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Microtubule organization in root cells of Medicago truncatula during development of an arbuscular mycorrhizal symbiosis with Glomus versiforme.

The colonization of plants by arbuscular mycorrhizal fungi has been shown to induce changes in cytoplasmic organization and morphology of root cells. Because of their role in a variety of cellular functions in plants, it is likely that microtubules are involved either in the signaling events leading to the establishment of the symbiosis or in changes in host cell morphology and cytoplasmic architecture. Recent studies of the arbuscular mycorrhizal symbiosis have shown that root cortical cells reorganize their microtubules upon colonization. These studies, however, have focused primarily on the cells containing hyphal coils or arbuscules and did not include descriptions of microtubule changes in adjacent cells. To probe further into the potential role of the microtubule cytoskeleton in the establishment of arbuscular mycorrhizal symbiosis, we examined the three-dimensional arrangement of microtubules in roots of the model legume Medicago truncatula colonized by the arbuscular mycorrhizal fungus Glomus versiforme by indirect immunofluorescence and confocal microscopy. Our data show extensive remodeling of the microtubule cytoskeleton from the early stages of arbuscule development until arbuscule collapse and senescence. While confirming some of the microtubule patterns shown in other mycorrhizal systems, our results also reveal that cortical cells adjacent to those containing arbuscules or adjacent to intercellular hyphae reorganize their microtubules. This indicates that the cortical cells initiate the modification of their cytoskeleton prior to entry of the fungus and is consistent with signal exchange between the symbionts prior to fungal penetration of the cells.

Cell Wall↗

The origin of eukaryotes is suggested as the symbiosis of pyrococcus into gamma-proteobacteria by phylogenetic tree based on gene content.

Attempts were made to define the relationship among the three domains (eukaryotes, archaea, and eubacteria) using phylogenetic tree analyses of 16S rRNA sequences as well as of other protein sequences. Since the results are inconsistent, it is implied that the eukaryotic genome has a chimeric structure. In our previous studies, the origin of eukaryotes to be the symbiosis of archaea into eubacteria using the whole open reading frames (ORF) of many genomes was suggested. In these studies, the species participating in the symbiosis were not clarified, and the effect of gene duplication after speciation (in-paralog) was not addressed. To avoid the influence of the in-paralog, we developed a new method to calculate orthologous ORFs. Furthermore, we separated eukaryotic in-paralogs into three groups by sequence similarity to archaea, eubacteria (other than alpha-proteobacteria), and alpha-proteobacteria and treated them as individual organisms. The relationship between the three ORF groups and the functional classification was clarified by this analysis. The introduction of this new method into the phylogenetic tree analysis of 66 organisms (4 eukaryotes, 13 archaea, and 49 eubacteria) based on gene content suggests the symbiosis of pyrococcus into gamma-proteobacteria as the origin of eukaryotes.

Eukaryotic Cells↗

Arbuscular mycorrhizal symbiosis changes the colonization pattern of Acacia tortilis spp. Raddiana rhizosphere by two strains of rhizobia.

The aim of the study was to assess the effect of the mycorrhizosphere of A. tortillis spp. raddiana mycorrhized with Glomus intraradices on the root nodulation by Sinorhizobium terangae (ORS 1009) and/or Mesorhizobium plurifarium (ORS 1096) in two different culture substrates (sandy soil and sand). The endomycorrhizal fungus only stimulated plant growth in the sandy soil. Moreover, arbuscular mycorrhizal infection enhanced the nodulation process in both culture substrates. Beside the stimulatory effects of the mycorrhizosphere on both rhizobia development, fungal symbiosis induces two different dynamics of each bacterial strains in the sand-grown plants. These results suggest specific relationships could occur during the development of the tripartite symbiosis, at physiological and molecular level. From a practical point of view, the role of arbuscular mycorrhizas in improving nodulation and N2 fixation is universally recognized. The fungal symbiosis could modify the development of bacterial inoculants along the root systems. This effect is of particular interest in the controlled inoculation of selected rhizobia.

Acacia↗

The role of PHB metabolism in the symbiosis of rhizobia with legumes.

