Regulation Of Symbiosis Research Articles

TML1 AND TML2 SYNERGISTICALLY REGULATE NODULATION AND AFFECT ARBUSCULAR MYCORRHIZA IN MEDICAGO TRUNCATULA.

Two symbiotic processes, nodulation and arbuscular mycorrhiza, are primarily controlled by the plant's need for nitrogen (N) and phosphorus (P), respectively. Autoregulation of Nodulation (AON) and Autoregulation of Mycorrhization (AOM) both negatively regulate their respective processes and share multiple components - plants that make too many nodules usually have higher AM fungal root colonization. The protein TML (TOO MUCH LOVE) was shown to function in roots to maintain susceptibly to rhizobial infection under low N conditions and control nodule number through AON in Lotus japonicus . M. truncatula has two sequence homologs: Mt TML1 and Mt TML2. We report the generation of stable single and double mutants harboring multiple allelic variations in MtTML1 and MtTML2 using CRISPR-Cas9 targeted mutagenesis and screening of a transposon mutagenesis library. Plants containing single mutations in Mt TML1 or Mt TML2 produced 2-3 times the nodules of wild-type plants whereas plants containing mutations in both genes displayed a synergistic effect, forming 20x more nodules compared to wild type plants. Examination of expression and heterozygote effects suggest genetic compensation may play a role in the observed synergy. Plants with mutations in both TMLs only showed mild increases in AM fungal root colonization at later timepoints in our experiments, suggesting these genes may also play a minor role in AM symbiosis regulation. The mutants created will be useful tools to dissect the mechanism of synergistic action of Mt TML1 and Mt TML2 in M. truncatula symbiosis with beneficial microbes.

Insights into links between redox cycling of dissolved organic matter ranked by molecular weight and methanogen-bacteria symbiosis-driven methane production

Molecular weight (MW) of dissolved organic matter (DOM) governs its redox capacity, playing pivotal roles in methanogen-bacteria symbiosis-driven CH4 production. However, the effect of redox capacity of DOM ranked by MW on these symbiotic associations during anaerobic digestion have never been investigated. The electron–donating (EDC) and –accepting capacity (EAC) of DOM with different MW were quantified, elucidating their impacts on bacteria-methanogen symbiosis-driven CH4 production. By contrast, DOM with 7000 > MW > 14,000 Da constituted the primary contribution to EAC, with an average contribution of 44.63 %. DOM with MW > 14,000 Da emerged as the predominant contributor to EDC, with an average contribution of 49.10 %. Random forest showed that EAC/EDC of DOM ranked by MW was the important factors for methanogenesis by driving shifts in microbial symbiotic relationships. 46 genera (relative abundance of 69.55 %) of microorganisms exhibited robust associations with EAC/EDC. EDC of DOM with 3500 < MW < 7000 Da exerted positive effect on CH4 by modulating the corporation of Caldicoprobacter, norank_o__TSCOR001-H18, norank_o__MBA03 and Methanobrevibacter. EDC of DOM (7000 < MW < 14,000 Da) promotes CH4 production by regulating cooperation of Corynebacterium, Pseudomonas and Methanosarcina, Methanothermus. EDC of DOM (MW > 14,000 Da) enhances CH4 production by modulating cooperation of Ureibacillus, Treponema and methanomassiliicoccus, methanogenium. EAC of DOMs were negatively correlated with CH4. This study broadens our knowledge on the intricate process of methanogenesis and holds significant importance in developing a microbial symbiosis regulation strategy based on electron transfer system.

Evolution of small molecule-mediated regulation of arbuscular mycorrhiza symbiosis.

