Impact of agropolymers on seed germination, root development, and plant-microorganism interactions

Impacts d’agropolymères sur la germination, le développement racinaire, et les interactions plantes-microorganismes. Les travaux réalisés au cours de la thèse sont de nature exploratoire et visent à déterminer les effets physiologiques de la gomme de guar (Cyamopsis tetragonoloba) et trois de ses dérivés fonctionnalisés, sur la germination des semences, le développement racinaire des plantes et leur capacité à interagir avec un microorganisme symbiotique.Arabidopsis thaliana a été choisie comme plante modèle pour déterminer si le développement des plantes est modifié lorsqu’elles sont soumises à ces molécules, et explorer les mécanismes impliqués. Une analyse fine des modifications architecturales a été menée, via des études de microscopie et l’utilisation de plantes transgéniques. Des développements racinaires contrastés ont été mis en évidence pour deux des agropolymères fonctionnalisés, avec un raccourcissement de la racine primaire et des modifications du nombre de racines latérales. Les plantes traitées par ces deux agropolymères présentent une augmentation de la production d’espèces réactives de l’oxygène (ROS) qui traduisent un état de stress des plantes. Ces macromolécules polysaccharidiques ne semblent pas avoir d’impact sur les nutritions carbonée et azotée des plantes.Parallèlement aux recherches entreprises avec A. thaliana, des travaux ont été menés sur l’arbre tropical actinorhizien Casuarina glauca. Cette espèce d’intérêt pour certains pays du Sud peut se développer dans les sols pauvres en raison de son association symbiotique avec l’actinobactérie fixatrice d’azote Frankia. Les résultats obtenus mettent en évidence que les polymères induisent sur C. glauca des phénotypes racinaires identiques à ceux observés avec A. thaliana. Pour ce qui concerne l’interaction symbiotique, selon l’agropolymère testé et les concentrations choisies, des impacts positifs ou négatifs ont été observés sur le processus de nodulation résultant de l’interaction avec Frankia. L’ensemble de ces travaux fournit des éléments préliminaires qui devraient permettre de promouvoir, dans le futur, ces agropolymères dans le milieu agricole comme biostimulant, afin de tendre vers une agriculture plus durable et responsable.Mots clefs : Agropolymères – germination – développement racinaire – interactions plantes / microorganismes – Arabidopsis thaliana – Casuarina glaucaImpact of agropolymers on seed germination, root development, and plant-microorganism interactionsThe work performed during this thesis is of an exploratory nature and aims at providing insight to the physiological effects of guar (Cyamopsis tetragonoloba) gum polymer and three functionalized derivatives on seed germination, root development and a symbiotic plant-microorganism interaction.Arabidopsis thaliana was chosen as model plant to determine whether plant development was modified by the presence of these polymers and to explore the mechanisms involved. A detailed microscopic analysis of the root architecture was carried out using transgenic plants. Contrasted root development was evident for two of the functionalized agropolymers. Primary roots were shorter and lateral roots distribution was modified. Biochemical analyses revealed that the plants treated by these two agropolymers exibited an increase of reactive oxygen species (ROS) production in root tissues indicating that plants are in stress conditions. Such polysaccharides did not seem to have any effects on carbon and nitrogen plant nutrition.In parallel, studies were performed on the tropical actinorhizal tree Casuarina glauca. This species is of great environmental value in mitigating desertification since is able to grow in poor tropical soils because of its symbiotic interaction with the nitrogen-fixing actinobacteria Frankia. Treatment of C. glauca with the polymers induced root phenotypes that were very similar to those observed with A. thaliana. Treatment with agropolymers resulted in positive or negative effects on the nodulation process during the symbiotic interaction between C. glauca and FrankiaTaken together this work provides preliminary evidence supporting the value of the use agropolymers as agricultural biostimulants, facilitating the development of sustainable agriculture.Key words: Agropolymers – germination – root development – plants-microormanisms interactions – Arabidopsis thaliana – Casuarina glauc

Recent advances in actinorhizal symbiosis signaling

Nitrogen and phosphorus availability are frequent limiting factors in plant growth and development. Certain bacteria and fungi form root endosymbiotic relationships with plants enabling them to exploit atmospheric nitrogen and soil phosphorus. The relationships between bacteria and plants include nitrogen-fixing Gram-negative proteobacteria called rhizobia that are able to interact with most leguminous plants (Fabaceae) but also with the non-legume Parasponia (Cannabaceae), and actinobacteria Frankia, which are able to interact with about 260 species collectively called actinorhizal plants. Fungi involved in the relationship with plants include Glomeromycota that form an arbuscular mycorrhizal (AM) association intracellularly within the roots of more than 80 % of land plants. Increasing numbers of reports suggest that the rhizobial association with legumes has recycled part of the ancestral program used by most plants to interact with AM fungi. This review focuses on the most recent progress made in plant genetic control of root nodulation that occurs in non-legume actinorhizal plant species

Elucidating the Effect of Endophytic Entomopathogenic Fungi on Bread Wheat Growth through Signaling of Immune Response-Related Hormones

