| 孙煜炜,蒋林,李蕾,等.海绵共附生菌Streptomyces olivaceus LHW2444的分离鉴定及其抗青枯菌活性代谢产物研究[J].中国海洋药物,2023,42(4):1-7. |
| 海绵共附生菌Streptomyces olivaceus LHW2444的分离鉴定及其抗青枯菌活性代谢产物研究 |
| Marine sponge symbiotic Streptomyces olivaceus LHW2444: isolation, identification, and active metabolites against Ralstonia solanacearum |
| 投稿时间：2022-03-29 修订日期：2022-04-18 |
| DOI：10.13400/j.cnki.cjmd.2023.04.012 |
| 中文关键词: 青枯菌 海绵 放线菌 代谢产物 抗菌活性 |
| English Keywords:Ralstonia solanacearum marine sponge actinomycetes metabolites antibacterial activity |
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| 中文摘要: |
| 目的 从海绵中分离培养放线菌,筛选具有抗青枯菌活性的菌株,分离鉴定活性代谢产物。方法 以来源于南海西沙永兴岛附近海域的海绵Leucetta chagosensis为实验材料,采用三种选择性分离培养基分离海绵的共附生放线菌,利用16S rRNA序列分析对各菌株进行种属鉴定。对各菌株的发酵提取物进行抗青枯菌活性筛选,采用硅胶柱层析、Sephadex LH-20凝胶柱层析和高效液相色谱法对筛选到的活性菌株的发酵产物进行分离、纯化,运用核磁共振(NMR)、质谱(MS)等手段,鉴定化合物的结构。采用微量稀释法测定化合物的最小抑菌浓度(MIC)。结果 分离培养海绵共附生放线菌16株,筛选得到一株具有较好抗青枯菌活性的菌株Streptomyces olivaceus LHW2444,并从该菌株的发酵产物中分离鉴定2个吡咯类化合物、1个苯并二恶茂类化合物、2个吡喃酮类化合物,分别为pyrrole-2-carboxamide(1)、pyrrole-2-carboxylic acid(2)、1,3-benzodioxole-2-one-4-carboxylamide(3)、germicidin B(4)、germicidin C(5),化合物1-5为首次从该种属放线菌分离得到。首次发现pyrrole-2-carboxylic acid对青枯菌具有较强抑制作用,MIC值为8 μg/mL。结论 海绵共附生菌Streptomyces olivaceus LHW2444是潜在的植物青枯病生防菌,pyrrole-2-carboxylic acid是抗青枯菌的活性代谢产物。 |
| English Summary: |
| Objective To isolate and culture actinomycetes from marine sponges, screen for strains with activities against Ralstonia solanacearum, and identify the active metabolites. Methods The marine sponge Leucetta chagosensis from adjacent waters of the Yongxing Island of Xisha, South China Sea was used as experimental material. Actinomycetes isolated by using three selective isolation culture medium were identified by 16S rRNA sequence analysis. The fermentation broth of each strain was screened for activities against R. solanacearum. Silica gel column chromatography, Sephadex LH-20 gel column chromatography and high performance liquid chromatography were used to isolate and purify compounds from the fermented products of the active strain. The structures of the compounds were identified by means of nuclear magnetic resonance (NMR) and mass spectrometry (MS). The minimal inhibitory concentration (MIC) of the compounds were determined by microdilution method. Results 16 actinomycetes strains were isolated and cultured, and Streptomyces olivaceus LHW2444 with activity against R. solanacearum was obtained. Two pyrroles, one benzodioxole and two pyrones were isolated and identified from fermentation products of the strain LHW2444, and their structures were elucidated as pyrrole-2-carboxamide (1), pyrrole-2-carboxylic acid (2), 1,3-benzodioxole-2-one-4-carboxylamide (3), germanidin B (4) and germicidin C (5). Compounds 1-5 were isolated from Streptomyces olivaceus for the first time. Pyrrole-2-carboxylic acid was found to show strong inhibitory effect on R. solanacearum for the first time, with a MIC value of 8 μg/mL. Conclusion Marine sponge symbiotic Streptomyces olivaceus LHW2444 is a potential biocontrol actinomycetes against plant bacterial wilt, and pyrrole-2-carboxylic acid is an active metabolite against R. solanacearum. |
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