The arbuscular mycorrhizal (AM) symbiosis formed by most extant land plants with symbiotic fungi evolved 450 Ma. AM promotes plant growth by improving mineral nutrient and water uptake, while the symbiotic fungi obtain carbon in return. A number of plant genes regulating the steps leading to an efficient symbiosis have been identified; however, our understanding of the metabolic processes involved in the symbiosis and how they were wired to symbiosis regulation during plant evolution remains limited. Among them, the exchange of chemical signals, the activation of dedicated biosynthesis pathways and the production of secondary metabolites regulating late stages of the AM symbiosis begin to be well described across several land plant clades. Here, we review our current understanding of these processes and propose future directions to fully grasp the phylogenetic distribution and role played by small molecules during this ancient plant symbiosis. This article is part of the theme issue 'The evolution of plant metabolism'.

The Defective in Autoregulation (DAR) gene of Medicago truncatula encodes a protein involved in regulating nodulation and arbuscular mycorrhiza

BackgroundLegumes utilize a long-distance signaling feedback pathway, termed Autoregulation of Nodulation (AON), to regulate the establishment and maintenance of their symbiosis with rhizobia. Several proteins key to this pathway have been discovered, but the AON pathway is not completely understood.ResultsWe report a new hypernodulating mutant, defective in autoregulation, with disruption of a gene, DAR (Medtr2g450550/MtrunA17_Chr2g0304631), previously unknown to play a role in AON. The dar-1 mutant produces ten-fold more nodules than wild type, similar to AON mutants with disrupted SUNN gene function. As in sunn mutants, suppression of nodulation by CLE peptides MtCLE12 and MtCLE13 is abolished in dar. Furthermore, dar-1 also shows increased root length colonization by an arbuscular mycorrhizal fungus, suggesting a role for DAR in autoregulation of mycorrhizal symbiosis (AOM). However, unlike SUNN which functions in the shoot to control nodulation, DAR functions in the root.ConclusionsDAR encodes a membrane protein that is a member of a small protein family in M. truncatula. Our results suggest that DAR could be involved in the subcellular transport of signals involved in symbiosis regulation, but it is not upregulated during symbiosis. DAR gene family members are also present in Arabidopsis, lycophytes, mosses, and microalgae, suggesting the AON and AOM may use pathway components common to other plants, even those that do not undergo either symbiosis.

Genetically Modified Legume Plants as a Basis for Studying the Signal Regulation of Symbiosis with Nodule Bacteria

The development of legume–rhizobial symbiosis results in the formation of nitrogen-fixing root nodules. In response to rhizobial molecules, Nod factors, signal transduction is mediated by the interaction of activated receptors with downstream signaling proteins. Previously, some new regulators of the signal pathway, such as phospholipases D, which regulate the level of phosphatidic acid (PA), as well as mitogen-activated protein kinases (MAPKs), have been identified in legumes. Since PA is an important signal messenger, we tested the hypothesis that increasing the level of proteins involved in the reversible binding of PA in plant tissues may have a positive effect on symbiosis. Our findings showed that overexpression of MtSPHK1-PA, encoding the PA-binding domain of sphingosine kinase 1 (SPHK1), stimulated plant growth and nodule development in legume plants. Furthermore, the influence of MAPK6 on the development of symbiosis was studied. Using genetic engineering methods, we increased MAPK6 transcriptional activity in transgenic roots, leading to an increase in the number of nodules and the biomass of pea plants. Therefore, new approaches to obtain plants with an increased efficiency of symbiosis were tested. We report here that both genes that encode signaling proteins may be used as potential targets for future modification using biotechnological approaches.

Genetically modified legume plants as a basis for studying the signal regulation of symbiosis with nodule bacteria