The carbon storage polymer poly-beta-hydroxybutyrate (PHB) is a potential biodegradable alternative to plastics, which plays a key role in the cellular metabolism of many bacterial species. Most species of rhizobia synthesize PHB but not all species accumulate it during symbiosis with legumes; the reason for this remains unclear, although it was recently shown that a metabolic mutant of a nonaccumulating species retains the capacity to store PHB in symbiosis. Although the precise roles of PHB metabolism in these bacteria during infection, nodulation, and nitrogen fixation are not determined, the elucidation of these roles will influence our understanding of the metabolic nature of the symbiotic relationship. This review explores the progress that was made in determining the biochemistry and genetics of PHB metabolism. This includes the elucidation of the PHB cycle, variations in PHB metabolism among rhizobial species, and the implications of these variations, while proposing a model for the role of PHB metabolism and storage in symbiosis.

Fabaceae↗

Consequences of symbiosis for food web dynamics.

Basic Lotka-Volterra type models in which mutualism (a type of symbiosis where the two populations benefit both) is taken into account, may give unbounded solutions. We exclude such behaviour using explicit mass balances and study the consequences of symbiosis for the long-term dynamic behaviour of a three species system, two prey and one predator species in the chemostat. We compose a theoretical food web where a predator feeds on two prey species that have a symbiotic relationships. In addition to a species-specific resource, the two prey populations consume the products of the partner population as well. In turn, a common predator forages on these prey populations. The temporal change in the biomass and the nutrient densities in the reactor is described by ordinary differential equations (ODE). Since products are recycled, the dynamics of these abiotic materials must be taken into account as well, and they are described by odes in a similar way as the abiotic nutrients. We use numerical bifurcation analysis to assess the long-term dynamic behaviour for varying degrees of symbiosis. Attractors can be equilibria, limit cycles and chaotic attractors depending on the control parameters of the chemostat reactor. These control parameters that can be experimentally manipulated are the nutrient density of the inflow medium and the dilution rate. Bifurcation diagrams for the three species web with a facultative symbiotic association between the two prey populations, are similar to that of a bi-trophic food chain; nutrient enrichment leads to oscillatory behaviour. Predation combined with obligatory symbiotic prey-interactions has a stabilizing effect, that is, there is stable coexistence in a larger part of the parameter space than for a bi-trophic food chain. However, combined with a large growth rate of the predator, the food web can persist only in a relatively small region of the parameter space. Then, two zero-pair bifurcation points are the organizing centers. In each of these points, in addition to a tangent, transcritical and Hopf bifurcation a global heteroclinic bifurcation is emanating. This heteroclinic cycle connects two saddle equilibria where the predator is absent. Under parameter variation the period of the stable limit cycle goes to infinity and the cycle tends to the heteroclinic cycle. At this global bifurcation point this cycle breaks and the boundary of the basin of attraction disappears abruptly because the separatrix disappears together with the cycle. As a result, it becomes possible that a stable two-nutrient-two-prey population system becomes unstable by invasion of a predator and eventually the predator goes extinct together with the two prey populations, that is, the complete food web is destroyed. This is a form of over-exploitation by the predator population of the two symbiotic prey populations. When obligatory symbiotic prey-interactions are modelled with Liebig's minimum law, where growth is limited by the most limiting resource, more complicated types of bifurcations are found. This results from the fact that the Jacobian matrix changes discontinuously with respect to a varying parameter when another resource becomes most limiting.

Algorithms↗

The expression of a symbiosis-regulated gene in eucalypt roots is regulated by auxins and hypaphorine, the tryptophan betaine of the ectomycorrhizal basidiomycete Pisolithus tinctorius.

A full-length cDNA coding for a symbiosis-regulated transcript, EgHypar, was isolated by differential screening from a Eucalyptus globulus bicostata--Pisolithus tinctorius ectomycorrhiza. The sequence of this clone revealed a protein with an estimated molecular mass of 25.5 kDa that exhibited a high degree of homology (66%) with plant auxin-induced glutathione-S-transferases. Expression of the EgHypar gene in seedlings was confined largely in roots and it is drastically increased by ectomycorrhiza development. The concentration of EgHypar transcripts was similarly up-regulated in roots incubated in media supplemented with P. tinctorius cell-free extracts, indole-3-acetic acid, 2,4-dichlorophenoxyacetic acid or hypaphorine (tryptophan betaine), the major indolic compound secreted by P. tinctorius. The latter fungal alkaloid concomitantly induced a decrease in root hair elongation in eucalypt seedlings. Up-regulation of EgHypar expression by auxins and fungal metabolites suggests that this symbiosis-regulated gene could be involved in the morphological changes taking place in plants roots upon symbiosis development. To our knowledge, these results provide the first molecular evidence that gene expression of the host plant is altered by molecules produced by the ectomycorrhizal mycobiont.

