Physiologia Plantarum

Photosynthesis is the only yield-related trait that has not yet been substantially improved by plant breeding. The limited results of previous attempts to increase yield via improvement of photosynthetic pathways suggest that more knowledge is still needed to achieve this goal. To learn more about the genetic and physiological basis of high photosynthetic light-use efficiency (LUE) at high irradiance, we study Hirschfeldia incana. Here, we compare the transcriptomic response to high light of H. incana with that of three other members of the Brassicaceae, Arabidopsis thaliana, Brassica rapa, and Brassica nigra, which have a lower photosynthetic LUE. First, we built a high-light, high-uniformity growing environment in a climate-controlled room. Plants grown in this system developed normally and showed no signs of stress during the whole growth period. Then we compared gene expression in low and high-light conditions across the four species, utilizing a panproteome to group homologous proteins efficiently. As expected, all species actively regulate genes related to the photosynthetic process. An in-depth analysis on the expression of genes involved in three key photosynthetic pathways revealed a general trend of lower gene expression in high-light conditions. However, H. incana distinguishes itself from the other species through higher expression of certain genes in these pathways, either through constitutive higher expression, as for LHCB8, ordinary differential expression, as for PSBE, or cumulative higher expression obtained by simultaneous expression of multiple gene copies, as seen for LHCA6. These differentially expressed genes in photosynthetic path-ways are interesting leads to further investigate the exact relationship between gene expression, protein abundance and turnover, and ultimately the LUE phenotype. In addition, we can also exclude thousands of genes from "explaining" the phenotype, because they do not show differential expression between both light conditions. Finally, we deliver a transcriptomic resource of plant species fully grown under, rather than briefly exposed to, a very high irradiance, supporting efforts to develop highly efficient photosynthesis in crop plants.

Macroautophagy is known for long as essential for the degradation and the recycling of different macromolecules in eukaryotes. However how important is autophagy for nitrogen management at the whole plant level and for plant biomass and yield productivity in unstressed and well feed plants needed further investigation. In this study, we used both autophagy knock-out mutants and autophagy over-expressors that constitutively produce numerous autophagosomes. These mutants and over-expressors were cultivated using hydroponic system to observe and compare their phenotypes under sufficient nitrate supply, and when submitted after a while to strict nitrate starvation. The shift from nitrate sufficient condition to nitrate starvation allowed us to determine how autophagy defective or stimulated lines can use their own nitrogen resources to complete their cycle. Unexpectedly we observed that irrespective of the nitrate conditions, both mutants and over-expressors exhibited early leaf senescence phenotypes relative to wild type. While autophagy mutants exhibited strong defect for N remobilisation and seed production irrespective of nitrate condition, the better performance of autophagy-over expressors for N remobilisation and seeds production was only significant under sufficient nitrate supply, i.e. when autophagy was not naturally stimulated by nitrate limitation. Interestingly, comparisons of genotypes showed that the nitrogen pool used for seed filling originated from rosette leaves, as if rosette and seeds were used as communicating vessels independently of the stem and pod connecting organs. Altogether, results show that autophagy is a master player in nitrogen management at the whole plant level that controls yield production and leaf senescence.

Summary * The SUC/SUT sucrose transporters belong to a family of active H+/sucrose symporters, with a role of SUC2 in active apoplasmic phloem loading to drive long-distance phloem transport of sucrose in Arabidopsis. However, the cooperation with the symplasmic pathway for phloem loading remains unclear. * In this study, we explored the consequences of reducing either apoplasmic or symplasmic pathways of phloem loading. We compared a series of lines with modified expression of SUC2 gene, and we analyzed the effects on plant growth, sugar accumulation in source and sink organs, phloem transport, and gene expression. * Our data revealed that a modified expression of SUC2 impacted apoplasmic sucrose levels in source leaves but did not impact phloem transport, as might be expected, while increasing foliar storage of carbohydrates. This response differed from lines in which symplasmic communications between phloem cells was disrupted by the over-expression of a plasmodesmata-associated protein, NHL26. * Altogether, our studies indicate an unexpected effect of SUC2 for apoplasmic sucrose levels in source leaves, together with SUC1, and suggest a feedback regulation on foliar storage. This data sheds new light on the interplay between symplasmic and apoplasmic pathways for sugar loading and the consequences on leaf water flows. Summary statementThe mechanisms that coordinate apoplasmic and symplasmic loading pathways, and their effects on foliar carbon storage, remain largely unexplored. Surprisingly, the sucrose transporter SUC2 plays a significant role in maintaining sucrose levels in the apoplasm, shedding light on how apoplasmic sugar levels and water flows can interact for phloem loading.

