Plant-Microbe Symbiosis
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Rhizobia–diatom symbiosis fixes missing nitrogen in the ocean

Nitrogen (N2) fixation in oligotrophic surface waters is the main source of new nitrogen (N) to the ocean1 and plays a key role in fueling the biological carbon pump2. Oceanic N2 fixation is almost exclusively attributed to cyanobacteria, even though genes encoding nitrogenase, the enzyme fixing N2 into ammonia, are widespread among marine bacteria and archaea3-5. Little is known about these non-cyanobacterial N2-fixers and direct proof that they can fix N in the ocean is missing. Here we report the discovery of a non-cyanobacterial N2-fixing symbiont, Candidatus Tectiglobus diatomicola, which provides its diatom host with fixed-N in return for photosynthetic carbon. The N2-fixing symbiont belongs to the order Rhizobiales and its association with a unicellular diatom expands the known hosts for this order beyond the well-known N2-fixing rhizobia-legume symbioses on land6. Our results show that the rhizobia-diatom symbiosis can contribute as much fixed-N as cyanobacterial N2-fixers in the tropical North Atlantic, and that they may be responsible for N2 fixation in the vast regions of the ocean where cyanobacteria are too rare to account for the measured rates.
Jean-Michel Ané's insight:

Eew... The use of the term "rhizobia" here seems inappropriate to me. To call something "rhizobia" one need to demonstrate that it forms a root nodule symbiosis. Not all Rhizobiales (like Agrobacterium) are rhizobia!

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Plant-Microbe Symbiosis
Beneficial associations between plants and microbes
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Dissection of rhizosphere microbiome and exploiting strategies for sustainable agriculture

The rhizosphere microbiome plays critical roles in plant growth and provides promising solutions for sustainable agriculture. While the rhizosphere microbiome frequently fluctuates with the soil environment, recent studies have demonstrated that a small proportion of the microbiome is consistently assembled in the rhizosphere of a specific plant genotype regardless of the soil condition, which is determined by host genetics. Based on these breakthroughs, which involved exploiting the plant-beneficial function of the rhizosphere microbiome, we propose to divide the rhizosphere microbiome into environment-dominated and plant genetic-dominated components based on their different assembly mechanisms. Subsequently, two strategies to explore the different rhizosphere microbiome components for agricultural production are suggested, that is, the precise management of the environment-dominated rhizosphere microbiome by agronomic practices, and the elucidation of the plant genetic basis of the plant genetic-dominated rhizosphere microbiome for breeding microbiome-assisted crop varieties. We finally present the major challenges that need to be overcome to implement strategies for modulating these two components of the rhizosphere microbiome.

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Prevalence, diversity and applications potential of nodules endophytic bacteria: a systematic review

Legumes are renowned for their distinctive biological characteristic of forming symbiotic associations with soil bacteria, mostly belonging to the Rhizobiaceae familiy, leading to the establishment of symbiotic root nodules. Within these nodules, rhizobia play a pivotal role in converting atmospheric nitrogen into a plant-assimilable form. However, it has been discerned that root nodules of legumes are not exclusively inhabited by rhizobia; non-rhizobial endophytic bacteria also reside within them, yet their functions remain incompletely elucidated. This comprehensive review synthesizes available data, revealing that Bacillus and Pseudomonas are the most prevalent genera of nodule endophytic bacteria, succeeded by Paenibacillus, Enterobacter, Pantoea, Agrobacterium, and Microbacterium. To date, the bibliographic data available show that Glycine max followed by Vigna radiata, Phaseolus vulgaris and Lens culinaris are the main hosts for nodule endophytic bacteria. Clustering analysis consistently supports the prevalence of Bacillus and Pseudomonas as the most abundant nodule endophytic bacteria, alongside Paenibacillus, Agrobacterium, and Enterobacter. Although non-rhizobial populations within nodules do not induce nodule formation, their presence is associated with various plant growth-promoting properties (PGPs). These properties are known to mediate important mechanisms such as phytostimulation, biofertilization, biocontrol, and stress tolerance, emphasizing the multifaceted roles of nodule endophytes. Importantly, interactions between non-rhizobia and rhizobia within nodules may exert influence on their leguminous host plants. This is particularly shown by co-inoculation of legumes with both types of bacteria, in which synergistic effects on plant growth, yield, and nodulation are often measured. Moreover these effects are pronounced under both stress and non-stress conditions, surpassing the impact of single inoculations with rhizobia alone.

