Establishment Of Symbiosis Research Articles

Abscisic acid supports colonization of Eucalyptus grandis roots by the mutualistic ectomycorrhizal fungus Pisolithus microcarpus.

The pathways regulated in ectomycorrhizal (EcM) plant hosts during the establishment of symbiosis are not as well understood when compared to the functional stages of this mutualistic interaction. Our study used the EcM host Eucalyptus grandis to elucidate symbiosis-regulated pathways across the three phases of this interaction. Using a combination of RNA sequencing and metabolomics we studied both stage-specific and core responses of E. grandis during colonization by Pisolithus microcarpus. Using exogenous manipulation of the abscisic acid (ABA), we studied the role of this pathway during symbiosis establishment. Despite the mutualistic nature of this symbiosis, a large number of disease signalling TIR-NBS-LRR genes were induced. The transcriptional regulation in E. grandis was found to be dynamic across colonization with a small core of genes consistently regulated at all stages. Genes associated to the carotenoid/ABA pathway were found within this core and ABA concentrations increased during fungal integration into the root. Supplementation of ABA led to improved accommodation of P. microcarpus into E. grandis roots. The carotenoid pathway is a core response of an EcM host to its symbiont and highlights the need to understand the role of the stress hormone ABA in controlling host-EcM fungal interactions.

The primary transcriptome of hormogonia from a filamentous cyanobacterium defined by cappable-seq.

Hormogonia are motile filaments produced by many filamentous cyanobacteria that function in dispersal, phototaxis and the establishment of nitrogen-fixing symbioses. The gene regulatory network promoting hormogonium development is initiated by the hybrid histidine kinase HrmK, which in turn activates a sigma factor cascade consisting of SigJ, SigC and SigF. In this study, cappable-seq was employed to define the primary transcriptome of developing hormogonia in the model filamentous cyanobacterium Nostoc punctiforme ATCC 29133 in both the wild-type, and sigJ, sigC and sigF mutant strains 6 h post-hormogonium induction. A total of 1544 transcriptional start sites (TSSs) were identified that are associated with protein-coding genes and are expressed at levels likely to lead to biologically relevant transcripts in developing hormogonia. TSS expression among the sigma-factor deletion strains was highly consistent with previously reported gene expression levels from RNAseq experiments, and support the current working model for the role of these genes in hormogonium development. Analysis of SigJ-dependent TSSs corroborated the presence of the previously identified J-Box in the -10 region of SigJ-dependent promoters. Additionally, the data presented provides new insights on sequence conservation within the -10 regions of both SigC- and SigF-dependent promoters, and demonstrates that SigJ and SigC coordinate complex co-regulation not only of hormogonium-specific genes at different loci, but within an individual operon. As progress continues on defining the hormogonium gene regulatory network, this data set will serve as a valuable resource.

The Lotus japonicus ROP3 Is Involved in the Establishment of the Nitrogen-Fixing Symbiosis but Not of the Arbuscular Mycorrhizal Symbiosis.

Legumes form root mutualistic symbioses with some soil microbes promoting their growth, rhizobia, and arbuscular mycorrhizal fungi (AMF). A conserved set of plant proteins rules the transduction of symbiotic signals from rhizobia and AMF in a so-called common symbiotic signaling pathway (CSSP). Despite considerable efforts and advances over the past 20 years, there are still key elements to be discovered about the establishment of these root symbioses. Rhizobia and AMF root colonization are possible after a deep cell reorganization. In the interaction between the model legume Lotus japonicus and Mesorhizobium loti, this reorganization has been shown to be dependent on a SCAR/Wave-like signaling module, including Rho-GTPase (ROP in plants). Here, we studied the potential role of ROP3 in the nitrogen-fixing symbiosis (NFS) as well as in the arbuscular mycorrhizal symbiosis (AMS). We performed a detailed phenotypic study on the effects of the loss of a single ROP on the establishment of both root symbioses. Moreover, we evaluated the expression of key genes related to CSSP and to the rhizobial-specific pathway. Under our experimental conditions, rop3 mutant showed less nodule formation at 7- and 21-days post inoculation as well as less microcolonies and a higher frequency of epidermal infection threads. However, AMF root colonization was not affected. These results suggest a role of ROP3 as a positive regulator of infection thread formation and nodulation in L. japonicus. In addition, CSSP gene expression was neither affected in NFS nor in AMS condition in rop3 mutant. whereas the expression level of some genes belonging to the rhizobial-specific pathway, like RACK1, decreased in the NFS. In conclusion, ROP3 appears to be involved in the NFS, but is neither required for intra-radical growth of AMF nor arbuscule formation.

