Exploring apoplast reorganization in the nodules of Lotus corniculatus L. growing on old Zn-Pb calamine wastes.

Nodulation and symbiotic nitrogen fixation are important factors that determine legume growth. A pot experiment was carried out to determine the effects of Zn-Pb contamination on nodule apoplast (cell walls and intercellular spaces) of bird's foot trefoil (Lotus corniculatus L.) that spontaneously colonized old calamine wastes. The plants were grown in pots filled with sterile calamine substrate (M, metal treated) or expanded clay (NM, untreated) and inoculated with calamine-derived Lotus-nodulating Bradyrhizobium liaoningense. Apoplast reorganization in the nodules was examined using specific dyes for cellulose, pectin and lignin detection, and immuno-histochemical techniques based on monoclonal antibodies against xyloglucan (Lm25), pectins (Jim5 and Jim7), and structural proteins (arabinogalactan protein - Lm14 and extensin - Jim12). Microscopic analysis of metal-treated nodules revealed changes in the apoplast structure and composition of nodule cortex tissues and infected cells. Wall thickening was accompanied by intensified deposition of cellulose, xyloglucan, esterified pectin, arabinogalactan protein and extensin. The metal presence redirected also lignin and suberin deposition in the walls of the nodule cortex tissues. Our results showed reorganization of the apoplast of cortex tissues and infected cells of Lotus nodules under Zn-Pb presence. These changes in the apoplast structure and composition may have created actual barriers for the toxic ions. For this reason, they can be regarded as an element of legume defense strategy against metal stress that enables effective functioning of L. corniculatus-rhizobia symbiosis on Zn-Pb polluted calamine tailings.

Structure Analysis and Study of Biological Activities of Condensed Tannins from Bruguiera gymnorhiza (L.) Lam and Their Effect on Fresh-Cut Lotus Roots.

Bruguiera gymnorhiza (L.) Lam is a mangrove plant that spread in many parts of the world. Though mangrove plant polyphenols have been reported to exhibit many biological activities, little is known about mangrove plant tannins. To explore the application value of tannins from B. gymnorhiza, analyses on the structure and biological activity of condensed tannins (CTs) from Bruguiera gymnorhiza (L.) Lam were carried out. The results from 13C nuclear magnetic resonance (13C-NMR) and reversed-phase, high-performance liquid chromatography (RP-HPLC) showed that the CTs were dominated by procyanidins, with a small quantity of prodelphinidins and propelargonidins; and that the monomeric constituents of B. gymnorhiza tannins were catechin/epicatechin, gallocatechin/epigallocatechin and afzelechin/epiafzelechin. The CTs were reversible and mixed competitive inhibitors of tyrosinase and the 50% inhibiting concentration (IC50) was estimated to be 123.90 ± 0.140 µg/mL. The antioxidant activities of CTs from B. gymnorhiza leaves were evaluated, the IC50 for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzo-thiazoline-6-sulfonic acid diammonium salt) (ABTS) scavenging activities were 88.81 ± 0.135 and 105.03 ± 0.130 µg/mL, respectively, and the ferric ion reducing antioxidant power (FRAP) value was 1052.27 ± 4.17 mgAAE/g. In addition, the results from fresh-keeping assays on fresh-cut lotus root reveal that CTs from B. gymnorhiza had excellent effects on inhibiting the activities of polyphenol oxidase (PPO) and peroxidase (POD), protecting fresh-cut lotus root from the oxidation of total phenolics and malondialdehyde (MDA) content and slowing the increase in total phenol content (TPC) at 4 °C during the whole storage period. Therefore, CTs showed good effects against the browning of fresh-cut lotus root. Together, these results suggested that B. gymnorhiza CTs are promising antibrowning agents for fresh-cut fruits.

Contrasting response of two Lotus corniculatus L. accessions to combined waterlogging-saline stress.