Entomopathogenic fungi (EF) provide a potent biocontrol tool; also, their endophytic behavior has broadened their contribution to integrated pest management (IPM) and crop production. In this work, Beauveria bassiana and Metarhizium brunneum were applied to bread wheat (Triticum aestivum) seedlings to elucidate how fungal colonization influences plant growth and the relative expression of 24 genes involved in hormonal syntheses and plant immune mechanisms. A preliminary assay was used to determine the time needed for fungal colonization and assess its effect on wheat growth. Then, plant material collected at various times after inoculation (viz., 2, 8, 20, and 36 h and 9 and 15 days) was used to investigate gene expression by quantitative reverse transcription PCR (RT-qPCR). During the colonization time, B. bassiana and M. brunneum caused strong downregulation of most genes associated with plant immunity and the synthesis of hormones like auxin, cytokinin, and gibberellin. This effect was concomitant with a slowdown of endophytic-colonization-related plant growth until 19 days postinoculation (dpi). However, the wheat started to recover at 15 dpi, simultaneously with upregulation of auxin- and gibberellin-related genes. The results suggest that the EF trigger induced systemic resistance rather than acquired systemic resistance during early plant-microbe cross talk in wheat. Also, they confirm that the hormone and immune responses of wheat triggered by EF inoculation influenced plant growth, which can be useful with a view to optimizing management of these microorganisms for sustainable agriculture. IMPORTANCE Microbial control of insect and mite pests is a key tool to develop integrated pest management (IPM) and sustainable agriculture. Entomopathogenic fungi (EF) may have associations with the plants, playing additional ecological roles in the rhizosphere, in the phylloplane, and as plant endophytes. Beauveria bassiana 04/01TIP and Metarhizium brunneum 01/58Su are two strains that showed very good results either in pest control or plant growth promotion and would be good candidates to develop mycoinsecticides as an alternative to pesticides. However, deep knowledge about their interaction with the plant would let farmers optimize their use and understand the plant response, enhancing and promoting their broader contribution to IPM and crop production

Biotechnological strategies for studying actinorhizal symbiosis in Casuarinaceae: transgenesis and beyond

International audienceSince the recovery of the first transgenic plant in the early 1980s, plant transformation technologies have enabled advances in many aspects of basic plant science, including nitrogen-fixing root endosymbioses. Using the biological vectors Agrobacterium tumefaciens and A. rhizogenes, gene constructs have been successfully introduced in the actinorhizal tree species Casuarina glauca, thereby paving the way for functional analysis of the key genes involved in the symbiotic process with the actinobacterium Frankia. In recent years, not only studies of gene promoters in transgenic Casuarinaceae, but also the use of RNA interference to down-regulate genes of interest, have provided new insights into the early stages of the interaction between the root system and the actinobacterium. Opportunities offered by recent developments in genome editing technologies based on the engineered nucleases ZFNs (zinc-finger nucleases), TALENs (transcription activator-like effector nucleases) and RNA-guided CRISPR-Cas (clustered regularly interspaced short palindromic repeats-associated protein/Cas) will be briefly presented

Prokaryotes and evolution

In natural world, no organism exists in absolute isolation, and thus every organism must interact with the environment and other organisms. Next-generation sequencing technologies are increasingly revealing that most of the cells in the environment resist cultivation in the laboratory and several prokaryotic divisions have no known cultivated representatives. Based on this, we hypothesize that species that live together in the same ecosystem are more or less dependent upon each other and are very large in diversity and number, outnumbering those that can be isolated in single-strain laboratory culture. In natural environments, bacteria and archaea interact with other organisms (viruses, protists, fungi, animals, plants, and human) in complex ecological networks, resulting in positive, negative, or no effect on one or another of the interacting partners. These interactions are sources of ecological forces such as competitive exclusion, niche partitioning, ecological adaptation, or horizontal gene transfers, which shape the biological evolution. In this chapter, we review the biological interactions involving prokaryotes in natural ecosystems, including plant, animal, and human microbiota, and give an overview of the insights into the evolution of living beings. We conclude that studies of biological interactions, including multipartite interactions, are sources of novel knowledge related to the biodiversity of living things, the functioning of ecosystems, the evolution of the cellular world, and the ecosystem services to the living beings

Interactions between soil compositions and the wheat root microbiome under drought stress : from an in silico to in planta perspective

As wheat (Triticum aestivum) is an important staple food across the world, preservation of stable yields and increased productivity are major objectives in breeding programs. Drought is a global concern because its adverse impact is expected to be amplified in the future due to the current climate change. Here, we analyzed the effects of edaphic, environmental, and host factors on the wheat root microbiomes collected in soils from six regions in Belgium. Amplicon sequencing analysis of unplanted soil and wheat root endosphere samples indicated that the microbial community variations can be significantly explained by soil pH, microbial biomass, wheat genotype, and soil sodium and iron levels. Under drought stress, the biodiversity in the soil decreased significantly, but increased in the root endosphere community, where specific soil parameters seemingly determine the enrichment of bacterial groups. Indeed, we identified a cluster of drought-enriched bacteria that significantly correlated with soils compositions. Interestingly, integration of a functional analysis further revealed a strong correlation between the same cluster of bacteria and β-glucosidase and osmoprotectant proteins, two functions known to be involved in coping with drought stress. By means of this in silico analysis, we identified amplicon sequence variants (ASVs) that could potentially protect the plant from drought stress and validated them in planta. Yet, ASVs based on 16S rRNA sequencing data did not completely distinguish individual isolates because of their intrinsic short sequences. Our findings support the efforts to maintain stable crop yields under drought conditions through implementation of root microbiome analyses