Mitigation of salt stress in lettuce by a biostimulant that protects the root absorption zone and improves biochemical responses.

Horticultural crops constantly face abiotic stress factors such as salinity, which have intensified in recent years due to accelerated climate change, significantly affecting their yields and profitability. Under these conditions, it has become necessary to implement effective and sustainable solutions to guarantee agricultural productivity and food security. The influence of BALOX®, a biostimulant of plant origin, was tested on the responses to salinity of Lactuca sativa L. var. longifolia plants exposed to salt concentrations up to 150 mM NaCl, evaluating different biometric and biochemical properties after 25 days of treatment. Control plants were cultivated under the same conditions but without the biostimulant treatment. An in situ analysis of root characteristics using a non-destructive, real-time method was also performed. The salt stress treatments inhibited plant growth, reduced chlorophyll and carotenoid contents, and increased the concentrations of Na+ and Cl- in roots and leaves while reducing those of Ca2+. BALOX® application had a positive effect because it stimulated plant growth and the level of Ca2+ and photosynthetic pigments. In addition, it reduced the content of Na+ and Cl- in the presence and the absence of salt. The biostimulant also reduced the salt-induced accumulation of stress biomarkers, such as proline, malondialdehyde (MDA), and hydrogen peroxide (H2O2). Therefore, BALOX® appears to significantly reduce osmotic, ionic and oxidative stress levels in salt-treated plants. Furthermore, the analysis of the salt treatments' and the biostimulant's direct effects on roots indicated that BALOX®'s primary mechanism of action probably involves improving plant nutrition, even under severe salt stress conditions, by protecting and stimulating the root absorption zone.

Agroindustrial By-Products as a Source of Biostimulants Enhancing Responses to Abiotic Stress of Horticultural Crops.

Together with other abiotic stresses such as drought and high temperatures, salt stress is one of the most deleterious environmental factors affecting plant development and productivity, causing significant crop yield reductions. The progressive secondary salinisation of irrigated farmland is a problem as old as agriculture but is aggravated and accelerated in the current climate change scenario. Plant biostimulants, developed commercially during the last decade, are now recognised as innovative, sustainable agronomic tools for improving crop growth, yield, plant health and tolerance to abiotic stress factors such as water and soil salinity. Biostimulants are a disparate collection of biological extracts, natural and synthetic organic compounds or mixtures of compounds, inorganic molecules and microorganisms, defined by the positive effects of their application to crops. The growing interest in biostimulants is reflected in the increasing number of scientific reports published on this topic in recent years. However, the processes triggered by the biostimulants and, therefore, their mechanisms of action remain elusive and represent an exciting research field. In this review, we will mainly focus on one specific group of biostimulants, protein hydrolysates, generally produced from agricultural wastes and agroindustrial by-products-contributing, therefore, to more sustainable use of resources and circular economy-and primarily on the consequences of their application on the abiotic stress resistance of horticultural crops. We will summarise data in the scientific literature describing the biostimulants' effects on basic, conserved mechanisms activated in response to elevated salinity and other abiotic stress conditions, such as the control of ion transport and ion homeostasis, the accumulation of osmolytes for osmotic adjustment, or the activation of enzymatic and non-enzymatic antioxidant systems to counteract the induced secondary oxidative stress. The collected information confirms the positive effects of biostimulants on crop tolerance to abiotic stress by enhancing morphological, physiological and biochemical responses, but also highlights that more work is needed to further establish the molecular mechanisms underlying biostimulants' effects.

Salt Tolerance of Sea Flax (Linum maritimum L.), a Rare Species with Conservation Interest in Eastern Spain.

Seldom found in saltmarshes, Linum maritimum is a halophyte of great conservation interest in the eastern Iberian Peninsula. Although the species has been reported in different plant communities, there is no information on its range of salinity tolerance or mechanisms of response to environmental stress factors. In this study, L. maritimum plants were subjected to increasing salt concentrations in controlled conditions in a greenhouse. After six months of watering with salt solutions, only plants from the control, 50 mM and 100 mM NaCl treatment groups survived, but seeds were produced only in the first two. Significant differences were found between the plants from the various treatment groups in terms of their growth parameters, such as plant height, fresh weight, and the quantity of flowers and fruits. The main mechanism of salt tolerance is probably related to the species' ability to activate K+ uptake and transport to shoots to partly counteract the accumulation of toxic Na+ ions. A biochemical analysis showed significant increases in glycine betaine, flavonoids and total phenolic compounds, highlighting the importance of osmotic regulation and antioxidant compounds in the salt tolerance of Linum maritimum. These findings have implications for the conservation of the species, especially under changing climatic conditions that may lead to increased soil salinity in its Mediterranean distribution area.

