Lotus Tenuis Research Articles

An efficiently biological nitrogen fixation of non-native Lotus tenuis justifies its key role in the flooding Pampas (Argentina)

Forage production is a major challenge in the Salado River basin, Argentina (also denominated “Flooding Pampas”), the largest livestock raising area. The successful naturalization of the European legume Lotus tenuis (native from Mediterranean area) has been of productive and environmental relevance to the region. The main objective of this work was to quantify the efficiency of L. tenuis through its ability to fix nitrogen biologically and thus strengthening their strategic role in the region. The 15N natural abundance method was used to evaluate the Biological Nitrogen Fixation (BNF) of L. tenuis in soil plots. Non-leguminous weed species in the same plots were utilized as reference plants. Simultaneously, evaluations were carried out in the greenhouse and the relative reproducibility of the field experiments. The results showed that L. tenuis promotion was accompanied by an increase in forage quality improving the Crude Protein (CP) content and the Digestibility of Dry Matter (DDM). Moreover, its contribution to N inputs to the system through BNF in symbiosis with native rhizobia was about 80%, a highly relevant percentage for these constrained agroecosystems where the presence of native legumes is very scarce. In conclusion, this study supports the hypothesis that L. tenuis promotion plays a strategic role in ecosystem sustainability in the Salado River basin, especially in soil constrained areas. Together with previously published data, including information on ruminant management, the results obtained contribute to supporting the idea that the naturalized legume plays a strategic role in the sustainability of marginal agricultural soils.

Inclusion of Lotus tenuis in beef cattle systems in the Argentinian flooding pampa as an enteric methane mitigation strategy

ABSTRACT While concerns about greenhouse gas (GHG) emissions and animal welfare persist, livestock farming offers unique ecosystem services. In Argentina, where livestock significantly contributes to GHG emissions, particularly enteric methane (CH4), urgent action is required to develop effective mitigation strategies while maintaining animal performance. This study assesses the potential of the inclusion of Lotus tenuis in natural grasslands to mitigate CH4 emissions and improve forage quality in the Salado River basin, Argentina. Twenty-four beef heifers (361 ± 19 kg LW) were assigned to two treatments with contrasting botanical composition (LOT: > 80% Lotus tenuis and NG: > 60% grasses). LOT showed better forage quality and reduced CH4 emissions (−17% gCH4/day, p < 0.01; Ym: - 7 to −30%, p < 0.05) compared to NG. However, the condensed tannin content in LOT was insufficient to conclusively attribute any impact on CH4 emissions. This study underscores the potential of promoted Lotus tenuis pastures in the Salado River basin to enhance forage quality and mitigate enteric CH4 emissions in free-ranging cattle systems, albeit additional research is necessary to fully comprehend how this practice interacts within the ecosystem and optimise its benefits.

Plant colonizers of a mercury contaminated site: trace metals and associated rhizosphere bacteria

Background and aimsMercury (Hg) contamination poses severe human and environmental health risks. We aimed to evaluate the colonization of Hg-contaminated sites by native plants and the prokaryotic composition of rhizosphere soil communities of the dominant plant species.MethodsA field study was conducted at a Hg-contaminated site in Romania. Metal concentrations in soil and plant samples were analyzed using portable X-ray fluorescence spectrometry. The prokaryotic composition of rhizosphere soil communities was determined through 16S rRNA amplicon sequencing and community functionality was predicted through PICRUSt2.ResultsSite-specific trace metal distribution across the site drove plant species distribution in the highly contaminated soil, with Lotus tenuis and Diplotaxis muralis associated with higher Hg concentrations. In addition, for the bacterial communities in the rhizosphere soil of D. muralis, there was no observable decrease in alpha diversity with increasing soil Hg levels. Notably, Actinomycetota had an average of 24% relative abundance in the rhizosphere communities that also tested positive for the presence of merA, whereas in the absence of merA the phylum’s relative abundance was approximately 2%. merA positive rhizosphere communities also displayed an inferred increase in ABC transporters.ConclusionsThe results suggest a dependence of species-wise plant survival on local trace metal levels in soil, as well as an intricate interplay of the latter with rhizosphere bacterial diversity. Knowledge of these interdependencies could have implications for phytoremediation stakeholders, as it may allow for the selection of plant species and appropriate soil microbial inoculates with elevated Hg tolerance.

Multiple bHLH/MYB-based protein complexes regulate proanthocyanidin biosynthesis in the herbage of Lotus spp.