The development of legume-rhizobium symbiosis is based on signal exchange between partners, which leads to the formation of nitrogen-fixing root nodules. Under the influence of rhizobial signal molecules, the Nod factors, the signal transduction cascade is activated, where the LysM-type receptor kinases and a complex of intracellular regulators, a significant part of which are still unknown, can play an important role.&#x0D; Using transcriptomic and proteomic analysis, we searched for new regulators of the signal pathway in pea Pisum sativum L., which are activated under the influence of Nod factors. Phospholipases C and D, GTPases, calcium-dependent protein kinases, and mitogen-activated protein kinases (MAPKs) have been identified among such novel regulators. The influence of one of such regulators, the MAPK6, on the development of symbiosis was studied in more detail. Using genetic engineering approaches, we increased the transcriptional activity of MAPK6 in transgenic roots, which led to an increase in the number of nodules and the biomass of pea plants. A similar effect was also found for the homologous MAPK6 gene in another legume Medicago truncatula, which has a type of nodulation similar to that one in pea. New approaches have been developed to obtain stable pea transformants with enhanced level of MAPK6 transcription using the constitutive p35S promoter. A comprehensive study of such plants inoculated with rhizobia was carried out. In addition, the approaches for genome editing of pea plants have been designed using the CRISPR/Cas system, when the MAPK6 gene was used as a target. Thus, the possibility of using genetic engineering methods to obtain plants with increased symbiosis efficiency was investigated.

Coupled carbon and nitrogen cycling regulates the cnidarian–algal symbiosis

Efficient nutrient recycling underpins the ecological success of cnidarian-algal symbioses in oligotrophic waters. In these symbioses, nitrogen limitation restricts the growth of algal endosymbionts in hospite and stimulates their release of photosynthates to the cnidarian host. However, the mechanisms controlling nitrogen availability and their role in symbiosis regulation remain poorly understood. Here, we studied the metabolic regulation of symbiotic nitrogen cycling in the sea anemone Aiptasia by experimentally altering labile carbon availability in a series of experiments. Combining 13C and 15N stable isotope labeling experiments with physiological analyses and NanoSIMS imaging, we show that the competition for environmental ammonium between the host and its algal symbionts is regulated by labile carbon availability. Light regimes optimal for algal photosynthesis increase carbon availability in the holobiont and stimulate nitrogen assimilation in the host metabolism. Consequently, algal symbiont densities are lowest under optimal environmental conditions and increase toward the lower and upper light tolerance limits of the symbiosis. This metabolic regulation promotes efficient carbon recycling in a stable symbiosis across a wide range of environmental conditions. Yet, the dependence on resource competition may favor parasitic interactions, explaining the instability of the cnidarian-algal symbiosis as environmental conditions in the Anthropocene shift towards its tolerance limits.

Genomic conservation and putative downstream functionality of the phosphatidylinositol signalling pathway in the cnidarian-dinoflagellate symbiosis.

The mutualistic cnidarian-dinoflagellate symbiosis underpins the evolutionary success of stony corals and the persistence of coral reefs. However, a molecular understanding of the signalling events that lead to the successful establishment and maintenance of this symbiosis remains unresolved. For example, the phosphatidylinositol (PI) signalling pathway has been implicated during the establishment of multiple mutualistic and parasitic interactions across the kingdoms of life, yet its role within the cnidarian-dinoflagellate symbiosis remains unexplored. Here, we aimed to confirm the presence and assess the specific enzymatic composition of the PI signalling pathway across cnidaria and dinoflagellates by compiling 21 symbiotic anthozoan (corals and sea anemones) and 28 symbiotic dinoflagellate (Symbiodiniaceae) transcriptomic and genomic datasets and querying genes related to this pathway. Presence or absence of PI-kinase and PI-phosphatase orthologs were also compared between a broad sampling of taxonomically related symbiotic and non-symbiotic species. Across the symbiotic anthozoans analysed, there was a complete and highly conserved PI pathway, analogous to the pathway found in model eukaryotes. The Symbiodiniaceae pathway showed similarities to its sister taxon, the Apicomplexa, with the absence of PI 4-phosphatases. However, conversely to Apicomplexa, there was also an expansion of homologs present in the PI5-phosphatase and PI5-kinase groups, with unique Symbiodiniaceae proteins identified that are unknown from non-symbiotic unicellular organisms. Additionally, we aimed to unravel the putative functionalities of the PI signalling pathway in this symbiosis by analysing phosphoinositide (PIP)-binding proteins. Analysis of phosphoinositide (PIP)-binding proteins showed that, on average, 2.23 and 1.29% of the total assemblies of anthozoan and Symbiodiniaceae, respectively, have the potential to bind to PIPs. Enrichment of Gene Ontology (GO) terms associated with predicted PIP-binding proteins within each taxon revealed a broad range of functions, including compelling links to processes putatively involved in symbiosis regulation. This analysis establishes a baseline for current understanding of the PI pathway across anthozoans and Symbiodiniaceae, and thus a framework to target future research.

