Inoculation of Herbaspirillum seropedicae strain SmR1 increases biomass in maize roots DKB 390 variety in the early stages of plant development

Cunha, Elisandra Triches da; Pedrolo, Ana Marina; Bueno, Jéssica Cavalheiro Ferreira; Pereira, Tomás Pelizzaro; Soares, Cláudio Roberto Fonsêca Sousa; Arisi, Ana Carolina Maisonnave

doi:10.1007/s00203-022-02986-8

Cited by 3 publications

(6 citation statements)

References 43 publications

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“…Furthermore, Herbaspirillum strains themselves have been shown to be important members of microbiomes. Specifically, Herbaspirillum strains promote plant growth and fix nitrogen for not only maize roots but other plant hosts. − Taken together, Hro has great potential for engineering the microbiome and acting as a potential chassis for understanding plant–microbe interactions.…”

Section: Resultsmentioning

confidence: 99%

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for Studying Plant–Microbe Interactions

van Schaik,

Li,

Cheadle

et al. 2023

ACS Synth. Biol.

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Sustainably enhancing crop production is a global necessity to meet the escalating demand for staple crops while sustainably managing their associated carbon/nitrogen inputs. Leveraging plant-associated microbiomes is a promising avenue for addressing this demand. However, studying these communities and engineering them for sustainable enhancement of crop production have remained a challenge due to limited genetic tools and methods. In this work, we detail the development of the Maize Root Microbiome ToolKit (MRMTK), a rapid Modular Cloning (MoClo) toolkit that only takes 2.5 h to generate desired constructs (5400 potential plasmids) that replicate and express heterologous genes in Enterobacter ludwigii strain AA4 (Elu), Pseudomonas putida strain AA7 (Ppu), Herbaspirillum robiniae strain AA6 (Hro), Stenotrophomonas maltophilia strain AA1 (Sma), and Brucella pituitosa strain AA2 (Bpi), which comprise a model maize root synthetic community (SynCom). In addition to these genetic tools, we describe a highly efficient transformation protocol (107–109 transformants/μg of DNA) 1 for each of these strains. Utilizing this highly efficient transformation protocol, we identified endogenous Expression Sequences (ES; promoter and ribosomal binding sites) for each strain via genomic promoter trapping. Overall, MRMTK is a scalable and adaptable platform that expands the genetic engineering toolbox while providing a standardized, high-efficiency transformation method across a diverse group of root commensals. These results unlock the ability to elucidate and engineer plant–microbe interactions promoting plant growth for each of the 5 bacterial strains in this study.

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Section: Resultsmentioning

confidence: 99%

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for Studying Plant–Microbe Interactions

van Schaik,

Li,

Cheadle

et al. 2023

ACS Synth. Biol.

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“…The diazotrophic bacterium H. seropedicae, can affect the nitrogen metabolism of the plant host [ 25 , 26 , 48 , 49 , 50 ]. Using transcriptomic and proteomic approaches, this study allowed us to better understand how inoculation can modulate nitrogen acquisition and metabolism.…”

Section: Discussionmentioning

confidence: 99%

“…Brazil is the second larger exporter of maize in the world, with around 16 million hectares of cultivated area [ 21 ]. Selected strains of H. seropedicae had shown a promising effect in promoting plant growth and protection, being a potential candidate for microbial formulation in maize and other grasses such as grass pastures [ 22 , 23 , 24 , 25 , 26 , 27 ].…”

Section: Introductionmentioning

confidence: 99%

Multiomic Approaches Reveal Hormonal Modulation and Nitrogen Uptake and Assimilation in the Initial Growth of Maize Inoculated with Herbaspirillum seropedicae