Amino Acid Sequence↗

Arbuscular mycorrhizal symbiosis and alleviation of osmotic stress. New perspectives for molecular studies.

Water deficit is considered one of the most important abiotic factors limiting plant growth and yield in many areas on earth. Several eco-physiological studies have demonstrated that the arbuscular mycorrhizal (AM) symbiosis often results in altered rates of water movement into, through and out of the host plants, with consequent effects on tissue hydration and plant physiology. It is now accepted that the contribution of AM symbiosis to plant drought tolerance is the result of accumulative physical, nutritional, physiological and cellular effects. This review considers several aspects that should be investigated at a molecular level in order to gain a whole understanding of the different mechanisms by which the AM symbiosis protects the host plants against the detrimental effects of water deficit.

Aquaporins↗

Effect of two AMF life strategies on the tripartite symbiosis with Bradyrhizobium japonicum and soybean.

This study is the first in assessing the effect of soil disturbance on the contribution of arbuscular mycorrhizal fungi (AMF) with different life-history strategies to the tripartite symbiosis with soybeans and Bradyrhizobium japonicum (Kirchner) Jordan. We hypothesized that Gigaspora margarita Becker and Hall would be more affected by soil disturbance than Glomus clarum Nicol. and Schenck, and consequently, the tripartite symbiosis would develop more rapidly and lead to greater N(2) fixation in the presence of the latter. Soil pasteurization allowed the establishment of treatments with individual AMF species and soil disturbance enabled the development of contrasting root colonization potentials. In contrast, the colonization potential of B. japonicum was kept the same in all treatments. Soil disturbance significantly reduced root colonization by both AMF, with Gi. margarita being considerably more affected than G. clarum. Furthermore, the tripartite symbiosis progressed faster with G. clarum, and at 10 days after plant emergence, there was 30% more nodules when G. clarum was present compared to that when the bacterial symbiont alone was present. At flowering, the absence of soil disturbance stimulated N(2) fixation by 17% in mycorrhizal plants. However, this response was similar for both AMF.

Bradyrhizobium↗

Fungal and plant gene expression in arbuscular mycorrhizal symbiosis.

Arbuscular mycorrhizas (AMs) are a unique example of symbiosis between two eukaryotes, soil fungi and plants. This association induces important physiological changes in each partner that lead to reciprocal benefits, mainly in nutrient supply. The symbiosis results from modifications in plant and fungal cell organization caused by specific changes in gene expression. Recently, much effort has gone into studying these gene expression patterns to identify a wider spectrum of genes involved. We aim in this review to describe AM symbiosis in terms of current knowledge on plant and fungal gene expression profiles.

Gene Expression Regulation, Fungal↗

Transconjugants of Agrobacterium radiobacter harbouring sym genes of Rhizobium galegae can form an effective symbiosis with Medicago sativa.

It is known that the Rhizobium galegae genomes contain megaplasmids. The suicide vector pSUP2111 with nifH gene of R. meliloti was introduced into the strains CIAM 0703 and CIAM 0711 of R. galegae inducing effective nodules on Galega orientalis plants. The formation of self-transmissible megaplasmids was observed. The megaplasmid transfer into non-nodulating R. meliloti mutants resulted in partial complementation of the nodulation defect in recipient strains though only one transconjugant showed the nitrogen-fixing activity in symbiosis with alfalfa and another one in symbiosis with G. orientalis plants. Among the Agrobacterium strains harbouring R. galegae megaplasmids there were four classes of transconjugants: (1) Nod+ Fix- in symbiosis with goat's rue plants (three strains); (2) Nod+ Fix- on Medicago sativa (two strains); (3) Nod+ Fix+ on M. sativa (five strains); (4) Nod- with both plant hosts (11 strains).

Acetylene↗

Cooperation strategies, signals and symbiosis.