Lipopolysaccharide O-antigen molecular and supramolecular modifications of plant root microbiota are pivotal for host recognition

Lipopolysaccharides, the major outer membrane components of Gram-negative bacteria, are crucial actors of the host-microbial dialogue. They can contribute to the establishment of either symbiosis or bacterial virulence, depending on the bacterial lifestyle. Plant microbiota shows great complexity, promotes plant health and growth and assures protection from pathogens. How plants perceive LPS from plant-associated bacteria and discriminate between beneficial and pathogenic microbes is an open and urgent question. Here, we report on the structure, conformation, membrane properties and immune recognition of LPS isolated from the Arabidopsis thaliana root microbiota member Herbaspirillum sp. Root189. The LPS consists of an O-methylated and variously acetylated D-rhamnose containing polysaccharide with a rather hydrophobic surface. Plant immunology studies in A. thaliana demonstrate that the native acetylated O-antigen shields the LPS from immune recognition whereas the O-deacylated one does not. These findings highlight the role of Herbaspirillum LPS within plant-microbial crosstalk, and how O-antigen modifications influence membrane properties and modulate LPS host recognition.

Strain-level analysis reveals the vertical microbial transmission during the life cycle of bumblebee

BackgroundMicrobial acquisition and development of the gut microbiota impact the establishment of a healthy host-microbes symbiosis. Compared with other animals, the eusocial bumblebees and honeybees possess a simple, recurring, and similar set of gut microbiota. However, all bee gut phylotypes have high strain-level diversity. Gut communities of different bee species are composed of host-specific groups of strains. The variable genomic regions among strains of the same species often confer critical functional differences, such as carbon source utilization, essential for the natural selection of specific strains. The annual bumblebee colony founded by solitary queens enables tracking the transmission routes of gut bacteria during development stages.ResultsHere, we first showed the changes in the microbiome of individual bumblebees across their holometabolous life cycle. Some core gut bacteria persist throughout different stages of development. Gut microbiota of newly emerged workers always resembles those of their queens, suggesting a vertical transmission of strains from queens to the newborn workers. We then follow the dynamic changes in the gut community by comparing strain-level metagenomic profiles of queen-worker pairs longitudinally collected across different stages of the nest development. Species composition of both queen and worker shifts with the colony’s growth, and the queen-to-worker vertical inheritance of specific strains was identified. Finally, comparative metagenome analysis showed clear host-specificity for microbes across different bee hosts. Species from honeybees often possess a higher level of strain variation, and they also exhibited more complex gene repertoires linked to polysaccharide digestion. Our results demonstrate bacterial transmission events in bumblebee, highlighting the role of social interactions in driving the microbiota composition.ConclusionsBy the community-wide metagenomic analysis based on the custom genomic database of bee gut bacteria, we reveal strain transmission events at high resolution and the dynamic changes in community structure along with the colony development. The social annual life cycle of bumblebees is key for the acquisition and development of the gut microbiota. Further studies using the bumblebee model will advance our understanding of the microbiome transmission and the underlying mechanisms, such as strain competition and niche selection.DndA2Lw_mPA4b2NMU2mTwyVideo

Arbuscular mycorrhizae and silicon alleviate arsenic toxicity by enhancing soil nutrient availability, starch degradation and productivity in Cajanus cajan (L.) Millsp.

