骏枣内生生防细菌的分离、筛选与鉴定

选择交城骏枣(Zizyphus jujube cv.Jiaocheng Junzao)作为分离内生细菌的材料,从中共分离到18株内生菌株,运用平板对峙法从中筛选出5株对大枣病原菌及部分植物病原菌有拮抗作用的内生细菌。抑菌试验结果表明:筛选出的5株细菌对刺盘胞菌Colletotrichum gloeosporides、链格孢菌Alternaria alternata、尖孢镰刀菌Fusarium oxysporium、烟草赤星病菌Alternarial longipes、稻瘟病菌Pyricularia oryzae、人参立枯病菌Rhizoctonia solani、人参菌核病菌Sclerotinia schinseng和小麦根腐病菌Bipolaris sorokiniana均有一定的抑菌活性。对这5株内生细菌进行了形态特征观察和生理生化鉴定。     

Effects of Nitrogen and Phosphorus on the Growth of Microorganisms Associated with Apple Replant Disease and on Apple Seedlings Grown in Soil Infested with these Microorganisms*

The growth of fungi causing apple replant disease (ARD) was inhibited by the addition of N and P to the growing medium. The population of bacteria antagonistic to ARD-causing fungi was significantly increased in the growing medium supplemented with N 400 P 400 mg/l or greater. The application of nitrogen alone or in combination with phosphorus to soil infested with fungi or bacteria that cause apple replant disease significantly increased seedling height. The addition of P alone to these infested soils did not have any effect on seedling height. Significant increases in seedling height occurred with N applications when seedlings were grown in soil to which bacteria that are antagonistic to fungi causing ARD had been added. These results suggest that the application of N, with or without P, to apple replant soils may suppress the growth of ARD-causing fungi or bacteria and promote the growth of antagonistic bacteria.     

A new antitumor antibiotic, chounghwamycin A. I. Taxonomy, isolation and characterization

Chounghwamycin A, a new antitumor antibiotic produced by a strain of Streptomyces sp. No. PL-D-5, was isolated and characterized. It appeared to belong to the actinomycin group of antibiotics from physico-chemical studies and has an empirical formula of C63H88N11O21. The antibiotic is extractable into an organic solvent from the fermentation broth, possessing potent antileukemic activity against P388 mouse leukemia in mice and antimicrobial activity against Gram-positive bacteria with MIC values about 0.1-0.4 microgram/ml, but showed no activity on Gram-negative bacteria, yeast and fungi tested.     

地锦草内生细菌多样性、拮抗及促生特性测定

【背景】植物内生细菌既能抑制病原菌对植物的侵染,也具有促生作用。分离具有拮抗和促生活性的内生细菌可为开发微生物菌肥提供理论依据。【目的】筛选内生细菌中的优势拮抗、促生菌种资源。【方法】以地锦草为材料,采用4种分离培养基分离该植物内生细菌,通过形态特征以及16S r RNA基因序列分析,鉴定内生细菌的分类归属。采用平板对峙法,测定内生细菌对棉花立枯丝核菌(Rhizoctonia solani)、小麦赤霉病菌(Fusarium graminearum)、玉米小斑病菌(Bipolaris maydis)的拮抗活性。通过固氮、解磷、产吲哚乙酸(Indole Acetic Acid,IAA)、产铁载体能力等指标初步检测地锦草内生细菌的促生活性。【结果】共分离到133株内生细菌,分属于4门5纲8目13科25属,其中变形菌门(Proteobacteria)为优势门(52.63%),优势属为芽孢杆菌属(Bacillus),占15.79%。发现有8株菌相似性小于98.65%,可能为潜在新物种。拮抗活性结果表明,22株菌有不同程度的抑菌作用,其中菌株DHL56、DHN17、DHP3、DHP8对这3种病原菌...     