The development of approaches to create new symbiotic systems

Plants interact with a wide range of soil microorganisms, while interactions with symbiotic microorganisms are the most important for them. Symbiosis with nitrogen-fixing nodule bacteria provides a significant advantage in the existence of plants in nitrogen-poor soils. Since only plants from the Fabales and Rosales (a single representative Parasponia) enter into symbiosis with nodule bacteria, an idea about expanding the number of plants entering into such interactions became popular. To solve the problem of constructing new symbiotic systems, it is necessary to provide recognition of the symbiont (by transferring receptor genes into non-legume plants), morphogenesis of the nodule, and its infection.&#x0D; We have managed to introduce the genes encoding receptors to surface components of rhizobia into the non-legume plants, and it provided increased colonization by nodule bacteria. It may improve the growth and development of non-legume plants and ensure their greater resistance to phytopathogens. Genome analysis of non-legume plant hop Humulus lupulus showed the existence of genes involved in symbiosis regulation, but some important regulators such as gene encoding NIN transcription factor were lost. Using genetic engineering approaches, the hop plants carrying the gene encoding NIN transcription factor were created. Analysis of these plants may provide important information about regulation of organogenesis in non-legume plants.&#x0D; The work was financially supported by RSF 21-16-00106.

Stable isotope tracing reveals compartmentalized nitrogen assimilation in scleractinian corals

Corals form symbiotic relationships with dinoflagellate algae of the family Symbiodiniaceae, bacteria, and other microbes. Central to that relationship is the regulation of nutrition flux between the animal host and the photosynthetic Symbiodiniaceae that it is reliant on for the majority of metabolic needs. Nitrogen availability controls the growth and density of Symbiodiniaceae within coral tissues and has been proposed to play a role in host derived symbiosis regulation. Warming ocean temperatures and subsequent increases in dissolved organic carbon can potentially increase nitrogen fixation and lead to bleaching. We investigated the importance of nitrogen metabolism in vivo with LC-MS based stable isotope tracing using nubbins from three species of Hawaiian coral, the more heat tolerant Montipora capitata and Porites compressa and the more heat sensitive Pocillopora acuta, that were collected from reefs in Kāne’ohe Bay, O’ahu. In addition to 15N incorporation into nucleotides, amino acids, and urea cycle metabolites, we also observed significant isotopic labeling in dipeptides, supporting their previous identification as major heat stress response metabolites. Surprisingly, the dipeptides are highly enriched in 15N compared to free amino acids, which are the biosynthetic precursors for dipeptides. This suggests that there is a high turnover of dipeptide pools and distinct biosynthetic mechanisms that separately mediate amino acid and dipeptide production. These preliminary data show that nitrogen assimilation in the coral holobiont is likely compartmentalized, with rapid assimilation and quick dipeptide turnover occurring in one region of the holobiont and slow turnover of other nitrogen containing metabolites in other region(s).

Variation in symbiont density is linked to changes in constitutive immunity in the facultatively symbiotic coral, Astrangia poculata.

Scleractinian corals are essential ecosystem engineers, forming the basis of coral reef ecosystems. However, these organisms are in decline globally, in part due to rising disease prevalence. Most corals are dependent on symbiotic interactions with single-celled algae from the family Symbiodiniaceae to meet their nutritional needs, however, suppression of host immunity may be essential to this relationship. To explore immunological consequences of algal symbioses in scleractinian corals, we investigated constitutive immune activity in the facultatively symbiotic coral, Astrangia poculata. We compared immune metrics (melanin synthesis, antioxidant production and antibacterial activity) between coral colonies of varying symbiont density. Symbiont density was positively correlated to both antioxidant activity and melanin concentration, likely as a result of the dual roles of these pathways in immunity and symbiosis regulation. Our results confirm the complex nature of relationships between algal symbiosis and host immunity and highlight the need for nuanced approaches when considering these relationships.