O_LIAn efficient nitrate uptake system contributes to the improvement of crop nitrogen use efficiency under low nitrogen availability. The High Affinity nitrate Transport System (HATS) in plants is active in low external nitrate and is mediated by a two-component system [high affinity transporters NRT2 associated to a partner protein NRT3 (NAR2)]. C_LIO_LIIn Brachypodium, the model plant for C3 cereals, we investigated the role of BdNRT2A and BdNRT3.2 through various experimental approaches including gene expression profiling, functional characterisation in heterologous system, intracellular localization by imaging, and reverse genetics via gene silencing. C_LIO_LIExpression of BdNRT2.A and BdNRT3.2 genes in response to nitrate availability fits with the characteristics of the HATS components. Co-expression of BdNRT2A and BdNRT3.2 is required for an effective nitrate transport in the heterologous expression system Xenopus oocytes. Functional interaction between BdNRT2A-GFP and BdNRT3.2-RFP fusion proteins has been observed at the plasma membrane in Arabidopsis protoplasts in transient expression experiments. BdNRT3.2 appeared to be necessary for the plasma membrane localization of BdNRT2A. 15Nitrate influx measurements with bdnrt2a mutants (two amiRNA mutants and one NaN3 induced mutant with a truncated NRT2A protein), confirmed that BdNRT2A is a major contributor of the HATS in Brachypodium. C_LIO_LIDirected mutagenesis in BdNRT2A of a conserved Ser residue (S461) specific to monocotyledons has been performed to mimic a non-phosphorylated S461A or a constitutively phosphorylated S461D, in order to evaluate its potential role in the BdNRT2A and BdNRT3.2 interaction leading to plasma membrane targeting. Interestingly, the phosphorylation status of S461 did not modify the interaction, suggesting on a more complex mechanism. C_LIO_LIIn conclusion, our data show that BdNRT2A and BdNRT3.2 are the main components of the nitrate HATS activity in Brachypodium (Bd21-3) and allow an optimal growth in low N conditions. C_LI

This study tested a hypothesis that boron (B) supply alleviates aluminum (Al) toxicity by modifying auxin distribution in functionally different root zones. Auxin distribution and transport at various Al and B ratios were analyzed using the range of molecular and imaging techniques. Al stress resulted in increased auxin accumulation in root apical meristem (MZ) and transition zones (TZ) while reducing its content in elongation zone (EZ). This phenomenon was explained by reduction in basipetal auxin transport caused by Al blockage of PIN2 endocytosis, regulated at posttranscriptional level. This inhibition of PIN2 endocytosis was dependent on actin filaments and microtubules. B supply facilitated the endocytosis and exocytosis of PIN2 carriers via recycling endosomes conjugated with IAA to modify Al-induced auxin depletion in the EZ. However, disruption of auxin signaling with auxinole did not alleviate Al-induced inhibition of root growth. B supply alleviates Al-induced inhibition of root growth via restoring the endocytic recycling of PIN2 proteins involved in the basipetal (shootward) auxin transport, restoring Al-induced auxin depletion in the elongation zone. Short summaryAluminum-intensified PIN2 abundance, nontranscriptional, via repressing PIN2 endocytosis to block polar auxin transport, and this adverse effect could be alleviated by boron supply.

