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基于培养法和代谢条形码技术对甜菜(Beta vulgaris L.)微生物组进行表征,以高产方式分离具有促植物生长特性的细菌。

Culture-Dependent and Metabarcoding Characterization of the Sugar Beet ( L.) Microbiome for High-Yield Isolation of Bacteria with Plant Growth-Promoting Traits.

作者信息

Krstić Tomić Tamara, Atanasković Iva, Nikolić Ivan, Joković Nataša, Stević Tatjana, Stanković Slaviša, Berić Tanja, Lozo Jelena

机构信息

University of Belgrade, Faculty of Biology, 11000 Belgrade, Serbia.

University of Belgrade, Faculty of Biology, Centre for Biological Control and Plant, Growth Promotion, 11000 Belgrade, Serbia.

出版信息

Microorganisms. 2023 Jun 9;11(6):1538. doi: 10.3390/microorganisms11061538.

Abstract

The diversity of plant-associated bacteria is vast and can be determined by 16S rRNA gene metabarcoding. Fewer of them have plant-beneficial properties. To harness their benefits for plants, we must isolate them. This study aimed to check whether 16S rRNA gene metabarcoding has predictive power in identifying the majority of known bacteria with plant-beneficial traits that can be isolated from the sugar beet ( L.) microbiome. Rhizosphere and phyllosphere samples collected during one season at different stages of plant development were analyzed. Bacteria were isolated on rich unselective media and plant-based media enriched with sugar beet leaves or rhizosphere extracts. The isolates were identified by sequencing the 16S rRNA gene and tested in vitro for their plant-beneficial properties (stimulation of germination; exopolysaccharide, siderophore, and HCN production; phosphate solubilization; and activity against sugar beet pathogens). The highest number of co-occurring beneficial traits was eight, found in isolates of five species: , , , , and . These species were not detected by metabarcoding and have not previously been described as plant-beneficial inhabitants of sugar beets. Thus, our findings point out the necessity of a culture-dependent microbiome analysis and advocate for low-nutrient plant-based media for high-yield isolation of plant-beneficial taxa with multiple beneficial traits. A culture-dependent and -independent approach is required for community diversity assessment. Still, isolation on plant-based media is the best approach to select isolates for potential use as biofertilizers and biopesticides in sugar beet cultivation.

摘要

与植物相关的细菌种类繁多,可通过16S rRNA基因 metabarcoding来确定。其中具有植物有益特性的细菌较少。为了利用它们对植物的益处,我们必须将它们分离出来。本研究旨在检验16S rRNA基因 metabarcoding在鉴定大多数已知具有植物有益性状的细菌方面是否具有预测能力,这些细菌可从甜菜(Beta vulgaris L.)微生物群中分离得到。分析了在一个季节内植物发育不同阶段采集的根际和叶际样本。在富含营养的非选择性培养基以及添加了甜菜叶或根际提取物的植物基培养基上分离细菌。通过对16S rRNA基因进行测序来鉴定分离株,并在体外测试它们的植物有益特性(刺激发芽;胞外多糖、铁载体和HCN的产生;磷溶解;以及对甜菜病原体的活性)。共现有益性状数量最多为八个,在五个物种的分离株中发现:Pseudomonas fluorescens、Bacillus amyloliquefaciens、Burkholderia cepacia、Enterobacter cloacae和Stenotrophomonas maltophilia。这些物种未通过metabarcoding检测到,且此前未被描述为甜菜的植物有益栖息菌。因此,我们的研究结果指出了依赖培养的微生物组分析的必要性,并提倡使用低营养植物基培养基来高产分离具有多种有益性状的植物有益类群。群落多样性评估需要依赖培养和不依赖培养的方法。不过,在植物基培养基上进行分离是选择潜在用作甜菜种植中生物肥料和生物农药的分离株的最佳方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1abe/10302512/2172711fd0b0/microorganisms-11-01538-g001.jpg

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