Jean-Michel Ané's insight:

Someone should review the number of reviews published on this topic.

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On the path to [Fe-S] protein maturation: A personal perspective

On the path to [Fe-S] protein maturation: A personal perspective | Plant-Microbe Symbiosis | Scoop.it
Azotobacter vinelandii is a genetically tractable Gram-negative proteobacterium able to fix nitrogen (N2) under aerobic growth conditions. This narrative describes how biochemical-genetic approaches using A. vinelandii to study nitrogen fixation led to the formulation of the “scaffold hypothesis” for the assembly of both simple and complex [Fe-S] clusters associated with biological nitrogen fixation. These studies also led to the discovery of a parallel, but genetically distinct, pathway for maturation of [Fe-S] proteins that support central metabolic processes.

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Rhizobial‐induced phosphatase GmPP2C61A positively regulates soybean nodulation

Symbiotic nitrogen fixation (SNF) is crucial for legumes, providing them with the nitrogen necessary for plant growth and development. Nodulation is the first step in the establishment of SNF. However, the determinant genes in soybean nodulation and the understanding of the underlying molecular mechanisms governing nodulation are still limited. Herein, we identified a phosphatase, GmPP2C61A, which was specifically induced by rhizobia inoculation. Using transgenic hairy roots harboring GmPP2C61A::GUS, we showed that GmPP2C61A was mainly induced in epidermal cells following rhizobia inoculation. Functional analysis revealed that knockdown or knock-out of GmPP2C61A significantly reduced the number of nodules, while overexpression of GmPP2C61A promoted nodule formation. Additionally, GmPP2C61A protein was mainly localized in the cytoplasm and exhibited conserved phosphatase activity in vitro. Our findings suggest that phosphatase GmPP2C61A serves as a critical regulator in soybean nodulation, highlighting its potential significance in enhancing symbiotic nitrogen fixation.

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Proxiome assembly of the plant nuclear pore reveals an essential hub for gene expression regulation

The nuclear pore complex (NPC) is vital for nucleocytoplasmic communication. Recent evidence emphasizes its extensive association with proteins of diverse functions, suggesting roles beyond cargo transport. Yet, our understanding of NPC’s composition and functionality at this extended level remains limited. Here, through proximity-labelling proteomics, we uncover both local and global NPC-associated proteome in Arabidopsis, comprising over 500 unique proteins, predominantly associated with NPC’s peripheral extension structures. Compositional analysis of these proteins revealed that the NPC concentrates chromatin remodellers, transcriptional regulators and mRNA processing machineries in the nucleoplasmic region while recruiting translation regulatory machinery on the cytoplasmic side, achieving a remarkable orchestration of the genetic information flow by coupling RNA transcription, maturation, transport and translation regulation. Further biochemical and structural modelling analyses reveal that extensive interactions with nucleoporins, along with phase separation mediated by substantial intrinsically disordered proteins, may drive the formation of the unexpectedly large nuclear pore proteome assembly.

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How Can Fungus In Cameroon Help Farmers?

How Can Fungus In Cameroon Help Farmers? | Plant-Microbe Symbiosis | Scoop.it
A researcher from Cameroon is looking at how fungus could be used as fertilizer to improve plant production for farmers.

Arbuscular mycorrhizal fungi (AMF) supply water, phosphate and nitrogen to the host plant and in receive up to 20% of plant-fixed carbon in return — a useful symbiosis.

Astride Carole Djeuani, a lecturer and researcher at the University of Yaounde in Cameroon says it is important to research the AMF around plant roots, because they can be used as fertilizer to improve plant yields.