Arsenic (As) pollution of soil reduces the growth and reproductive potential of plants. Silicon (Si) and arbuscular mycorrhizal (AM) fungi play significant roles in alleviating adverse effects of As stress. However, studies are scant regarding alleviative effects of Si in pigeonpea (Cajanus cajan L. Millsp.) because legumes are considered low Si-accumulators. We investigated the individual as well as synergistic potential of Si with two AM species (M1-Claroideoglomus etunicatum and M2-Rhizoglomus intraradices) in modulating soil properties, thereby improving growth and productivity of pigeonpea genotype Pusa 2001 grown in AsV and AsIII challenged soils. Both As species hampered the establishment of AM symbiosis, thus, reducing nutrient uptake, growth and yield, with AsIII more toxic than AsV. Exogenously applied Si and AM species enhanced soil glomalin and phosphatases activity, hence decreased metal bioavailability in soil, increased plant nutrient acquisition, biomass and chlorophylls; with maximum benefits provided by M2, closely followed by Si and least by M1. These amendments boosted the activities of starch hydrolytic enzymes (α-, β-amylase, starch phosphorylase) in plants, along with a simultaneous increase in total soluble sugars (TSS). This enhanced sugar accumulation directly led to improved reproductive attributes, more efficiently by M2 and Si than by M1. Moreover, there was a substantial increase in proline biosynthesis due to significantly enhanced activities of its biosynthetic enzymes. Additionally, combined applications of Si and AM, especially +Si+M2, complemented each other where AM enhanced Si uptake, while Si induced mycorrhization, suggesting their mutual and beneficial roles in ameliorating metal(loid) toxicity and achieving sustainability in pigeonpea production under As stress.

Arbuscular mycorrhizal symbiosis enhances water stable aggregate formation and organic matter stabilization in Fe ore tailings

Organo-mineral association and water-stable aggregation in finely textured tailings are critically important to the eco-engineered soil formation from alkaline Fe ore tailings for sustainable mine site rehabilitation. Arbuscular mycorrhizal (AM) symbiosis plays important roles in soil aggregate formation and organic matter (OM) stabilization. However, it is unknown if AM symbiosis could enhance aggregate formation and OM stabilization in alkaline Fe ore tailings. The present study aimed to investigate the establishment of AM symbiosis and their role in tailing aggregate formation coupled with OM stabilization, as well as the underlying mechanisms. After initial eco-engineering (OM amendment and pioneer plant cultivation) to improve physicochemical conditions for plant survival, Sorghum spp. Hybrid cv. Silk inoculated with/without AM fungi (Glomus spp.) were cultivated in the tailings under glasshouse conditions for 14 weeks. The results indicated that AM fungi formed symbiotic association with Sorghum spp. plants, improved mineral nutrient (e.g., P) acquisition and root growth in the eco-engineered tailings. The AM symbiosis significantly improved aggregate formation. The association of organic carbon and nitrogen with tailing minerals of the aggregates was enhanced by the AM symbiosis. As revealed by synchrotron-based C 1 s near edge X-ray absorption fine structure (C 1 s NEXAFS) and Fe K edge X-ray absorption fine structure (Fe K edge XAFS) spectroscopy, the AM symbiosis favoured carboxyl and aromatic C association with secondary Fe-Si minerals, which may have been formed from AM driven mineral weathering. Overall, the study revealed that the AM symbiosis could not only improve the growth of pioneer plant species in the early eco-engineered tailings, but also advance soil formation through enhancing organic C and N sequestration and physical structure development via water-stable aggregation. These findings help to advance our understanding of the importance of AM symbiosis in the eco-engineering of tailings into functional soil (or technosols) for sustainable rehabilitation of Fe-ore tailings.