Waterlogging and salinity impair crop growth and productivity worldwide, with their combined effects being larger than the additive effects of the two stresses separately. Here, a common forage tetraploid Lotus corniculatus (cv. San Gabriel) and a diploid L. corniculatus accession, collected from a coastal area with high frequency of waterlogging-saline stress events, were evaluated for tolerance to waterlogging, salinity and these two stresses combined. We hypothesize that, due to its environmental niche, the diploid accession would show better adaptation to combined waterlogging-saline stress compared to the tetraploid L. corniculatus. Plants were evaluated under control conditions, waterlogging, salinity and a combined waterlogging-saline treatment for 33 days. Shoot and root growth were assessed, together with chlorophyll fluorescence and gas exchange measurements. Results showed that salinity and waterlogging effects were more severe for the tetraploid accession, with a larger effect being observed under the combined stress condition. Concentrations of Na+ , Cl- and K+ were measured in apical and basal leaves, and in roots. A larger accumulation of Na+ and Cl- was observed under both saline and combined stress treatments for the tetraploid L. corniculatus, for which ion toxicity effects were evident. The expression of CLC gene, coding for a Cl- transporter, was only increased in diploid L. corniculatus plants in response to the combined stress condition, suggesting that ion compartmentalization mechanisms were induced in this accession. Thus, this recently characterized L. corniculatus could be used for the introduction of new tolerance traits in other Lotus species used as forage.

Endogenous gibberellins affect root nodule symbiosis via transcriptional regulation of NODULE INCEPTION in Lotus japonicus.

Legumes and nitrogen-fixing rhizobial bacteria establish root nodule symbiosis, which is orchestrated by several plant hormones. Exogenous addition of biologically active gibberellic acid (GA) is known to inhibit root nodule symbiosis. However, the precise role of GA has not been elucidated because of the trace amounts of these hormones in plants and the multiple functions of GAs. Here, we found that GA signaling acts as a key regulator in a long-distance negative-feedback system of root nodule symbiosis called autoregulation of nodulation (AON). GA biosynthesis is activated during nodule formation in and around the nodule vascular bundles, and bioactive GAs accumulate in the nodule. In addition, GA signaling induces expression of the symbiotic transcription factor NODULE INCEPTION (NIN) via a cis-acting region on the NIN promoter. Mutants with deletions of this cis-acting region have increased susceptibility to rhizobial infection and reduced GA-induced CLE-RS1 and CLE-RS2 expression, suggesting that the inhibitory effect of GAs occurs through AON. This is supported by the GA-insensitive phenotypes of an AON-defective mutant of HYPERNODULATION ABERRANT ROOT FORMATION1 (HAR1) and a reciprocal grafting experiment. Thus, endogenous GAs induce NIN expression via its GA-responsive cis-acting region, and subsequently the GA-induced NIN activates the AON system to regulate nodule formation.

Effect of postharvest oxalic acid application on enzymatic browning and quality of lotus (Nelumbo nuciferaGaertn.) root slices.

Post-cut surface browning is one of the major constraints for shelf-life extension of lotus root slices. In the present study, lotus roots slices were treated with 0, 5 and 10 mmol L-1 oxalic acid and stored at 20 ± 1 °C for 5 days. Results showed that 10 mmol L-1 oxalic acid treated lotus slices exhibited reduced browning, superoxide anion, hydrogen peroxide, electrolyte leakage and malondialdehyde content than control. The 10 mmol L-1 treated slices had better visual quality and higher ascorbic acid and total phenolic contents. In addition, 10 mmol L-1 treated slices showed reduced total bacterial count along with lower soluble quinones, peroxidase and polyphenol oxidase activities in contrast to control. Similarly, 10 mmol L-1 treatment showed higher superoxide dismutase, catalase and ascorbate peroxidase activities as compared to control. In conclusion, 10 mmol L-1 oxalic acid application could be considered suitable to delay post-cut browning of lotus root slices.

Differentiation between physical and chemical effects of oil presence in freshly spiked soil during rhizoremediation trial.

