Rhizobium-legume symbiosis is a specific, coordinated interaction that results in the formation of a root nodule, where biological nitrogen fixation occurs. NADPH oxidases, or Respiratory Burst ...Oxidase Homologs (RBOHs) in plants, are enzymes that generate superoxide (O
). Superoxide produces other reactive oxygen species (ROS); these ROS regulate different stages of mutualistic interactions. For example, changes in ROS levels are thought to induce ROS scavenging, cell wall remodeling, and changes in phytohormone homeostasis during symbiotic interactions. In common bean (Phaseolus vulgaris), PvRbohB plays a key role in the early stages of nodulation.
In this study, to explore the role of PvRbohB in root nodule symbiosis, we analyzed transcriptomic data from the roots of common bean under control conditions (transgenic roots without construction) and roots with downregulated expression of PvRbohB (by RNA interference) non-inoculated and inoculated with R. tropici. Our results suggest that ROS produced by PvRBOHB play a central role in infection thread formation and nodule organogenesis through crosstalk with flavonoids, carbon metabolism, cell cycle regulation, and the plant hormones auxin and cytokinin during the early stages of this process.
Our findings provide important insight into the multiple roles of ROS in regulating rhizobia-legume symbiosis.
Desert plants are hypothesized to survive the environmental stress inherent to these regions in part thanks to symbioses with microorganisms, and yet these microbial species, the communities they ...form, and the forces that influence them are poorly understood. Here we report the first comprehensive investigation of the microbial communities associated with species of Agave, which are native to semiarid and arid regions of Central and North America and are emerging as biofuel feedstocks. We examined prokaryotic and fungal communities in the rhizosphere, phyllosphere, leaf and root endosphere, as well as proximal and distal soil samples from cultivated and native agaves, through Illumina amplicon sequencing. Phylogenetic profiling revealed that the composition of prokaryotic communities was primarily determined by the plant compartment, whereas the composition of fungal communities was mainly influenced by the biogeography of the host species. Cultivated A. tequilana exhibited lower levels of prokaryotic diversity compared with native agaves, although no differences in microbial diversity were found in the endosphere. Agaves shared core prokaryotic and fungal taxa known to promote plant growth and confer tolerance to abiotic stress, which suggests common principles underpinning Agave–microbe interactions.
Non-specific lipid transfer proteins (LTPs) constitute a large protein family in plants characterized by having a tunnel-like hydrophobic cavity, which allows them to transfer different lipid ...molecules. LTPs have been studied in various model plants including those of agronomic interest. Recent studies have demonstrated that LTPs play key functions in both biotic and abiotic stress. In plants-pathogen interaction, they act as either positive or negative regulators of defense responses. However, little is known about the roles of LTPs in symbiotic interactions, especially in root nodule symbiosis. Here, we performed a broad genome analysis of LTP family members in legumes and other important model plants, focusing on their possible roles in legume-rhizobium symbiosis.
In silico
analysis showed that legumes contain large LTP families, with at least 70 LTP members clustered into four clades. Although the structures of LTP genes and proteins are conserved among species, differences were observed between clades from different species. LTPs are widely expressed in different plant tissues. In general, genes of the
LTP1
and
LTP2
classes are highly expressed in shoot and reproductive tissues in all analyzed species. Furthermore, genes of the different classes are also expressed in roots inoculated with rhizobia and nodules of legumes. RT-qPCR expression profile analysis of seven
PvLTP
genes in common bean (
Phaseolus vulgaris
) revealed that these genes are differentially expressed during the early and late stages of nodulation and they are genetically regulated by
PvRbohA
. These findings provide insight into the putative roles of LTP family members in legume-rhizobium symbiosis and their possible interactions with RBOH-dependent ROS production.