Symbiotic interactions between a newly identified native mycorrhizal fungi complex and the endemic tree Argania spinosa mediate growth, photosynthesis, and enzymatic responses under drought stress conditions

Water deficit or drought is the most important abiotic stress limiting plant growth performance and plant community development; this is typical in the Mediterranean area where plants are often severely and permanently water limited. Such is the case of the argan tree (Argania spinosa (L.) Skeels), one of the tree species most affected by desertification and global warming. To advance knowledge on how this tree can withstand drought stress, inoculation with a native complex of arbuscular mycorrhizal fungi (AMF), composed mainly of the genus Glomus, was studied in connection with a set of growth and physiological parameters. Under controlled conditions, inoculated and non-inoculated argan seedlings were grown for 3 months under three water regimens: 25%, 50%, and 75% relative to the field capacity of used soil substrate. The results showed that the argan tree had different growth abilities to develop and withstand the various applied water limitations. The AMF complex stimulated the growth and mineral nutrition of argan seedlings under the different imposed levels of water deficiency. Relative water content (RWC) in leaves, water potential, and stomatal conductance in argan leaves showed a general improvement in inoculated seedlings compared to non-inoculated ones. Soluble sugar and proline contents significantly increased in non-inoculated seedlings compared with inoculated seedlings under water-limiting conditions (25%). Similarly, oxidative enzyme (catalase, peroxidase, superoxide dismutase) activity increased significantly in drought-stressed seedlings. Non-inoculated seedlings showed the highest accumulation of these enzymes. Moreover, mycorrhizal symbiosis establishment positively correlated with argan tree seedlings in terms of growth, mineral nutrition, soluble sugar and proline contents, and enzymes activities. The main results from the current study suggest that AMF improve the ability of A. spinosa to tolerate drought by enhancing mineral nutrition and the transport of high levels of water by enhancing the RWC and water potential in leaves. Finally, the alleviation of the destructive effects of reactive oxygen species was modulated by enzymatic scavenging activity. Hence, the use of AMF in the technical process of argan seedlings production is highly recommended in different ecofriendly restoration strategies, with the aim of producing high quality seedlings capable of tolerating drought stress.

Can orchid mycorrhizal fungi be persistently harbored by the plant host?

The environmental distribution of non-obligate orchid mycorrhizal (OM) symbionts belonging to the ‘rhizoctonia’ complex remains elusive. Some of these fungi, indeed, are undetectable in soil outside the host rhizosphere. A manipulation experiment was performed to assess the importance of neighbouring non-orchid plants and soil as possible reservoirs of OM fungi for Spiranthes spiralis , a widespread photosynthetic European terrestrial orchid species. Fungi of S. spiralis roots were identified by DNA metabarcoding before and 4 months after the removal of the surrounding vegetation and soil. Although such a treatment significantly affected fungal colonization of newly-formed orchid roots, most OM fungi were consistently associated with the host roots. Frequency patterns in differently aged roots suggest that these fungi colonize new orchid roots from either older roots or other parts of the same plant, which may thus represent an environmental source for the subsequent establishment of the OM symbiosis. • We addressed the importance of vegetation and soil as reservoirs of orchid mycobionts. • Removal of other plants and soil impacted Spiranthes spiralis root fungal community. • Most orchid mycorrhizal fungi were consistently associated with the host roots. • Some fungi may colonize mature orchid roots from other parts of the same plant. • Our findings have implications for the interpretation of orchid distribution.

Medicago SPX1 and SPX3 regulate phosphate homeostasis, mycorrhizal colonization, and arbuscule degradation.

To acquire sufficient mineral nutrients such as phosphate (Pi) from the soil, most plants engage in symbiosis with arbuscular mycorrhizal (AM) fungi. Attracted by plant-secreted strigolactones (SLs), the fungi colonize the roots and form highly branched hyphal structures called arbuscules inside inner cortex cells. The host plant must control the different steps of this interaction to maintain its symbiotic nature. However, how plants sense the amount of Pi obtained from the fungus, and how this determines the arbuscule lifespan, are far from understood. Here, we show that Medicago truncatula SPX-domain containing proteins SPX1 and SPX3 regulate root Pi starvation responses, in part by interacting with PHOSPHATE RESPONSE REGULATOR2, as well as fungal colonization and arbuscule degradation. SPX1 and SPX3 are induced upon Pi starvation but become more restricted to arbuscule-containing cells upon the establishment of symbiosis. This induction in arbuscule-containing cells is associated with the presence of cis-regulatory AW-boxes and transcriptional regulation by the WRINKLED1-like transcription factor WRI5a. Under Pi-limiting conditions, SPX1 and SPX3 facilitate the expression of the SL biosynthesis gene DWARF27, which could help explain the increased fungal branching in response to root exudates. Later, in arbuscule-containing cells, SPX1 and SPX3 redundantly control arbuscule degradation. Thus, SPX proteins play important roles as phosphate sensors to maintain a beneficial AM symbiosis.