"Today the damages caused by the application of chemicals in agriculture are very obvious, so hopefully, the strains that I would have isolated and multiplied after screening tests in the laboratory, will serve as a fertilizer factory that I will make available to farmers," she says, adding that the idea is to add these AMFs with biochar and compost to fertilize the plants.
Jean-Michel Ané's insight:

Inspiring work from @astride_car with @spununderground on arbuscular mycorrhizal fungi highlighted in @Forbes

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Biochar is colonized by select arbuscular mycorrhizal fungi in agricultural soils

Biochar is colonized by select arbuscular mycorrhizal fungi in agricultural soils | Plant-Microbe Symbiosis | Scoop.it
Arbuscular mycorrhizal fungi (AMF) colonize biochar in soils, yet the processes governing their colonization and growth in biochar are not well characterized. Biochar amendment improves soil health by increasing soil carbon, decreasing bulk density, and improving soil water retention, all of which can increase yield and alleviate environmental stress on crops. Biochar is often applied with nutrient addition, impacting mycorrhizal communities. To understand how mycorrhizas explore soils containing biochar, we buried packets of non-activated biochar in root exclusion mesh bags in contrasting agricultural soils. In this greenhouse experiment, with quinoa (Chenopodium quinoa) as the host plant, we tested impacts of mineral nutrient (as manure and fertilizer) and biochar addition on mycorrhizal colonization of biochar. Paraglomus appeared to dominate the biochar packets, and the community of AMF found in the biochar was a subset (12 of 18) of the virtual taxa detected in soil communities. We saw differences in AMF community composition between soils with different edaphic properties, and while nutrient addition shifted those communities, the shifts were inconsistent between soil types and did not significantly influence the observation that Paraglomus appeared to selectively colonize biochar. This observation may reflect differences in AMF traits, with Paraglomus previously identified only in soils (not in roots) pointing to predominately soil exploratory traits. Conversely, the absence of some AMF from the biochar implies either a reduced tendency to explore soils or an ability to avoid recalcitrant nutrient sources. Our results point to a selective colonization of biochar in agricultural soils.

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Paraglomus likes this biochar

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Efficiently recording and processing data from arbuscular mycorrhizal colonization assays using AMScorer and AMReader

Efficiently recording and processing data from arbuscular mycorrhizal colonization assays using AMScorer and AMReader | Plant-Microbe Symbiosis | Scoop.it
Arbuscular mycorrhizal (AM) fungi engage with land plants in a widespread, mutualistic endosymbiosis which provides their hosts with increased access to nutrients and enhanced biotic and abiotic stress resistance. The potential for reducing fertiliser use and improving crop resilience has resulted in rapidly increasing scientific interest. Microscopic quantification of the level of AM colonization is of fundamental importance to this research, however the methods for recording and processing these data are time-consuming and tedious. In order to streamline these processes, we have developed AMScorer, an easy-to-use Excel spreadsheet, which enables the user to record data rapidly during from microscopy-based assays, and instantly performs the subsequent data processing steps. In our hands, AMScorer has more than halved the time required for data collection compared to paper-based methods. Subsequently, we developed AMReader, a user-friendly R package, which enables easy visualization and statistical analyses of data from AMScorer. These tools require only limited skills in Excel and R, and can accelerate research into AM symbioses, help researchers with variable resources to conduct research, and facilitate the storage and sharing of data from AM colonization assays. They are available for download at https://github.com/EJarrattBarnham/AMReader, along with an extensive user manual.

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Les sols et leurs fonctions | Colloques, conférences et débats | Encourager la vie scientifique

Les sols et leurs fonctions | Colloques, conférences et débats | Encourager la vie scientifique | Plant-Microbe Symbiosis | Scoop.it
Les sols sont le support de la vie terrestre et le substrat de la végétation. Ce sont des écosystèmes complexes et fragiles qui contribuent à la qualité de notre environnement. Leur étude est, par essence, pluridisciplinaire et se situe au carrefour de la géologie, de la physique, de la chimie, de la biologie, de l'agriculture et de la climatologie. Leurs caractéristiques physicochimiques et biologiques conditionnent la nature de la végétation, la qualité et le rendement des cultures. Les pratiques de l'agriculture intensive et industrielle les appauvrissent considérablement dans nombre de régions du globe, y compris dans notre pays et il convient d'en prendre conscience et de tenter d'y remédier. Les sols contribuent aussi au stockage et au piégeage du gaz carbonique, au travers de la minéralisation de la matière organique, et sont donc un puits de carbone, mais ils peuvent aussi, dans certaines conditions, en libérer et devenir une source supplémentaire de ce gaz à effet de serre. Leur gestion est donc un facteur important à maitriser dans les efforts pour atténuer le changement climatique .

