放线菌产生各种有用物质的初步探索

从云南若干地区采集土样、12个湖泊采集湖底泥和水样。用多种培养基分离其中的放线菌。采用不同方法筛选了纤维蛋白溶酶等各种有用物质的产生菌。并对不同来源不同属的放线菌产生各种有用物质的规律作了初步探讨。     

青海高寒草甸土壤放线菌区系研究

2001～2002年从海北高寒草甸生态系统采集土样,用不同方法从中分离放线菌300余株,根据其形态和分类特征,分别归入小单孢菌属(Micromonospora)、诺卡氏菌属(Nocardia)、糖多孢菌属(Saccharopolyspora)、原小单孢菌属(Promicromonospora)和链霉菌属(Streptomyces),并将链霉菌归入7个类群。同时对230株中温菌和110株低温菌的部分酶活性及其对真菌和细菌的拮抗性进行了测定,发现链霉菌不仅具有许多酶活性,而且对真菌和细菌有拮抗性。     

拉鲁湿地自然保护区放线菌组成分析及生物活性测定

探究拉鲁湿地自然保护区的放线菌组成及其抑菌和酶活性,为放线菌新药物先导化合物和高活性酶的筛选提供资源。从拉鲁湿地自然保护区不同土壤类型、不同优势植被采集25份土样。用分散差速离心法分离了拉鲁湿地中温放线菌和低温放线菌。从中温放线菌中选择15株代表菌株进行了初步分类鉴定。采用打孔法检测了其对2株细菌和4株病原真菌的抑菌活性。结果表明：（1）拉鲁湿地放线菌数量从水生环境向陆地生态系统递增,中温放线菌数量显著多于低温放线菌;（2）拉鲁湿地土壤中分离到链霉菌属、小单孢菌属、诺卡氏菌属、马杜拉菌属、小链孢菌属5个放线菌属。其中以链霉菌属和小单孢菌属为优势属。链霉菌属以金色类群、白孢类群和粉红孢类群为主,小单孢菌分离到黄橙类群和黑褐类群;（3）供试菌株分解纤维素能力较强,分解蛋白质活性较低,具有抗菌活性的菌株很少,且抗菌活性较弱;（4）供试菌株耐毒性物质的能力较强。这些菌可用于毒害有机物污染物的处理。     

云南若干地区土壤放线菌区系及资源考察* Ⅷ.滇中高原的放线菌资源

从云南省的鸡足山,紫金山和安宁温泉等地采集了不同植被、不同海拔高度的土样,用四种培养基分离放线菌。按常规方法进行鉴定,用不同方法筛选产生纤维蛋白溶酶等有用菌。对滇中高原地区放线菌的区系组成及产生有用代谢产物的规律进行了探讨。     

云南高原湖泊水生放线菌的研究VIII．马杜拉放线菌属的鉴定

从云南省程海湖泥样品中分离到12株马杜拉放线菌。其中的两株分别定为新种：程海马杜拉放线菌(Actinomadura chenghaicnsis sp. Nvo.)和绿黄马杜拉放线菌(Actinomadtsraviridoflava sp. Nov.)。     

不同红树林地区老鼠簕内生放线菌的分离及其环境适应性

【目的】比较不同红树林地区的老鼠簕内生放线菌的地理分布,了解内生放线菌与其所处环境的相关性。【方法】分别从5个不同地点的红树林采集老鼠簕全株植物,采用9种分离培养基,从植株不同部位分离内生放线菌,用16S rRNA基因序列分析鉴定到属,用添加不同NaCl浓度的ISP 2液体培养基进行耐盐度测试,用无氮基础培养基进行固氮活性测试。【结果】共分离得到内生放线菌52株,其中从叶、茎和根部分别获得5株、2株和45株,花和果中未分离到。52株内生放线菌分别属于小单孢菌属(47株),链霉菌属(3株),疣孢菌属(1株)和继生菌属(1株)。48株菌表现出耐盐或嗜盐特征,其中18株最高耐盐度20%,4株不能在无盐条件下生长,12株菌可在含有3.3%NaCl的培养基上生长良好。4株菌可在无氮培养基下生长。【结论】对47株内生小单孢菌的地理分布分析表明,老鼠簕内生小单孢菌的类群因不同地理位置有很大差异。耐盐和固氮活性测试结果表明了老鼠簕内生放线菌对环境的适应性。     

