Champacyclin,a New Cyclic Octapeptide from Streptomyces Strain C42 Isolated from the Baltic Sea

New isolates of Streptomyces champavatii were isolated from marine sediments of the Gotland Deep (Baltic Sea), from the Urania Basin (Eastern Mediterranean), and from the Kiel Bight (Baltic Sea). The isolates produced several oligopeptidic secondary metabolites, including the new octapeptide champacyclin (1a) present in all three strains. Herein, we report on the isolation, structure elucidation and determination of the absolute stereochemistry of this isoleucine/leucine (Ile/Leu = Xle) rich cyclic octapeptide champacyclin (1a). As 2D nuclear magnetic resonance (NMR) spectroscopy could not fully resolve the structure of (1a), additional information on sequence and configuration of stereocenters were obtained by a combination of multi stage mass spectrometry (MSⁿ) studies, amino acid analysis, partial hydrolysis and subsequent enantiomer analytics with gas chromatography positive chmical ionization/electron impact mass spectrometry (GC-PCI/EI-MS) supported by comparison to reference dipeptides. Proof of the head-to-tail cyclization of (1a) was accomplished by solid phase peptide synthesis (SPPS) compared to an alternatively side chain cyclized derivative (2). Champacyclin (1a) is likely synthesized by a non-ribosomal peptide synthetase (NRPS), because of its high content of (d)-amino acids. The compound (1a) showed antimicrobial activity against the phytopathogen Erwinia amylovora causing the fire blight disease of certain plants.     

苹果树腐烂病内生拮抗放线菌A-2的鉴定及其活性评价

以苹果树腐烂病菌Valsa mali为靶标菌,从健康苹果树枝条中获得一株高效内生放线菌A-2。采用生长速率法、对峙培养法和玻片法测定了菌株A-2的抑菌谱及对苹果树腐烂病菌菌丝生长和孢子萌发的影响;利用离体枝条烫伤接种法测定了菌株A-2对腐烂病的防治效果。结果表明:A-2发酵滤液对腐烂病菌的抑制率达90%以上,对其他9种常见的植物病原真菌也具有良好的拮抗作用;菌株A-2可导致腐烂病菌菌丝畸形、分支增多和局部膨大;不同稀释倍数的A-2发酵滤液对苹果离体枝条腐烂病均有明显的防效,接种病原菌前后24 h和同时施用500倍发酵滤液的防效均达70%以上。根据培养特征、生理生化特性和16S rDNA 序列分析,将菌株A-2鉴定为卡伍尔链霉菌Streptomyces cavourensis。     

可可链霉菌182-2活性代谢产物对烟草赤星病菌的作用

【目的】明确可可链霉菌（Streptomyces cacaoi）182-2的活性代谢产物——KA08对烟草赤星病菌（Alternaria alternata）的抑菌活性及其机理,为后续农用抗生素产品开发和应用提供科学依据。【方法】采用菌丝湿重法和培养皿中萌发法,测定KA08对烟草赤星病菌菌丝生长和孢子萌发的抑制作用;采用电导率法和紫外吸收法测定其对细胞膜渗透性、麦角甾醇含量、脂质过氧化程度、可溶性蛋白含量的影响。【结果】KA08对烟草赤星病菌菌丝生长和孢子萌发均有强烈的抑制作用,同时,还可造成菌丝节间缩短、扭曲,芽管顶端膨大,并产生大量囊状膨大物。处理24 h时,对孢子萌发的抑菌中浓度为177.53 µg•mL-1。抑菌机理研究发现KA08能导致菌丝体内电解质泄漏,细胞膜上麦角甾醇含量显著降低,脂质过氧化产物—丙二醛含量明显升高,并显著抑制菌丝体内蛋白质含量。【结论】农抗KA08对烟草赤星病菌具有显著抑制作用,主要通过抑制菌丝细胞膜上麦角甾醇合成,引发细胞膜脂质过氧化而使病菌细胞膜受损、通透性增加,从而导致菌丝生长受阻。细胞膜是KA08的主要作用位点之一。     

An Endophytic Actinomycete, Streptomyces sp. AOK-30, Isolated from Mountain Laurel and Its Antifungal Activity

To survey endophytic actinomycetes as potential biocontrol agents against fungal diseases of mountain laurel, young plants of mountain laurel (Kalmia latifolia L.) were used as an isolation source. From a total of 73 actinomycetes isolates obtained from leaves, stems and roots of test plants, Streptomyces sp. AOK-30 was selected, because i) it had a broad and intense antimicrobial spectrum against various yeasts and fungal pathogens of Ericaceae, ii) it grew on the multiplication and rooting media for tissue culture for this plant, and iii) the tissue-cultured seedlings that had been treated with this isolate became resistant to Pestalotia disease without any adverse growth effects such as malformation, dwarfing, discolorization and defoliation. This isolate was identified as Streptomyces sp., based on cultural, physiological and morphological characteristics. The present results indicate that isolate AOK-30 is a potential biocontrol agent against diseases of mountain laurel. Received 10 September 2002/ Accepted in revised form 22 November 2002     

S-921菌株发酵液对泡桐腐烂病菌作用的研究

室内实验结果表明,金色链霉菌S-921菌株对泡桐腐烂病菌具有很强的抗生作用,将两者于PDA平板培养基上共同培养8天,出现直径3cm的抑菌圈。经S-921菌株发酵液浸泡处理的泡桐腐烂病病原菌菌丝体,会出现大量液泡,原生质凝集,菌丝尖膨大等异常形态,并失去致病力。S-921菌株发酵液(原液)对泡桐腐烂病的治疗效果,以刮除病班后涂抹发酵液的效果最佳,治愈率达100％。     