Roles of three different plasma membrane aquaporins (PIPs) in leaf-level gas exchange of Arabidopsis thaliana were examined using single, double and triple knockout mutants and compared to the Columbia-0 wild type (WT) plants. Since multiple Arabidopsis PIPs are implicated in conducting carbon dioxide across membranes, we focused on identifying whether the examined isoforms affect photosynthesis, either mediated through the control of stomatal conductance to water vapour (gs) or mesophyll conductance of CO2 (gm) or a combination of both. In two separate studies, we grew Arabidopsis plants in a low humidity environment and under high humidity conditions. We found that the contribution of functional PIPs to gs was larger under conditions of low air humidity when the evaporative demand was high, whereas any effect of lacking PIP function was minimal under higher humidity conditions. The pip2;4 knockout mutants had 44% higher gs than the WT under low humidity conditions, which in turn resulted in an increased photosynthetic rate (Anet). AtPIP2;4 is thus likely to be involved in maintaining a positive water balance and high water use efficiency through mediation of transmembrane water flow. The lack of functional AtPIP2;5 on the other hand did not affect gs, but reduced gm indicating a possible role in regulating CO2 membrane permeability. This potential regulatory function was indeed confirmed by subsequent stopped flow measurements of yeast expressing AtPIP2;5.

K+ and NO3- are the major forms of potassium and nitrogen that are absorbed by the roots of most terrestrial plants. In this study, we observed that the close relationship between NO3- and K+ homeostasis was mediated by nitrate transporter1 (NRT1.1) in Arabidopsis. The nrt1.1 mutants lacking NRT1.1 function showed disturbed K+ uptake and root-to-shoot allocation, especially under K+-limited conditions, and had a yellow-shoot sensitive phenotype on K+-limited medium. The K+ uptake and root-to-shoot allocation of these mutants were partially rescued by expressing NRT1.1 in the root epidermis-cortex and central vasculature by using Sultr1;2 and PHO1 promoters, respectively. Furthermore, two-way analysis of variance based on the K+ content in nrt1.1-1/akt1, nrt1.1-1/hak5-3, nrt1.1-1/kup7, and nrt1.1-1/skor-2 double mutants and their corresponding single mutants and wild-type plants revealed physiological interactions between NRT1.1 and K+ channels located in the root epidermis-cortex and central vasculature. Taken together, these data suggest that the expression of NRT1.1 in the root epidermis-cortex coordinates with K+ uptake channels to improve K+ uptake, whereas its expression in the root central vasculature coordinates with the channels loading K+ into the xylem to facilitate K+ allocation from the roots to the shoot.

Salt tolerant is strongly related to potassium (K+) retention in plant tissues under salt stress conditions. However, it is unclear for different Echinacea species. So, mechanistic basis of four Echinacea species (i.e. Echinacea purpurea, Echinacea angustifolia, Echinacea pallida, and Echinacea sanguinea) to salinity stress tolerance, and K+ retention were assessed in the present study. Non-invasive microelectrode ion flux measuring, DHAR and MDHAR activities, and pharmacological measurements were performed based on the standard methods. Ion flux measurements revealed higher K+ efflux in E. pallida and E. sanguinea species compared to the E. purpurea and E. angustifolia species in the elongation zone. Higher salinity-induced H+ efflux was found in the elongation zone than mature zone. However, E. angustifolia and E. purpurea had more Ca2+ influx compared to E. pallida and E. sanguinea species. Net K+ efflux decreased (> 90%) in the presence of TEA and GdCl3. Increasing of Ca2+ uptake and K+ loss in four Echinacea species roots were found in the presence of 0.3 mM Cu/Ascorbate (Cu/Asc). The significant role of H+-ATPase in H+ efflux was demonstrated by Sodium orthovanadate. Ultimately, the physiological properties of Echinacea species have a critical role in salinity-resistant/sensitive differences. Future scientific understanding of Echinacea species physiognomies may be necessary for better understanding of the plant behavior to salinity stress. One-sentence summaryHigher K+ efflux in E. pallida and E. sanguinea species as a result of NaCl and ROS act as a metabolic switch to save energy for adaptations and repairs in salinity stress conditions.