L'objectif de cette séance, commune avec l'Académie d'agriculture de France est de faire un point des connaissances sur quelques aspects de la science des sols et sur les enjeux qui s'y rattachent.
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Chickpea (Cicer arietinum) PHO1 family members function redundantly in Pi transport and root nodulation 

Phosphorus (P), a macronutrient, plays key roles in plant growth, development, and yield. Phosphate (Pi) transporters (PHTs) and PHOSPHATE1 (PHO1) are central to Pi acquisition and distribution. Potentially, PHO1 is also involved in signal transduction under low P. The current study was designed to identify and functionally characterize the PHO1 gene family in chickpea (CaPHO1s). Five CaPHO1 genes were identified through a comprehensive genome-wide search. Phylogenetically, CaPHO1s formed two clades, and protein sequence analyses confirmed the presence of conserved domains. CaPHO1s are expressed in different plant organs including root nodules and are induced by Pi-limiting conditions. Functional complementation of atpho1 mutant with three CaPHO1 members, CaPHO1, CaPHO1;like, and CaPHO1;H1, independently demonstrated their role in root to shoot Pi transport, and their redundant functions. To further validate this, we raised independent RNA-interference (RNAi) lines of CaPHO1, CaPHO1;like, and CaPHO1;H1 along with triple mutant line in chickpea. While single gene RNAi lines behaved just like WT, triple knock-down RNAi lines (capho1/like/h1) showed reduced shoot growth and shoot Pi content. Lastly, we showed that CaPHO1s are involved in root nodule development and Pi content. Our findings suggest that CaPHO1 members function redundantly in root to shoot Pi export and root nodule development in chickpea.

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Genomic and metabolomic profiling reveal Streptomyces rochei S32 contributes to plant growth by nitrogen fixation and production of bioactive substances

S. rochei S32 significantly improved the growth of wheat and tomato. The shoot length (24.7%) and root length (25.3%) of wheat (400-fold dilution of cell-free fermentation filtrate) were increased, and the root length of tomato (200-fold dilution) was prolonged (40.9%), and the field yield was also increased. S. rochei S32 showed antagonistic activity against multiple pathogenic fungi, especially Macropoma kawatsukai. The bacterial genome contains an 8,041,158-bp chromosome and two plasmids. A total of 7486 annotated genes were classified into 31 Gene Ontology functional categories. Genomic analysis revealed the potential for the production of indole-3-acetic acid, fungal cell wall hydrolases, antibiotics (e.g., candicidin, streptothricin, borrellin, albaflavenone), and siderophores. Thirty-nine phytohormones and 2205 secondary metabolites were detected, including indole-3-acetic acid, phytosphingosine, acivicin, and corynebactin. Normal bacterial growth occurred on a nitrogen-free medium.

Jean-Michel Ané's insight:

The evidence supporting nitrogen fixation in this bacterium is quite weak...

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Soybean CEP6 Signaling Peptides Positively Regulate Nodulation

Soybean CEP6 Signaling Peptides Positively Regulate Nodulation | Plant-Microbe Symbiosis | Scoop.it
Nodulation is the most efficient nitrate assimilation system in the ecosystem, while excessive fertilization has an increased nitrate inhibition effect; deciphering the nitrate signal transduction mechanism in the process is of the utmost importance. In this study, genome-wide analyses of the GmCEP genes were applied to identify nodulation-related CEP genes; 22 GmCEP family members were identified, while GmCEP6 was mainly expressed in nodules and significantly responded to nitrate treatment and rhizobium infection, especially in later stages. Overexpression and CRISPR-Cas9 were used to validate its role in nodulation. We found that GmCEP6 overexpression significantly increased the nodule number, while GmCEP6 knock-out significantly decreased the nodule number, which suggests that GmCEP6 functions as a positive regulator in soybean nodulation. qRT-PCR showed that alterations in the expression of GmCEP6 affected the expression of marker genes in the Nod factor signaling pathway. Lastly, the function of GmCEP6 in nitrate inhibition of nodulation was analyzed; nodule numbers in the GmCEP6-overexpressed roots significantly increased under nitrogen treatments, which suggests that GmCEP6 functions in the resistance to nitrate inhibition. The study helps us understand that GmCEP6 promotes nodulation and participates in the regulation of nitrate inhibition of nodulation, which is of great significance for high efficiency utilization of nitrogen in soybeans.
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Root mucilage nitrogen for rhizosphere microorganisms under drought