农抗120对苹果树腐烂病菌的作用研究

试验表明:农抗120单用,和农抗120加助剂B,对苹果树腐烂病菌菌丝的生长,均有明显的抑制作用。最小抑菌浓度为250μg/ml和125μg/ml;最小杀菌浓度为2000μg/ml和1000μg/ml。其杀菌作用机理是使病菌体的原生质凝集,小分子物质外流,形成大量空泡,菌体重量剧减。病菌在该浓度中浸泡12小时后便失去致病力。涂药试验:先刮除病斑,再用农抗120 1000μg/ml或农抗120加助剂B500μg/ml涂抹处理,16天后复发率仍为零,不刮除病斑,只用刀划刻病枝再涂药液,4天后病斑又继续扩展。     

中生菌素对水稻白叶枯病的防治技术研究

实验证明，中生菌素２００倍液对水稻白叶枯病的防治效果达８９％，相当于化学农药１０％叶枯净１５０倍和２５％川化０１８５００倍药液处理。大田最佳施药时机为病株率５～１０％的发病初期，连续防治２～３次可有效控制病情，千粒重提高１７～１８．７％。秧田于３～４叶期和移栽前５天各喷中生菌素１５０倍液一次，可推迟大田发病期４５天，防效高达９０．５７％。同时节省防治成本７０～８０％。     

牛筋草内生菌砖红链霉菌CSF09杀虫成分鉴定及活性初步研究

为了从微生物中筛选天然杀虫活性化合物,从牛筋草中分离到1株具有杀虫活性的内生放线菌砖红链霉菌Streptomyces lateritius CSF09。采用大孔吸附树脂提取和硅胶柱层析技术,从CSF09菌株发酵液中分离到1个杀虫活性成分,通过质谱和核磁共振波谱鉴定了其化学结构,并初步测定了该化合物对3龄粘虫幼虫和家蝇成虫的杀虫活性。结果表明:该菌株发酵液中的杀虫活性成分为4-甲氧基水杨醛,发酵单位约为13 mg/L;4-甲氧基水杨醛对家蝇和粘虫均表现出一定的触杀活性,其中对家蝇的LD50值为1.51 μg/头,对粘虫毒力较弱,以10 μg/头剂量处理时校正死亡率仅为36.6%;4-甲氧基水杨醛杀虫活性的突出特性是作用迅速,其击倒中时（KT50）大多在5 min以内。研究表明,4-甲氧基水杨醛具有一定的卫生杀虫剂开发应用潜力。     

抗农作物病原真菌的Streptomyces sp.IIR21菌株分离及其克拉维烷基因簇分析

从神农架烟草种植地土壤中分离出1株对7株农作物病原真菌具有抗性活性菌株,命名为ⅡR21菌株。采用琼脂块法测定,该菌株对稻瘟病菌、立枯丝核病菌、腐霉病菌、禾谷镰刀病菌和黄色镰刀病菌具有较强的抑菌活性。对该菌株的生理生化特性和16SrRNA基因序列同源性比对,初步鉴定ⅡR21菌株属于链霉菌属。经过有机溶剂萃取、硅胶柱层析、薄层层析和制备型高效液相色谱对ⅡR21菌株发酵液进行分离纯化,获得了1个纯度为90.34%的具有抗稻瘟病菌168菌株活性的化合物单体。采用高通量测序技术对ⅡR21菌株进行全基因组测序,通过全基因组序列分析,ⅡR21菌株基因组中包含克拉维烷的合成基因簇,并推测出克拉维烷的合成代谢途径。     

Soil suppressiveness and functional diversity of the soil microflora in organic farming systems

Arable fields of 10 organic farms from different locations in The Netherlands were sampled in three subsequent years. The soil samples were analysed for disease suppressiveness against Rhizoctonia solani AG2.2IIIB in sugar beet, Streptomyces scabies in radish and Verticillium longisporum in oilseed rape. In addition, a variety of microbial, chemical and physical soil characteristics were assessed. All data were correlated by multiple regression and multivariate analyses with the objective to find correlations between soil suppressiveness and biotic or abiotic soil characteristics. Significant differences in soil suppressiveness were found between the fields for all three diseases. Multiple regression indicated a significant correlation between suppressiveness against Rhizoctonia and the number of antagonistic Lysobacter spp., as well as with % active fungi and bacterial diversity. Grass-clover stimulated Rhizoctonia suppression as well as the presence of antagonistic Lysobacter spp. (mainly L. antibioticus and L. gummosus) in clay soils. Streptomyces suppression correlated with the number of antagonistic Streptomyces spp., % of active fungi and bacterial population size. The presence of antagonistic Streptomyces spp. correlated with a high fungal/bacterial biomass ratio. Verticillium suppression was only measured in 2004 and 2005, due to the inconsistent suppressiveness along the years. Nevertheless, a significant correlation with pH, potential nitrogen mineralization and bacterial biomass was found. Bacterial and fungal PCR-denaturing gel electrophoresis fingerprinting of bacterial and fungal communities, in general, did not significantly correlate with disease suppression. Highly significant explanatory factors of the composition of the dominating bacterial and fungal populations were % lutum, pH, C/N quotient, biomass and growth rate of bacteria. Additionally, the % of organic matter and years of organic farming were explaining significantly the composition of the bacterial population.Thus, significant correlations between several soil characteristics and suppressiveness of different soil-borne pathogens were found. For two of the three pathogens, suppression correlated with biotic soil characteristics combined with the presence of specific bacterial antagonists. Probably the soil suppressiveness measured in the organic fields is a combined effect of general and specific disease suppression.