Mycorrhizae Helper Bacteria: Unlocking Their Potential as Bioenhancers of Plant–Arbuscular Mycorrhizal Fungal Associations

The dynamic interactions of plants and arbuscular mycorrhizal fungi (AMF) that facilitate the efficient uptake of minerals from soil and provide protection from various environmental stresses (biotic and abiotic) are now also attributed to a third component of the symbiosis. These are the less investigated mycorrhizae helper bacteria (MHB), which constitute a dense, active bacterial community, tightly associated with AMF, and involved in the development and functioning of AMF. Although AMF spores are known to host several bacteria in their spore walls and cytoplasm, their role in promoting the ecological fitness and establishment of AMF symbiosis by influencing spore germination, mycelial growth, root colonization, metabolic diversity, and biocontrol of soil borne diseases is now being deciphered. MHB also promote the functioning of arbuscular mycorrhizal symbiosis by triggering various plant growth factors, leading to better availability of nutrients in the soil and uptake by plants. In order to develop strategies to promote mycorrhization by AMF, and particularly to stimulate the ability to utilize phosphorus from the soil, there is a need to decipher crucial metabolic signalling pathways of MHB and elucidate their functional significance as mycorrhiza helper bacteria. MHB, also referred to as AMF bioenhancers, also improve agronomic efficiency and formulations using AMF along with enriched population of MHB are a promising option. This review covers the aspects related to the specificity and mechanisms of action of MHB, which positively impact the formation and functioning of AMF in mycorrhizal symbiosis, and the need to advocate MHB as AMF bioenhancers towards their inclusion in integrated nutrient management practices in sustainable agriculture.

Suppression of LjBAK1-mediated immunity by SymRK promotes rhizobial infection in Lotus japonicus

An important question in biology is how organisms can associate with different microbes that pose no threat (commensals), pose a severe threat (pathogens), and those that are beneficial (symbionts). The root nodule symbiosis serves as an important model system for addressing such questions in the context of plant-microbe interactions. It is now generally accepted that rhizobia can actively suppress host immune responses during the infection process, analogous to the way in which plant pathogens can evade immune recognition. However, much remains to be learned about the mechanisms by which the host recognizes the rhizobia as pathogens and how, subsequently, these pathways are suppressed to allow establishment of the nitrogen-fixing symbiosis. In this study, we found that SymRK (Symbiosis Receptor-like Kinase) is required for rhizobial suppression of plant innate immunity in Lotus japonicus. SymRK associates with LjBAK1 (BRASSINOSTEROID INSENSITIVE 1-Associated receptor Kinase 1), a well-characterized positive regulator of plant innate immunity, and directly inhibits LjBAK1 kinase activity. Rhizobial inoculation enhances the association between SymRK and LjBAK1 in planta. LjBAK1 is required for the regulation of plant innate immunity and plays a negative role in rhizobial infection in L. japonicus. The data indicate that the SymRK-LjBAK1 protein complex serves as an intersection point between rhizobial symbiotic signaling pathways and innate immunity pathways, and support that rhizobia may actively suppress the host's ability to mount a defense response during the legume-rhizobium symbiosis.

