云南大学云南省微生物研究所放线菌实验室简介

30多年来,放线菌实验室以放线菌资源的收集,保存,分类为主要研究内容。重点进行云南及周边地区、我国西部极端环境放线菌的分离,保存,发酵,制备提取物供协作单位进行生物活性物质产生菌的筛选,有效菌种的系统进化与多相分类。自1983年以来,承担国家基金重大项目、重点项目、面上项目,国家科技部973项目、重大基础研究前期专项,省重点项目,国家和省国际合作项目20多项,发现放线菌新科一个(阎氏菌科Yani-aceae),新属9个(双孢放线菌属Actinobispora,链单孢菌属Streptomonospo-ra,阎氏菌属Yania,产丝菌属Myceligenerans,姜氏菌属Jiangella,碱…     

昆虫共生放线菌多样性、功能及其活性次生代谢产物研究进展

作为一类具有重要经济价值的微生物资源,已发现的放线菌所产的生物活性物质占到所有微生物生物活性物质的70%,其中包括抗生素、免疫抑制剂和酶抑制剂等。从昆虫体内分离放线菌将会是发现新菌种和新化合物的重要渠道。本文主要阐述了昆虫共生放线菌的多样性及功能,总结了昆虫共生放线菌的活性次生代谢产物的种类及其应用进展情况,最后对昆虫共生放线菌代谢活性物质研究与开发中存在的问题提出了对策。     

河西走廊酒泉地区盐碱土壤中可培养放线菌多样性

为了探究甘肃河西走廊酒泉地区盐碱土壤中可培养放线菌种群多样性信息,采用稀释平板涂布法对采集到的4个盐碱土样进行分离培养纯化,并基于16S rRNA基因序列对分离的放线菌株进行系统发育分析。结果显示:根据菌落的表型差异从中共分离到63株放线菌菌株,它们分属于7个亚目、11个科、24个属;其中Streptomyces属为河西走廊酒泉地区盐碱土壤中可培养放线菌的优势菌群,共分离到24株,占放线菌种群总数的38.1%,并且有22株放线菌与已知菌序列的相似性低于98%,占放线菌总数量的34.9%。表明,河西走廊酒泉地区盐碱土壤中的放线菌资源丰富,类群具有多样性。     

荒漠放线菌的资源分布、多样性及分离方法研究进展

荒漠具有高度干旱、植被稀少和强辐射等特征,是极端放线菌资源的宝库。近年来,多种荒漠来源的放线菌新分类单元和新结构活性天然产物被报道,引起了人们的极大兴趣。了解荒漠放线菌的群落结构特征,选择有效的分离方法是挖掘荒漠放线菌物种资源的关键。本文综述了全球主要荒漠区放线菌的资源分布与多样性,归纳分析了荒漠放线菌的主要分离方法及存在问题,并对未来的研究工作进行了展望。     

嗜盐放线菌的研究进展

嗜盐放线菌是极端微生物的重要组成部分 ,也是一类极具应用前景的微生物资源。就嗜盐菌的分类标准、嗜盐放线菌的分离、分布及系统学研究的历史、现状及其发展趋势、应用前景进行了概述。同时还讨论了嗜盐放线菌的分离及其生理学、分类学研究中存在的问题及困难     

新疆特殊环境微生物资源的研究与发展

新疆特殊环境微生物资源丰富、代谢类型多样。目前,已发现了放线菌新属2个,放线菌新种24个,根瘤菌新属1个,嗜盐古生菌2个,食用真菌195种,药用真菌97种。通过对新疆特殊环境微生物资源的分析,论述了新疆特殊环境微生物资源研发对策,明确了重点研发领域为新疆特殊环境极端环境微生物研究、新疆特色药用真菌研究和新疆维吾尔医药用植物内生菌的研究,同时建立新疆特殊环境微生物资源库及功能基因库和生物信息系统,从而实现新疆特殊环境微生物资源最大限度的开发、保护及持续利用。     

