Search Results (14)

Global warming-induced climate change haunts the world, posing a critical threat to plant health and crop production. Overusing chemical fertilizers and pesticides poses a significant threat to soil health. Ceanothus velutinus (snowbrush) is a drought-tolerant, actinorhizal native plant found in the Intermountain West region of the US that harbors many plant growth-promoting rhizobacteria (PGPR). In this study, we evaluated the effects of PGPR CK-06, CK-22, CK-44, and CK-50 from C. velutinus on the growth and development of two tall fescue genotypes: (i) a lawn-type tall fescue blend and (ii) an endophyte-free forage-type tall fescue known as Armory. Tall fescue plants were grown in field soil and sand mix in pots and treated twice with 5 mL of bacterial inoculum. Two isolates, CK-06 and CK-22, significantly increased tiller numbers (p < 0.05) in the lawn-type tall fescue blend, and all isolates showed a significant increase in fresh and dry weight compared to the control. Isolate CK-22 significantly increased the tiller number and fresh and dry weight of the forage-type tall fescue Armory compared to the control. Isolates CK-44 and CK-50 tested positive for sulfur-oxidizing properties, and CK-44 was able to restore the sulfur content in sulfur-deficient soil compared to the control. Full article

Atmospheric CO₂ levels have been increasing, and these changes may result in differential adaptive responses in both genera and species and highlight the need to increase carbon sequestration. Ecophysiological and morphological responses of four early-successional deciduous species were examined under ambient CO₂ (aCO₂, 400 ppm) and elevated CO₂ (eCO₂, 800 ppm) treatments. The four species, all of which are used in restoration, were Alnus viridis subsp. crispa (Ait.) Turrill (green alder), A. incana subsp. rugosa (Du Roi) R.T. Clausen (speckled alder), Betula populifolia (Marshall) (gray birch), and B. papyrifera (Marshall) (white birch); all are from the same phylogenetic family, Betulaceae. We examined biochemical efficiencies, gas exchange, chlorophyll fluorescence, chlorophyll concentrations, foliar nitrogen (N), and growth traits. A general linear model, analysis of variance, was used to analyze the functional carbon efficiency and growth differences, if any, among genera, species, and provenances (only for growth traits). The alders had greater biochemical efficiency traits than birches, and alders upregulated these traits, whereas birches mostly downregulated these traits in response to eCO₂. In response to eCO₂, assimilation either remained the same or was upregulated for alders but downregulated for birches. Stomatal conductance was downregulated for all four species in response to eCO₂. Intrinsic water use efficiency was greater for alders than for birches. Alders exhibited a consistent upregulation of stem dry mass and height growth, whereas birches were somewhat lower in height and stem dry mass in response to eCO₂. Foliar N played an important role in relation to ecophysiological traits and had significant effects relative to genus (alders > birches) and CO₂ (aCO₂ > eCO₂), and a significant genus × CO₂ interaction, with alders downregulating foliar N less than did birches. Covariate analysis examining carbon efficiency traits in relation to foliar N showed clear functional responses. Both species in both genera were consistent in their ecophysiological and morphological responses to CO₂ treatments. There was supporting evidence that assimilation was sink-driven, which is related to a plant organ’s ability to continue to grow and incorporate assimilates. The alders used in this study are actinorhizal, and the additional available foliar N, paired with increased stem dry mass sink activity, appeared to be driving upregulation of the carbon efficiencies and growth in response to eCO₂. Alders’ greater carbon efficiencies and carbon sequestration in impoverished soils demonstrate that alders, as opposed to birches, should be used to accelerate ecological restoration in a world of increasing atmospheric CO₂. Full article

Nitrogen is an essential element needed for plants to survive, and legumes are well known to recruit rhizobia to fix atmospheric nitrogen. In this widely studied symbiosis, legumes develop specific structures on the roots to host specific symbionts. This review explores alternate nodule structures and their functions outside of the more widely studied legume–rhizobial symbiosis, as well as discussing other unusual aspects of nodulation. This includes actinorhizal-Frankia, cycad-cyanobacteria, and the non-legume Parasponia andersonii-rhizobia symbioses. Nodules are also not restricted to the roots, either, with examples found within stems and leaves. Recent research has shown that legume–rhizobia nodulation brings a great many other benefits, some direct and some indirect. Rhizobial symbiosis can lead to modifications in other pathways, including the priming of defence responses, and to modulated or enhanced resistance to biotic and abiotic stress. With so many avenues to explore, this review discusses recent discoveries and highlights future directions in the study of nodulation. Full article