Touch, feel, heal. The use of hospital green spaces and landscape as sensory-therapeutic gardens: a case study in a university clinic.

It has been documented that patients with mental or physical disabilities can benefit from being placed within the setting of a natural environment. Consequently, the concept of creating spaces that can enhance health preservation or patient recovery, while also augmenting environmental and aesthetic value, has merged as a contemporary discourse. Green areas around hospitals can offer a great opportunity to incorporate healing gardens to benefit their patients and not only. The aim of this paper is to propose a design for a sensory-therapeutic garden based on key principles derived from selected academic literature, focusing on the application of these principles in a healthcare setting in Cluj-Napoca, Romania. The design was informed also by onsite data collection and analysis, and it aims to create a healing landscape that addresses the needs of patients, healthcare providers, and visitors. This study seeks to augment the discourse in the field by demonstrating the practical application of key therapeutic garden design principles in a specific context and how these principles impacted the design process.

Responses of different invasive and non-invasive ornamental plants to water stress during seed germination and vegetative growth.

Biological invasions represent a major threat to natural ecosystems. A primary source of invasive plants is ornamental horticulture, which selects traits related to invasiveness. This study evaluated the responses to water stress during germination and vegetative growth of six species used as ornamental or medicinal plants. Three of them are recognised as invasive weeds in many world areas. Seeds were exposed to increasing concentrations of polyethylene glycol (PEG) mimicking drought stress, and young plants in the vegetative growth stage were subjected to two levels of water stress. Results indicated that in the absence of stress in control conditions, the most competitive species were those reported as weeds, namely Bidens pilosa L., Oenothera biennis L., and Centaurea cyanus L., the last regarding germination velocity. Under stress, only two species, Limonium sinuatum (L.) Mill. and C. cyanus, maintained germination at -1 MPa osmotic potential, but in the recovery experiment, an osmopriming effect of PEG was observed. The most tolerant species during growth were two natives in the Mediterranean region, L. sinuatum and Lobularia maritima (L.) Desv., both accumulating the highest proline concentrations. The sixth species studied, Echinacea purpurea (L.) Moench., proved to be more susceptible to stress in the two developmental stages. This study reveals that the most significant traits associated with invasiveness were related to germination, especially in the absence of stress.

Evaluation of Drought Responses in Two Tropaeolum Species Used in Landscaping through Morphological and Biochemical Markers.

One of the most important challenges horticultural crops confront is drought, particularly in regions such as the Mediterranean basin, where water supplies are usually limited and will become even scarcer due to global warming. Therefore, the selection and diversification of stress-tolerant cultivars are becoming priorities of contemporary ornamental horticulture. This study explored the impact of water stress on two Tropaeolum species frequently used in landscaping. Young plants obtained by seed germination were exposed to moderate water stress (half the water used in the control treatments) and severe water stress (complete withholding of irrigation) for 30 days. Plant responses to these stress treatments were evaluated by determining several growth parameters and biochemical stress markers. The latter were analysed by spectrophotometric methods and, in some cases, by non-destructive measurements using an optical sensor. The statistical analysis of the results indicated that although the stress responses were similar in these two closely related species, T. minus performed better under control and intermediate water stress conditions but was more susceptible to severe water stress. On the other hand, T. majus had a stronger potential for adaptation to soil water scarcity, which may be associated with its reported expansion and naturalisation in different regions of the world. The variations in proline and malondialdehyde concentrations were the most reliable biochemical indicators of water stress effects. The present study also showed a close relationship between the patterns of variation of flavonoid and chlorophyll contents obtained by sensor-based and spectrophotometric methods.

Effect of Water Deficit on Germination, Growth and Biochemical Responses of Four Potentially Invasive Ornamental Grass Species.

