利用人造生态系统(EcoFABs)评估 Brachypodium distachyon 根瘤中的水平基因转移。

IF 3.9 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Applied and Environmental Microbiology Pub Date : 2024-11-20 Epub Date: 2024-11-04 DOI:10.1128/aem.01505-24
Shweta Priya, Silvia Rossbach, Thomas Eng, Hsiao-Han Lin, Peter F Andeer, Jenny C Mortimer, Trent R Northen, Aindrila Mukhopadhyay
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引用次数: 0

摘要

水平基因转移(HGT)是根瘤菌圈微生物之间基因转移的主要过程。然而,由于模拟根瘤菌圈内条件的复杂性,研究 HGT 仍具有挑战性。人造生态系统(EcoFABs)已被用于研究植物相关环境中的若干复杂过程。在这里,我们展示了 EcoFABs 是研究和测量根瘤菌圈中 HGT 频率的有效工具。我们首次展示了通过三亲共轭体系在 Brachypodium distachyon 根瘤菌圈的 EcoFAB 中进行基因转移的情况,该系统使用假单胞菌 Putida KT2440 作为供体和受体细菌菌株,供体含有可移动且不可自我传播的质粒。我们观察到,质粒在根瘤菌圈中的转移频率可能取决于植物的发育阶段以及根渗出物的成分和数量。质粒转移的频率也随着供体细胞数量的增加而增加。我们展示了质粒从 P. putida 转移到另一种 B. distachyon 根定植菌 Burkholderia sp. OAS925 的过程,显示了根圈微生物群落中的 HGT。环境胁迫也影响了根圈中不同物种和菌属之间 HGT 的速率和效率。这项研究提供了一个稳健的工作流程,当工程质粒可能被引入田间或其他植物相关环境时,可用于评估工程质粒在根瘤菌圈中的转移。 重要意义我们报告了使用 EcoFABs 研究根瘤菌圈环境中的 HGT 过程。它凸显了 EcoFABs 在重现动态根瘤菌圈条件方面的潜力,以及其在研究植物与微生物相互作用方面的多功能性。这项研究还强调了研究影响 HGT 频率的参数的重要性。植物发育阶段、营养条件、供体细胞数量和环境压力等因素都会影响根圈微生物群落内的基因转移。这项研究为今后研究了解工程质粒在田间环境中的命运和移动铺平了道路。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Assessing horizontal gene transfer in the rhizosphere of Brachypodium distachyon using fabricated ecosystems (EcoFABs).

Horizontal gene transfer (HGT) is a major process by which genes are transferred between microbes in the rhizosphere. However, examining HGT remains challenging due to the complexity of mimicking conditions within the rhizosphere. Fabricated ecosystems (EcoFABs) have been used to investigate several complex processes in plant-associated environments. Here we show that EcoFABs are efficient tools to examine and measure HGT frequency in the rhizosphere. We provide the first demonstration of gene transfer via a triparental conjugation system in the Brachypodium distachyon rhizosphere in an EcoFAB using Pseudomonas putida KT2440 as both donor and recipient bacterial strain with the donor containing a mobilizable and non-self-transmissible plasmid. We observed that the frequency of plasmid transfer in the rhizosphere is potentially dependent on the plant developmental stage and the composition and amount of root exudates. The frequency of plasmid transfer also increased with higher numbers of donor cells. We demonstrate the transfer of plasmid from P. putida to another B. distachyon root colonizer, Burkholderia sp. OAS925, showing HGT within a rhizosphere microbial community. Environmental stresses also influenced the rate and efficiency of HGT in the rhizosphere between different species and genera. This study provides a robust workflow to evaluate transfer of engineered plasmids in the rhizosphere when such plasmids are potentially introduced in a field or other plant-associated environments.IMPORTANCEWe report the use of EcoFABs to investigate the HGT process in a rhizosphere environment. It highlights the potential of EcoFABs in recapitulating the dynamic rhizosphere conditions as well as their versatility in studying plant-microbe interactions. This study also emphasizes the importance of studying the parameters impacting the HGT frequency. Several factors such as plant developmental stages, nutrient conditions, number of donor cells, and environmental stresses influence gene transfer within the rhizosphere microbial community. This study paves the way for future investigations into understanding the fate and movement of engineered plasmids in a field environment.

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来源期刊
Applied and Environmental Microbiology
Applied and Environmental Microbiology 生物-生物工程与应用微生物
CiteScore
7.70
自引率
2.30%
发文量
730
审稿时长
1.9 months
期刊介绍: Applied and Environmental Microbiology (AEM) publishes papers that make significant contributions to (a) applied microbiology, including biotechnology, protein engineering, bioremediation, and food microbiology, (b) microbial ecology, including environmental, organismic, and genomic microbiology, and (c) interdisciplinary microbiology, including invertebrate microbiology, plant microbiology, aquatic microbiology, and geomicrobiology.
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