Main conclusionThe complexes involving MYBPA2, TT2b, and TT8 proteins are the critical regulators of ANR and LAR genes to promote the biosynthesis of proanthocyanidins in the leaves of Lotus spp.The environmental impact and health of ruminants fed with forage legumes depend on the herbage's concentration and structure of proanthocyanidins (PAs). Unfortunately, the primary forage legumes (alfalfa and clover) do not contain substantial levels of PAs. No significant progress has been made to induce PAs to agronomically valuable levels in their edible organs by biotechnological approaches thus far. Building this trait requires a profound knowledge of PA regulators and their interplay in species naturally committed to accumulating these metabolites in the target organs. Against this background, we compared the shoot transcriptomes of two inter-fertile Lotus species, namely Lotus tenuis and Lotus corniculatus, polymorphic for this trait, to search for differentially expressed MYB and bHLH genes. We then tested the expression of the above-reported regulators in L. tenuis x L. corniculatus interspecific hybrids, several Lotus spp., and different L. corniculatus organs with contrasting PA levels. We identified a novel MYB activator and MYB-bHLH-based complexes that, when expressed in Nicotiana benthamiana, trans-activated the promoters of L. corniculatus anthocyanidin reductase and leucoanthocyanidin reductase 1 genes. The last are the two critical structural genes for the biosynthesis of PAs in Lotus spp. Competition between MYB activators for the transactivation of these promoters also emerged. Overall, by employing Lotus as a model genus, we refined the transcriptional network underlying PA biosynthesis in the herbage of legumes. These findings are crucial to engineering this trait in pasture legumes.

Lotus tenuis maintains high arbuscular mycorrhizal diversity in grasslands regardless of soil properties or management

This study aimed to evaluate the root colonization and diversity of arbuscular mycorrhizal fungi (AMF) associated with Lotus tenuis grown in natural and L. tenuis-promoted grasslands (by herbicide application, which consists in removing above-ground biomass to favor the establishment and growth of L. tenuis) in the Salado River Basin (Argentina). Soils ranged from saline and/or sodic to neutral and non-saline. AMF colonization was slightly higher in natural grasslands than in L. tenuis-promoted sites (93% and 86%, respectively). A total of 22 AMF species were identified in root-associated soil. The Glomeraceae was the only family present in all sites, and Funneliformis mosseae was the only species found in all the sites studied, regardless of the soil properties and management. Claroideoglomus etunicatum and Septoglomus constrictum were present in all sites except in one of the L. tenuis-promoted sites studied. The PCR-Single-Strand Conformation Polymorphism analysis showed that amplicons of AMF-Glomeraceae did not cluster in a site-specific or management-specific way. The sequences obtained were highly similar to sequences of Glomeraceae detected by morphological spore taxonomy. L. tenuis promotion by herbicide application did not negatively affect AMF colonization or diversity. L. tenuis contributed to maintaining AMF diversity through high root colonization, regardless of soil properties or management. This should be considered for the maintenance of L. tenuis and AMF-associated communities.

Shaking off the blow: plant adjustments during submergence and post-stress growth in Lotus forage species.

Flooding significantly hampers global forage production. In flood-prone regions, Lotus tenuis and Lotus corniculatus are common forage legumes, yet little is known about their responses to partial or complete submergence. To address this, we evaluated 10 Lotus accessions subjected to 11days of either partial or complete submergence, analysing growth traits related to tolerance and recovery after de-submergence. Principal component analyses revealed that submergence associated growth parameters were linked to L. corniculatus accessions, whereas recovery was associated with L. tenuis accessions. Notably, in L. tenuis , recovery from complete submergence positively correlated with leaf mass fraction but negatively with root mass fraction, showing an opposite pattern than in L. corniculatus . Encouragingly, no trade-off was found between inherent growth capacity and submergence tolerance (both partial and complete) or recovery ability, suggesting genetic selection for increased tolerance would not compromise growth potential. L. tenuis exhibited accessions with both partial and complete submergence tolerance, making them versatile for flood-prone environments, whereas L. corniculatus accessions were better suited for partial submergence. These findings offer valuable insights to enhance forage production in flood-prone areas and guide the selection of appropriate Lotus accessions for specific flood conditions.