Ornamental plant species introduced into new environments can exhibit an invasive potential and adaptability to abiotic stress factors. In this study, the drought stress responses of four potentially invasive ornamental grass species (Cymbopogon citratus, Cortaderia selloana, Pennisetum alopecuroides and P. setaceum) were analysed. Several seed germination parameters were determined under increasing polyethylene glycol (PEG 6000) concentrations. Additionally, plants in the vegetative stage were subjected to intermediate and severe water stress treatments for four weeks. All species registered high germination rates in control conditions (no stress treatment), even at high PEG concentrations, except C. citratus, which did not germinate at -1 MPa osmotic potential. Upon applying the water stress treatments, P. alopecuroides plants showed the highest tolerance, and C. citratus appeared the most susceptible to drought. Stress-induced changes in several biochemical markers (photosynthetic pigments, osmolytes, antioxidant compounds, root and shoot Na+ and K+ contents), highlighted different responses depending on the species and the stress treatments. Basically, drought tolerance seems to depend to a large extent on the active transport of Na+ and K+ cations to the aerial part of the plants, contributing to osmotic adjustment in all four species and, in the case of the most tolerant P. alopecuroides, on the increasing root K+ concentration under water deficit conditions. The study shows the invasive potential of all species, except C. citratus, in dry areas such as the Mediterranean region, especially in the current climate change scenario. Particular attention should be given to P. alopecuroides, which is widely commercialised in Europe as ornamental.

Use of a Biostimulant to Mitigate the Effects of Excess Salinity in Soil and Irrigation Water in Tomato Plants.

Global warming is linked to progressive soil salinisation, which reduces crop yields, especially in irrigated farmland on arid and semiarid regions. Therefore, it is necessary to apply sustainable and effective solutions that contribute to enhanced crop salt tolerance. In the present study, we tested the effects of a commercial biostimulant (BALOX®) containing glycine betaine (GB) and polyphenols on the activation of salinity defense mechanisms in tomato. The evaluation of different biometric parameters and the quantification of biochemical markers related to particular stress responses (osmolytes, cations, anions, oxidative stress indicators, and antioxidant enzymes and compounds) was carried out at two phenological stages (vegetative growth and the beginning of reproductive development) and under different salinity conditions (saline and non-saline soil, and irrigation water), using two formulations (different GB concentrations) and two doses of the biostimulant. Once the experiments were completed, the statistical analysis revealed that both formulations and doses of the biostimulant produced very similar effects. The application of BALOX® improved plant growth and photosynthesis and assisted osmotic adjustment in root and leaf cells. The biostimulant effects are mediated by the control of ion transport, reducing the uptake of toxic Na+ and Cl- ions and favoring the accumulation of beneficial K+ and Ca2+ cations, and a significant increase in leaf sugar and GB contents. BALOX® significantly reduced salt-induced oxidative stress and its harmful effects, as evidenced by a decrease in the concentration of oxidative stress biomarkers, such as malondialdehyde and oxygen peroxide, which was accompanied by the reduction of proline and antioxidant compound contents and the specific activity of antioxidant enzymes with respect to the non-treated plants.

Effect of Recurrent Salt and Drought Stress Treatments on the Endangered Halophyte Limonium angustebracteatum Erben

Limonium angustebracteatum is an endemic halophyte from the Spanish Mediterranean coastal salt marshes. To investigate this species' ability to cope with recurrent drought and salt stress, one-year-old plants were subjected to two salt stress treatments (watering with 0.5 and 1 M NaCl solutions), one water stress treatment (complete irrigation withholding), or watered with non-saline water for the control, across three phases: first stress (30 days), recovery from both stresses (15 days), and second stress (15 days). Growth and biochemical parameters were determined after each period. The plants showed high salt tolerance but were sensitive to water deficit, as shown by the decrease in leaf fresh weight and water content, root water content, and photosynthetic pigments levels in response to the first water stress; then, they were restored to the respective control values upon recovery. Salt tolerance was partly based on the accumulation of Na+, Cl- and Ca2+ in the roots and predominantly in the leaves; ion levels also decreased to control values during recovery. Organic osmolytes (proline and total soluble sugars), oxidative stress markers (malondialdehyde and H2O2), and antioxidant compounds (total phenolic compounds and flavonoids) increased by various degrees under the first salt and water stress treatments, and declined after recovery. The analysed variables increased again, but generally to a lesser extent, during the second stress phase, suggesting the occurrence of stress acclimation acquired by the activation of defence mechanisms during the first stress period.

Transgressive Biochemical Response to Water Stress in Interspecific Eggplant Hybrids.