嗜酸丝状放线菌的选择性分离与多样性

摘要:【目的】针对酸性土壤中的嗜酸丝状放线菌,建立有效的选择性分离方法,并了解其多样性。【方法】用不同的样品预处理方式和分离培养基,并添加不同的抑制剂进行分离;根据放线菌的菌落数和出菌率确定最佳分离方法组合。采用最佳分离方法对从江西采集的17份酸性土壤样品进行分离;根据培养特征对分离菌株进行分群,进一步通过对各类群的显微形态观察和pH梯度生长实验确定代表菌株;对代表菌株进行16S rRNA基因序列分析研究其多样性。【结果】嗜酸丝状放线菌的最佳分离方法为：土壤样品经分散差速离心预处理后,涂布添加了放线菌酮、制霉菌素和萘啶酮酸（各50 mg/L）的GTV培养基。用此方法共分离到放线菌369株,归为10个不同的颜色类群,其中6.6%为严格嗜酸放线菌,72.4%为中度嗜酸放线菌,21.0%为耐酸放线菌。52株嗜酸放线菌代表菌株分布于放线菌目中的12个属：链霉菌属(Streptomyces)、小单孢菌属(Micromonospora) 、诺卡氏菌属(Nocardia)、野野村菌属(Nonomuraea) 、韩国生工属(Kribbella) 、小双孢菌属(Microbispora)、马杜拉菌属(Actinomadura)、拟无枝菌酸菌属(Amycolatopsis)、指孢囊菌属(Dactylosporangium)、伦茨氏菌属(Lentzea)、游动四孢菌属(Planotetraspora) 和链嗜酸菌属(Streptacidiphilus),其中链霉菌分离菌株在系统发育树上形成12个不同的进化类群。【结论】所建立的选择性分离方法可用于土壤嗜酸丝状放线菌的高效分离;江西酸性土壤含有丰富多样的嗜酸丝状放线菌种属。     

绿色糖单孢菌-木素过氧化物酶的发酵工艺研究

本研究以一株中度耐热耐碱放线菌--绿色糖单孢菌(Saccharomonospora viridis)为研究对象,用16 L发酵罐对该菌进行了木素过氧化物酶(lignin peroxidases, LiP)的诱导发酵,确定了最适的产酶工艺条件:接种量为10%,C/N为1∶3,搅拌速度为250 r/min,通气量为5 L/min,通过控制通气量和调整搅拌转速,使溶氧维持在35%以上,此条件下绿色糖单孢菌较摇瓶实验提前将近24 h达到产酶高峰,酶活最高可达0.41 U/ml;同时在发酵罐中测定该菌株的生长曲线和代谢曲线以确定其发酵代谢规律.     

微波处理对嗜碱和嗜盐海洋放线菌分离效果的影响

【目的】研究微波处理对于分离嗜碱和嗜盐海洋放线菌的效果。【方法】用微波处理7份海泥样品,梯度稀释后涂布于3种分离培养基,分离具有嗜碱和嗜盐特性的海洋放线菌。【结果】微波处理后的7份样品中,4份样品中嗜碱海洋稀有放线菌和3份样品的嗜盐海洋稀有放线菌数量极显著提高;7份样品中的嗜碱、嗜盐海洋小单孢菌属、游动放线菌属、诺卡氏菌属等稀有放线菌数量均有显著增加,不同样品中新分离到链孢菌属、小双孢菌属、链孢囊菌属及其他未鉴定的海洋稀有放线菌,分离到属的数量提高了1-4个。【结论】微波处理不仅显著提高嗜碱和嗜盐海洋放线菌的分离数量,而且明显增加了海洋稀有放线菌的分离种类。     

以形态、细胞壁和醌化学组份鉴别放线菌的属

将经典的形态分类法与化学分类法(细胞壁与醌的化学分析)相结合,对从我国不同地区分离到的22株放线菌进行了分类鉴定。它们分别属于类诺卡氏菌属(Nocardioides)、拟诺卡氏菌属(Nocardiopsis)、小多孢菌属(Micropolyspora)、马杜拉放线菌属(Actinomadura)、游动放线菌属(Actinoplanes)和小单孢菌属(Micromonospora)等6个属,13个种。     