The authors of models concerning cooperation found an interesting application point in mutualism and symbiosis. Phenomena which are to be found in symbiosis are integrated by the various models in relation to the 'Prisoner's Dilemma': 'cost-benefit' ratio components, signals, neighbourhood interaction or reciprocity. Reciprocal altruism is found in symbiosis, but others strategies seem more to reflect human societies, where choice is unrestricted. It is very difficult to give a value to cost and benefit equilibriums. The biological environment interferes and a model based on a dominant strategy is often limited or insufficient to report on the living system's evolution.

Animals↗

Dynamics of arbuscular mycorrhizal symbiosis in heavy metal phytoremediation: meta-analytical and conceptual perspectives.

To estimate dynamics of arbuscular mycorrhizal (AM) symbiosis in heavy metal (HM) phytoremediation, we conducted a literature survey and correlated HM uptake and relative plant growth parameters from published data. After estimating AM feedback responses for these parameters at low and high soil-HM concentration intervals, we determined that the roles of AM symbiosis are characterized by (1) an increased HM phytoextraction via mycorrhizospheric 'Enhanced Uptake' at low soil-HM concentrations, and (2) a reduced HM bioavailability via AM fungal 'Metal-Binding' processes at high soil-HM levels, hence resulting in increased plant biomass and enhanced plant tolerance through HM stress-avoidance. We present two conceptual models which illustrate the important compromise between plant growth, plant HM uptake and HM tolerance, and further emphasize the importance of AM symbiosis in buffering the soil environment for plants under such stress conditions.

Biodegradation, Environmental↗

Recognition events in AM symbiosis: analysis of fungal gene expression at the early appressorium stage.

Arbuscular mycorrhizal symbiosis is induced upon a series of recognition events involving the reorganization of both plant and fungal cellular programs culminating in the formation of appressoria on the epidermal root cells. In this work we monitored for the first time the genetic changes occurring in the fungal partner during early appressorium development. We established an in vitro system of Glomus mosseae and Petroselinum crispum for studying appressorium formation and found that after 120 h first appressoria developed in the root epidermis. We have constructed a fungal subtractive suppressive library enriched in genes up-regulated at this stage. Our aim was to identify early signaling events during plant recognition leading to appressoria formation. The library contains 375 clones with an average size of 500 bp. From these, 200 clones were sequenced and most of them represent gene fragments with no known homologues (63%) and therefore putative new genes specific to the mycorrhiza symbiosis. Reverse-Northern blot and RT-PCR analyses confirmed that ca. 30% of the genes present in the library were up-regulated upon plant induction after 120 h. Among the genes with homologues in other organisms we found several genes common to other plant-microbe interactions including some genes related to Ca2+-dependent signaling. The up-regulation of these genes opens the possibility that Ca2+ plays a role in the early stages of mycorrhiza formation as it has been found in other plant-microbe interactions such as the Rhizobium symbiosis or the Magnaporthe grisea/rice pathogenic interaction.

Blotting, Northern↗

Symbiosis-enhanced gene expression in cnidarian-algal associations: cloning and characterization of a cDNA, sym32, encoding a possible cell adhesion protein.

Mutualistic endosymbioses between two partners are complex associations that are regulated by the genetic interactions of the partners. One important marine symbiosis is that between various cnidarians, such as corals and anemones, and their photosynthetic algal symbionts. We have been interested in characterizing cnidarian host genes that are expressed as a function of the symbiotic state, using the temperate sea anemone Anthopleura elegantissima as a model. In this study, we report on symbiosis-enhanced expression and synthesis of sym32 in anemones. We characterized the full-length sym32 cDNA, obtained by RT-PCR, and demonstrated, by semi-quantitative RT-PCR, that sym32 transcript was much more abundant in symbiotic than in non-symbiotic host anemone RNA. Further, using immunoblots, we determined that an antibody made to a sym32 fusion protein labeled a 32 kD band much more strongly in symbiotic compared to non-symbiotic anemone protein homogenates. Databank searches revealed that the sym32 deduced amino acid sequence shares significant homology with the fasciclin I (Fas I) family of homophilic cell adhesion proteins, present in a variety of organisms ranging from bacteria to humans. This strong homology with the Fas I family suggests that sym32 is involved in regulation of the symbiosis by mediating cell-cell interactions.

Amino Acid Sequence↗

Metabolic symbiosis at the origin of eukaryotes.

