Factors governing attachment of Rhizobium leguminosarum to legume roots at acid, neutral, and alkaline pHs.

IF 5 2区 生物学 Q1 MICROBIOLOGY mSystems Pub Date : 2024-09-17 Epub Date: 2024-08-21 DOI:10.1128/msystems.00422-24
Jack D Parsons, Clare R Cocker, Alison K East, Rachel M Wheatley, Vinoy K Ramachandran, Farnusch Kaschani, Markus Kaiser, Philip S Poole
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Abstract

Rhizobial attachment to host legume roots is the first physical interaction of bacteria and plants in symbiotic nitrogen fixation. The pH-dependent primary attachment of Rhizobium leguminosarum biovar viciae 3841 to Pisum sativum (pea) roots was investigated by genome-wide insertion sequencing, luminescence-based attachment assays, and proteomic analysis. Under acid, neutral, or alkaline pH, a total of 115 genes are needed for primary attachment under one or more environmental pH, with 22 genes required for all. These include components of cell surfaces and membranes, together with enzymes that construct and modify them. Mechanisms of dealing with stress also play a part; however, exact requirements vary depending on environmental pH. RNASeq showed that knocking out the two transcriptional regulators required for attachment causes massive changes in the bacterial cell surface. Approximately half of the 54 proteins required for attachment at pH 7.0 have a role in the later stages of nodule formation. We found no evidence for a single rhicadhesin responsible for alkaline attachment, although sonicated cell surface fractions inhibited root attachment at alkaline pH. Our results demonstrate the complexity of primary root attachment and illustrate the diversity of mechanisms involved.

Importance: The first step by which bacteria interact with plant roots is by attachment. In this study, we use a combination of insertion sequencing and biochemical analysis to determine how bacteria attach to pea roots and how this is influenced by pH. We identify several key adhesins, which are molecules that enable bacteria to stick to roots. This includes a novel filamentous hemagglutinin which is needed at all pHs for attachment. Overall, 115 proteins are required for attachment at one or more pHs.

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豆科根瘤菌在酸性、中性和碱性 pH 值条件下附着在豆科植物根部的因素。
根瘤菌对寄主豆科植物根部的附着是细菌和植物在共生固氮过程中的第一次物理相互作用。通过全基因组插入测序、基于发光的附着试验和蛋白质组分析,研究了豆科根瘤菌(Rhizobium leguminosarum biovar viciae 3841)与豌豆(Pisum sativum)根的初级附着对 pH 值的依赖性。在酸性、中性或碱性 pH 值条件下,一种或多种环境 pH 值条件下的原生附着共需要 115 个基因,其中 22 个基因是全部需要的。这些基因包括细胞表面和细胞膜的组成成分,以及构建和改造细胞表面和细胞膜的酶。处理压力的机制也发挥了作用;不过,具体要求因环境 pH 值的不同而不同。RNASeq 研究表明,敲除附着所需的两个转录调节因子会导致细菌细胞表面发生巨大变化。在 pH 值为 7.0 时附着所需的 54 个蛋白质中,约有一半在结核形成的后期阶段发挥作用。尽管超声处理的细胞表面分馏物会抑制根在碱性 pH 值下的附着,但我们没有发现任何证据表明有单一的根瘤鞘蛋白负责碱性附着。我们的研究结果证明了原生根附着的复杂性,并说明了相关机制的多样性:重要意义:细菌与植物根系相互作用的第一步是附着。在这项研究中,我们结合插入测序和生化分析,确定了细菌如何附着在豌豆根上,以及这种附着如何受到 pH 值的影响。我们发现了几种关键的粘附素,它们是使细菌能够粘附到根部的分子。其中包括一种新型丝状血凝素,它在所有 pH 值下都需要进行附着。总体而言,有 115 种蛋白质需要在一种或多种 pH 值条件下进行附着。
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来源期刊
mSystems
mSystems Biochemistry, Genetics and Molecular Biology-Biochemistry
CiteScore
10.50
自引率
3.10%
发文量
308
审稿时长
13 weeks
期刊介绍: mSystems™ will publish preeminent work that stems from applying technologies for high-throughput analyses to achieve insights into the metabolic and regulatory systems at the scale of both the single cell and microbial communities. The scope of mSystems™ encompasses all important biological and biochemical findings drawn from analyses of large data sets, as well as new computational approaches for deriving these insights. mSystems™ will welcome submissions from researchers who focus on the microbiome, genomics, metagenomics, transcriptomics, metabolomics, proteomics, glycomics, bioinformatics, and computational microbiology. mSystems™ will provide streamlined decisions, while carrying on ASM''s tradition of rigorous peer review.
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