稀有放线菌分离方法*

稀有放线菌是生物活性物质的重要来源。从样品预处理,抑制剂的选择,噬菌体的使用,碳源的选择及培养基的设计等各个方面介绍了稀有放线菌分离方法及作者的经验。     

稀有放线菌分离方法

稀有放线菌是生物活性物质的重要来源。从样品预处理,抑制剂的选择,噬菌体的使用,碳源的选择及培养基的设计等各个方面介绍了稀有放线菌分离方法及作者的经验。     

阔苞菊内生放线菌的分离鉴定和抗菌活性研究

为了分离鉴定阔苞菊内生放线菌并研究其抑菌活性,通过与高氏一号培养基和1/10ATCC CL-173培养基的对比试验,选用改进的HV培养基从阔苞菊中分离纯化出48株内生放线菌;对其16S rDNA进行PCR扩增及基因测序,结果显示其中79.17%为链霉菌属菌株;对所提取放线菌基因组DNA进行BOX-PCR多样性指纹分析,结果与16Sr DNA序列分析的结果一致;利用滤纸片法测量其抑菌活性,其中菌株KBJ37、KBJ52、KBJ114对诺氏布丘氏菌、气单胞菌、乡间布丘氏菌、小牛葡萄球菌有抑制效果,仅有KBJ114对酿酒酵母有抑制效果;通过检测编码非核糖体多肽合成酶(NRPS)和聚酮化合物合酶基因Ⅰ、Ⅱ(PKS-Ⅰ和PKS-Ⅱ)的序列,来判断阔苞菊内生放线菌合成生物活性物质的潜力,结果显示97.92%的放线菌拥有合成生物活性物质的潜力。以上研究表明阔苞菊内生放线菌具有较为广谱的抑菌活性,且绝大多数都具有合成生物活性物质的潜力,为进一步探明阔苞菊内生放线菌资源多样性及药用价值提供了理论依据。     

嗜碱放线菌的分离方法

嗜碱放线菌是一类嗜好碱性生活环境的特殊资源微生物。近年来,从嗜碱放线菌中发现多个新分类单元和结构新颖、活性独特的天然产物,由此引起人们的极大兴趣。有效的选择性分离方法是收集与挖掘嗜碱放线菌物种资源的关键。本文分析了嗜碱放线菌的生理特点,归纳了嗜碱放线菌的分离方法及存在问题,并探讨了未来的发展趋势。     

酸性和碱性土壤新分离的放线菌中线型和环型质粒的检测

从湖南、云南采集的19份酸性土壤样品分离到50株放线菌,在pH4.5和pH7.0的培养基上都能生长.从新疆、青海、云南的14份碱性土壤样品中分离到38株放线菌,在pH10.0和pH7.0的培养基上均能生长.因此,这些菌株属于中度嗜酸或中度嗜碱放线菌.利用脉冲电泳技术和普通凝胶电泳在其中12株放线菌中检测到线型质粒,其中3株放线菌中检测到环型质粒.这是首次在中度嗜酸和中度嗜碱的放线菌中报道线型和环型质粒.     

青海盐碱环境中具抗肿瘤活性放线菌的筛选和多样性研究

从我国青海省采集盐碱土样或泥样,用添加1.0~3.0mol/L NaCl的GPY琼脂培养基和ISP2琼脂培养基分离到145株典型放线菌菌株。采用6种肿瘤细胞株对分离菌株的发酵产物进行体外筛选,得到26株抗肿瘤活性阳性菌株(17.9%),19株为拟诺卡氏属(Nocardiopsis)菌株,7株为链霉菌属(Streptomyces)菌株。在抗肿瘤活性、形态特征、生理生化特性和全细胞水解物氨基酸组分分析等实验结果的基础上,选取差异较大的8株抗肿瘤活性阳性菌株进行16S rRNA基因序列测定和系统发育多样性分析。结果表明,2株属于链霉菌属(Streptomyces)的1个已知物种和1个潜在新种;6株属于拟诺卡氏菌属(Nocardiopsis),可能代表该属的4个新种。研究表明青海盐碱环境中存在产生抗肿瘤活性物质的重要放线菌资源,也提示其中蕴藏着较丰富的微生物多样性。     

新疆青海中度嗜盐放线菌生物多样性初步研究

从我国新疆、青海等地采集数份盐碱土样或泥样,采用淀粉-酪素琼脂培养基、甘油天门冬酰胺琼脂培养基、土壤浸汁琼脂培养基分别从中分离到8株、32株中度嗜盐放线菌菌株。经形态、生理学特性与全细胞壁氨基酸组分分析结果比较,选取其中的14株进行16S rDNA序列分析。就物种多样性而言,新疆、青海分离到中度嗜盐放线菌分布至少有3个科,5个属。其中有拟诺卡氏菌科（Nocardiopsaceae）的拟诺卡氏菌属(Nocardiopsis)和链单孢菌属（Streptomonospora）;假诺卡氏菌科(Pseudonocardiaceae)的普氏菌属(Prauserella)和糖单孢菌属(Saccharomonospora);链霉菌科(Streptomycetaceae)的链霉菌属(Streptomyces)。就地区分布来讲,新疆分离到的中度嗜盐放线菌种类要远高于青海。     

嗜碱放线菌生理学特性的初步研究*

对从新疆、青海采集的盐碱土样中分离获得的29株嗜碱放线菌(AlkAlipbilic actinomyeetes)及1株模式菌株Nocardiopsis prasina的pH实验,不同碱性物质KOH、K2CO3、NaOH、Na2CO3对生长的影响,及对NaCl、KCl耐受性进行了研究。结果发现,少数嗜碱放线菌对Na^ 有一定的依赖性,对K^ 碱性物质敏感;一些嗜碱放线菌对CO3^2-敏感,NaCl、KCl对一些嗜碱放线菌的生长有抑制作用;4种碱性物质对Nocardiopsis菌株生长无影响,且它们可耐受高浓度NaCl、KCl。因此提出高碱性环境中是否存在K^ 、CO3^2-专一性依赖的嗜碱放线菌的推测。     

New genus-specific primers for the PCR identification of novel isolates of the genus Streptomonospora

The halophilic actinomycete genus Streptomonospora, forming a distinct branch in the 16S rRNA gene phylogenetic tree adjacent to the genera Nocardiopsis and Thermobifida, was first proposed by Cui et al. to accommodate the species type Streptomonospora salina. During a biodiversity and taxonomic study on halophilic filamentous actinomycetes from a saline lake in western China, numerous new halophilic actinomycetes strains were isolated. To confirm whether they are members of the genus Streptomonospora, one set of genus-specific oligonucleotides was designed which allows rapid detection of members of the genus Streptomonospora by means of PCR amplification. The genus specificity of these primers was validated with reference strains as well as with wild-type isolates, which exhibited morphological characteristics common to this genus.     