Cell surface carbohydrates of symbiotic dinoflagellates and their role in the establishment of cnidarian\u2013dinoflagellate symbiosis

Symbiodiniaceae algae are often photosymbionts of reef-building corals. The establishment of their symbiosis resembles a microbial infection where eukaryotic pattern recognition receptors (e.g. lectins) are thought to recognize a specific range of taxon-specific microbial-associated molecular patterns (e.g. glycans). The present study used the sea anemone, Exaiptasia diaphana and three species of Symbiodiniaceae (the homologous Breviolum minutum, the heterologous-compatible Cladocopium goreaui and the heterologous-incompatible Fugacium kawagutii) to compare the surface glycomes of three symbionts and explore the role of glycan–lectin interactions in host–symbiont recognition and establishment of symbiosis. We identified the nucleotide sugars of the algal cells, then examined glycans on the cell wall of the three symbiont species with monosaccharide analysis, lectin array technology and fluorescence microscopy of the algal cell decorated with fluorescently tagged lectins. Armed with this inventory of possible glycan moieties, we then assayed the ability of the three Symbiodiniaceae to colonize aposymbiotic E. diaphana after modifying the surface of one of the two partners. The Symbiodiniaceae cell-surface glycome varies among algal species. Trypsin treatment of the alga changed the rate of B. minutum and C. goreaui uptake, suggesting that a protein-based moiety is an essential part of compatible symbiont recognition. Our data strongly support the importance of D-galactose (in particular β-D-galactose) residues in the establishment of the cnidarian–dinoflagellate symbiosis, and we propose a potential involvement of L-fucose, D-xylose and D-galacturonic acid in the early steps of this mutualism.

Arbuscular Mycorrhizal Fungi Symbiosis in Durum Wheat (Triticum durum Desf.) under No-tillage and Tillage Practices in a Semiarid Region

Background: Arbuscular mycorrhizal fungi (AMF) are root symbionts that improve host plant growth and resilience against biotic and abiotic stresses allowing a sustain plant production particularly under harsh conditions. Methods: The objective of this study is to compare the effect of conventional tillage and no-tillage practices on AMF symbiosis with durum wheat (Triticum durum Desf) through the evaluation of root colonization and AMF spore density in the soil of three different sites in a semi-arid region in Algeria. Two sites were conducted under rain-fed conditions and one was irrigated.Result: Mycorrhizal root colonization varied according to the site and the tillage practice, while spore density differed between sites. Spore density was higher in sites under rain-fed conditions suggesting that water deficit stimulate sporulation, while root colonization seems to be limited by drought. No-tillage improved root colonization rates by 54.3% compared to conventional tillage system and this improvement, particularly the arbuscular percentage, was higher under drought conditions. In addition, root colonization rates showed a positive correlation with the organic matter content and pH in the soil and a negative correlation with the available phosphorus in the soil. These results indicate that no-tillage enhance the establishment of AMF symbiosis with durum wheat under semi-arid conditions.

Can Acropora tenuis larvae attract native Symbiodiniaceae cells by green fluorescence at the initial establishment of symbiosis?

Most corals acquire symbiodiniacean symbionts from the surrounding environment to initiate symbiosis. The cell densities of Symbiodiniaceae in the environment are usually low, and mechanisms may exist by which new coral generations attract suitable endosymbionts. Phototaxis of suitable symbiodiniacean cells toward green fluorescence in corals has been proposed as one such mechanism. In the present study, we observed the phototaxis action wavelength of various strains of Symbiodiniaceae and the fluorescence spectra of aposymbiotic Acropora tenuis larvae at the time of endosymbiont uptake. The phototaxis patterns varied among the Symbiodiniaceae species and "native" endosymbionts-commonly found in Acropora juveniles present in natural environments; that is, Symbiodinium microadriaticum was attracted to blue light rather than to green light. Another native endosymbiont, Durusdinium trenchii, showed no phototaxis specific to any wavelength. Although the larvae exhibited green and broad orange fluorescence under blue-violet excitation light, the maximum green fluorescence peak did not coincide with that of the phototaxis action spectrum of S. microadriaticum. Rather, around the peak wavelength of larval green fluorescence, this native endosymbiont showed slightly negative phototaxis, suggesting that the green fluorescence of A. tenuis larvae may not play a role in the initial attraction of native endosymbionts. Conversely, broad blue larval fluorescence under UV-A excitation covered the maximum phototaxis action wavelength of S. microadriaticum. We also conducted infection tests using native endosymbionts and aposymbiotic larvae under red LED light that does not excite visible larval fluorescence. Almost all larvae failed to acquire S. microadriaticum cells, whereas D. trenchii cells were acquired by larvae even under red illumination. Thus, attraction mechanisms other than visible fluorescence might exist, at least in the case of D. trenchii. Our results suggest that further investigation and discussion, not limited to green fluorescence, would be required to elucidate the initial attraction mechanisms.

