Antimicrobial Activity of Specialized Metabolites of Soil Chitinolytic Streptomycetes

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-fifty strains of streptomycetes have been isolated from gray and dark gray forest soils, as well as from typical chernozem. The isolated strains were analyzed in vitro for antimicrobial activity on nutrient media and in gray forest soil against 23 collection pathogenic test cultures of fungi and bacteria. Four biologically active isolates with a wide action spectrum have been identified and deposited at the All-Russia Collection of Industrial Microorganisms of the National Research Center “Kurchatov Institute” under the following numbers: Streptomyces xiamenensis TB VKPM As-2204, Streptomyces anulatus TG VKPM As-2203, Streptomyces sindenensis TK VKPM As-2205, and Streptomyces flavovirens TT VKPM As-2202. A study of the effect of presowing treatment of wheat seeds with 15-day culture liquids of S. xiamenensis TB VKPM As-2204, S. anulatus TG VKPM As-2203, and S. sindenensis TK VKPM As-2205 for germination rates and levels of infection with F. graminearum has revealed that they inhibited the growth of a pathogenic fungus and improved the seed vigor and germination of wheat. The resulting strains of soil actinomycetes can be used in biotechnology for creating new bioinoculants to combat phytopathogenic bacteria and fungi. The strains can also be used for stimulating the plant growth, as well as for soil bioremediation in organic farming. Based on the HPLC method (high-performance liquid chromatography), we have identified specialized antimicrobial metabolites of monosporous strain suspensions. The identified 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), and Cyclo(phenylalanyl-prolyl). The identified antiseptics are Cetrimonium and Carvone. The identified phytohormone is auxin indole-3-acetic acid (IAA). Observation of the dynamics of development of the introduced actinobacteria in soil samples has shown a high activity of streptomycetes that use chitin. Analysis of the diversity of the prokaryotic complex of the studied soil samples based on high-throughput sequencing of the conserved region of the 16S rRNA gene has revealed its controlling role in the microbial community during the introduction of S. xiamenensis TB VKPM As-2204, which is determined by its antibiotic-forming activity.

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土壤甲壳素溶解链霉菌特殊代谢物的抗菌活性
摘要 从灰色和深灰色的森林土壤以及典型的 Chernozem 中分离出了 150 株链霉菌。对分离出的菌株进行了体外抗菌活性分析,以确定其在营养培养基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。已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,编号如下:厦门链霉菌(Streptomyces xiamenensis)TB VKPM As-2204,安努拉链霉菌(Streptomyces anulatus)TG VKPM As-2203,辛登链霉菌(Streptomyces sindenensis)TK VKPM As-2205,黄链霉菌(Streptomyces flavovirens)TT VKPM As-2202。研究发现,用厦门链霉菌 TB VKPM As-2204、安氏链霉菌 TG VKPM As-2203 和辛登链霉菌 TK VKPM As-2205 的 15 天培养液对小麦种子进行播前处理,可抑制病原真菌的生长,提高小麦的种子活力和发芽率。由此产生的土壤放线菌菌株可用于生物技术,以创造新的生物接种剂来对抗植物病原菌和真菌。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。根据高效液相色谱法(HPLC),我们鉴定出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)和环(苯丙氨酰)。已鉴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。通过对 16S rRNA 基因保守区的高通量测序,对所研究土壤样本中原核复合菌的多样性进行了分析,结果表明,在引进 S. xiamenensis TB VKPM As-2204 的过程中,原核复合菌在微生物群落中起到了控制作用,这是由其抗生素形成活性决定的。
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