Petroleum contamination and its remediation via plant-based solutions have got increasing attention by environmental scientists and engineers. In the current study, the physiological and growth responses of two diesel-tolerant plant species (tolerance limit: 1500-2000 mg/kg), Italian ryegrass (Lolium multiflorum) and Birdsfoot trefoil (Lotus corniculatus), have been investigated in vegetable oil- and diesel oil-amended soils. A long-term (147-day) greenhouse pot experiment was conducted to differentiate the main focus of the study: physical and chemical effects of oil (vegetable and diesel) in freshly spiked soils via evaluating the plant performance and hydrocarbon degradation. Moreover, plant performance was evaluated in terms of seed germination, plant shoot biomass, physiological parameters, and root biomass. Addition of both diesel oil and vegetable oil in freshly spiked soils showed deleterious effects on seedling emergence, root/shoot biomass, and chlorophyll content of grass and legume plants. Italian ryegrass showed more sensitivity in terms of germination rate to both vegetable and diesel oil as compared to non-contaminated soils while Birdsfoot trefoil reduced the germination rate only in diesel oil-impacted soils. The results of the current study suggest that both physical and chemical effects of oil pose negative effects of plant growth and root development. This observation may explain the phenomenon of reduced plant growth in aged/weathered contaminated soils during rhizoremediation experiments.

Multi-omic and physiologic approach to understand Lotus japonicus response upon exposure to 3,4 dimethylpyrazole phosphate nitrification inhibitor.

Nitrogen fertilization is a major force in global greenhouse gases emissions and causes environmental contamination through nitrate leaching. The use of nitrification inhibitors has been proven successful to mitigate these effects. However, there is an increasing concern about the undesired effects that their potential persistence in the soil or accumulation in plants may provoke. In this study, we first exposed Lotus japonicus plants to high amounts of 3,4 dimethylpyrazole phosphate (DMPP) and 2-(N-3,4-dimethyl-1H-pyrazol-1-yl) succinic acid isomeric mixture (DMPSA) nitrification inhibitors. Exposure to doses higher than 1 mg·L-1 provoked DMPP accumulation mostly in the aerial part, while DMPSA was only detected from 10 mg·L-1 and nearly no translocation. To evaluate the effect that DMPP accumulation in leaves may provoke on plant performance we combined a transcriptome, proteome, and physiological analysis in plants treated with 10 mg/ L of DMPP. This treatment provoked changes in the expression of 229 genes and 59 proteins. Overall, we evidence that when DMPP accumulates in leaves it induces stress responses, notably provoking changes in cell redox balance, hormone signaling, protein synthesis and turnover and carbon and nitrogen metabolism.

Chemical compounds from Dictyostelium discoideum repel a plant-parasitic nematode and can protect roots.

Slime mold species in the genus Dictyostelium are considered to have a close relationship with non-parasitic nematodes; they are sympatric in soils and can exhibit interspecific competition for food. We investigated whether this relationship extends to a plant-parasitic nematode that is active in the rhizosphere and has broad host specificity, damaging crops worldwide. Using a novel assay to examine the interaction between the cellular slime mold, Dictyostelium discoideum, and the plant-parasitic nematodes, Meloidogyne spp., we found that cellular slime molds can repel plant parasitic nematodes. Specifically, the repulsion activity was in response to chemical compounds released by cellular slime mold fruiting bodies. Under laboratory conditions, these soluble chemical extracts from fruiting bodies of D. discoideum showed repulsion activity strong enough to protect plant roots. The fruiting body cell extracts repelled but were not toxic to the plant-parasitic nematodes.

Transcriptome profiling reveals an IAA-regulated response to adventitious root formation in lotus seedling.

Adventitious roots (ARs) of lotus (Nelumbonucifera Gaertn.) play a critical role in water and nutrient uptake. We found that exogenously applied 10-µM indole-3-acetic acid (IAA) promoted the formation of ARs, while 150-µM IAA significantly inhibited the emergence of ARs. However, little is known about these different responses to various concentrations of IAA at the molecular level. This study, therefore, examined the gene expression profiling in four libraries treated with 10- and 150-µM IAA based on the high-throughout tag sequencing technique. Approximately 2.4×107 clean tags were obtained after the removal of low-quality tags from each library respectively, among which about 10% clean tags were unambiguous tag-mapped genes to the reference genes. We found that some genes involved in auxin metabolism showed a similar tendency for expression in the A/CK and C/CK libraries, while three genes were enhanced their expression only in the A/CK libraries. Two transcription factors including B3 domain-containing protein At2g36080-like and trihelix transcription factor were up-regulated for transcriptional level in the A/C libraries. The expressions of six important genes related to AR formation were significantly different in the A/CK and C/CK libraries. In summary, this study provides a comprehensive understanding of gene expression regulated by IAA involved in AR formation in lotus.