The mutualistic interaction between gram-negative soil bacteria and the roots of legumes leads to the establishment of nodules, where atmospheric nitrogen is fixed. Nodulation is a multistep process ...with numerous essential players. Among these are reactive oxygen species (ROS), which are mainly generated by Nicotinamide Adenine Dinucleotide Phosphate (NADPH) oxidases. In plants, these enzymes are known as respiratory burst oxidase homologs (RBOHs). In legumes, these proteins are encoded by a multigene family with members that are differentially expressed in various tissues and organs at distinct developmental stages. RBOHs have critical roles at several stages of nodulation: in the early signaling pathway triggered by nodulation factors in the root hairs, during both the progression of infection threads and nodule ontogeny, and in nitrogen fixation and senescence. Data from the literature along with the analysis conducted here imply that legumes use different RBOHs for different stages of nodulation; these RBOHs belong to the same phylogenetic subgroup, even though they are not strictly orthologous. Accordingly, the regulation of activity of a given RBOH during the nodulation process probably varies among legumes.
Cactaceae represents one of the most species-rich families of succulent plants native to arid and semi-arid ecosystems, yet the associations Cacti establish with microorganisms and the rules ...governing microbial community assembly remain poorly understood. We analyzed the composition, diversity, and factors influencing above- and below-ground bacterial, archaeal, and fungal communities associated with two native and sympatric Cacti species: Myrtillocactus geometrizans and Opuntia robusta. Phylogenetic profiling showed that the composition and assembly of microbial communities associated with Cacti were primarily influenced by the plant compartment; plant species, site, and season played only a minor role. Remarkably, bacterial, and archaeal diversity was higher in the phyllosphere than in the rhizosphere of Cacti, while the opposite was true for fungi. Semi-arid soils exhibited the highest levels of microbial diversity whereas the stem endosphere the lowest. Despite their taxonomic distance, M. geometrizans and O. robusta shared most microbial taxa in all analyzed compartments. Influence of the plant host did only play a larger role in the fungal communities of the stem endosphere. These results suggest that fungi establish specific interactions with their host plant inside the stem, whereas microbial communities in the other plant compartments may play similar functional roles in these two species. Biochemical and molecular characterization of seed-borne bacteria of Cacti supports the idea that these microbial symbionts may be vertically inherited and could promote plant growth and drought tolerance for the fitness of the Cacti holobiont. We envision this knowledge will help improve and sustain agriculture in arid and semi-arid regions of the world.
The plant receptor-like-kinase subfamily CrRLK1L has been widely studied, and CrRLK1Ls have been described as crucial regulators in many processes in
(L.), Heynh. Little is known, however, about the ...functions of these proteins in other plant species, including potential roles in symbiotic nodulation. We performed a phylogenetic analysis of CrRLK1L subfamily receptors of 57 different plant species and identified 1050 CrRLK1L proteins, clustered into 11 clades. This analysis revealed that the CrRLK1L subfamily probably arose in plants during the transition from chlorophytes to embryophytes and has undergone several duplication events during its evolution. Among the
of legumes and
, protein structure, gene structure, and expression patterns were highly conserved. Some legume
genes were active in nodules. A detailed analysis of eight nodule-expressed genes in
L. showed that these genes were differentially expressed in roots at different stages of the symbiotic process. These data suggest that
are both conserved and underwent diversification in a wide group of plants, and shed light on the roles of these genes in legume-rhizobia symbiosis.
Microbial symbionts account for survival, development, fitness and evolution of eukaryotic hosts. These microorganisms together with their host form a biological unit known as holobiont. Recent ...studies have revealed that the holobiont of agaves and cacti comprises a diverse and structured microbiome, which might be important for its adaptation to drylands. Here, we investigated the functional signatures of the prokaryotic communities of the soil and the episphere, that includes the rhizosphere and phyllosphere, associated with the cultivated
and the native and sympatric
and
by mining shotgun metagenomic data. Consistent with previous phylogenetic profiling, we found that Proteobacteria, Actinobacteria and Firmicutes were the main represented phyla in the episphere of agaves and cacti, and that clustering of metagenomes correlated with the plant compartment. In native plants, genes related to aerobic anoxygenic phototrophy and photosynthesis were enriched in the phyllosphere and soil, while genes coding for biofilm formation and quorum sensing were enriched in both epiphytic communities. In the episphere of cultivated
fewer genes were identified, but they belonged to similar pathways than those found in native plants.