Thirty years after Margulis revived the endosymbiosis theory for the origin of mitochondria and chloroplasts, two novel symbiosis hypotheses for the origin of eukaryotes have been put forward. Both propose that eukaryotes arose through metabolic symbiosis (syntrophy) between eubacteria and methanogenic Archaea. They also propose that this was mediated by interspecies hydrogen transfer and that, initially, mitochondria were anaerobic. These hypotheses explain the mosaic character of eukaryotes (i.e. an archaeal-like genetic machinery and a eubacterial-like metabolism), as well as distinct eukaryotic characteristics (which are proposed to be products of symbiosis). Combined data from comparative genomics, microbial ecology and the fossil record should help to test their validity.

Archaea↗

Surface polysaccharide involvement in establishing the rhizobium-legume symbiosis.

When the rhizosphere is nitrogen-starved, legumes and rhizobia (soil bacteria) enter into a symbiosis that enables the fixation of atmospheric dinitrogen. This implies a complex chemical dialogue between partners and drastic changes on both plant roots and bacteria. Several recent works pointed out the importance of rhizobial surface polysaccharides in the establishing of the highly specific symbiosis between symbionts. Exopolysaccharides appear to be essential for the early infection process. Lipopolysaccharides exhibit specific roles in the later stages of the nodulation processes such as the penetration of the infection thread into the cortical cells or the setting up of the nitrogen-fixing phenotype. More generally, even if active at different steps of the establishing of the symbiosis, all the polysaccharide classes seem to be involved in complex processes of plant defense inhibition that allow plant root invasion. Their chemistry is important for structural recognition as well as for physico-chemical properties.

Carbohydrate Sequence↗

Six nonnodulating plant mutants defective for Nod factor-induced transcriptional changes associated with the legume-rhizobia symbiosis.

As the legume-rhizobia symbiosis is established, the plant recognizes bacterial-signaling molecules, Nod factors (NFs), and initiates transcriptional and developmental changes within the root to allow bacterial invasion and the construction of a novel organ, the nodule. Plant mutants defective in nodule initiation (Nod(-)) are thought to have defects in NF-signal transduction. However, it is unknown whether WT plants respond to NF-independent bacterial-derived signals or whether Nod(-) plant mutants show defects in global symbiosis-associated gene expression. To characterize plant gene expression in the establishment of the symbiosis, we used an Affymetrix oligonucleotide microarray representing 9,935 Medicago truncatula expressed sequences. We identified 46 sequences that are differentially expressed in plants exposed for 24 h to WT Sinorhizobium meliloti or to the invasion defective S. meliloti mutant, exoA. Eight of these genes encode nucleolar proteins, which are implicated in ribosome biogenesis. We also identified differentially expressed transcription factors, signaling components, defense response proteins, stress response proteins, and several previously uncharacterized genes. NF appears both necessary and sufficient to induce most changes. Six of seven Nod(-) M. truncatula mutants (nfp, dmi1, dmi2, dmi3, nsp1, and nsp2) showed no transcriptional response to S. meliloti, suggesting that the encoded proteins are required for initiating new transcription. The Nod(-) mutant hcl, however, exhibits a reduced transcriptional response to S. meliloti, indicating that the machinery responsible for initiating new transcription is at least partially functional in this mutant.

Lipopolysaccharides↗

A dual-genome Symbiosis Chip for coordinate study of signal exchange and development in a prokaryote-host interaction.

The soil-dwelling alpha-proteobacterium Sinorhizobium meliloti engages in a symbiosis with legumes: S. meliloti elicits the formation of plant root nodules where it converts dinitrogen to ammonia for use by the plant in exchange for plant photosynthate. To study the coordinate differentiation of S. meliloti and its legume partner during nodule development, we designed a custom Affymetrix GeneChip with the complete S. meliloti genome and approximately 10,000 probe sets for the plant host, Medicago truncatula. Expression profiling of free-living S. meliloti grown with the plant signal molecule luteolin in defined minimal and rich media or of strains altered in the expression of key regulatory proteins (NodD1, NodD3, and RpoN) confirms previous data and identifies previously undescribed regulatory targets. Analyses of root nodules show that this Symbiosis Chip allows the study of gene expression in both partners simultaneously. Our studies detail nearly 5,000 transcriptome changes in symbiosis and document complex transcriptional profiles of S. meliloti in different environments.

Bacterial Proteins↗