MtNF-YC6 and MtNF-YC11 are involved in regulating the transcriptional program of arbuscular mycorrhizal symbiosis.

Arbuscular mycorrhizal fungi are obligate symbionts that transfer mineral nutrients to host plants through arbuscules, a fungal structure specialized for exchange for photosynthetic products. MtNF-YC6 and MtNF-YC11, which encode the C subunits of nuclear factor Y (NF-Y) family in Medicago truncatula are induced specifically by arbuscular mycorrhizal symbiosis (AMS). A previous study showed that MtNF-YC6 and MtNF-YC11 are activated in cortical cells of mycorrhizal roots, but the gene functions were unknown. Herein, we identified both MtNF-YB17 and MtNF-YB12 as the interacting partners of MtNF-YC6 and MtNF-YC11 in yeast and plants. MtNF-YB17 was highly induced by AMS and activated in cortical cells only in mycorrhizal roots but MtNF-YB12 was not affected. The formation of B/C heterodimers led the protein complexes to transfer from the cytoplasm to the nucleus. Silencing MtNF-YC6 and C11 by RNA interference (RNAi) resulted in decreased colonization efficiency and arbuscule richness. Coincidently, genes associated with arbuscule development and degeneration in RNAi roots were also downregulated. In silico analysis showed CCAAT-binding motifs in the promoter regions of downregulated genes, further supporting the involvement of NF-Y complexes in transcriptional regulation of symbiosis. Taken together, this study identifies MtNF-YC6- or MtNF-YC11-containing protein complexes as novel transcriptional regulators of symbiotic program and provides a list of potential downstream target genes. These data will help to further dissect the AMS regulatory network.

Molecular Regulation of Arbuscular Mycorrhizal Symbiosis.

Plant-microorganism interactions at the rhizosphere level have a major impact on plant growth and plant tolerance and/or resistance to biotic and abiotic stresses. Of particular importance for forestry and agricultural systems is the cooperative and mutualistic interaction between plant roots and arbuscular mycorrhizal (AM) fungi from the phylum Glomeromycotina, since about 80% of terrestrial plant species can form AM symbiosis. The interaction is tightly regulated by both partners at the cellular, molecular and genetic levels, and it is highly dependent on environmental and biological variables. Recent studies have shown how fungal signals and their corresponding host plant receptor-mediated signalling regulate AM symbiosis. Host-generated symbiotic responses have been characterized and the molecular mechanisms enabling the regulation of fungal colonization and symbiosis functionality have been investigated. This review summarizes these and other recent relevant findings focusing on the molecular players and the signalling that regulate AM symbiosis. Future progress and knowledge about the underlying mechanisms for AM symbiosis regulation will be useful to facilitate agro-biotechnological procedures to improve AM colonization and/or efficiency.

Symbiotic regulation: How plants seek salvation in starvation

When plants are starved of phosphate, many rely on an ancient symbiosis with arbuscular mycorrhizal fungi to secure a critical supply. A new study demonstrates a molecular basis for the regulation of symbiosis by phosphate starvation.

Searching for regulators that interact with BELL1 transcription factor and control the legume-rhizobial symbiosis development

The development of nitrogen-fixing nodule, which is formed during legume-rhizobial symbiosis, requires the involvement of cell cycle regulators, phytohormones and homeodomain-containing transcription factors as well as other organogenesis processes. Along with homedomain-containing transcription factors from KNOX family, which participation in the control of nodule development has been studied recently, the role of transcription factors from BELL family in this process remains under-explored. At the same time, transcriptomic data for legumes shows an increase in the expression levels of genes encoding proteins of this family in the plant roots inoculated by Rhizobium. In this work we performed screening of сDNA library, which was generated from inoculated pea roots, using BELL1 protein in yeast two-hybrid system. As a result, we detected two interacting proteins, which are highly interesting for future examination. In the first case we discovered interaction between BELL1 and LysM-containing receptor-like kinase LYK9. Another identified regulator interacting with BELL1 became the glycine-rich protein A3, which may be involved in the regulation of defense reactions in plants and their resistance to phytopathogens. Transcriptomic analysis for pea roots has revealed high expression level of gene, which encodes this protein in the nodules, that may demonstrate its important role in symbiosis regulation.