The Nitrate Transporter Family Protein LjNPF8.6 Controls the N-Fixing Nodule Activity.

N-fixing nodules are new organs formed on legume roots as a result of the beneficial interaction with soil bacteria, rhizobia. The nodule functioning is still a poorly characterized step of the symbiotic interaction, as only a few of the genes induced in N-fixing nodules have been functionally characterized. We present here the characterization of a member of the Lotus japonicus nitrate transporter1/peptide transporter family, LjNPF8.6 The phenotypic characterization carried out in independent L. japonicus LORE1 insertion lines indicates a positive role of LjNPF8.6 on nodule functioning, as knockout mutants display N-fixation deficiency (25%) and increased nodular superoxide content. The partially compromised nodule functioning induces two striking phenotypes: anthocyanin accumulation already displayed 4 weeks after inoculation and shoot biomass deficiency, which is detected by long-term phenotyping. LjNPF8.6 achieves nitrate uptake in Xenopus laevis oocytes at both 0.5 and 30 mm external concentrations, and a possible role as a nitrate transporter in the control of N-fixing nodule activity is discussed.

Novel impacts of functionalized multi-walled carbon nanotubes in plants: promotion of nodulation and nitrogenase activity in the rhizobium-legume system.

The rhizobium-legume symbiosis system is critical for nitrogen-cycle balance in agriculture. However, the potential effects of carbon nanomaterials (CNMs) on this system remain largely unknown. Herein, we studied the effects of four carbon-based materials (activated carbon (AC), single-walled carbon nanotubes (SWCNTs), multi-walled carbon nanotubes (MWCNTs) and graphene oxide (GO)) on the rhizobium-legume symbiosis system consisting of Lotus japonicus and Mesorhizobium loti MAFF303099. Under non-symbiotic conditions, the bacterial growth and root development of plants were both clearly inhibited by SWCNTs and GO, while the elongation of plant stems was enhanced by MWCNTs to a certain degree. More importantly, only MWCNTs could increase the number of nodules and enhance the activity of nitrogenase in the rhizobium-plant interaction. Further analyses showed that the average number of nodules in plants treated with 100 µg mL-1 MWCNTs was significantly increased by 39% at 14 days post inoculation (dpi) and by 41% at 28 dpi. Meanwhile, the biological nitrogen fixation of the nodules was promoted by more than 10% under 100 µg mL-1 MWCNT treatment, which enhanced the above- and below-ground fresh biomass by 14% and 25% respectively at 28 dpi. Transmission electron microscopy images further indicated that MWCNTs penetrated the cell wall, and pierced through the cell membrane to be transmitted into the cytoplasm. In addition, gene expression analysis showed that the promotion of nodulation by MWCNTs was correlated with the up-regulation of certain genes involved in this signaling pathway. In particular, the expression of NIN, a crucial gene regulating the development of nodules, was significantly elevated 2-fold by MWCNTs at an early stage of nodulation. These findings are expected to facilitate the understanding and future utilization of MWCNTs in agriculture.

Oxyresveratrol and ascorbic acid O/W microemulsion: Preparation, characterization, anti-isomerization and potential application as antibrowning agent on fresh-cut lotus root slices.

The purpose of this study is to prepare an oxyresveratrol (Oxy) microemulsion (ME) with improved Oxy's solubility and stability and to investigate its antibrowning effects on fresh-cut lotus root slices. The formula of OxyME consisted of ethyl butyrate, Tween 80, PEG400, and water with w/w of 4%, 10.67%, 5.33%, and 80%, respectively. Encapsulating Oxy into OxyME greatly increased its solubility and stability compared with that of in water. Strong antibrowning effects were observed on fresh-cut lotus root slices treated with OxyME, even better than 4-hexylresorcinol. The addition of ascorbic acid (VC) into OxyME greatly improved the Oxy stability in long-term storage and antibrowning effects on fresh-cut lotus root slices. However, the simultaneous addition of calcium chloride and VC did not obviously improve the antibrowning effects compared with the addition of VC alone. These results indicated that Oxy+VCME may be suitable as an antibrowning agent for fresh-cut vegetables.

