Bipartite interactions, antibiotic production and biosynthetic potential of the Arabidopsis leaf microbiome.

Bipartite interactions, antibiotic production and biosynthetic potential of the Arabidopsis leaf microbiome.
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DOI:
10.1038/s41564-018-0200-0
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发表时间:
2018-08
影响因子:
28.3
通讯作者:
Vorholt JA
Vorholt JA
中科院分区:
生物学1区
文献类型:
--
作者:
Helfrich EJN;Vogel CM;Ueoka R;Schäfer M;Ryffel F;Müller DB;Probst S;Kreuzer M;Piel J;Vorholt JA

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植物在系统发育上具有多样性的微生物会影响植物的生长和健康。最近已经建立了具有代表性的模式植物细菌分离株的基因组测序培养集,包括拟南芥。这些资源提供了系统的相互作用筛选与基因组挖掘相结合的机会,以发现未确定特征的天然产品。在这里,我们报告了224个菌株的生物合成潜力,这些菌株是从拟南芥中分离出来的。基因组挖掘发现了1000多个预测的天然产物生物合成基因簇(BGC),其中数百个与MIBiG的特征BGC数据库相比是未知的。对于功能验证,我们使用了高通量筛选方法来监测超过50,000个二元菌株组合。我们观察到了725个抑制相互作用,其中26个菌株起到了大部分作用。结合成像质谱学和生物活性导向分离最有效的抑制剂,富含BGC的短杆菌。Leaf182,揭示了三种不同的天然产品支架,它们有助于观察到的抗生素活性。此外,基于基因组挖掘的策略导致了一种由反式AT聚酮合成酶衍生的抗生素Macbrevin的分离,它显示了前所未有的天然产品结构。我们的发现表明,叶圈是鉴定抗生素和具有不寻常支架的天然产物的有价值的环境。
Plants are colonized by phylogenetically diverse microorganisms that affect plant growth and health. Representative genome-sequenced culture collections of bacterial isolates from model plants, including Arabidopsis thaliana, have recently been established. These resources provide opportunities for systematic interaction screens combined with genome mining to discover uncharacterized natural products. Here, we report on the biosynthetic potential of 224 strains isolated from the A. thaliana phyllosphere. Genome mining identified more than 1,000 predicted natural product biosynthesis gene clusters (BGCs), hundreds of which are unknown compared to the MIBiG database of characterized BGCs. For functional validation, we used a high-throughput screening approach to monitor over 50,000 binary strain combinations. We observed 725 inhibitory interactions, with 26 strains contributing to the majority of these. A combination of imaging mass spectrometry and bioactivity-guided fractionation of the most potent inhibitor, the BGC-rich Brevibacillus sp. Leaf182, revealed three distinct natural product scaffolds that contribute to the observed antibiotic activity. Moreover, a genome mining-based strategy led to the isolation of a trans-AT polyketide synthase-derived antibiotic, macrobrevin, which displays an unprecedented natural product structure. Our findings demonstrate that the phyllosphere is a valuable environment for the identification of antibiotics and natural products with unusual scaffolds.
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