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从大麦根际土壤功能宏基因组中鉴定矿质磷酸盐溶解特性。

Characterization of mineral phosphate solubilization traits from a barley rhizosphere soil functional metagenome.

机构信息

Department of Science and Health, Institute of Technology Carlow, Carlow, Ireland.

出版信息

Microbiologyopen. 2013 Oct;2(5):717-24. doi: 10.1002/mbo3.110. Epub 2013 Jul 25.

Abstract

Mineral phosphate solubilization (MPS) microorganisms are important for their provision of orthophosphate anions for plant growth promotion activity in soil. In this study, we applied a functional metagenomic approach to identify this trait directly from the microbiome in barley rhizosphere soil that had not received P fertilizer over a 15-year period. A fosmid system was used to clone the metagenome of which 18,000 clones (~666 Mb of DNA) was screened for MPS. Functional assays and High Performance Liquid Chromatography analysis recognized gluconic acid production and MPS activity in the range 24.8-77.1 mmol/L and 27.6-38.16 μg/mL, respectively, when screened in an Escherichia coli host (at frequency of one MPS-positive clone hit per 114 Mb DNA tested). The MPS clones (with average insert size of ~37 kb) were analysed by 454 Roche sequencing and annotated. A number of genes/operons with homology to Phosphorous (P) uptake, regulatory and solubilization mechanisms were identified, linking the MPS function to the uncultivated microbiome present in barley rhizosphere soil.

摘要

矿质磷酸盐溶解(MPS)微生物对于其在土壤中提供植物生长促进活动所需的正磷酸盐阴离子非常重要。在这项研究中,我们应用了一种功能宏基因组学方法,直接从未施磷肥的大麦根际土壤微生物组中识别出这一特性,该土壤已经连续 15 年未施磷肥。我们使用了一种 fosmid 系统来克隆宏基因组,其中筛选了 18000 个克隆(约 666Mb 的 DNA),以检测 MPS 活性。功能测定和高效液相色谱分析在大肠杆菌宿主中检测到了葡萄糖酸的产生和 MPS 活性,范围分别为 24.8-77.1mmol/L 和 27.6-38.16μg/mL(在测试的每 114Mb DNA 中,一个 MPS 阳性克隆的频率为一次)。对 MPS 克隆(平均插入大小约为 37kb)进行了 454 罗氏测序和注释。鉴定出了一些与磷(P)吸收、调控和溶解机制具有同源性的基因/操纵子,将 MPS 功能与大麦根际土壤中未培养的微生物组联系起来。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6e44/3831634/395d6948c813/mbo30002-0717-f1.jpg

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