Glyphosate sprayed on the pre-existing vegetation reduces seedling emergence and growth of forage species

Seeding pastures or forage crops by no-tillage methods usually involves the spray of glyphosate to suppress the existing vegetation. While many studies found detrimentaleffects of glyphosate on seed germination and seedling growth of the subsequent crop, others found negligible effects. This study aimed to evaluate the effect of glyphosate spraying on germination, seedling emergence and seedling growth of four forage species: Trifolium repens, Lotus tenuis, Festuca arundinacea and Paspalum dilatatum. The experiment was carried out spraying glyphosate on the pre-existing vegetation and on bare soil 1, 30, 60 and 90 days before sowing, and a control treatmentsprayed with water. Glyphosate sprayed on pre-existing vegetation 1 to 60 days before seeding reduced emergence, while sprayed 1 to 30 or 1 to 60 days before seeding reduced seedlings belowground biomass and root length of all species and aboveground biomass of legumes respect to sprayed 90 days before seeding, sprayed on bare soil, and control treatment. This herbicide would remain active in the soil environment for at least 60 days after spraying when it was previously absorbed by plants, causing a severe damage to seedlings emergence and growth. Highlights: Glyphosate sprayed on pre-existing vegetation 1 to 60 days before seeding reduced seedling emergence of Trifolium repens, Lotus tenuis, Festuca arundinacea and Paspalum dilatatum. Glyphosate sprayed on pre-existing vegetation 1-30 or 1-60 days before seeding reduced seedlings belowground biomass and root length of all species and aboveground biomass of legumes Gyphosate sprayed 90 days before seeding or sprayed on bare soil did not affect seggling emergence and growth. Gyphosate would remain active in the soil environment for at least 60 days after spraying when it was previously absorbed by plants.

Arbuscular mycorrhizal fungi improve legume seedling performance as the competition with conspecific neighbouring adult plants decreases

In temperate grasslands, the balance between competitive and facilitative interactions between adult plants and conspecific seedlings can be affected by defoliation intensity, soil nutrient deficiency and arbuscular mycorrhizal fungi (AMF). Here, we investigated the effect of AMF on the interactions between Lotus tenuis Waldst. & Kit. seedlings grown in the presence or absence of conspecific adult plants defoliated at different intensities (0, 50 and 75%) in a P-deficient soil of the Salado River Basin (Argentina). Mycorrhizal seedlings grown near defoliated and non-defoliated adult plants produced less shoot dry weight (DW) than non-mycorrhizal counterparts, whereas mycorrhizal seedlings alone showed 258% and 504% more shoot DW and shoot P content, respectively than non-mycorrhizal counterparts. This was associated with higher growth and P uptake of mycorrhizal adult plants than non-mycorrhizal counterparts. The competition index between seedlings and adult plants showed competition interaction in mycorrhizal systems (from −2.6 to −1.9) and neutral interaction in non-mycorrhizal systems (near to zero) despite the defoliation intensity. We propose that moderate defoliation (50%) allows maintaining high mycorrhizal benefits in adult plants (110.1% and 168.8% mycorrhizal growth and P response, respectively), whereas high defoliation (75%) decreases mycorrhizal benefits (27.18% and 80.21% mycorrhizal growth and P response, respectively). We highlight AMF symbiosis as an important strategy of L. tenuis adult plants to regulate the growth of neighbouring conspecific seedlings in a P-deficient soil.

Phytochemistry and Antiviral Properties of Two Lotus Species ‎Growing in Egypt

Background: Lotus arabicus L and Lotus glaber Mill. belong to the family Fabaceae, and they grow in the wild in ‎Egypt and have different therapeutic uses in folk medicine. Objectives: This study aimed to evaluate the phytochemical profile and ‎antimicrobial properties of the methanolic extracts of two Lotus spp. growing in Egypt, L. ‎arabicus and L. glaber. Material and methods: Gas chromatography-mass spectrometry was used to identify the compounds of the extracts of two Lotus species. An MTT colorimetric assay and the disc diffusion method were performed to investigate the antiviral and antimicrobial activities of two lotus species, respectively. Results: The n-hexane and methanol extracts of L. ‎arabicuscontained high percentages of alkane hydrocarbons, such as 5-methyloctadecane, ‎while L. glaber contained dodecane. The major compounds in the methanol extract of L. ‎arabicus were hexadecanoic acid methyl ester and dodecanoic acid,2,3-bis(acetyloxy)propyl ‎ester. The major compounds in the methanol extract of L. glaber were palmitic acid and ‎lucenin 2. The indole alkaloid ditaine was found only in L. arabicus. This alkaloid was ‎identified for the first time in the genus Lotus. The antimicrobial properties of the extracts of ‎the two Lotus species showed that the n-hexane extract of ‎both Lotus species may have potential antifungal activity against Candida parapsilosis and ‎Aspergillus flavus. Moreover, the methanolic extracts of both Lotus species have potential ‎antiviral activity against the coxsackie B virus, but only the L. arabicus extract showed ‎activity against the hepatitis A virus.‎ Conclusion: Lotus arabicus might have potential antifungal or antiviral activity greater than L. glaber.