云南高原湖泊中高温放线菌四新种

在进行云南高原湖泊水生放线菌研究的过程中,分离到大量高温放线菌,按国内通用的方法进行鉴定,有4个新种。模式菌株Y86—8217,Y86—8100,Y86—8010,Y86—8048均保存于云南省微生物研究所。     

The occurrence and distribution of actinomycetes in lakes of the English Lake District

Investigation of the occurrence of mesophilic actinomycetes in the lakes of the English Lake District revealed their widespread distribution in the lacustrine environment. Although only low numbers of actinomycetes occurred in the water, high numbers were recovered from all the lake muds. Total numbers of actinomycetes in the muds correlated quite well with the lakes’ productivity status. High numbers of Micro-monospora, Streptomyces and nocardioform actinomycetes were isolated from all the lakes sampled. Low numbers of Streptosporangium were isolated from all the muds but strains of Actinomadura, Actinoplanes, Dactylosporangium, Microbispora and Thermomonospora were only encountered occasionally. Micromonospora was the numerically dominant genus isolated from all the lakes sampled. This dominance was even more striking in deeper layers of mud and this was thought to reflect a more resistant spore stage in Micromonospora than in either of the other two main genera.     

Isolation of actinomycetes from soil using extremely high frequency radiation

A new method employing extremely high frequencies (EHFs) is proposed for the selective isolation of actinomycetes from soil. The pretreatment of soil suspensions with EHF wavelengths of 5.6 and 7.1 mm led to a nonselective isolation of actinomycetes. At the same time, the irradiation of soil suspensions within wavelength bands of 3.8-5.8 and 8-11.5 mm considerably augmented the total number of isolated actinomycetes and increased the fraction of the isolated rare genera by 2 and 7 times, respectively. The rare actinomycete genera were represented by Actinomadura, Microtetraspora, Nonomuraea, Micromonospora, Amycolatopsis, Pseudonocardia, Saccharotrix, and Streptosporangium.     

Diversity of Aquatic Actinomycetes in Lakes of the Middle Plateau, Yunnan, China

A total of 749 sediment and water samples were collected from 12 lakes of the Middle Plateau of Yunnan from 1983 to 1993. The diversity and biological characteristics of the aquatic actinomycetes in these lakes were studied. Sixteen genera of actinomycetes were isolated from these samples. Micromonospores assumed a notable dominance (from 39 to 89%) in the actinomycete populations of these lake sediments. Streptomycetes were the second most abundant organisms. The diversity and counts of actinomycetes varied with the season. Thermophilic actinomycetes have a wide distribution in these lakes, but their counts were smaller. The cell wall compositions of certain Micromonospora and Streptomyces strains from an alkaline lake revealed an unusual combination of glycine and isomers of diaminopimelic acid. It seems that aquatic actinomycetes play a significant role in the decomposition of organic substances, including some toxic compounds such as phenol, in these lakes. It also appears that aquatic actinomycetes are one of the important resources for screening useful enzymes and metabolites.     

Isolation of Actinomycetes from Soil Using Extremely High Frequency Radiation

A new method employing extremely high frequencies (EHFs) is proposed for the selective isolation of actinomycetes from soil. The pretreatment of soil suspensions with EHF wavelengths of 5.6 and 7.1 mm led to a nonselective isolation of actinomycetes. At the same time, the irradiation of soil suspensions within wavelength bands of 3.8–5.8 and 8–11.5 mm considerably augmented the total number of isolated actinomycetes and increased the fraction of the isolated rare genera by 2 and 7 times, respectively. The rare actinomycete genera were represented by Actinomadura, Microtetraspora, Nonomuraea, Micromonospora, Amycolatopsis, Pseudonocardia, Saccharotrix, and Streptosporangium.     