Iron-induced nitric oxide leads to an increase in the expression of ferritin during the senescence of Lotus japonicus nodules.

Iron is an essential nutrient for legume-rhizobium symbiosis and accumulates abundantly in the nodules. However, the concentration of free iron in the cells is strictly controlled to avoid toxicity. It is known that ferritin accumulates in the cells as an iron storage protein. During nodule senescence, the expression of the ferritin gene, Ljfer1, was induced in Lotus japonicus. We investigated a signal transduction pathway leading to the increase of Ljfer1 in the nodule. The Ljfer1 promoter of L. japonicus contains a conserved Iron-Dependent Regulatory Sequence (IDRS). The expression of Ljfer1 was induced by the application of iron or sodium nitroprusside, which is a nitric oxide (NO) donor. The application of iron to the nodule increased the level of NO. These data strongly suggest that iron-induced NO leads to increased expression of Ljfer1 during the senescence of L. japonicus nodules.

Effects of 24-epibrassinolide on enzymatic browning and antioxidant activity of fresh-cut lotus root slices.

Fresh-cut lotus root slices were treated with 80nM 24-epibrassinolide (EBR) and then stored at 4°C for 8days to investigate the effects on cut surface browning. The results showed that EBR treatment reduced cut surface browning in lotus root slices and alleviated membrane lipid peroxidation as reflected by low malondialdehyde content and lipoxygenase activity. EBR treatment inhibited the activity of phenylalanine ammonia lyase and polyphenol oxidase, and subsequently decreased phenolics accumulation and soluble quniones formation. The treatment also stimulated the activity of peroxidase, catalase and ascorbate peroxidase and delayed the loss of ascorbic acid, which would help prevent membrane lipid peroxidation, as a consequence, reducing decompartmentation of enzymes and substrates causing enzymatic browning. These results indicate that EBR treatment is a promising attempt to control browning at cut surface of fresh-cut lotus root slices.

The alkaline tolerance in Lotus japonicus is associated with mechanisms of iron acquisition and modification of the architectural pattern of the root.

The response of fifty-four Lotus japonicus ecotypes, and of six selected ecotypes was investigated under alkaline conditions. Sensitive, but not tolerant ecotypes, showed interveinal chlorosis under all alkalinity conditions and high mortality under extreme alkalinity. Interveinal chlorosis was associated with Fe deficiency, as a reduced Fe2+ shoot content was observed in all sensitive ecotypes. In addition, some showed a decline in photosynthesis rate and PSII performance compared to the control. In contrast, some tolerant ecotypes did not change these parameters between treatments. Alkaline tolerance could be explained by a mechanism of Fe acquisition and a root structural modification. This conclusion was based on the fact that all tolerant, but not the sensitive ecotypes, presented high ferric reductase oxidase activity under alkaline stress compared to the control, and a Herringbone root pattern modification. On this basis, the analysis of these mechanisms of alkaline tolerance could be used in screening programs for the selection of new tolerant genotypes in the Lotus genus.

Expression of the CLE-RS3 gene suppresses root nodulation in Lotus japonicus.

Cell-to-cell communication, principally mediated by short- or long-range mobile signals, is involved in many plant developmental processes. In root nodule symbiosis, a mutual relationship between leguminous plants and nitrogen-fixing rhizobia, the mechanism for the autoregulation of nodulation (AON) plays a key role in preventing the production of an excess number of nodules. AON is based on long-distance cell-to-cell communication between roots and shoots. In Lotus japonicus, two CLAVATA3/ESR-related (CLE) peptides, encoded by CLE-ROOT SIGNAL 1 (CLE-RS1) and -RS2, act as putative root-derived signals that transmit signals inhibiting further nodule development through interaction with a shoot-acting receptor-like kinase HYPERNODULATION ABERRANT ROOT FORMATION 1 (HAR1). Here, an in silico search and subsequent expression analyses enabled us to identify two new L. japonicus CLE genes that are potentially involved in nodulation, designated as CLE-RS3 and LjCLE40. Time-course expression patterns showed that CLE-RS1/2/3 and LjCLE40 expression is induced during nodulation with different activation patterns. Furthermore, constitutive expression of CLE-RS3 significantly suppressed nodule formation in a HAR1-dependent manner. TOO MUCH LOVE, a root-acting regulator of AON, is also required for the CLE-RS3 action. These results suggest that CLE-RS3 is a new component of AON in L. japonicus that may act as a potential root-derived signal through interaction with HAR1. Because CLE-RS2, CLE-RS3 and LjCLE40 are located in tandem in the genome and their expression is induced not only by rhizobial infection but also by nitrate, these genes may have duplicated from a common gene.