Epichloë Fungal Endophytes-From a Biological Curiosity in Wild Grasses to an Essential Component of Resilient High Performing Ryegrass and Fescue Pastures.

The relationship between Epichloë endophytes found in a wide range of temperate grasses spans the continuum from antagonistic to mutualistic. The diversity of asexual mutualistic types can be characterised by the types of alkaloids they produce in planta. Some of these are responsible for detrimental health and welfare issues of ruminants when consumed, while others protect the host plant from insect pests and pathogens. In many temperate regions they are an essential component of high producing resilient tall fescue and ryegrass swards. This obligate mutualism between fungus and host is a seed-borne technology that has resulted in several commercial products being used with high uptake rates by end-user farmers, particularly in New Zealand and to a lesser extent Australia and USA. However, this has not happened by chance. It has been reliant on multi-disciplinary research teams undertaking excellent science to understand the taxonomic relationships of these endophytes, their life cycle, symbiosis regulation at both the cellular and molecular level, and the impact of secondary metabolites, including an understanding of their mammalian toxicity and bioactivity against insects and pathogens. Additionally, agronomic trials and seed biology studies of these microbes have all contributed to the delivery of robust and efficacious products. The supply chain from science, through seed companies and retailers to the end-user farmer needs to be well resourced providing convincing information on the efficacy and ensuring effective quality control to result in a strong uptake of these Epichloë endophyte technologies in pastoral agriculture.

The expression of LjROP4 is required for rhizobial infection in Lotus japonicus

Increasing evidence suggests that Rho of plant (ROP) GTPases play important roles in the rhizobium–legume symbiotic nodulation, but the molecular mechanisms of their regulation in symbiosis remain poorly understood. In this study, we showed that ROP4 in Lotus japonicus (LjROP4) is involved in the symbiotic interaction between L. japonicus and Mesorhizobium loti. Tissue expression analysis showed that LjROP4 expressed highly in the root. Histochemical staining analysis showed that after rhizobia inoculation, GUS reporter activity increased in the root vascular bundle, root tip, and lateral root primordia. During nodule development, GUS activity was detected in the cortex of nodule primordia and young nodules. In the mature nodules, GUS activity was detected only in the vascular bundle. Compared with the control, the overexpression of ROP4 and ROP4-CA produced much more pronounced root hair swelling and curling induced by M. loti. The infection event and nodule number noticeably increased, which was consistent with this promotion of root hair deformation. Moreover, RNA interference of LjROP4 produced opposite phenotypes. These data suggest that LjROP4 is required for root hair deformation during rhizobial infection. Thus, our study provides important information about root hair deformation responses induced by nod factors in the early stages of symbiotic interaction.

Identification of cells expressing two peptidoglycan recognition proteins in the gill of the vent mussel, Bathymodiolus septemdierum

In symbiotic systems in which symbionts are transmitted horizontally, hosts must accept symbionts from the environment while defending themselves against invading pathogenic microorganisms. How they distinguish pathogens from symbionts and how the latter evade host immune defences are not clearly understood. Recognition of foreign materials is one of the most critical steps in stimulating immune responses, and pattern recognition receptors (PRRs) play vital roles in this process. In this study, we focused on a group of highly conserved PRRs, peptidoglycan recognition proteins (PGRPs), in the deep-sea mussel, Bathymodiolus septemdierum, which harbours chemosynthetic bacteria in their gill epithelial cells. We isolated B. septemdierum PGRP genes BsPGRP-S and BsPGRP-L, which encode a short- and a long-type PGRP, respectively. The short-type PGRP has a signal peptide and was expressed in the asymbiotic goblet mucous cells in the gill epithelium, whereas the long-type PGRP was predicted to include a transmembrane domain and was expressed in gill bacteriocytes. Based on these findings, we hypothesize that the secreted and transmembrane PGRPs are engaged in host defence against pathogenic bacteria and/or in the regulation of symbiosis via different cellular localizations and mechanisms.