In a climate change scenario, crop tolerance to drought must be urgently improved, as it represents an increasingly critical stress reducing agricultural yields worldwide. Although most crops are relatively sensitive to water stress, many of their wild relatives are more tolerant and may be used to improve drought tolerance in our crops. In this study, the response to drought of eggplant (Solanum melongena), its close wild relatives S. insanum and S. incanum and their interspecific hybrids with S. melongena was assessed. The plants were subjected to two treatments for 18 days: control, with irrigation every four days, and drought, with complete interruption of irrigation. Morphological and biomass traits were measured, and physiological and biochemical responses were analysed using stress biomarkers such as proline, flavonoids, and total phenolic compounds. Oxidative stress was quantified by measuring malondialdehyde (MDA) content. As a result of the drought treatment, plant development and tissue water content were seriously affected. Generally, water deficit also caused significant increases in MDA, proline, flavonoids, and total phenolics compounds. Our results comparing parental accessions reveal a better response to drought in one of the S. insanum accessions. The hybrid between S. melongena and S. incanum displayed a better response than the other hybrids and even its parents. The results obtained here might be helpful for future eggplant breeding programmes aimed at improving drought tolerance.

Comparative Analysis of Tolerance to Salt Stress and Water Deficit in Two Invasive Weeds of the Genus Erigeron (Asteraceae).

Erigeron bonariensis and E. sumatrensis are two noxious weeds present in many parts of the world. Their tolerance to salinity and water deficit was analysed at the seed germination stage and during vegetative development. Seed germination was tested in solutions with different concentrations of NaCl and polyethylene glycol (PEG). Growth parameters, photosynthetic pigments, ion accumulation, and antioxidant mechanisms were analysed in plants that were subjected to increasing NaCl solutions, or severe water deficit by completely restricting irrigation. Seed germination was mostly affected by NaCl, but less by PEG in both species. E. bonariensis had a faster germination in all treatments and maintained a higher percentage of germination under the highest concentration of salt applied. Growth responses were similar in the two species, both being more affected by higher salt concentrations than by water deficit. The main differences in the responses of the two species to stress regard K+ and proline concentration. K+ in roots decreased under salt stress in E. sumatrensis, but remained constant in leaves, whereas in E. bonariensis increased in roots and leaves in salt-stressed plants. Proline concentration increased in all E. bonariensis plants under salt stress, but only in those under the highest salt concentration in E. sumatrensis. The results obtained indicate that the two species are relatively tolerant to water deficit and medium salinity but are susceptible to high NaCl concentrations.

Constitutive and Adaptive Traits of Environmental Stress Tolerance in the Threatened Halophyte Limonium angustebracteatum Erben (Plumbaginaceae).

Limonium angustebracteatum is a halophyte endemic to the E and SE Iberian Peninsula with interest in conservation. Salt glands represent an important adaptive trait in recretohalophytes like this and other Limonium species, as they allow the excretion of excess salts, reducing the concentration of toxic ions in foliar tissues. This study included the analysis of the salt gland structure, composed of 12 cells, 4 secretory and 8 accessory. Several anatomical, physiological and biochemical responses to stress were also analysed in adult plants subjected to one month of water stress, complete lack of irrigation, and salt stress, by watering with aqueous solutions of 200, 400, 600 and 800 mM NaCl. Plant growth was inhibited by the severe water deficit and, to a lesser extent, by high NaCl concentrations. A variation in the anatomical structure of the leaves was detected under conditions of salt and water stress; plants from the salt stress treatment showed salt glands sunken between epidermal cells, bordered by very large epidermal cells, whereas in those from the water stress treatment, the epidermal cells were heterogeneous in shape and size. In both, the palisade structure of the leaves was altered. Salt excretion is usually accompanied by the accumulation of salts in the foliar tissue. This was also found in L. angustebracteatum, in which the concentration of all ions analysed was higher in the leaves than in the roots. The increase of K+ in the roots of plants subjected to water stress was also remarkable. The multivariate analysis indicated differences in water and salt stress responses, such as the accumulation of Na and Cl, or proline, but K+ homeostasis played a relevant role in the mechanism of tolerance to both stressful conditions.

Recovery from Salinity and Drought Stress in the Perennial Sarcocornia fruticosa vs. the Annual Salicornia europaea and S. veneta.

Current agricultural problems, such as the decline of freshwater and fertile land, foster saline agriculture development. Salicornia and Sarcocornia species, with a long history of human consumption, are ideal models for developing halophyte crops. A greenhouse experiment was set up to compare the response of the perennial Sarcocornia fruticosa and the two annual Salicornia europaea and S. veneta to 30 days of salt stress (watering with 700 mM NaCl) and water deficit (complete withholding of irrigation) separate treatments, followed by 15 days of recovery. The three species showed high tolerance to salt stress, based on the accumulation of ions (Na+, Cl-, Ca2+) in the shoots and the synthesis of organic osmolytes. These defence mechanisms were partly constitutive, as active ion transport to the shoots and high levels of glycine betaine were also observed in non-stressed plants. The three halophytes were sensitive to water stress, albeit S. fruticosa to a lesser extent. In fact, S. fruticosa showed a lower reduction in shoot fresh weight than S. europaea or S. veneta, no degradation of photosynthetic pigments, a significant increase in glycine betaine contents, and full recovery after the water stress treatment. The observed differences could be due to a better adaptation of S. fruticosa to a drier natural habitat, as compared to the two Salicornia species. However, a more gradual stress-induced senescence in the perennial S. fruticosa may contribute to greater drought tolerance in this species.