Root mucilage nitrogen for rhizosphere microorganisms under drought | Plant-Microbe Symbiosis | Scoop.it
Nitrogen (N) is a crucial nutrient for the growth and activity of rhizosphere microorganisms, particularly during drought conditions. Plant root-secreted mucilage contains N that could potentially nourish rhizosphere microbial communities. However, there remains a significant gap in understanding mucilage N content, its source, and its utilization by microorganisms under drought stress. In this study, we investigated the impact of four maize varieties (DH02 and DH04 from Kenya, and Kentos and Keops from Germany) on the secretion rates of mucilage from aerial roots and explored the origin of mucilage N supporting microbial life in the rhizosphere. We found that DH02 exhibited a 96% higher mucilage secretion rate compared to Kentos, while Keops showed 114% and 89% higher secretion rates compared to Kentos and DH04, respectively. On average, the four maize varieties released 4 μg N per root tip per day, representing 2% of total mucilage secretion. Notably, the natural abundance of 15N isotopes increased (higher δ15N signature) with mucilage N release. This indicates a potential dilution of the isotopic signal from biological fixation of atmospheric N by mucilage-inhabiting bacteria as mucilage secretion rates increase. We proposed a model linking mucilage secretion to a mixture of isotopic signatures and estimated that biological N fixation may contribute to 45 - 75% of mucilage N per root tip. The N content of mucilage from a single maize root tip can support a bacterial population ranging from 107 to 1010 cells per day. In conclusion, mucilage serves as a significant N-rich resource for microbial communities in the rhizosphere during drought conditions.

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The soil microbiome modulates the sorghum root metabolome and cellular traits with a concomitant reduction of Striga infection

• The soil microbiome hinders Striga parasitism of sorghum roots
• A Striga-suppressive microbiome tweaks root exudate, aerenchyma, and suberin content
• Pseudomonas strain VK46 reduces haustorium formation by degrading syringic acid
• Arthrobacter strain VK49 increases sorghum endodermal suberization

Jean-Michel Ané's insight:

Exciting progress made in the battle against parasitic plants such as Striga using microbes!

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Heritable microbiome variation is correlated with source environment in locally adapted maize varieties

Beneficial interactions with microorganisms are pivotal for crop performance and resilience. However, it remains unclear how heritable the microbiome is with respect to the host plant genotype and to what extent host genetic mechanisms can modulate plant–microbiota interactions in the face of environmental stresses. Here we surveyed 3,168 root and rhizosphere microbiome samples from 129 accessions of locally adapted Zea, sourced from diverse habitats and grown under control and different stress conditions. We quantified stress treatment and host genotype effects on the microbiome. Plant genotype and source environment were predictive of microbiome abundance. Genome-wide association analysis identified host genetic variants linked to both rhizosphere microbiome abundance and source environment. We identified transposon insertions in a candidate gene linked to both the abundance of a keystone bacterium Massilia in our controlled experiments and total soil nitrogen in the source environment. Isolation and controlled inoculation of Massilia alone can contribute to root development, whole-plant biomass production and adaptation to low nitrogen availability. We conclude that locally adapted maize varieties exert patterns of genetic control on their root and rhizosphere microbiomes that follow variation in their home environments, consistent with a role in tolerance to prevailing stress.

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Check out this amazing paper on how maize genetics and environment influence the rhizosphere microbiome!

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Release of a ubiquitin brake activates OsCERK1-triggered immunity in rice

Plant pattern-recognition receptors perceive microorganism-associated molecular patterns to activate immune signalling1,2. Activation of the pattern-recognition receptor kinase CERK1 is essential for immunity, but tight inhibition of receptor kinases in the absence of pathogen is crucial to prevent autoimmunity3,4. Here we find that the U-box ubiquitin E3 ligase OsCIE1 acts as a molecular brake to inhibit OsCERK1 in rice. During homeostasis, OsCIE1 ubiquitinates OsCERK1, reducing its kinase activity. In the presence of the microorganism-associated molecular pattern chitin, active OsCERK1 phosphorylates OsCIE1 and blocks its E3 ligase activity, thus releasing the brake and promoting immunity. Phosphorylation of a serine within the U-box of OsCIE1 prevents its interaction with E2 ubiquitin-conjugating enzymes and serves as a phosphorylation switch. This phosphorylation site is conserved in E3 ligases from plants to animals. Our work identifies a ligand-released brake that enables dynamic immune regulation.