Biological N₂ fixation, a major pathway for new nitrogen (N) input to terrestrial ecosystems, largely determines the dynamics of ecosystem structure and functions under global change. Nevertheless, the responses of N₂ fixation to multiple global change factors remain poorly understood. Here, saplings of two N₂-fixing plant species, Alnus cremastogyne and Cajanus cajan, were grown at rural and urban sites, respectively, with the latter representing an environment with changes in multiple factors occurring simultaneously. Symbiotic N₂ fixation per unit of nodule was significantly higher at the urban site than the rural site for A. cremastogyne, but the rates were comparable between the two sites for C. cajan. The nodule investments were significantly lower at the urban site relative to the rural site for both species. Symbiotic N₂ fixation per plant increased by 31.2 times for A. cremastogyne, while that decreased by 88.2% for C. cajan at the urban site compared to the rural site. Asymbiotic N₂ fixation rate in soil decreased by 46.2% at the urban site relative to the rural site. The decrease in symbiotic N₂ fixation per plant for C. cajan and asymbiotic N₂ fixation in soil was probably attributed to higher N deposition under the urban conditions, while the increase in symbiotic N₂ fixation per plant for A. cremastogyne was probably related to the higher levels of temperature, atmospheric CO₂, and phosphorus deposition at the urban site. The responses of N₂ fixation to multiple global change factors and the underlying mechanisms may be divergent either between symbiotic and asymbiotic forms or among N₂-fixing plant species. While causative evidence is urgently needed, we argue that these differences should be considered in Earth system models to improve the prediction of N₂ fixation under global change. Full article

The present study aimed to use comparative genomics to explore the relationships between Frankia and actinorhizal plants using a data set made of 33 Frankia genomes. The determinants of host specificity were first explored for “Alnus-infective strains” (i.e., Frankia strains belonging to Cluster Ia). Several genes were specifically found in these strains, including an agmatine deiminase which could possibly be involved in various functions as access to nitrogen sources, nodule organogenesis or plant defense. Within “Alnus-infective strains”, Sp+ Frankia genomes were compared to Sp− genomes in order to elucidate the narrower host specificity of Sp+ strains (i.e., Sp+ strains being capable of in planta sporulation, unlike Sp− strains). A total of 88 protein families were lost in the Sp+ genomes. The lost genes were related to saprophytic life (transcriptional factors, transmembrane and secreted proteins), reinforcing the proposed status of Sp+ as obligatory symbiont. The Sp+ genomes were also characterized by a loss of genetic and functional paralogs, highlighting a reduction in functional redundancy (e.g., hup genes) or a possible loss of function related to a saprophytic lifestyle (e.g., genes involved in gas vesicle formation or recycling of nutrients). Full article

Climate change and the accelerated rate of population growth are imposing a progressive degradation of natural ecosystems worldwide. In this context, the use of pioneer trees represents a powerful approach to reverse the situation. Among others, N₂-fixing actinorhizal trees constitute important elements of plant communities and have been successfully used in land reclamation at a global scale. In this study, we have analyzed the transcriptome of the photosynthetic organs of Casuarina glauca (branchlets) to unravel the molecular mechanisms underlying salt stress tolerance. For that, C. glauca plants supplied either with chemical nitrogen (KNO₃⁺) or nodulated by Frankia (NOD⁺) were exposed to a gradient of salt concentrations (200, 400, and 600 mM NaCl) and RNA-Seq was performed. An average of ca. 25 million clean reads was obtained for each group of plants, corresponding to 86,202 unigenes. The patterns of differentially expressed genes (DEGs) clearly separate two groups: (i) control- and 200 mM NaCl-treated plants, and (ii) 400 and 600 mM NaCl-treated plants. Additionally, although the number of total transcripts was relatively high in both plant groups, the percentage of significant DEGs was very low, ranging from 6 (200 mM NaCl/NOD⁺) to 314 (600 mM NaCl/KNO₃⁺), mostly involving down-regulation. The vast majority of up-regulated genes was related to regulatory processes, reinforcing the hypothesis that some ecotypes of C. glauca have a strong stress-responsive system with an extensive set of constitutive defense mechanisms, complemented by a tight mechanism of transcriptional and post-transcriptional regulation. The results suggest that the robustness of the stress response system in C. glauca is regulated by a limited number of genes that tightly regulate detoxification and protein/enzyme stability, highlighting the complexity of the molecular interactions leading to salinity tolerance in this species. Full article