Biological Traits and Genetic Relationships Amongst Cultivars of Three Species of Tagetes (Asteraceae).

Marigolds (Tagetes spp.) are multifunctional flowering plants belonging to the Asteraceae family, well-known and widespread for their ornamental value and many other uses. In this study, morphological differences and genetic relationships among 21 cultivars of three species of marigold (Tagetes patula, T. erecta and T. tenuifolia) were analysed. Results have revealed obvious differences among genotypes, starting from the morphological characteristics of the seeds and their capacity to germinate to adult plant morphological characteristics, both between cultivars and species. The genotypic differences were manifested in considerable variation in the development of phenological stages and the main morphological traits of plants and flowers. PCA and hierarchical clustering analyses of morphological traits revealed a homogeneous grouping of cultivars within each species, except for Orion, belonging to T. patula, which was closer to T. erecta cultivars. A subset of 13 cultivars from the three species was subjected to SSR analysis, revealing considerable genetic diversity and good separation between T. patula on the one side and T. erecta and T. tenuifolia on the other. The observed heterozygosity was much lower than the expected heterozygosity, revealing a high degree of fixation. The results reveal that the three species evaluated have considerable morphological and genetic diversity, which has important implications for assessing genetic diversity, conserving germplasm and selecting parents for new breeding works in marigolds.

Responses to Salinity in Four Plantago Species from Tunisia.

The genus Plantago is particularly interesting for studying the mechanisms of salt tolerance in plants, as it includes both halophytes and glycophytes, as well as species adapted to xeric environments. In this study, the salt stress responses of two halophytes, P. crassifolia and P. coronopus, were compared with those of two glycophytes, P. ovata and P. afra. Plants obtained by seed germination of the four species, collected in different regions of Tunisia, were subjected to increasing salinity treatments for one month under greenhouse conditions. Morphological traits and biochemical parameters, such as ion accumulation and the leaf contents of photosynthetic pigments, osmolytes, oxidative stress markers and antioxidant metabolites, were measured after the treatments. Salt-induced growth inhibition was more pronounced in P. afra, and only plants subjected to the lowest applied NaCl concentration (200 mM) survived until the end of the treatments. The biochemical responses were different in the two groups of plants; the halophytes accumulated higher Na+ and proline concentrations, whereas MDA levels in their leaves decreased, indicating a lower level of oxidative stress. Overall, the results showed that P. coronopus and P. crassifolia are the most tolerant to salt stress, and P. afra is the most susceptible of the four species. Plantago ovata is also quite resistant, apparently by using specific mechanisms of tolerance that are more efficient than in the halophytes, such as a less pronounced inhibition of photosynthesis, the accumulation of higher levels of Cl- ions in the leaves, or the activation of K+ uptake and transport to the aerial part under high salinity conditions.

Responses to Salt Stress in Portulaca: Insight into Its Tolerance Mechanisms.

Climate change and its detrimental effects on agricultural production, freshwater availability and biodiversity accentuated the need for more stress-tolerant varieties of crops. This requires unraveling the underlying pathways that convey tolerance to abiotic stress in wild relatives of food crops, industrial crops and ornamentals, whose tolerance was not eroded by crop cycles. In this work we try to demonstrate the feasibility of such strategy applying and investigating the effects of saline stress in different species and cultivars of Portulaca. We attempted to unravel the main mechanisms of stress tolerance in this genus and to identify genotypes with higher tolerance, a procedure that could be used as an early detection method for other ornamental and minor crops. To investigate these mechanisms, six-week-old seedlings were subjected to saline stress for 5 weeks with increasing salt concentrations (up to 400 mM NaCl). Several growth parameters and biochemical stress markers were determined in treated and control plants, such as photosynthetic pigments, monovalent ions (Na+, K+ and Cl-), different osmolytes (proline and soluble sugars), oxidative stress markers (malondialdehyde-a by-product of membrane lipid peroxidation-MDA) and non-enzymatic antioxidants (total phenolic compounds and total flavonoids). The applied salt stress inhibited plant growth, degraded photosynthetic pigments, increased concentrations of specific osmolytes in both leaves and roots, but did not induce significant oxidative stress, as demonstrated by only small fluctuations in MDA levels. All Portulaca genotypes analyzed were found to be Na+ and Cl- includers, accumulating high amounts of these ions under saline stress conditions, but P. grandiflora proved to be more salt tolerant, showing only a small reduction under growth stress, an increased flower production and the lowest reduction in K+/Na+ rate in its leaves.