Differential proteome analysis of pea roots at the early stages of symbiosis with nodule bacteria

In this paper, we have analyzed changes in the proteomic spectrum of pea Pisum sativum L. roots during inoculation with rhizobial bacteria with the aim of revealing new regulators of symbiosis development. To study the changes in the proteome spectrum of pea roots, a differential twodimensional (2-D) electrophoresis was performed using fluorescent labels Cy2 and Cy5. The images obtained made it possible to identify differences between the control variant (uninoculated roots) and the root variant after inoculation with Rhizobium leguminosarum bv. viciae RCAM 1026 (24 hours after treatment). 20 proteins were revealed and identified, the synthesis of which was enhanced during the inoculation of pea roots by nodule bacteria. To identify the proteins, a mass spectrometric analysis of tryptic peptides was performed on a quadrupole-time-of-flight mass spectrometer combined with a high-performance liquid chromatograph. Among such proteins, the beta-subunit of the G protein and the disulfide isomerase/phospholipase C were first found, whose function can be related to the signal regulation of symbiosis. This indicates that G-proteins and phospholipases can play a key role in the development of early stages of symbiosis in peas. Further experiments are expected to show whether the beta-subunit of the G protein interacts with the receptors to Nod factors, and how this affects the further signaling. Other proteins that might be interesting were annexin D8 and D1, protein kinase interacting with calcinerin B, actin-binding protein profilin, GTP-binding protein Ran1. They may be involved in the regulation of reactions with calcium, the reorganization of the actin cytoskeleton and other important processes in plants. The study of the role of such regulatory proteins will later become the basis for understanding the complex system of signal regulation, which is activated in pea plants by interaction with nodule bacteria.

Identification of potential transcriptional regulators of actinorhizal symbioses in Casuarina glauca and Alnus glutinosa.

BackgroundTrees belonging to the Casuarinaceae and Betulaceae families play an important ecological role and are useful tools in forestry for degraded land rehabilitation and reforestation. These functions are linked to their capacity to establish symbiotic relationships with a nitrogen-fixing soil bacterium of the genus Frankia. However, the molecular mechanisms controlling the establishment of these symbioses are poorly understood. The aim of this work was to identify potential transcription factors involved in the establishment and functioning of actinorhizal symbioses.ResultsWe identified 202 putative transcription factors by in silico analysis in 40 families in Casuarina glauca (Casuarinaceae) and 195 in 35 families in Alnus glutinosa (Betulaceae) EST databases. Based on published transcriptome datasets and quantitative PCR analysis, we found that 39% and 26% of these transcription factors were regulated during C. glauca and A. glutinosa-Frankia interactions, respectively. Phylogenetic studies confirmed the presence of common key transcription factors such as NSP, NF-YA and ERN-related proteins involved in nodule formation in legumes, which confirm the existence of a common symbiosis signaling pathway in nitrogen-fixing root nodule symbioses. We also identified an actinorhizal-specific transcription factor belonging to the zinc finger C1-2i subfamily we named CgZF1 in C. glauca and AgZF1 in A. glutinosa.ConclusionsWe identified putative nodulation-associated transcription factors with particular emphasis on members of the GRAS, NF-YA, ERF and C2H2 families. Interestingly, comparison of the non-legume and legume TF with signaling elements from actinorhizal species revealed a new subgroup of nodule-specific C2H2 TF that could be specifically involved in actinorhizal symbioses. In silico identification, transcript analysis, and phylogeny reconstruction of transcription factor families paves the way for the study of specific molecular regulation of symbiosis in response to Frankia infection.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-014-0342-z) contains supplementary material, which is available to authorized users.