ANTIMICROBIAL ACTIVITY OF SPECIALISED METABOLITES OF SOIL STREPTOMYCETES-CHITINOLYTIC

G. A. Teregulova, N. A. Manucharova, N. A. Urazbakhtina, N. S. Zhemchuzhina, L. I. Yevtushenko, A. L. Stepanov
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Abstract

One hundred and f f y strains of streptomycetes were isolated from grey and dark grey forest soils, as well as from typical chernozem. T e isolated strains were analyzed in Vitro for antimicrobial activity on nutrient media and in grey forest soil against twenty-three collection pathogenic test cultures of fungi and bacteria. Four biologically active isolates with a wide spectrum of action were identif ed and deposited in the All-Russian Collection of industrial microorganisms of the National Research Centre Kurchatov Institute. T ey were identif ed and deposited under the following numbers : Streptomyces xiamenensis TB ВКПМ Ас-2204, Streptomyces anulatus TG ВКПМ Ас-2203, Streptomyces sindenensis TK ВКПМ Ас-2205, Streptomyces f avovirens TT ВКПМ АС-2202. A study of the ef ect of presowing treatment of wheat seeds by the 15-day cultural liquids of the strains S. xiamenensis TB ВКПМ Ac-2204, S. anulatus TG ВКПМ Ac-2203, S. sindenensis TK ВКПМ Ac-2205 on germination rates and infection with F. graminearum revealed that they inhibited the growth of a pathogenic fungus. Besides, they improved seed vigour and germination of wheat. T e resulting strains of soil actinomycetes can be used in biotechnology with an aim to create the new bioinoculants when dealing with phytopathogenic bacteria and fungi. T e strains can also be used for stimulating the plant growth, as well as for soil bioremediation in organic farming. T rough the application of the HPLC method, specialized antimicrobial metabolites of monosporic strain sus-pensions were identif ed. T e identif ed antibiotics are N-Butylbenzenesulfonamide, 1-(1H-Benzo[d]imidazol-2-yl)ethan-1-ol, 2-[(3S)-1-(Cyclohexylmethyl)-3-pyrrolidinyl]-1H-benzimidazole-5-carbonitrile, Cyclo(leucylprolyl), Cyclo(phenylalanyl-prolyl). T e identif ed antiseptics are Cetrimonium and Carvone. T e identif ed phytohormone is auxin indole-3-acetic acid (IAA). Observation of the dynamics of development of introduced actinobacteria under study in soil samples showed high activity of streptomycetes that use chitin. Analysis of the diversity of the prokaryotic complex of the studied soil samples with the application of the high-throughput sequencing of the conserved region of the 16S rRNA gene method revealed during the introduction of S. xiamenensis TB ВКПМ Ac-2204 that its controlling role in the microbial community is due to its antibiotic-forming activity.
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土壤链霉菌特殊代谢产物--几丁质溶解剂的抗菌活性
从灰色和深灰色森林土壤以及典型的切尔诺泽姆土壤中分离出了一百零一株链霉菌。对分离的菌株在营养培养基上和灰色森林土壤中对 23 种真菌和细菌致病试验培养物的抗菌活性进行了体外分析。鉴定出四种生物活性强、作用范围广的分离菌株,并将其保存在国家研究中心库尔恰托夫研究所的全俄工业微生物保藏中心。经鉴定,这些分离菌的编号如下厦门链霉菌(Streptomyces xiamenensis)TB ВКПМ Ас-2204,阿努拉图斯链霉菌(Streptomyces anulatus)TG ВКПМ Ас-2203,辛登链霉菌(Streptomyces sindenensis)TK ВКПМ Ас-2205,阿沃维伦链霉菌(Streptomyces f avovirens)TT ВКПМ АС-2202。用厦门链霉菌(S. xiamenensis TB)ВКПМ Ac-2204、阿努拉图斯链霉菌(S. anulatus TG)ВКПМ Ac-2203、辛德诺斯链霉菌(S. sindenensis TK)ВКПМ Ас-2205 的 15 天培养液对小麦种子进行播前处理的效果研究。它们抑制了病原真菌的生长。此外,它们还提高了小麦种子的活力和发芽率。所获得的土壤放线菌菌株可用于生物技术,目的是在处理植物病原菌和真菌时创造新的生物接种剂。这些菌株还可用于刺激植物生长,以及有机农业中的土壤生物修复。通过应用高效液相色谱法,鉴定了单孢菌株悬浮液中的特殊抗菌代谢物。鉴定出的抗生素有 N-丁基苯磺酰胺、1-(1H-苯并[d]咪唑-2-基)乙醇、2-[(3S)-1-(环己基甲基)-3-吡咯烷基]-1H-苯并咪唑-5-甲腈、环(亮氨酰)、环(苯丙氨酰)。已确定的杀菌剂是西曲铵和卡文酮。确定的植物激素是吲哚-3-乙酸(IAA)。对土壤样本中引入的放线菌发展动态的观察表明,利用几丁质的链霉菌活性很高。利用 16S rRNA 基因保守区的高通量测序方法分析了所研究土壤样本中原核复合菌的多样性,结果显示,在引进厦门链霉菌 TB ВКПМ Ac-2204 的过程中,其在微生物群落中的控制作用是由于其抗生素形成活性。
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