Efecto in vitro del extracto de Lotus corniculatus L. sobre nemátodos gastrointestinales bovinos / In vitro effect of extract of Lotus corniculatus L. on bovine gastrointestinal nematodes

Introducción: las parasitosis son consideradas como uno de los problemas más importantes que afectan la producción bovina, debido a la disminución de los parámetros productivos y reproductivos. Objetivos: determinar el efecto del extracto Lotus corniculatus L. in vitro sobre nemátodos gastrointestinales bovinos. Métodos: se usaron las hojas en estado seco de L. corniculatus con una edad de rebrote de 45 y 70 días, el extracto fue etanólico y se obtuvo mediante lixiviación, el extracto se conservó a temperatura de 4 °C, se utilizó el extracto puro y diluciones obedeciéndose a un intervalo de 1,5 mg/mL. Para determinar el efecto del extracto se trabajó mediante la técnica de eclosión de huevos (Egg hatch assay) para los nemátodos: Bunostomum sp, Strongyloides papilosus y Trichostrongylus colubriformis, tomando como mínimo eficaz un porcentaje del 60 por ciento. Se determinó la presencia de taninos. Resultados: se encontró una inhibición de eclosión de huevos S. papilus con la concentración máxima y la dilución 1 y 2, para el extracto de 45 días de rebrote, con porcentajes de 100 por ciento ,86 por ciento y 66,5 por ciento; el extracto con 70 días de rebrote mostró un porcentaje de inhibición de eclosión únicamente[y1] con la concentración máxima y la dilución 1, con 98,5 por ciento y 78,5 por ciento. Para el T. colubriformis la única dilución que no fue eficaz fue la dilución 4 del extracto de 70 días de rebrote con un porcentaje de 50,5 por ciento y para el Bunostomun spp fue eficaz el extracto con la concentración máxima y la dilución 1 para el extracto de 45 y 70 días de rebrote, con porcentajes de 100 por ciento y 69,5 por ciento para el de 45 días y 100 por ciento y 68,5 por ciento para el de 70 días. Conclusiones: el extracto de L. corniculatus de 45 y 70 días de rebrote mostró los resultados más satisfactorios en inhibición de eclosión de huevos de T. colubriformis(AU)

Introduction: The parasites are considered as one of the most important issues affecting cattle production due to the decline in productive and reproductive parameters. Objective: Determine the effect of Lotus corniculatus L. extract on in vitro bovine gastrointestinal nematodes. Methods: Leaves were used in the dry state with L. corniculatus regrowth age 45 to 70 days, and the ethanolic extract was obtained by leaching, the extract was stored at 4 °C, the pure extract was used and dilutions obeying a range of 1.5 mg/mL. To determine the effect of the extract worked through the technique of hatching eggs (Egg hatch assay) for nematodes: Bunostomum ssp, Trichostrongylus and Strongyloides papilosus, on the effective minimum percentage of 60 percent. The presence of tannins was determined. Results: Inhibition of egg hatching S. papilus with maximum concentration and dilution 1 and 2, to extract 45 days of regrowth, with percentages of 100 percent, 86 percent and 66.5 percent respectively was found; extract regrowth 70 days showed a percent inhibition of hatching only the maximum concentration and dilution 1,98.5 percent and 78.5 percent. For T. colubriformis the only was ineffective dilution was 4 dilution extract regrowth 70 days with a percentage of 50.5 percent and for Bunostomun spp extract was effective with a high concentration and dilution 1 for extract 45 and 70 days of regrowth, with rates of 100 percent and 69.5 percent for 45 days and 100 percent and 68.5 percent for 70 days. Conclusions: L. corniculatus extract 45 and 70 days of regrowth showed the most satisfactory results in inhibition of hatching eggs of T. colubriformis(AU)

Enhanced hyphal growth of arbuscular mycorrhizae by root exudates derived from high R/FR treated Lotus japonicus.