Dinoflagellate symbionts escape vomocytosis by host cell immune suppression.

Alveolata comprises diverse taxa of single-celled eukaryotes, many renowned for their ability to live inside animal cells. Notable examples are apicomplexan parasites and dinoflagellate symbionts, the latter of which power coral reef ecosystems. Although functionally distinct, they evolved from a common, free-living ancestor and must evade their hosts’ immune response for persistence. Both the initial cellular events that gave rise to this intracellular lifestyle and the role of host immune modulation in coral-dinoflagellate endosymbiosis are poorly understood. Here, we use a comparative approach in the cnidarian endosymbiosis model Aiptasia, which re-establishes endosymbiosis with free-living dinoflagellates every generation. We find that uptake of microalgae is largely indiscriminate, but non-symbiotic microalgae are expelled by vomocytosis, while symbionts induce host-cell innate immune suppression and form a LAMP1-positive niche. We demonstrate that exogenous immune stimulation results in symbiont expulsion and conversely, inhibition of canonical toll-like receptor (TLR) signaling enhances infection of host animals. Our findings indicate that symbiosis establishment is dictated by local innate immune suppression, to circumvent expulsion and promote niche formation. This work provides insight into the evolution of the cellular immune response and key steps involved in mediating endosymbiotic interactions.

Auxin Response Factor 2 (ARF2), ARF3, and ARF4 Mediate Both Lateral Root and Nitrogen Fixing Nodule Development in Medicago truncatula.

Auxin Response Factors (ARFs) constitute a large family of transcription factors that mediate auxin-regulated developmental programs in plants. ARF2, ARF3, and ARF4 are post-transcriptionally regulated by the microRNA390 (miR390)/trans-acting small interference RNA 3 (TAS3) module through the action of TAS3-derived trans-acting small interfering RNAs (ta-siRNA). We have previously reported that constitutive activation of the miR390/TAS3 pathway promotes elongation of lateral roots but impairs nodule organogenesis and infection by rhizobia during the nitrogen-fixing symbiosis established between Medicago truncatula and its partner Sinorhizobium meliloti. However, the involvement of the targets of the miR390/TAS3 pathway, i.e., MtARF2, MtARF3, MtARF4a, and MtARF4b, in root development and establishment of the nitrogen-fixing symbiosis remained unexplored. Here, promoter:reporter fusions showed that expression of both MtARF3 and MtARF4a was associated with lateral root development; however, only the MtARF4a promoter was active in developing nodules. In addition, up-regulation of MtARF2, MtARF3, and MtARF4a/b in response to rhizobia depends on Nod Factor perception. We provide evidence that simultaneous knockdown of MtARF2, MtARF3, MtARF4a, and MtARF4b or mutation in MtARF4a impaired nodule formation, and reduced initiation and progression of infection events. Silencing of MtARF2, MtARF3, MtARF4a, and MtARF4b altered mRNA levels of the early nodulation gene nodulation signaling pathway 2 (MtNSP2). In addition, roots with reduced levels of MtARF2, MtARF3, MtARF4a, and MtARF4b, as well as arf4a mutant plants exhibited altered root architecture, causing a reduction in primary and lateral root length, but increasing lateral root density. Taken together, our results suggest that these ARF members are common key players of the morphogenetic programs that control root development and the formation of nitrogen-fixing nodules.

N-Acetyl-d-Glucosamine-Binding Lectin in Acropora tenuis Attracts Specific Symbiodiniaceae Cell Culture Strains.