The mining of the oil sands region of Canada’s boreal forest creates disturbed land with elevated levels of salts. Understanding how native plants respond to salt stress is critical in reclaiming these lands. The native species, Alnus alnobetula subsp. crispa forms nitrogen-fixing nodules with Frankia, and ectomycorrhizae with a number of fungal species. These relationships may make the plant particularly well suited for restoring disturbed land. We inoculated A. alnobetula subsp. crispa with Frankia and Hebeloma crustiliniforme and exposed the plants to 0, 50, or 100 mM NaCl for seven weeks. Frankia-inoculated plants had increased biomass regardless of salt exposure, even though salt exposure reduced nitrogen fixation and reduced the efficiency of nitrogen-fixing nodules. The nitrogen-fixing symbiosis also decreased leaf stress and increased root phosphatase levels. This suggests that N-fixing plants not only have increased nitrogen nutrition but also have increased access to soil phosphorus. Mycorrhizae did not affect plant growth but did reduce nodule numbers and nodule efficiency. These results suggest that the nitrogen-fixing trait is more critical than mycorrhizae. While salt stress inhibits nitrogen-fixing symbiosis, plants still benefit from nitrogen fixation when exposed to salt. Full article

Actinorhizal plants have been regarded as promising species in the current climate change context due to their high tolerance to a multitude of abiotic stresses. While combined salt-heat stress effects have been studied in crop species, their impact on the model actinorhizal plant, Casuarina glauca, has not yet been fully addressed. The effect of single salt (400 mM NaCl) and heat (control at 26/22 °C, supra optimal temperatures at 35/22 °C and 45/22 °C day/night) conditions on C. glauca branchlets was characterised at the physiological level, and stress-induced metabolite changes were characterised by mass spectrometry-based metabolomics. C. glauca could withstand single salt and heat conditions. However, the harshest stress condition (400 mM NaCl, 45 °C) revealed photosynthetic impairments due to mesophyll and membrane permeability limitations as well as major stress-specific differential responses in C and N metabolism. The increased activity of enzymatic ROS scavengers was, however, revealed to be sufficient to control the plant oxidative status. Although C. glauca could tolerate single salt and heat stresses, their negative interaction enhanced the effects of salt stress. Results demonstrated that C. glauca responses to combined salt-heat stress could be explained as a sum of the responses from each single applied stress. Full article

Nitrogen is essential for the growth of plants. The ability of some plant species to obtain all or part of their requirement for nitrogen by interacting with microbial symbionts has conferred a major competitive advantage over those plants unable to do so. The function of certain flavonoids (a group of secondary metabolites produced by the plant phenylpropanoid pathway) within the process of biological nitrogen fixation carried out by Rhizobium spp. has been thoroughly researched. However, their significance to biological nitrogen fixation carried out during the actinorhizal and arbuscular mycorrhiza–Rhizobium–legume interaction remains unclear. This review catalogs and contextualizes the role of flavonoids in the three major types of root endosymbiosis responsible for biological nitrogen fixation. The importance of gaining an understanding of the molecular basis of endosymbiosis signaling, as well as the potential of and challenges facing modifying flavonoids either quantitatively and/or qualitatively are discussed, along with proposed strategies for both optimizing the process of nodulation and widening the plant species base, which can support nodulation. Full article

Legumes and actinorhizal plants are capable of forming root nodules symbiosis with rhizobia and Frankia bacteria. All these nodulating species belong to the nitrogen fixation clade. Most likely, nodulation evolved once in the last common ancestor of this clade. NIN (NODULE INCEPTION) is a transcription factor that is essential for nodulation in all studied species. Therefore, it seems probable that it was recruited at the start when nodulation evolved. NIN is the founding member of the NIN-like protein (NLP) family. It arose by duplication, and this occurred before nodulation evolved. Therefore, several plant species outside the nitrogen fixation clade have NLP(s), which is orthologous to NIN. In this review, we discuss how NIN has diverged from the ancestral NLP, what minimal changes would have been essential for it to become a key transcription controlling nodulation, and which adaptations might have evolved later. Full article

Nitrogen-fixing Actinobacteria of the genus Frankia can be subdivided into four phylogenetically distinct clades; members of clusters one to three engage in nitrogen-fixing root nodule symbioses with actinorhizal plants. Mur enzymes are responsible for the biosynthesis of the peptidoglycan layer of bacteria. The four Mur ligases, MurC, MurD, MurE, and MurF, catalyse the addition of a short polypeptide to UDP-N-acetylmuramic acid. Frankia strains of cluster-2 and cluster-3 contain two copies of murC, while the strains of cluster-1 and cluster-4 contain only one. Phylogenetically, the protein encoded by the murC gene shared only by cluster-2 and cluster-3, termed MurC1, groups with MurC proteins of other Actinobacteria. The protein encoded by the murC gene found in all Frankia strains, MurC2, shows a higher similarity to the MurC proteins of plants than of Actinobacteria. MurC2 could have been either acquired via horizontal gene transfer or via gene duplication and convergent evolution, while murC1 was subsequently lost in the cluster-1 and cluster-4 strains. In the nodules induced by the cluster-2 strains, the expression levels of murC2 were significantly higher than those of murC1. Thus, there is clear sequence divergence between both types of Frankia MurC, and Frankia murC1 is in the process of being replaced by murC2, indicating selection in favour of murC2. Nevertheless, protein modelling showed no major structural differences between the MurCs from any phylogenetic group examined. Full article