新疆泥火山产酶嗜盐放线菌的筛选及多样性

[目的]了解新疆乌苏泥火山嗜盐放线菌及其产酶功能多样性.[方法]分别采用含有5%与10%NaCl的5种分离培养基,稀释平板涂布法对泥火山土壤样品进行分离;利用五种筛选培养基定性检测酶活性;在形态特征、耐盐性实验及16S rDNA基因测序的基础上进行系统发育学分析.[结果]获得嗜盐放线菌43株,极端嗜盐放线菌3株.4株嗜盐放线菌产脂肪酶,30株产半乳糖苷酶,27株产淀粉酶,6株产酯酶,4株产纤维素酶,1株同时产4种酶.系统发育学分析结果表明其中24株为拟诺卡氏菌属(Nocardiopsis),1株为链霉菌属(Streptomyces).产两种酶的菌株10006与Nocardiopsis exhalans(AY03600)相似性为96.64%(小于97%),可能是潜在的新种.[结论]本研究表明新疆乌苏泥火山中存在大量的产半乳糖苷酶及淀粉酶的嗜盐放线菌,所分离到的拟诺卡氏菌属产酶多样性比较高,并且潜藏着新的微生物资源.     

Evolution of bacterial flora from a subterranean saline well by graduated salinity changes in enrichment media

The bacterial flora of samples taken from a subterranean saline well was enriched (1) by periodic salinity increase and (2) by periodic salinity decrease, both at 25° and 35°C. During the enrichment process, bacterial flora, including halotolerant, marine, moderately and extremely halophilic bacteria, were enumerated. Results were similar at both temperatures. Marine bacteria and moderately halophilic microorganisms were the most favoured groups, predominating between 3 and 30% (w/v) salt content; extremely halophilic bacteria also appeared when salinity reached 30% (w/v) and halotolerant bacteria were poorly represented. The taxonomic distribution of 125 selected strains, chosen at random from counting media, was essentially similar to those from other hypersaline environments.     

Biodegradation of organic pollutants by halophilic bacteria and archaea

Hypersaline environments are important for both surface extension and ecological significance. As all other ecosystems, they are impacted by pollution. However, little information is available on the biodegradation of organic pollutants by halophilic microorganisms in such environments. In addition, it is estimated that 5% of industrial effluents are saline and hypersaline. Conventional nonextremophilic microorganisms are unable to efficiently perform the removal of organic pollutants at high salt concentrations. Halophilic microorganisms are metabolically different and are adapted to extreme salinity; these microorganisms are good candidates for the bioremediation of hypersaline environments and treatment of saline effluents. This literature survey indicates that both the moderately halophilic bacteria and the extremely halophilic archaea have a broader catabolic versatility and capability than previously thought. A diversity of contaminating compounds is susceptible to be degraded by halotolerant and halophile bacteria. Nevertheless, significant research efforts are still necessary in order to estimate the true potential of these microorganisms to be applied in environmental processes and in the remediation of contaminated hypersaline ecosystems. This effort should be also focused on basic research to understand the overall degradation mechanism, to identify the enzymes involved in the degradation process and the metabolism regulation.     

Biodiversity of poly-extremophilic Bacteria: Does combining the extremes of high salt, alkaline pH and elevated temperature approach a physico-chemical boundary for life?

Bacterial microorganisms that grow optimally at Na⁺ concentrations of 1.7 M, or the equivalent of 10% (w/v) NaCl, and greater are considered to be extreme halophiles. This review focuses on the correlation between the extent of alkaline pH and elevated temperature optima and the extent of salt tolerance of extremely halophilic eubacteria; the focus is on those with alkaline pH optima, above 8.5, and elevated temperature optima, above 50°C. If all three conditions are required for optimal growth, these microorganisms are termed "poly-extremophiles". However, only a very few extreme halophiles able to grow optimally under alkaline conditions as well as at elevated temperatures have been isolated so far. Therefore the question is: do the combined extreme growth conditions of the recently isolated poly-extremophiles, i.e., anaerobic halophilic alkalithermophiles, approach a physico-chemical boundary for life? These poly-extremophiles are of interest, as their adaptive mechanisms give insight into organisms' abilities to survive in environments which were previously considered prohibitive to life, as well as to possible properties of early evolutionary and extraterrestrial life forms.