Soybean plants are salinity (NaCl) sensitive, with their yield significantly decreased under moderately saline conditions. GmSALT3 is the dominant gene underlying a major QTL for salt tolerance in soybean. GmSALT3 encodes a transmembrane protein belonging to the plant cation/proton exchanger (CHX) family. It is currently unknown through which molecular mechanism(s) the ER-localised GmSALT3 contributes to salinity tolerance, as its localisation excludes direct involvement in ion exclusion. In order to gain insights into potential molecular mechanism(s), we used RNA-seq analysis of roots from two soybean NILs (Near Isogenic Lines); NIL-S (salt-sensitive, Gmsalt3) and NIL-T (salt-tolerant, GmSALT3), grown under control and saline conditions (200 mM NaCl) at three time points (0h, 6h, and 3 days). Gene ontology (GO) analysis showed that NIL-T has greater responses aligned to oxidation reduction. ROS were shown less abundant and scavenging enzyme activity was higher in NIL-T, consistent with the RNA-seq data. Further analysis indicated that genes related to calcium signalling, vesicle trafficking and Casparian strip (CS) development were upregulated in NIL-T following salt treatment. We propose that GmSALT3 improves the ability of NIL-T to cope with saline stress through preventing ROS overaccumulation in roots, and potentially modulating Ca2+ signalling, vesicle trafficking and formation of diffusion barriers. HighlightRNA-seq analysis revealed that GmSALT3, which confers improved salt tolerance on soybean, improves reactive oxygen species detoxification in roots.

Understanding comprehensive mechanisms of the downregulation of photosynthesis induced by accumulation of non-structural carbohydrates (NSCs) is essential for the future food security.x Despite numerous studies, whether NSCs accumulation directly affects steady-state maximum photosynthesis and photosynthetic induction, as well as underlying gene expression profiles, remains unknown so far. We evaluated the relationship between photosynthetic capacity and NSCs accumulation induced by cold-girdling, sucrose feeding, and low nitrogen treatment in Glycine max and Phaseolus vulgaris. In G. max, changes in transcriptome profiles were further investigated focusing on physiological processes of photosynthesis and NSCs accumulation. NSCs accumulation decreased maximum photosynthetic capacity and delayed photosynthetic induction in both species. In G. max, such photosynthetic downregulation was explained by coordinated downregulation of photosynthetic genes involved in Calvin cycle, Rubisco activase, photochemical reactions, and stomatal opening. Furthermore, sink-source imbalance may have triggered a change in the balance of sugar-phosphate translocators in chloroplast membranes, which may have promoted starch accumulation in chloroplasts. Our findings provided an overall picture of the photosynthetic downregulation and NSCs accumulation in G. max, demonstrating that the photosynthetic downregulation is triggered by NSCs accumulation and cannot be explained simply by N deficiency. One Sentence SummaryAccumulation of nonstructural carbohydrates directly induced both downregulation and delayed induction of photosynthesis by coordinated transcriptomic changes in photosynthetic genes in Glycine max.

O_LISoil salinity induces osmotic stress and ion toxicity in plants, detrimentally affecting their growth and development. Potato (Solanum tuberosum) faces yield reductions due to salt stress. The mechanisms of salt stress resilience, especially in adventitious roots, remain unknown. C_LIO_LIWe investigated the resilience of three potato cultivars - Desiree, Innovator, and Mozart - by studying their physiological and transcriptomic responses to salt stress. C_LIO_LIOur findings reveal that under salt stress, the growth of stolons and stolon node roots is similarly reduced unlike tubers, even though they are physically connected. Surprisingly, tubers accumulate Cl- but not Na+ under salt stress, suggesting an active Na+ exclusion mechanism. Innovator showed the lowest suberin and lignin deposition before salt stress and higher K+ leakage, leading to a stronger initial stress response with high ABA content and a distinct transcriptomic pattern. Nevertheless, Innovator was the most resilient, displaying lower growth, salt-tolerance index and tuber yield reduction. Transcriptomic analysis revealed several K+/Na+ channel genes which might regulate ions homeostasis during salt stress, in particular in Innovator. C_LIO_LIAltogether, we conclude that acclimation ability, rather than initial protection of roots against salt, prevails in long term salt-stress resilience of potato. C_LI