Diversity of antifungal actinomycetes in various vegetative soils of Korea

Diversity of actinomycetes and their antifungal activities against some plant pathogenic fungi were examined in various vegetative soils from 14 different sites in the western part of Korea. Actinomycete counts ranged from 1.17 x 10(6) to 4.20 x 10(6) cfu x g(-1) dried soil. A total of 1510 actinomycetes were isolated from the soil samples. Streptomyces was predominant in soils with a pH range of 5.1-6.5, 9.1-13.0% moisture, and 9.1-11.0% organic matter. Most Micromonospora, Dactylosporangium, and Streptosporangium were distributed in soils with pH 4.0-5.0, 2.0-9.0% moisture, and 4.0-7.0% organic matter. Actinomadura and nocardioform actinomycetes were abundant in soils with pH 4.0-5.0 and 13.1-20.0% moisture and with 9.1-11.0 and 4.0-7.0% organic matter, respectively. Populations of Streptomyces were predominant in all the soils, but were highest in grassland and lowest in mountain-forest soils. Micromonospora was most abundant in pepper-field soil and nocardioform actinomycetes were highest in rice paddy field soil. Dactylosporangium was predominant in lake-mud sediments and pepper-field soil, Streptosporangium in lake-mud sediments, and Actinomadura in mountain-forest soil. Antifungal actinomycetes were abundant in orchard soil and lake mud. More than 50% of antifungal isolates from most soils were classified as genus Streptomyces. Actinomycete isolates that showed strong antifungal activity against Alternaria mali, Colletotrichum gloeosporioides, Fusarium oxysporum f.sp. cucumerinum, and Rhizoctonia solani were predominant in pepper-field soils, whereas those against Magnaporthe grisea and Phytophthora capsici were abundant in radish-field soils.     

Haloalkaliphilic actinomycetes in soils of Mongolian desert steppes

The abundance of actinomycetes isolated from the soils of Mongolian desert steppes varies from several thousand to hundreds of thousands of CFU/g soil, depending on soil type and isolation medium. Eight actinomycete genera have been found in these soils: Streptomyces, Micromonospora, Saccharopolyspora, Actinomadura, Microtetraspora, Thermomonospora, Nocardia, and Dactylosporangium. The streptomycete complexes of brown desert-steppe and gray-brown desert alkaline soils include halophilic, alkaliphilic, and haloalkaliphilic species that grow most successfully on the media with a salt concentration of 5% and pH 8–9.     

不同放线菌属的化学与分子分类

随着科学的发展与新技术在分类学中不断地应用,放线菌分类学已从经典的形态分类转向化学分类(细胞壁化学组份,磷酸类脂,枝菌酸及甲基萘醌等).现在有些国家又开展了分子分类.本实验室自80年代始开展了放线菌化学分类,建立了上述化学指征的分析方法.自90年代起,又开展了分子分类,DNA-DNA杂交、23S rRNA寡核甘酸序列分析.近来,许多人用16S rRNA部分序列区分微生物不同的基因种.作者选用了23S rRNA部分序列区分放线菌的不同属种.现将研究结果简报如下:1 材料和方法1.1菌种菌株10,13,23,C_(43),350,41,53,4650及N分离自云南省土壤中.C_(51)及3306来自日本微生物菌种保藏中心.     

Rare genera of actinomycetes as potential producers of new antibiotics

A literature survey covering more than twenty-three thousand bioactive microbial products including eight thousand antiinfectives demonstrated the increasing relevance of the so called 'rare' actinomycetes as a source of new antibiotics. Past and present efforts in the isolation of rare actinomycetes have enriched the Biosearch Italia Strain Collection with more than twenty thousand strains, showing that, when selective isolation methods are developed and extensively applied, some genera, such as Actinomadura, Actinoplanes, Micromonospora, Microtetraspora, are not rare at all and can be recovered from many soil samples. The current focus is on the isolation of members of Streptosporangiaceae family, given their promising chemical diversity.     

山西运城盐湖放线菌区系研究

从山西运城盐湖分离到中、高温嗜碱或耐碱放线菌120株,对其进行了分类鉴定。根据形态和分类特征（细胞壁化学成分）,分别归入拟诺卡氏菌属（Nocardiopsis）、诺卡氏菌科（Nocediaceae）、小多孢菌属（Micromonospora）、马杜拉放线菌属（Actinomadura）及链霉菌属（Streptomyces）,并将链霉菌归入了11个类群。