Red/Far Red (R/FR) sensing positively influences the arbuscular mycorrhizal (AM) symbiosis of both legume and nonlegume plants through jasmonic acid (JA) and strigolactone signaling. We previously reported that root exudates obtained from high R/FR-grown plants contained more strigolactone than low R/FR-grown plants. To determine whether JA and JA derivatives were secreted from roots, we investigated the expression levels of JA-responsive genes in L. japonicus Miyakojima MG20 plants treated with root exudates prepared from either high or low R/FR light-treated plants. The root exudates from high R/FR light-treated plants were found to enhance the expression levels of JA-responsive genes significantly. Moreover, exogenous JA increased AM fungal hyphal elongation as did the root exudates derived from high R/FR-grown L. japonicus plants. We conclude that increased JA accumulation and secretion into root exudates from high R/FR light-grown plants is the best explanation for increased colonization and enhanced mycorrhization under these conditions.

Salt effects on functional traits in model and in economically important Lotus species.

A common stress on plants is NaCl-derived soil salinity. Genus Lotus comprises model and economically important species, which have been studied regarding physiological responses to salinity. Leaf area ratio (LAR), root length ratio (RLR) and their components, specific leaf area (SLA) and leaf mass fraction (LMF) and specific root length (SRL) and root mass fraction (RMF) might be affected by high soil salinity. We characterised L. tenuis, L. corniculatus, L. filicaulis, L. creticus, L. burtii and L. japonicus grown under different salt concentrations (0, 50, 100 and 150 mm NaCl) on the basis of SLA, LMF, SRL and RMF using PCA. We also assessed effects of different salt concentrations on LAR and RLR in each species, and explored whether changes in these traits provide fitness benefit. Salinity (150 mm NaCl) increased LAR in L. burtii and L. corniculatus, but not in the remaining species. The highest salt concentration caused a decrease of RLR in L. japonicus Gifu, but not in the remaining species. Changes in LAR and RLR would not be adaptive, according to adaptiveness analysis, with the exception of SLA changes in L. corniculatus. PCA revealed that under favourable conditions plants optimise surfaces for light and nutrient acquisition (SLA and SRL), whereas at higher salt concentrations they favour carbon allocation to leaves and roots (LMF and RMF) in detriment to their surfaces. PCA also showed that L. creticus subjected to saline treatment was distinguished from the remaining Lotus species. We suggest that augmented carbon partitioning to leaves and roots could constitute a salt-alleviating mechanism through toxic ion dilution.

Integrated multi-omics analysis supports role of lysophosphatidylcholine and related glycerophospholipids in the Lotus japonicus-Glomus intraradices mycorrhizal symbiosis.

Interaction of plant roots with arbuscular mycorrhizal fungi (AMF) is a complex trait resulting in cooperative interactions among the two symbionts including bidirectional exchange of resources. To study arbuscular mycorrhizal symbiosis (AMS) trait variation in the model plant Lotus japonicus, we performed an integrated multi-omics analysis with a focus on plant and fungal phospholipid (PL) metabolism and biological significance of lysophosphatidylcholine (LPC). Our results support the role of LPC as a bioactive compound eliciting cellular and molecular response mechanisms in Lotus. Evidence is provided for large interspecific chemical diversity of LPC species among mycorrhizae with related AMF species. Lipid, gene expression and elemental profiling emphasize the Lotus-Glomus intraradices interaction as distinct from other arbuscular mycorrhizal (AM) interactions. In G. intraradices, genes involved in fatty acid (FA) elongation and biosynthesis of unsaturated FAs were enhanced, while in Lotus, FA synthesis genes were up-regulated during AMS. Furthermore, FAS protein localization to mitochondria suggests FA biosynthesis and elongation may also occur in AMF. Our results suggest the existence of interspecific partitioning of PL resources for generation of LPC and novel candidate bioactive PLs in the Lotus-G. intraradices symbiosis. Moreover, the data advocate research with phylogenetically diverse Glomeromycota species for a broader understanding of the molecular underpinnings of AMS.