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This study aims at examining and confirming the patterns of phenetic relationships and the levels of variations within and among the species of Lotus L., 1753 in Egypt by using morphometric analysis techniques. We have evaluated 24 morphological characters from about 300 herbarium specimens representing 19 species of Lotus that are currently recognized. Based on numerical analyses of macromorphological characters (cluster analysis, principal coordinate analysis and principal component analysis), 19 species of Lotus were recognized from Egypt. These species were clustered in six species-specific groups: (I) Lotus halophilus Boiss. &amp; Spruner, L. angustissimus L., L. glinoides Delile and L. schimperi Steud. ex Boiss., (II) Lotus glaber Mill. and L. palustris Willd., (III) Lotus polyphyllos E.D. Clarke, L. creticus L. and L. cytisoides L., (IV) Lotus gebelia Vent., L. lanuginosus Vent. and L. arenarius Brot., (V) Lotus edulis L., L. tetragonolobus L. and L. conjugatus L. and (VI) Lotus ornithopodioides L., L. peregrinus L., L. arabicus L. and L. hebranicus Hochst. ex Brand. As a result of this study, we proposed that some characters, not previously examined in detail, showed significant characters in species delimitation: pod length, seed dimensions, features of upper and lower leaflets, calyx, length of corolla, length of style, numbers of flowers and ovules.

Lotus tenuis and Schedonorus arundinaceus co-culture exposed to defoliation and water stress

The present study aimed to investigate the effect of defoliation frequency (low and high) and water stress (excess or deficit) on biomass production, P and N nutrition, and symbiosis with native soil microorganisms on a Lotus tenuis and Schedonorus arundinaceus co-culture in a pot experiment. Combined effects of defoliation frequency and water stress affected plant accumulated shoot biomass. L. tenuis root biomass decreased in response to defoliation and water stress, while S. arundinaceus root biomass was similar between non-defoliated and defoliated plants, at all water levels. Low and high frequencies of defoliation in a waterlogged soil can be considered the most stressful scenario for L. tenuis and S. arundinaceus co-culture. Colonization of arbuscular mycorrhizal fungi in L. tenuis roots and dark septate endophytes colonization in S. arundinaceus roots were affected by both factors, whereas arbuscular mycorrhizal colonization in S. arundinaceus was affected only by water stress. Both plants tolerated defoliation and water stress due to the interaction between the translocation of nutrients and carbon compounds from roots to shoots, and P and N absorption (plus N2 fixation in L. tenuis). Highlights: Both plants tolerated defoliation and water stress due to the interaction between the translocation of nutrients and carbon compounds from roots to shoots, and P and N absorption (plus N2 fixation in tenuis). Low and high frequencies of defoliation in a waterlogged soil can be considered the most stressful scenario for tenuis and S. arundinaceus co-culture. Defoliation frequency increased AM colonization in plant roots under well watered and water deficit conditions. arundinaceus roots were co-colonized by AM fungi and DSE. Promoting the presence of tenuis through low defoliation frequency would improve forage yield and quality with the maintenance of AM symbiosis in legume–grass communities.

Metabolic Profiling and Metabolite Correlation Network Analysis Reveal That Fusarium solani Induces Differential Metabolic Responses in Lotus japonicus and Lotus tenuis against Severe Phosphate Starvation.