O_LIThe promoting effect of gibberellin (GA) on primary-root elongation is well-documented in several plant species, yet its influence in others, including tomato (Solanum lycopersicum), remains unclear. C_LIO_LIThe role of GA in primary-root elongation has been studied in tomato using the GA-deficient mutants gib-1 and ga20-oxidase (ga20ox1) and various growth systems, including Dark (D)-root and D-shoot plates. C_LIO_LIGA application to these mutants following germination on vermiculite, promoted primary-root elongation. However, when the roots grew deeper into the dark environment the hormone had no effect. RNA-seq analysis of dark-grown roots, treated with GA, revealed typical transcriptional responses, but the output for cell expansion remained unaffected. When dark-grown roots were illuminated deep in the ground, the hormone promoted their elongation. The results suggest that activation of Phytochrome B (PhyB) in the root, by red light, is essential for GA-induced elongation. C_LIO_LIWe propose that GA promotes tomato root elongation after germination, when roots are exposed to low light underground and this contributes to rapid seedling establishment. As roots penetrate deeper into the soil, insensitivity to GA due to the lack of light may be important for sustained root growth under fluctuating water availability, given that water deficiency suppresses GA accumulation. C_LI

Halophytes are salt-tolerant plants that grow in soil or waters of high salinity. Schrenkiella parvula is one of the halophyte plants that grow around Tuz (Salt) Lake, TURKEY that can survive at 600 mM NaCl. Intriguingly, S. parvula belongs to the same Brassicaceae family as the model plant Arabidopsis thaliana, and its genome is 90% homologous to the Arabidopsis genome. Here, we performed proteomic analysis and physiological studies on the roots of S. parvula seedlings cultivated under a moderate salt condition at 100 mM NaCl. Surprisingly, under 100 mM NaCl conditions, the primary roots elongated much faster than under NaCl-free conditions, although up to 200 mM those were reduced. On the other hand, iso-osmotic mannitol did not promote primary root elongation, suggesting a specific response to NaCl. Epidermal cell elongation was promoted in the elongation zone, but meristem size and DNA replication were decreased. In addition, root hair formation and lateral root elongation were suppressed at moderate salinity. Compared with A. thaliana, the cell death and ROS increase of root tip meristem cells under 100 mM NaCl condition were significantly lower in S. parvula seedlings. The size and starch content of sedimentary amyloplasts/statoliths in columella cells decreased, and gravitropism of primary roots was partially reduced. Gene expression analyses showed that the expression of auxin response and biosynthesis genes IAA1, IAA2, TAA1 and YUC8 were repressed and the SOS1 gene was upregulated two-fold in roots grown under moderate salt conditions. Proteomic analysis showed that co-chaperone and activator of HSPs such as Hop2 and Aha1 domain-containing protein orthologs were upregulated. Moreover, several secondary metabolic process-related proteins, antioxidant proteins, stress response proteins and proline catabolic process-related proteins were also increased. In contrast, enzymes associated with root hair elongation and nucleotide and protein syntheses were downregulated. These changes in auxin-related physiological responses, root architecture, lower ROS signaling, and stress-related protein expression promote primary root penetration into lower-salinity deeper soils as an adaptation of S. parvula.

Despite established understanding of plant physiological responses to elevated [CO2], the underlying genes are poorly understood. Soybean transcriptomics previously identified a GATA transcription factor, involved in carbon and nitrogen metabolism, as responsive to elevated [CO2]. Supported by in silico modeling, we therefore hypothesized that this gene plays a previously unrecognized role in responding to elevated [CO2]. Wildtype and a T-DNA insertion line of Arabidopsis thaliana for GNC (GATA, Nitrate Inducible, Carbon Metabolism Involved) were grown under three treatments: sustained ambient [CO2], sustained elevated [CO2], and transfer from ambient to elevated [CO2], to assess changes in their physiology, biochemistry, and transcriptome. Photosynthetic and biomass responses to elevated [CO2] and transfer [CO2] in plants lacking GNC were significantly weaker than WT. A lag of 25-73 hrs in transcriptomic responses after transfer to elevated [CO2] was consistent with indirect sensing, presumably via sugar signals. The breakdown of the gene expression network around GNC was most pronounced in the transfer treatment and suggests targets for further study of interactions between elevated [CO2] and sulfur and nitrogen metabolism. This work provides a case study of a CO2-responsive transcription factor that may be a compelling target for adapting crops to future growing conditions after further characterization. Summary StatementA GATA transcription factor modulates plant metabolic and productivity responses to elevated CO2.