Jean-Michel Ané's insight:

Fantastic paper. No doubt that this will be relevant to symbiosis too.

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Plant–fungus symbiosis: One receptor to switch on the green light

Arbuscular mycorrhiza, an ancient symbiosis with soil fungi, support mineral nutrition in most plants. How roots recognize such symbiotic fungi has long been debated. Recent research identifies a Medicago truncatula receptor as a key player in triggering symbiont accommodation responses.
Jean-Michel Ané's insight:

Nice and clear commentary from @magiovan and Andrea Genre

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Computer vision models enable mixed linear modeling to predict arbuscular mycorrhizal fungal colonization using fungal morphology

Computer vision models enable mixed linear modeling to predict arbuscular mycorrhizal fungal colonization using fungal morphology | Plant-Microbe Symbiosis | Scoop.it
The presence of Arbuscular Mycorrhizal Fungi (AMF) in vascular land plant roots is one of the most ancient of symbioses supporting nitrogen and phosphorus exchange for photosynthetically derived carbon. Here we provide a multi-scale modeling approach to predict AMF colonization of a worldwide crop from a Recombinant Inbred Line (RIL) population derived from Sorghum bicolor and S. propinquum. The high-throughput phenotyping methods of fungal structures here rely on a Mask Region-based Convolutional Neural Network (Mask R-CNN) in computer vision for pixel-wise fungal structure segmentations and mixed linear models to explore the relations of AMF colonization, root niche, and fungal structure allocation. Models proposed capture over 95% of the variation in AMF colonization as a function of root niche and relative abundance of fungal structures in each plant. Arbuscule allocation is a significant predictor of AMF colonization among sibling plants. Arbuscules and extraradical hyphae implicated in nutrient exchange predict highest AMF colonization in the top root section. Our work demonstrates that deep learning can be used by the community for the high-throughput phenotyping of AMF in plant roots. Mixed linear modeling provides a framework for testing hypotheses about AMF colonization phenotypes as a function of root niche and fungal structure allocations.

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Exploring overlooked growth-promoting mechanisms by plant-associated bacteria

Agriculture-oriented microbiome studies try to develop microbiota beneficial to their plant hosts. This positive goal extends to the soil quality driving plant growth and disease resistance. In research aimed at identifying the causes of this beneficial action, a common interpretation is that microbes will synthesize metabolites useful to their hosts. This view assumes that important microbial metabolites are exported for use by their hosts. Yet, this seems unlikely for essential metabolites, without a counterpart imported from the plants, as the corresponding syntheses would often involve the consumption of resources without explicit benefit to the microbes. Illustrating this function with the example of Bacilli of the Subtilis clade, we emphasize here that the most direct access to the contents of microbial cells is through cell lysis, a phenomenon often linked to the process of sporulation. This process also releases macromolecules that are digested in the environment, releasing key metabolites such as queuine, an important base analog present in the anticodon of some transfer RNAs. This overlooked importance of cell lysis could also be a major cause of the ubiquitous presence of bacteriophages in microbiota.

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Azotosporobacter soli gen. nov., sp. nov., a novel nitrogen-fixing bacterium isolated from paddy soil