Root fungal endophytes are essential mediators of plant nutrition under mild stress conditions. However, variations in the rhizosphere environment, such as nutrient depletion, could result in a stressful situation for both partners, shifting mutualistic to nonconvenient interactions. Mycorrhizal fungi and dark septate endophytes (DSEs) have demonstrated their ability to facilitate phosphate (Pi) acquisition. However, few studies have investigated other plant–fungal interactions that take place in the root environment with regard to phosphate nutrition. In the present research work, we aimed to analyze the effect of extreme Pi starvation and the fungal endophyte Fusarium solani on the model Lotus japonicus and the crop L. tenuis. We conducted metabolomics analysis based on gas chromatography-mass spectrometry (GC-MS) on plant tissues under optimal conditions, severe Pi starvation and F.solani presence. By combining statistical and correlation network analysis strategies, we demonstrated the differential outcomes of the two plant species against the combination of treatments. The combination of nutritional stress and Fusarium presence activated significant modifications in the metabolism of L. japonicus affecting the levels of sugars, polyols and some amino acids. Our results display potential markers for further inspection of the factors related to plant nutrition and plant–fungal interactions.

Configuration of daily grazing and searching of growing beef cattle in grassland: observational study

Many of the studies in Campos grasslands focus on management aspects such as the control of herbage allowance, and application of nutrients and/or overseeding with legumes. However, there is little literature on how the Campos grassland resource is utilised, especially regarding the grazing pattern and the relationship between pasture quantity and quality on daily grazing activities. The study of the ingestive behaviour in species-rich and heterogeneous native grasslands during daylight hours, and understanding how animals prioritise quality or quantity of intake in relation to pasture attributes, are important to comprehend the ingestive-digestive processes modulating the energy intake of animals and to achieve a better grazing management. Therefore, the objective was to describe and quantify the daily grazing behaviour of growing cattle grazing native pasture with different structures as a result of different management practices, and study the relationship of pasture attributes and intake through multivariate analysis. The study was carried out at the Faculty of Agronomy, Paysandú, Uruguay. Treatments were native grassland, overseeding with Trifolium pratense and Lotus tenuis + phosphorus, and native pasture + nitrogen-phosphorus. Grazing activities were discriminated into grazing, searching (defined when animals take 1–2 bites in one feeding station and then change to another feeding station and so on), ruminating and idling. The probability of time allocated to each activity was continuously measured during daylight hours (0700–1930) and was related to pasture structure and forage quality using regression tree models, while the bite rate was determined every 2 h. The diurnal pattern of growing cattle showed grazing and searching sessions, followed by ruminating and idling sessions. The length of sessions (as the probability of time allocated to each activity) varied throughout the day. The grazing probability was greater during afternoon than morning and midday (0.74 vs 0.45 vs 0.46, respectively), and it was associated with higher bite rate (34.2 bites/min). Regression tree models showed different grazing, searching and ruminating strategies according to pasture attributes. During the morning, animals modified grazing, searching, ruminating and idling strategies according to bite rate, crude protein in diet and herbage allowance. At midday, they only adjusted ruminating and idling, while during afternoon sessions, grazing activities were modified by pasture quantity attributes such as herbage mass and herbage allowance. By controlling the herbage allowance, herbage mass and pasture height, animals prioritise quality in the morning and quantity in the afternoon, integrating and modifying the grazing-searching and ruminating-idling pattern.

Prospecting phosphate solubilizing bacteria in alkaline-sodic environments reveals intra-specific variability in Pantoea eucalypti affecting nutrient acquisition and rhizobial nodulation in Lotus tenuis

A bioprospecting study in alkaline-sodic soils of the Argentinean flooding pampa was performed in order to identify and characterize rhizospheric bacteria associated to Lotus tenuis plants, capable of solubilizing phosphate under a broad range of alkaline-sodic conditions. Our analysis, supported by repetitive BOX element based PCR and 16S rRNA sequences, identified 74 strains. All of them belong to the Phylum Proteobacteria, specifically to the order Enterobacteriales, and Pseudomonadales, suggesting that in this environment, broad pH-range P-solubilizing bacteria (BRPSB) associated to L. tenuis, are grouped within a narrow taxonomic range. A subsequent objective was to focus in a subgroup of BRPSB strains belonging to the Pantoea eucalypti species (MA66, P63, P76, P163, P173 and a formerly identified isolate, M91) that also produced siderophores, indol-acetic acid and showed in vitro compatibility with the native rhizobial strain Mesorhizobium sanjuanii BSA136. Growth promoting effects of these P. eucalypti strains on L. tenuis plants in alkaline-sodic soils in symbiosis with the above mentioned rhizobial strain were analyzed. Despite all the P. eucalypti BRPSB strains exhibited the above-mentioned features, they exerted differential effects on plant growth and dry matter allocation to the nodules. Plants inoculated with P. eucalypti M91 displayed a superior capability to accumulate nitrogen, phosphorus and zinc. On the contrary, nodules dry matter allocation, and mineral nutrient accumulation in L. tenuis plants were negatively affected by P. eucalypti P76 compared with M91. Results hereby presented highlight the complexity of plant-microbe interactions and reveal that growth-promoting effects of P-solubilizing P. eucalypti strains cannot be predicted only on the basis of their in vitro PGPR features, complementary in planta assays being necessary for efficient strain selection. This study provides valuable information for biofertilization of L. tenuis plants in the flooding pampa.