The relationship between nitrogen (N) sources and photosynthetic capacity of leaf differs between species. However, the leaf anatomical variabilities related to photosynthesis (A) of shrubs under different forms of N remain imperfectly known. Here, Lonicera Japonica (a shrub) was grown hydroponically in the presence of three forms of N (sole NH4+, 50%/50% NH4+/NO3- and sole NO3-). A and photosynthetic N use efficiency significantly decreased under sole NH4+ supply, in parallel with down-regulated stomatal conductance (gs), mesophyll conductance (gm), and electron transfer rate (J). Up to the total A decline of 41.28% in sole NH4+ supply (compare with sole NO3-), the gm attributed to 60.3% of the total limitations. Besides, the decreased internal air space explained the increase of gas-phase resistance, and the increased liquid-phase resistance in sole NH4+ supply was ascribed to the thicker cell wall thickness (Tcw) and decreased chloroplasts exposed surface area per unit leaf area (Sc/S). The discrepancy of Sc/S could be interpreted by the altered chloroplasts numbers and the distance between adjacent chloroplasts (Dchl-chl). These results indicate the alteration of Tcw and chloroplast numbers were the main causes of the difference in gm in coping with varied N sources. HighlightCell wall and chloroplast variability determining the mesophyll conductance under different nitrogen forms

O_LIRoot hair growth is tuned in response to the environment surrounding plants. While most of previous studies focused on the enhancement of root hair growth during nutrient starvation, few studies investigated the root hair response in the presence of excess nutrients. C_LIO_LIWe report that the post-embryonic growth of wild-type Arabidopsis plants is strongly suppressed with increasing nutrient availability, particularly in the case of root hair growth. We further used gene expression profiling to analyze how excess nutrient availability affects root hair growth, and found that RHD6 subfamily genes, which are positive regulators of root hair growth, are down-regulated in this condition. C_LIO_LIOn the other hand, defects in GTL1 and DF1, which are negative regulators of root hair growth, cause frail and swollen root hairs to form when excess nutrients are supplied. Additionally, we observed that the RHD6 subfamily genes are mis-expressed in gtl1-1 df1-1. Furthermore, overexpression of RSL4, an RHD6 subfamily gene, induces swollen root hairs in the face of a nutrient overload, while mutation of RSL4 in gtl1-1 df1-1 restore root hair swelling phenotype. C_LIO_LIIn conclusion, our data suggest that GTL1 and DF1 prevent unnecessary root hair formation by repressing RSL4 under excess nutrient conditions. C_LI

Identification of signaling pathways that control C4 photosynthesis development is essential for introducing the C4 pathway into C3 crops. Species with dual photosynthesis in their life cycle are interesting models to study such regulatory mechanisms. The species used here Halimocnemis mollissima Bunge, belonging to the Caroxyleae tribe, displays C3 photosynthesis in its cotyledons and a NAD-ME subtype of C4 photosynthesis in the First leaves (FLs) onwards. We explored the long-distance signaling pathways that are probably implicated in the shoot-root coordination associated with the manifestation of the C4 traits, including efficient resource usage by comparing the mRNA content of hypocotyls before and after the C4 first leaves formation. Histological examination showed the presence of C3 anatomy in cotyledons and C4 anatomy in the FLs. Our transcriptome analyses verified the performance of the NAD-ME subtype of C4 in FLs and revealed differential transcript abundance of several potential mobile regulators and their associated receptors or transporters in two developmentally different hypocotyls of H. mollissima Bunge. These differentially expressed genes (DEGs) belong to diverse functional groups, including various transcription factor (TF) families, phytohormones metabolism, and signaling peptides, part of which could be related to hypocotyl development. Our findings support the higher nitrogen and water use efficiency associated with C4 photosynthetic and provide insights into the coordinated above- and under-ground tissue communication during the developmental transition of C3 to C4 photosynthesis in this species.