Isolation of Bacillus strains from the rhizosphere of cereals and in vitro screening for antagonism against phytopathogenic, food-borne pathogenic and spoilage micro-organisms

Bacillus strains were isolated from the rhizosphere of cereals in order to be used as natural biocontrol agents (BCAs). They were screened for antagonism in vitro against various test micro-organisms. The isolates showing antagonism were identified to species level. A combination of techniques was employed for the isolation of Bacillus species. Using the direct method, only one of the 25 isolates screened showed antagonistic properties. This strain (IFS-01) was identified by means of API test strips and the ATB Plus computer programme. It proved to be Bacillus subtilis and consequently has been designated as Bacillus subtilis IFS-01. This strain produced either a broad spectrum antimicrobial compound or several compounds with different activities. The fungi and Gram-positive bacteria were more sensitive to the antagonistic isolate than the Gram-negative bacteria. A Bacillus strain producing BCAs which can be used as biopesticides or organic preservatives has been isolated and identified.     

Antagonistic effect of Erwinia herbicola on in vitro spore germination and germ tube elongation of Botrytis cinerea and Penicillium expansum

Interaction between Erwinia herbicola and Botrytis cinerea and Penicillium expansum was studied in liquid culture. The results show that the bacteria directly inhibited spore germination of both fungi, especially during the first hours of the paired cultivation. The distinct taxis of bacteria to spores and germ tubes was frequently followed by their lysis. It is likely that bacteria act also by competition for nutrients. The rate of antagonistic activity of the bacteria against both fungi depended on their concentration in the mixture. Formation of chlamydospore-like structures at the apical end of B. cinerea germ tube suggests induction of a defence mechanism of this fungus while in unfavourable conditions.     

Antagonistic activity of Penicillium oxalicum Corrie and Thom,Penicillium decumbens Thom and Trichoderma harzianum Rifai isolates against fungi,bacteria and insects in vitro

The antibiotic activity of 70 isolates belonging to the genera Aspergillus, Penicillium, Fusarium, Alternaria and Trichoderma was tested as preliminary screening. The highest activity was obtained with three Penicillium oxalicum isolates, one Penicillium decumbens isolate and the Trichoderma harzianum isolate. After that, we chose these five isolates in order to carry out other studies with bacteria, fungi and insects. Extracts from these isolates were obtained. The extracts were tested for antibiotic activity with positive results, which implies that metabolite production is involved in this antagonistic effect. The highest activity was shown by T. harzianum and P. oxalicum extracts, but there was high variability among P. oxalicum isolates.     

Antibiotic activity of pyrenomycetes under submerged conditions

Twenty-three pyrenomycete species were tested for antibiotic activity in submerged cultures. When they were screened against bacteria and fungi, 15 showed positive results. Among these,Eutypa acharii, Diaporthe pustulata, Melanconis flavovirens andCamarops microspora were quite promising against bacteria and/or fungi. An antibacterial antibiotic fromCamarops microspora was partially purified and characterized. The author is thankful to UNESCO for the fellowship granted for this work (Long-term postgraduate course on Modern Problems in Biology).     

Characterization and partial purification of a broad spectrum antibiotic AS-48 produced by Streptococcus faecalis

Streptococcus faecalis S-48 produces a broad spectrum antibiotic, active against Gram-positive and Gram-negative bacteria. This substance is produced in solid and liquid media and also in a defined basal medium. It is sensitive to protease, pronase, or trypsin, heating at 70 degrees C, and alkaline pH, but resistant to treatment with lipase, lysozyme, alkaline phosphatase, DNAase, RNAase, acidic or neutral pHs, and also lower temperatures (60 degrees C). Several organic solvents cause precipitation, but not inactivation. This antibiotic has been partially purified by gel filtration and further ion-exchange chromatography. Its molecular weight has been estimated close to 2000. The biological activity of this antagonistic substance against the selected indicator strains, Streptococcus faecalis S-47 and Escherichia coli U-9, is bactericidal. The characterization of this substance, initially classified as a bacteriocin, indicates that it is an antibiotic of peptidic nature. The significance of antibiotic occurrence in group D of the genus Streptococcus is also discussed.     

