodide and MCF7 cell mitochondrial activity, is significantly lower than the antimicrobial activity. In tests with membrane models, trichoplaxin shows high affinity for anionic prokaryote-like membranes with good fit in kinetic studies. Conversely, there is a low affinity for neutral eukaryote-like membranes and absence of a dose dependent response. With high selectivity for bacterial cells and no homologous sequence in the UniProt, trichoplaxin is a new potential lead compound for development of broad-spectrum antibacterial drugs.     

胆道疾病患者343份胆汁细菌培养与药敏分析

了解胆汁中细菌分布及药敏情况,以指导临床合理用药。对343例胆道疾病手术患者抽取胆汁进行常规培养及应用英国OXID血液增菌瓶和ATB自动细菌鉴定统对标本进行细菌鉴定和药敏分析。343份胆汁中共培养出10种113株细菌(不包括厌养菌)阳性率为37.4%,主要菌种有大肠埃希菌、肺炎克雷伯菌、粪肠球菌、阴沟肠杆菌、铜绿假单胞菌比率较高。药敏结果显示,革兰阴性杆菌对亚胺培南、头孢三代、阿米卡星、庆大霉素敏感性较好;革兰阳性球菌对万古霉素、亚胺培南、利福平敏感性较好。胆道感染多由肠道细菌逆行感染引起的,为此,根据药敏试验结果指导临床用药是非常重要的。     

测定绿脓杆菌分裂指数反映生长势头的初步研究

本文以细胞生物学理论为基础,细菌细胞发育周期形态和群体细菌分裂相数量的变化为依据,研究了测定绿脓杆菌DI方法。建立了数学模型,分析了生长过程中DI动态。结果表明,DI可较准确地反映细菌生长势头。实验采用同龄细菌选择和同步培养技术,免疫荧光染色,显微镜下直接观察分裂相细菌,并计算其占总菌数的比例。在对数期前DI已上升,开始后迅速达高峰,接近稳定期时已下降到较低水平。与生长速度比较,两者动态基本一致,但DI高峰较早出现77分钟。认为DI与生长速度关系密切,是与细菌生命活动物质交换的三流学说相伴随的一种特殊信息流。     

Dimethylsulphopropionate (DMSP) and proline from the surface of the brown alga Fucus vesiculosus inhibit bacterial attachment

It was demonstrated previously that polar and non-polar surface extracts of the brown alga Fucus vesiculosus collected during winter from the Kiel Bight (Germany) inhibited bacterial attachment at natural concentrations. The present study describes the bioassay-guided identification of the active metabolites from the polar fraction. Chromatographic separation on a size-exclusion liquid chromatography column and bioassays identified an active fraction that was further investigated using nuclear magnetic resonance spectroscopy and mass spectrometry. This fraction contained the metabolites dimethylsulphopropionate (DMSP), proline and alanine. DMSP and proline caused the anti-attachment activity. The metabolites were further quantified on the algal surface together with its associated boundary layer. DMSP and proline were detected in the range 0.12-1.08 ng cm(-2) and 0.09-0.59 ng cm(-2), respectively. These metabolites were tested in the concentration range from 0.1 to 1000 ng cm(-2) against the attachment of five bacterial strains isolated from algae and sediment co-occurring with F. vesiculosus. The surface concentrations for 50% inhibition of attachment of these strains were always <0.38 ng cm(-2) for DMSP and in four cases <0.1 ng cm(-2) for proline, while one strain required 1.66 ng cm(-2) of proline for 50% inhibition. Two further bacterial strains that had been directly isolated from F. vesiculosus were also tested, but proved to be the least sensitive. This study shows that DMSP and proline have an ecologically relevant role as surface inhibitors against bacterial attachment on F. vesiculosus.