This study assessed the reproductive success of a temperate dioecious shrub, Canada buffaloberry, Shepherdia canadensis (L.) Nutt., in central Alberta, Canada, by examining the effects of spatial patterns and overstory canopy on flower and fruit production. S. canadensis is more abundant and productive (more fruit) at forest edges and in forest gaps, suggesting a dependence on higher light conditions than is typical of late-seral forests. We used path analysis to demonstrate that flower and fruit production exhibited density-dependent effects at a scale of 50 m² around focal female plants. Fruit production was positively affected by male intraspecific density (pollen supply) and negatively affected by female intraspecific density (pollen competition), but not correlated with overall intraspecific density. The effects of sex-differentiated density are partly due to pollinator responses to male plant density. Flower production was positively affected by overall intraspecific density. A pollen supplementation trial doubled fruit production relative to a control, demonstrating that local male density (pollen availability) and pollinator activity can limit fruit production in S. canadensis. Canopy cover was negatively related to both flower and total fruit production, with approximately one-third (34%) of the total effect of canopy on fruit production due to the effect of canopy on flower production. The commonly observed negative association between canopy cover and fruit production in buffaloberry, therefore, is partly a result of the reduction first in flower number and second in fruit set. This study clarifies the mechanisms associated with the often-noted observation, but not previously assessed at the level of individuals, that reproductive output in S. canadensis is density dependent, limited by canopy cover through reductions in both flowering and fruit set, and pollinator limited. These findings hold implications for managing animal species that depend on the fruit of S. canadensis and suggest future directions for research on dioecious and actinorhizal species. Full article

Casuarina glauca displays high levels of salt tolerance, but very little is known about how this tree adapts to saline conditions. To understand the molecular basis of C. glauca response to salt stress, we have analyzed the proteome from branchlets of plants nodulated by nitrogen-fixing Frankia Thr bacteria (NOD⁺) and non-nodulated plants supplied with KNO₃ (KNO₃⁺), exposed to 0, 200, 400, and 600 mM NaCl. Proteins were identified by Short Gel, Long Gradient Liquid Chromatography coupled to Tandem Mass Spectrometry and quantified by Sequential Window Acquisition of All Theoretical Mass Spectra -Mass Spectrometry. 600 proteins were identified and 357 quantified. Differentially Expressed Proteins (DEPs) were multifunctional and mainly involved in Carbohydrate Metabolism, Cellular Processes, and Environmental Information Processing. The number of DEPs increased gradually with stress severity: (i) from 7 (200 mM NaCl) to 40 (600 mM NaCl) in KNO₃⁺; and (ii) from 6 (200 mM NaCl) to 23 (600 mM NaCl) in NOD⁺. Protein–protein interaction analysis identified different interacting proteins involved in general metabolic pathways as well as in the biosynthesis of secondary metabolites with different response networks related to salt stress. Salt tolerance in C. glauca is related to a moderate impact on the photosynthetic machinery (one of the first and most important stress targets) as well as to an enhancement of the antioxidant status that maintains cellular homeostasis. Full article

Most field-grown plants are surrounded by microbes, especially from the soil. Some of these, including bacteria, fungi and nematodes, specifically manipulate the growth and development of their plant hosts, primarily for the formation of structures housing the microbes in roots. These developmental processes require the correct localization of the phytohormone auxin, which is involved in the control of cell division, cell enlargement, organ development and defense, and is thus a likely target for microbes that infect and invade plants. Some microbes have the ability to directly synthesize auxin. Others produce specific signals that indirectly alter the accumulation of auxin in the plant by altering auxin transport. This review highlights root–microbe interactions in which auxin transport is known to be targeted by symbionts and parasites to manipulate the development of their host root system. We include case studies for parasitic root–nematode interactions, mycorrhizal symbioses as well as nitrogen fixing symbioses in actinorhizal and legume hosts. The mechanisms to achieve auxin transport control that have been studied in model organisms include the induction of plant flavonoids that indirectly alter auxin transport and the direct targeting of auxin transporters by nematode effectors. In most cases, detailed mechanisms of auxin transport control remain unknown. Full article