Target Range of Zwittermicin A, an Aminopolyol Antibiotic from Bacillus cereus

Zwittermicin A is a novel antibiotic produced by Bacillus cereus UW85, which suppresses certain plant diseases in the laboratory and in the field. We developed a rapid method for large-scale purification of zwittermicin A and then studied the in vitro activity of zwittermicin A against bacteria, fungi, and protists. Zwittermicin A was highly active against the Oomycetes and their relatives, the algal protists, and had moderate activity against diverse Gram-negative bacteria and certain Gram-positive bacteria as well as against a wide range of plant pathogenic fungi. Zwittermicin A was more active against bacteria and fungi at pH 7–8 than at pH 5–6. When zwittermicin A was combined with kanosamine, another antibiotic produced by B. cereus, the two acted synergistically against Escherichia coli and additively against Phytophthora medicaginis, an Oomycete. The results indicate that there are diverse potential applications of this new class of antibiotic. Received: 1 December 1997 / Accepted: 9 January 1998     

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。     

Thermomonospora sp. T-SA-125 and its production of a growth promoting antibiotic

Thermomonospora sp. T-SA-125 is a true thermophilic actinomycete isolated from a soil sample collected from the Saudi Arabian desert. It is characterized by the formation of single spores at the tips of dichotomously branched aerial mycelium and differs from Thermomonospora curvata and T. viridis in certain aspects. It produces a basic water-soluble antibiotic which is active against Gram-positive bacteria, moderately active against Gram-negative bacteria and inactive against fungi. At high concentrations, this antibiotic, stimulated the growth of both Hordeum coleoptile and lettuce hypocotyl.     

苦豆子根瘤内生细菌分离及其对棉花枯萎病的生物防治效果测定

对从塔里木盆地苦豆子中分离得到的内生细菌进行皿内涂布拮抗实验、对峙培养法拮抗实验、胞外分泌物拮抗性测定和盆栽控病实验等研究,结果表明塔里木盆地苦豆子根瘤中存在大量的棉花枯萎病菌拮抗性内生细菌。皿内涂布法筛选结果表明60株苦豆子根瘤内生细菌中有48株相对抑菌率超过50%。对峙培养法对48株拮抗性内生细菌进一步筛选的结果表明,对棉花枯萎病菌抑菌距离超过20 mm的菌株有40株。40株拮抗性内生细菌胞外分泌物对棉花枯萎病菌的抑菌距离超过5 mm的有26株。KDRE12和KDRE41对棉花枯萎病的盆栽控病平均防效分别为67.11%和72.65%,具有应用潜力。     

A new antifungal antibiotic produced byStreptomyces galbus

A streptomycete producing an antibiotic having antifungal and antibacterial activity was isolated from a soil sample of West Bengal. It was characterized and identified as Streptomyces galbus. The antibiotic was isolated from the fermented broth by treatment with activated charcoal and purified by chromatography on alumina and paper. It is a colourless, odourless, hygroscopic, amorphous compound. Although homogeneous by paper and thin-layer chromatography, the possibility of the presence of two components was indicated by gel filtration. Its physico-chemical characteristics and UV-absorption spectrum suggest a non-polyenic nature. It is active against a wide variety of fungi and bacteria. It has some phytotoxic effect, but is relatively non-toxic to rats.     

Cytosporone E: racemic synthesis and preliminary antibacterial testing

The antibiotic cytosporone E (isolated from the broth of the endophytic fungi CR 200 (Cytospora sp.) and CR 146 (Diaporthe sp.)) was synthesized as a racemic mixture. The key step in the synthesis is the Meyers ortho-alkylation of a chiral aromatic oxazoline. Preliminary antibiotic activity shows antibiosis against Gram-positive bacteria but not Gram-negative bacteria as previously reported.     