Salt stress on Lotus tenuis triggers cell wall polysaccharide changes affecting their digestibility by ruminants

Lotus tenuis is a glycophytic forage legume (Fabaceae) used in feeding ruminants that can grow under salinity and waterlogging stresses. Plants obtained in controlled conditions were affected negatively in their growth by the effect of salt. Results from sequential extraction of plant cell wall polysaccharides and chemical characterization were related to those from nutritional parameters used to assess ruminants feedstuffs (Van Soest detergent system). Shoots and leaves were analyzed, and the most important differences were found for shoots. The salt-stressed shoots gave lower values of neutral detergent fiber and acid detergent fiber; they produced higher amounts of reserve α-glucans, and hemicelluloses (xyloglucans and glucuronoxylans from primary and secondary cell walls, respectively) and pectins, leaving less material resistant to extraction. This effect was clearly confirmed by an in vitro gas production assay. In addition, observations by light microcopy (LM) and transmission electron microscopy (TEM), showed in some tissues thicker walls and more opened cell wall structures in regard to control samples, which could allow easier access of degrading enzymes in the rumen. Although the plant biomass of Lotus tenuis produced under salt stress was lower, its quality as forage improved due to production of increased quantities of more digestible polysaccharides.

Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter

Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter

Post-reclamation microbial diversity and functions in hexachlorocyclohexane (HCH) contaminated soil in relation to spontaneous HCH tolerant vegetation

The toxicity, volatility and persistence of the obsolete organochlorine pesticide hexachlorocyclohexane (HCH), makes reclamation of contaminated areas a priority for the health and welfare of neighboring human communities. Microbial diversity and functions and their relation to spontaneous vegetation in post-excavation situations, are essential indicators to consider in bioaugmentation or microbe-assisted phytoremediation strategies at field scale. Our study aimed to evaluate the effects of long-term HCH contamination on soil and plant-associated microbial communities, and whether contaminated soil has the potential to act as a bacterial inoculum in post-excavation bioremediation strategies. To scrutinize the role of vegetation, the potential nitrogen fixation of free-living and symbiotic diazotrophs of the legume Lotus tenuis was assessed as a measure of nutrient cycling functions in soil under HCH contamination. Potential nitrogen fixation was generally not affected by HCH, with the exception of lower nifH gene counts in excavated contaminated rhizospheres, most probably a short-term HCH effect on early bacterial succession in this compartment. HCH shaped microbial communities in long-term contaminated bulk soil, where we identified possible HCH tolerants such as Sphingomonas and Altererythrobacter. In L. tenuis rhizosphere, microbial community composition was additionally influenced by plant growth stage. Sphingobium and Massilia were the bacterial genera characteristic for HCH contaminated rhizospheres. Long-term HCH contamination negatively affected L. tenuis growth and development. However, root-associated bacterial community composition was driven solely by plant age, with negligible HCH effect. Results showed that L. tenuis acquired possible HCH tolerant bacteria such as the Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium clade, Sphingomonas, Massilia or Pantoea which could simultaneously offer plant growth promoting (PGP) benefits for the host. Finally, we identified an inoculum with possibly HCH tolerant, PGP bacteria transferred from the contaminated bulk soil to L. tenuis roots through the rhizosphere compartment, consisting of Mesorhizobium loti, Neorhizobium galegae, Novosphingobium lindaniclasticum, Pantoea agglomerans and Lysobacter bugurensis.