O_LIBryophyta (mosses) are a basal group of plants that lack stomata in their haploid form, as well as developed vascular tissue and a hydrophobic cuticle. Consequently, these plants are classified as poikilohydric, meaning poor control over water loss and are often assumed to reach equilibrium with ambient humidity. This classification does not fully align with the diverse strategies observed in mosses. C_LIO_LIWe studied gas exchange in 14 species from Albuquerque and Boston, USA, under controlled dehydration conditions. C_LIO_LIOur results revealed significant variation in transpiration rates, cell wall equilibrium humidity, and desiccation times across species. These differences could not be explained by tissue water storage relative to the transpiring surface area, suggesting that water loss is not entirely passive. Additionally, species with better water control also presented traits of an avoidance strategy, including elastic tissues, high capacitance, and less negative osmotic potential, suggesting an adaptive constraint. C_LIO_LIThese findings point to a basal, non-stomatal mechanism of water loss control through cell membranes and/or cell walls. Potentially, this mechanism is homologous to the non-stomatal control recently identified in angiosperms, which induces unsaturated conditions in the substomatal cavities. Bryophyta presents a valuable non-stomatal model for further investigating this mechanism and its evolutionary significance. C_LI

The remarkable capacity of plants to tolerate and accumulate tremendous amount of nickel is a complex adaptative trait that appeared independently in more than 700 species distributed in about fifty families. Nickel hyperaccumulation is thus proposed as a model to investigate the evolution of complex traits in plants. However, the mechanisms involved in nickel hyperaccumulation are still poorly understood in part because comparative transcriptomic analyses struggle to identify genes linked to this trait from a wide diversity of species. In this work, we have implemented a methodology based on the quantification of the expression of orthologous groups and phylogenetic comparative methods to identify genes which expression is correlated to the nickel hyperaccumulation trait. More precisely, we performed de novo transcriptome assembly and reads quantification for each species on its own transcriptome using available RNA-Seq datasets from 15 nickel hyperaccumulator and non-accumulator species. Assembled contigs were associated to orthologous groups built using proteomes predicted from completed plant genome sequences. We then analyzed the transcription profiles of 5953 orthologous groups from distant species using a phylogenetic ANOVA. We identified 31 orthologous groups with an expression shift associated with nickel hyperaccumulation. These orthologous groups correspond to genes that have been previously implicated in nickel accumulation, and to new candidates involved in this trait. We thus believe that this method can be successfully applied to identify genes linked to other complex traits from a wide diversity of species.

O_LIDiazotrophic cyanobacteria fix both atmospheric carbon (C) and nitrogen (N) into biomass, but the two assimilation pathways are not compatible. Species like Anabaena sp. PCC 7120 physically separates C and N assimilation in different cell types. Even if separated, they are strongly intertwined, as N assimilation relies on the C skeletons and reducing power from photosynthesis, that in turn depends on N rich molecules as pigments and proteins. C_LIO_LIWhereas the two pathways have been extensively studied individually, here we investigate their interaction by analysing photosynthetic properties upon exposure to changes in light, CO2 and N availability, including the contribution of photosynthetic electron fluxes. C_LIO_LIGrowth depended on the availability of both light and CO2, while the N2 fixation activity mainly on the C supply. Upon diazotrophic conditions, the total photosynthetic electron transport activity increased, with a modified contribution of different electron pathways. A mutant strain affected in the vehiculation of fixed N between cell types showed that the modulation of photosynthesis depended on the metabolic connection between assimilation pathways. C_LIO_LIOverall, data showed that the regulation of photosynthetic electron fluxes is a major component of the synergic metabolic relationship between C and N assimilation pathways upon dynamic environmental conditions. C_LI