A nitrogen-fixing strain designated SG130T was isolated from paddy soil in Fujian Province, China. Strain SG130T was Gram-staining-negative, rod-shaped, and strictly anaerobic. Strain SG130T showed the highest 16S rRNA gene sequence similarities with the type strains Dendrosporobacter quercicolus DSM 1736T (91.7%), Anaeroarcus burkinensis DSM 6283T (91.0%) and Anaerospora hongkongensis HKU 15T (90.9%). Furthermore, the phylogenetic and phylogenomic analysis also suggested strain SG130T clustered with members of the family Sporomusaceae and was distinguished from other genera within this family. Growth of strain SG130T was observed at 25–45 °C (optimum 30 °C), pH 6.0–9.5 (optimum 7.0) and 0–1% (w/v) NaCl (optimum 0.1%). The quinones were Q-8 and Q-9. The polar lipids were phosphatidylserine (PS), phosphatidylethanolamine (PE), glycolipid (GL), phospholipid (PL) and an unidentified lipid (UL). The major fatty acids (> 10%) were iso-C13:0 3OH (26.6%), iso-C17:1 (15.6%) and iso-C15:1 F (11.4%). The genomic DNA G + C content was 50.7%. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain SG130T and the most closely related type strain D. quercicolus DSM 1736T (ANI 68.0% and dDDH 20.3%) were both below the cut-off level for species delineation. The average amino acid identity (AAI) between strain SG130T and the most closely related type strain D. quercicolus DSM 1736T was 63.2%, which was below the cut-off value for bacterial genus delineation (65%). Strain SG130T possessed core genes (nifHDK) involved in nitrogen fixation, and nitrogenase activity (106.38 μmol C2H4 g−1 protein h−1) was examined using the acetylene reduction assay. Based on the above results, strain SG130T is confirmed to represent a novel genus of the family Sporomusaceae, for which the name Azotosporobacter soli gen. nov., sp. nov. is proposed. The type strain is SG130T (= GDMCC 1.3312T = JCM 35641T).
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Novel rapid screening assay to incorporate complexity and increase throughput in early-stage plant biological testing

Novel rapid screening assay to incorporate complexity and increase throughput in early-stage plant biological testing | Plant-Microbe Symbiosis | Scoop.it
The search for new biological products with a positive impact on crop performance is typically initiated by laboratory based in vitro assays. However, live plants and their associated microbes are often removed from in vitro testing assays as a way to reduce biological complexity (variation) and facilitate molecular techniques in the pursuit of uncovering mode-of-action (MoA) mechanisms. Nevertheless, when studying biological candidates intended for use in agriculture, it is essential to incorporate this complexity and validate mechanisms under conditions as close to in situ as possible in order to understand the capacities and MoA of the biologicals in the intended application environments. To address this paradox, we have developed a high-capacity early-stage plant assay that incorporates a live non-sterile plant while also enabling molecular MoA investigations, and that can be conducted in laboratories without greenhouse facilities. The high-capacity design features plants grown in 8-chamber transparent boxes to allow for multiplex imaging and increased biological replicates for greater statistical power. The transparent box design allows the visualization of shoots, roots, tagged-microbes, or visible substrates, and further non-destructive access to shoots or roots for sampling. The boxes are held in racks that hold eight plant boxes during growth in a 19 by 17 cm space, further increasing the throughput to >670 plants per m2 and easing the logistical challenges of plant assays. Furthermore, the box can support various levels of microbial complexity with the option to select the plant growth medium that meets experimental objectives, as well as using sterile or non-sterile seeds. A script-based post-imaging quantification was developed to automate image processing and allow for individual plant readings, further enabling increased statistical confidence. As proof of concept, we use the high-capacity plant system to evaluate the biocontrol potential of Pseudomonas protegens and the biostimulation potential of Pseudomonas koreensis, and are in both cases able to show statistically significant differing plant biomass between treatments under these closer-to-nature conditions. We further demonstrate that the high-capacity plant system is suitable for paired molecular investigations by performing metabolomics and qPCR DNA quantification directly from the plant box to explore in situ chemical MoA, as well as confirm the survival of the P. protegens strains to validate their role in the improved plant phenotype. In conclusion, the study presents a modular high-capacity plant assay system that enables increased throughput functional testing of microbial biocontrol and biostimulant candidates in planta. This novel assaying system saves time, reduces human error, provides quantitative and non-destructive in planta data, and can be used in laboratories without greenhouse facilities. We therefore believe it provides a potent early-stage testing option that bridges in vitro and greenhouse testing, and will expedite the discovery of superior next-generation biological products in agriculture.

Jean-Michel Ané's insight:

Interesting setup

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Advancing nitrogen use efficiency in cereal crops: A comprehensive exploration of genetic manipulation, nitrogen dynamics, and plant nitrogen assimilation

• Increase of nitrogen use efficiency (NUE) through management and genetic manipulation is an urgent need.

• The review highlights the current trend of using genetic insights from plant systems biology to promote cereal crop breeding strategies that reduce reliance on fertilizer without sacrificing yield.

• This review explores the role of nitrogen in plants, its assimilation and transport, and new approaches to genetically improving cereal crops.