Impact of Water Level on Species Quantity and Composition Grown from the Soil Seed Bank of the Inland Salt Marsh: An Ex-Situ Experiment

The near elimination of inland salt marshes in Central Europe occurred throughout the 19th and 20th centuries, and the currently remaining marshes exist in a degraded condition. This work examines the impact of groundwater level on the growth of plants from a seed bank obtained from a degraded salt marsh in proximity to still existing one through an ex-situ experiment. An experimental tank was set up with the sample seed bank experiencing differing levels of water level. There were 1233 specimens of 44 taxa grown from the seed bank, of which 5 species were abundant, and 10 species are considered as halophytes. Only Lotus tenuis from halophytes was more abundant, and only five species of halophytes were represented by more than three individuals. The water level has a significant impact on the number of species (based on linear regression analysis) as well as species distribution among different water level treatments (a non-metric multidimensional analysis (nMDS) followed by linear regression). The results show a strong negative relationship between the average water level and the number of species. The water level did not affect the species composition of halophytes, but differences in individual species abundances were found among the halophytes. The species Bupleurum tenuissimum, Crypsis schoenoides, Melilotus dentatus, and Plantago maritima grew on the drier and non-inundated soils. Tripolium pannonicum, Spergularia maritima, and Lotus tenuis grew on both wet and dry soils. Trifolium fragiferum and Bolboschoenus maritimus were found in places with water stagnant at the soil level. Pulicaria dysenterica grew in inundated soil.

Germinación de semillas de Lotus tenuis: efecto del tratamiento pregerminativo, tiempo de almacenamiento y rol de la cubierta dura a largo plazo

Se evaluó el efecto de la escarificación mecánica en dos momentos luego de la cosecha sobre la germinación de semillas de 23 genotipos de Lotus tenuis Waldst et Kit almacenadas en condiciones de laboratorio sin escarificar y el rol de las cubiertas duras sobre dicha respuesta a largo plazo. Se usó un DCA con tres factores [genotipos, tratamiento pregerminativo (E=escarificado; NE=ausencia de escarificado) y tiempo de almacenamiento (t1=0,25 años; t2=22,5 años)] (n=4). Se calculó el porcentaje de germinación acumulada (Gac%) y de semillas duras (Sd%). Para t2 se calculó el índice de velocidad germinativa (IVG). Se aplicó ANOVA no paramétrico (Kruskal-Wallis) y comparación de medianas. Para t2 se analizaron correlación y regresión simple. La interacción triple resultó significativa (p ≤0,01) para ambos parámetros. Gac% tuvo valores mayores en el tratamiento E en t1 y menores en NE en t2. El factor genotipo resultó significativo (p≤0,01) para el E en t2. Sd% tuvo valores mayores en el tratamiento NE en t1 y menores en E independientemente del tiempo de almacenamiento. El factor genotipo resultó significativo (p≤0,01) para NE en t2. La correlación Sd% vs. Gac% e IVG resultó significativa (p≤0,01) y positiva. Las regresiones simples Sd% vs. Gac% e IVG resultaron significativas (p≤0,01). El estudio demostró que la escarificación y el tiempo de almacenamiento afectan de manera positiva y negativa, respectivamente, al comportamiento germinativo de los genotipos estudiados, que el ablandamiento de la cubierta dura es variable entre genotipos y que dicha cubierta estaría vinculada con la conservación de la capacidad germinativa en el largo plazo.

Submergence tolerance and recovery in Lotus: Variation among fifteen accessions in response to partial and complete submergence

Several Lotus species are perennial forage legumes which tolerate waterlogging, but knowledge of responses to partial or complete shoot submergence is scant. We evaluated the responses of 15 Lotus accessions to partial and complete shoot submergence and variations in traits associated with tolerance and recovery after de-submergence. Accessions of Lotus tenuis, L. corniculatus, L. pedunculatus and L. japonicus were raised for 43 d and then subjected to aerated root zone (control), deoxygenated stagnant root zone with shoots in air (stagnant), stagnant root zone with partial (75 %) and complete submergence of shoots, for 7 d. The recovery ability from complete submergence was also assessed. We found inter- and intra-specific variations in the stem extension responses (i.e. promoted or restricted compared to controls) depending on water depth. Eight of 15 accessions promoted the stem extension when in partial submergence, while three of those eight (all L. tenuis accessions) had a restricted stem extension when under complete submergence. Two accessions (belonging to L. corniculatus and L. penduculatus species) also promoted the stem extension under complete submergence. The accessions that attained better recovery in terms of leaves produced after de-submergence, were those that had high leaf and root sugar concentration at de-submergence, and high thickness and persistence of gas films on leaves during submergence (all L. tenuis accessions). We conclude that all Lotus accessions were able to tolerate 7 d of partial and complete shoot submergence, despite adopting different stem extension responses.