Many corals establish symbiosis with Symbiodiniaceae cells from surrounding environments, but very few Symbiodiniaceae cells exist in the water column. Given that the N-acetyl-d-glucosamine-binding lectin ActL attracts Symbiodiniaceae cells, we hypothesized that corals must attract Symbiodiniaceae cells using ActL to acquire them. Anti-ActL antibody inhibited acquisition of Symbiodiniaceae cells, and rearing seawater for juvenile Acropora tenuis contained ActL, suggesting that juvenile A. tenuis discharge ActL to attract these cells. Among eight Symbiodiniaceae cultured strains, ActL attracted NBRC102920 (Symbiodinium tridacnidorum) most strongly followed by CS-161 (Symbiodinium tridacnidorum), CCMP2556 (Durusdinium trenchii), and CCMP1633 (Breviolum sp.); however, it did not attract GTP-A6-Sy (Symbiodinium natans), CCMP421 (Effrenium voratum), FKM0207 (Fugacium sp.), and CS-156 (Fugacium sp.). Juvenile polyps of A. tenuis acquired limited Symbiodiniaceae cell strains, and the number of acquired Symbiodiniaceae cells in a polyp also differed from each other. The number of Symbiodiniaceae cells acquired by juvenile polyps of A. tenuis was correlated with the ActL chemotactic activity. Thus, ActL could be used to attract select Symbiodiniaceae cells and help Symbiodiniaceae cell acquisition in juvenile polyps of A. tenuis, facilitating establishment of symbiosis between A. tenuis and Symbiodiniaceae cells.

Dual oxidase enables insect gut symbiosis by mediating respiratory network formation

Most animals harbor a gut microbiota that consists of potentially pathogenic, commensal, and mutualistic microorganisms. Dual oxidase (Duox) is a well described enzyme involved in gut mucosal immunity by the production of reactive oxygen species (ROS) that antagonizes pathogenic bacteria and maintains gut homeostasis in insects. However, despite its nonspecific harmful activity on microorganisms, little is known about the role of Duox in the maintenance of mutualistic gut symbionts. Here we show that, in the bean bug Riptortus pedestris, Duox-dependent ROS did not directly contribute to epithelial immunity in the midgut in response to its mutualistic gut symbiont, Burkholderia insecticola Instead, we found that the expression of Duox is tracheae-specific and its down-regulation by RNAi results in the loss of dityrosine cross-links in the tracheal protein matrix and a collapse of the respiratory system. We further demonstrated that the establishment of symbiosis is a strong oxygen sink triggering the formation of an extensive network of tracheae enveloping the midgut symbiotic organ as well as other organs, and that tracheal breakdown by Duox RNAi provokes a disruption of the gut symbiosis. Down-regulation of the hypoxia-responsive transcription factor Sima or the regulators of tracheae formation Trachealess and Branchless produces similar phenotypes. Thus, in addition to known roles in immunity and in the formation of dityrosine networks in diverse extracellular matrices, Duox is also a crucial enzyme for tracheal integrity, which is crucial to sustain mutualistic symbionts and gut homeostasis. We expect that this is a conserved function in insects.

Are strigolactones a key in plant–parasitic nematodes interactions? An intriguing question

Plant parasitic nematodes (PPNs) are among the most important pests in agriculture. Chemical inputs are widely used for their control; however, the negative impact of these agrochemicals on environmental and human health is a current concern. Biological control and interventions on rhizosphere signaling are promising ecofriendly alternatives for managing these pests in the field. Nevertheless, the molecular mechanisms involved in plant-PPN interaction need to be unraveled in order to develop appropriate management strategies. Strigolactones (SLs) are phytohormones that are exuded from roots, acting as signaling molecules in the rhizosphere. They are important cues in the establishment of arbuscular mycorrhizal and Rhizobium-legume symbioses. Recently, it has been shown that SLs can also affect the interaction between plants and certain PPN species; however, data are scarce and ambiguous. Some studies propose that SLs positively regulate PPN species performance acting as attractants to roots, or by inhibiting plant defense responses. On the contrary, other studies suggest that SLs could negatively regulate PPN performance by reducing abscisic acid, or by promoting plant interactions with beneficial soil microorganisms. In the present Opinion paper, we discuss these controversial results and propose future research challenges to develop new management strategies against these harmful PPN species.