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Decoding the microbiome for sustainable agriculture

Decoding the microbiome for sustainable agriculture | Plant-Microbe Symbiosis | Scoop.it
Root-associated microbiota profoundly affect crop health and productivity. Plants can selectively recruit beneficial microbes from the soil and actively balance microbe-triggered plant-growth promotion and stress tolerance enhancement. The cost associated with this is the root-mediated support of a certain number of specific microbes under nutrient limitation. Thus, it is important to consider the dynamic changes in microbial quantity when it comes to nutrient condition-induced root microbiome reassembly. Quantitative microbiome profiling (QMP) has recently emerged as a means to estimate the specific microbial load variation of a root microbiome (instead of the traditional approach quantifying relative microbial abundances) and data from the QMP approach can be more closely correlated with plant development and/or function. However, due to a lack of detailed-QMP data, how soil nutrient conditions affect quantitative changes in microbial assembly of the root-associated microbiome remains poorly understood. A recent study quantified the dynamics of the soybean root microbiome, under unbalanced fertilization, using QMP and provided data on the use of specific synthetic communities (SynComs) for sustaining crop productivity. In this editorial, we explore potential opportunities for utilizing QMP to decode the microbiome for sustainable agriculture.

Jean-Michel Ané's insight:

Very good review

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Characterization of arbuscular mycorrhizal fungal species associating with Zea mays

Characterization of arbuscular mycorrhizal fungal species associating with Zea mays | Plant-Microbe Symbiosis | Scoop.it
Taxonomic identification of arbuscular mycorrhizal (AM) fungal spores extracted directly from the field is sometimes difficult because spores are often degraded or parasitized by other organisms. Single-spore inoculation of a suitable host plant allows for establishing monosporic cultures of AM fungi. This study aimed to propagate AM fungal spores isolated from maize soil using single spores for morphological characterization. First, trap cultures were established to trigger the sporulation of AM fungal species. Second, trap cultures were established with individual morphotypes by picking up only one spore under a dissecting microscope and transferring it to a small triangle of sterilized filter paper, which was then carefully inoculated below a root from germinated sorghum seeds in each pot and covered with a sterile substrate. All pots were placed in sunbags and maintained in a plant growth room for 120 days. Spores obtained from single spore trap cultures from each treatment, maize after oats (MO), maize after maize (MM), maize after peas (MP), and maize after soybean (MS), were extracted using the sieving method. Healthy spores were selected for morphological analysis. Direct PCR was conducted by crushing spores in RNAlater and applying three sets of primer pairs: ITS1 × ITS4, NS31 × AML2, and SSUmcf and LSUmBr. Nucleotide sequences obtained from Sanger sequencing were aligned on MEGA X. The phylogenetic tree showed that the closest neighbors of the propagated AM fungal species belonged to the genera Claroideoglomus, Funneliformis, Gigaspora, Paraglomus, and Rhizophagus. The morphological characteristics were compared to the descriptive features of described species posted on the INVAM website, and they included Acaulospora cavernata, Diversispora spurca, Funneliformis geosporus, Funneliformis mosseae, Gigaspora clarus, Gigaspora margarita, Glomus macrosporum, Paraglomus occultum, and Rhizophagus intraradices. These findings can provide a great contribution to crop productivity and sustainable management of the agricultural ecosystem. Also, the isolate analyzed could be grouped into efficient promoters of growth and mycorrhization of maize independent of their geographical location.

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Regulatory roles of the second messenger c-di-GMP in beneficial plant-bacteria interactions

The rhizosphere system of plants hosts a diverse consortium of bacteria that confer beneficial effects on plant, such as plant growth-promoting rhizobacteria (PGPR), biocontrol agents with disease-suppression activities, and symbiotic nitrogen fixing bacteria with the formation of root nodule. Efficient colonization in planta is of fundamental importance for promoting of these beneficial activities. However, the process of root colonization is complex, consisting of multiple stages, including chemotaxis, adhesion, aggregation, and biofilm formation. The secondary messenger, c-di-GMP (cyclic bis-(3′-5′) dimeric guanosine monophosphate), plays a key regulatory role in a variety of physiological processes. This paper reviews recent progress on the actions of c-di-GMP in plant beneficial bacteria, with a specific focus on its role in chemotaxis, biofilm formation, and nodulation.

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