Covalently linked kanamycin – Ciprofloxacin hybrid antibiotics as a tool to fight bacterial resistance

To address the growing problem of antibiotic resistance, a set of 12 hybrid compounds that covalently link fluoroquinolone (ciprofloxacin) and aminoglycoside (kanamycin A) antibiotics were synthesized, and their activity was determined against both Gram-negative and Gram-positive bacteria, including resistant strains. The hybrids were antagonistic relative to the ciprofloxacin, but were substantially more potent than the parent kanamycin against Gram-negative bacteria, and overcame most dominant resistance mechanisms to aminoglycosides. Selected hybrids were 42–640 fold poorer inhibitors of bacterial protein synthesis than the parent kanamycin, while they displayed similar inhibitory activity to that of ciprofloxacin against DNA gyrase and topoisomerase IV enzymes. The hybrids showed significant delay of resistance development in both E. coli and B. subtilis in comparison to that of component drugs alone or their 1:1 mixture. More generally, the data suggest that an antagonistic combination of aminoglycoside-fluoroquinolone hybrids can lead to new compounds that slowdown/prevent the emergence of resistance.     

Interactions between anaerobic cellulolytic bacteria and fungi in the presence of Methanobrevibacter smithii

A mixed inoculum of cellulolytic rumen bacteria depressed straw degradation by a mixed culture of cellulolytic fungi grown in the presence of Methanobrevibacter smithii. The inhibitory effect appeared to be caused by Ruminococcus albus strain JI and R. flavefaciens strain 007. Ruminococcus albus strain J1 also depressed straw degradation by the fungi, but R. albus strain SY3 and three strains of Bacteroides (Fibrobacter) succinogenes tested showed little or no inhibitory activity. It seems that some ruminococci show competitive or antagonistic activity towards certain rumen fungi.     

Production of heptaene antifungal antibiotic by Streptomyces purpeofuscus CM 1261

A strain of Streptomyces purpeofuscus CM 1261 isolated from a sample of compost collected locally was found to possess strong antagonistic activity against 4 human pathogenic fungi i.e., Candida albicans, Aspergillus niger, Microsporum gypseum and Trichophyton sp. The active antifungal compound produced by it was found to be a heptaene group of polyene antifungal antibiotic.     

Antagonism between Bacteria and Fungi on Decomposing Aquatic Plant Litter

Bacterial and fungal decomposers of aquatic plant litter may exhibit either synergistic or antagonistic interactions, which are likely to influence microbial growth as well as the decomposition of litter and, eventually, the carbon metabolism of aquatic systems. To elucidate such interactions, we inoculated decomposing Phragmites culms in microcosms with fungal isolates and with natural communities of bacteria and fungi in different combinations. The development of fungal and bacterial biomass and the carbon dynamics were studied during several months of degradation. The results show a bilateral antagonistic relationship between bacteria and fungi. After 3 months, fungal biomass accumulation was approximately 12 times higher in the absence than in the presence of bacteria. Bacterial biomass accumulation was about double in the absence of fungi compared to when fungi were present. Similar interactions developed between a natural assemblage of bacteria and five different fungal strains isolated from Phragmites litter (three identified hyphomycetes and two unidentified strains). Despite the great difference in biomass development between the treatments, the carbon metabolism was similar regardless of whether fungi and/or bacteria were present alone or in coexistence. We suggest that the antagonism between bacteria and fungi is an important controlling factor for microbial colonization and growth on aquatic plant litter.     

Antagonism between bacteria and fungi: substrate competition and a possible tradeoff between fungal growth and tolerance towards bacteria

Bacteria and fungi often share a common substrate, and their spatial proximity in many environments has lead to either synergistic or antagonistic interactions. In this paper, the interaction of bacterial and fungal decomposers from an aquatic environment was studied. We found indications of a tradeoff between fungal growth and tolerance towards bacteria. Fungal strains growing best in absence of bacteria were most severely affected by bacterial presence, while those less suppressed during co-existence with bacteria had lower maximal growth rates in bacterial absence. Additionally, we show that the antagonism between bacteria and fungi is connected to competition for substrate, but that this competition can be drastically altered if fungi are given an opportunity to establish before inoculation of bacteria. Established fungi out-competed bacteria, and gained higher biomass than in simultaneously inoculated treatments with higher substrate concentrations.