showed a depletion in several genes belonging to carbon metabolism, secondary metabolite biosynthesis and xenobiotic degradation suggesting that its lower microbial diversity might be linked to functional losses. However, this species also showed an enrichment in biofilm and quorum sensing in the epiphytic compartments, and evidence for nitrogen fixation in the rhizosphere. Aerobic anoxygenic phototrophic markers were represented by Rhizobiales (
) and Rhodospirillales (
) in the phyllosphere, while photosystem genes were widespread in Bacillales and Cyanobacteria. Nitrogen fixation and biofilm formation genes were mostly related to Proteobacteria. These analyses support the idea of niche differentiation in the rhizosphere and phyllosphere of agaves and cacti and shed light on the potential mechanisms by which epiphytic microbial communities survive and colonize plants of arid and semiarid ecosystems. This study establishes a guideline for testing the relevance of the identified functional traits on the microbial community and the plant fitness.
Abstract
One of the most important physicochemical factors that affect cell growth and development is pH, and living organisms have developed specific mechanisms to adapt to media with variable pH ...values. Most fungi posses a specific mechanism for such adaptation: the Pal/Rim pathway. To analyze the different metabolic processes regulated by this pathway, and its possible relationships with other physiological regulatory mechanisms, we analyzed the phenotype of a mutant in the PALB/RIM13 gene of the phytopathogenic fungus Ustilago maydis. The mutant displayed important alterations in the synthesis and organization of the cell wall and was affected in its response to stress, revealing its relationship with the MAPKC pathway involved in maintaining the integrity of the cell wall, and the stress response pathway, but not with the HOG pathway. An important observation was that the mutant, in contrast to the wild-type strain, was unable to maintain a constant intracellular pH, suggesting that probably the main function of the Pal/Rim pathway, in collaboration with other regulatory mechanisms, is to maintain a constant intracellular pH, despite the changes occurring in the environment.
Reactive oxygen species (ROS) are generated by NADPH oxidases known as respiratory burst oxidase homologs (RBOHs) in plants. ROS regulate various cellular processes, including the mutualistic ...interactions between legumes and nitrogen-fixing bacteria or arbuscular mycorrhizal (AM) fungi. Rboh is a multigene family comprising nine members (RbohA-I) in common bean (Phaseolus vulgaris). The RNA interference-mediated silencing of RbohB (PvRbohB-RNAi) in this species diminished its ROS production and greatly impaired nodulation. By contrast, the PvRbohB-RNAi transgenic roots showed early hyphal root colonization with enlarged fungal hypopodia; therefore, we proposed that PvRbohB positively regulates rhizobial infection (Rhizobium tropici) and inhibits AM colonization by Rhizophagus irregularis in P. vulgaris.
To corroborate this hypothesis, an RNA-Seq transcriptomic analysis was performed to identify the differentially expressed genes in the PvRbohB-RNAi roots inoculated with Rhizobium tropici or Rhizophagus irregularis. We found that, in the early stages, root nodule symbioses generated larger changes of the transcriptome than did AM symbioses in P. vulgaris. Genes related to ROS homeostasis and cell wall flexibility were markedly upregulated in the early stages of rhizobial colonization, but not during AM colonization. Compared with AM colonization, the rhizobia induced the expression of a greater number of genes encoding enzymes involved in the metabolism of auxins, cytokinins, and ethylene, which were typically repressed in the PvRbohB-RNAi roots.
Our research provides substantial insights into the genetic interaction networks in the early stages of rhizobia and AM symbioses with P. vulgaris, as well as the differential roles that RbohB plays in processes related to ROS scavenging, cell wall remodeling, and phytohormone homeostasis during nodulation and mycorrhization in this legume.
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