Irineu

Soares

et al. 2022

Plants

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Herbaspirillum seropedicae is an endophytic bacterium that can fix nitrogen and synthesize phytohormones, which can lead to a plant growth-promoting effect when used as a microbial inoculant. Studies focused on mechanisms of action are crucial for a better understanding of the bacteria-plant interaction and optimization of plant growth-promoting response. This work aims to understand the underlined mechanisms responsible for the early stimulatory growth effects of H. seropedicae inoculation in maize. To perform these studies, we combined transcriptomic and proteomic approaches with physiological analysis. The results obtained eight days after inoculation (d.a.i) showed increased root biomass (233 and 253%) and shoot biomass (249 and 264%), respectively, for the fresh and dry mass of maize-inoculated seedlings and increased green content and development. Omics data analysis, before a positive biostimulation phenotype (5 d.a.i.) revealed that inoculation increases N-uptake and N-assimilation machinery through differentially expressed nitrate transporters and amino acid pathways, as well carbon/nitrogen metabolism integration by the tricarboxylic acid cycle and the polyamine pathway. Additionally, phytohormone levels of root and shoot tissues increased in bacterium-inoculated-maize plants, leading to feedback regulation by the ubiquitin-proteasome system. The early biostimulatory effect of H. seropedicae partially results from hormonal modulation coupled with efficient nutrient uptake-assimilation and a boost in primary anabolic metabolism of carbon–nitrogen integrative pathways.

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“…The SynCom strains used in this study were Stenotrophomonas maltophilia AA1 (Sma), The empty vectors used for optimizing the transformation protocol were cloned using the NEB Q5 mutagenesis kit and Gibson cloning using NEB HiFi master mix starting from the plasmid pBBR122. Annotation of pBBR122 using plannotate (75), revealed that the Kan gene split pBBR122's origin of replication into two parts (Supplementary Figure S4). In order to In order to screen the genome of each strain for functional ESs, the base vectors pBBR122-CmR-sfGFP and pBBR122-KanR-sfGFP were created from pBBR122-CmR, pBBR122-KanR and pYTK0047 using Gibson assembly using NEB HiFi Master Mix.…”

Section: Bacterial Strains Media and Growth Conditionsmentioning

confidence: 99%

“…The length of each C plasmid is included in Supplementary Table1. The plasmids that were used to clone the plasmids created in this study can be found in Supplementary TableS3(20,55,(74)(75)(76)(77)(78).…”

mentioning

confidence: 99%

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for studying Plant-Microbe Interactions

Schaik

Cheadle

et al. 2023

Preprint

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Sustainably enhancing crop production is a necessity given the increasing demands for staple crops and their associated carbon/nitrogen inputs. Plant-associated microbiomes offer one avenue for addressing this demand; however, studying these communities and engineering them has remained a challenge due to limited genetic tools and methods. In this work, we detail the development of the Maize Root ToolKit (MRTK); a rapid Modular Cloning (MoClo) toolkit that only takes 2.5 hours to generate desired constructs (5400 potential plasmids) that replicate and express heterologous genes in Enterobacter ludwigii strain AA4 (Elu), Pseudomonas putida AA7 (Ppu), Herbaspirillum robiniae strain AA6 (Hro), Stenotrophomonas maltophilia strain AA1 (Sma) and Brucella pituitosa strain AA2 (Bpi) which comprise a model maize root synthetic community (SynCom). In addition to these genetic tools, we describe a highly efficient transformation protocol (10^7-10^9 transformants/ug of DNA) for each of these strains. Utilizing this highly efficient transformation protocol, we identified endogenous expression sequences for each strain (ES; promoter and ribosomal binding sites) via genomic promoter trapping. Overall, the MRTK is a scalable platform that expands the genetic engineering toolbox while providing a standardized, high efficiency transformation method that can be implemented across a diverse group of root commensals. These results unlock the ability to elucidate and engineer plant-microbe interactions promoting plant growth for each of the 5 bacterial strains in this study.

show abstract

Inoculation of Herbaspirillum seropedicae strain SmR1 increases biomass in maize roots DKB 390 variety in the early stages of plant development

Cited by 3 publications

References 43 publications

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for Studying Plant–Microbe Interactions

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for Studying Plant–Microbe Interactions

Multiomic Approaches Reveal Hormonal Modulation and Nitrogen Uptake and Assimilation in the Initial Growth of Maize Inoculated with Herbaspirillum seropedicae

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for studying Plant-Microbe Interactions

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