Responses to Increased Salinity and Severe Drought in the Eastern Iberian Endemic Species Thalictrum maritimum (Ranunculaceae), Threatened by Climate Change.

Thalictrum maritimum is an endangered, endemic species in East Spain, growing in areas of relatively low salinity in littoral salt marshes. A regression of its populations and the number of individuals has been registered in the last decade. This study aimed at establishing the causes of this reduction using a multidisciplinary approach, including climatic, ecological, physiological and biochemical analyses. The climatic data indicated that there was a direct negative correlation between increased drought, especially during autumn, and the number of individuals censused in the area of study. The susceptibility of this species to water deficit was confirmed by the analysis of growth parameters upon a water deficit treatment applied under controlled greenhouse conditions, with the plants withstanding only 23 days of complete absence of irrigation. On the other hand, increased salinity does not seem to be a risk factor for this species, which behaves as a halophyte, tolerating in controlled treatments salinities much higher than those registered in its natural habitat. The most relevant mechanisms of salt tolerance in T. maritimum appear to be based on the control of ion transport, by (i) the active transport of toxic ions to the aerial parts of the plants at high external salinity-where they are presumably stored in the leaf vacuoles to avoid their deleterious effects in the cytosol, (ii) the maintenance of K+ concentrations in belowground and aboveground organs, despite the increase of Na+ levels, and (iii) the salt-induced accumulation of Ca2+, particularly in stems and leaves. This study provides useful information for the management of the conservation plans of this rare and endangered species.

Effects of Drought and Salinity on Two Commercial Varieties of Lavandula angustifolia Mill.

Global warming is not only affecting arid and semi-arid regions but also becoming a threat to agriculture in Central and Eastern European countries. The present study analyzes the responses to drought and salinity of two varieties of Lavandula angustifolia cultivated in Romania. Lavender seedlings were subjected to one month of salt stress (100, 200, and 300 mM NaCl) and water deficit (complete withholding of irrigation) treatments. To assess the effects of stress on the plants, several growth parameters and biochemical stress markers (photosynthetic pigments, mono and divalent ions, and different osmolytes) were determined in control and stressed plants after the treatments. Both stress conditions significantly inhibited the growth of the two varieties, but all plants survived the treatments, indicating a relative stress tolerance of the two varieties. The most relevant mechanisms of salt tolerance are based on the maintenance of foliar K+ levels and the accumulation of Ca2+ and proline as a functional osmolyte in parallel with increasing external salinities. Under water stress, significant increases of Na+ and K+ concentrations were detected in roots, indicating a possible role of these cations in osmotic adjustment, limiting root dehydration. No significant differences were found when comparing the stress tolerance and stress responses of the two selected lavender varieties.

Insights on Salt Tolerance of Two Endemic Limonium Species from Spain.

We have analysed the salt tolerance of two endemic halophytes of the genus Limonium, with high conservation value. In the present study, seed germination and growth parameters as well as different biomarkers-photosynthetic pigments, mono and divalent ion contents-associated to salt stress were evaluated in response to high levels of NaCl. The study was completed with an untargeted metabolomics analysis of the primary compounds including carbohydrates, phosphoric and organic acids, and amino acids, identified by using a gas chromatography and mass spectrometry platform. Limonium albuferae proved to be more salt-tolerant than L. doufourii, both at the germination stage and during vegetative growth. The degradation of photosynthetic pigments and the increase of Na+/K+ ratio under salt stress were more accentuated in the less tolerant second species. The metabolomics analysis unravelled several differences between the two species. The higher salt tolerance of L. albuferae may rely on its specific accumulation of fructose and glucose under high salinity conditions, the first considered as a major osmolyte in this genus. In addition, L. albuferae showed steady levels of citric and malic acids, whereas the glutamate family pathway was strongly activated under stress in both species, leading to the accumulation of proline (Pro) and γ-aminobutyric acid (GABA).

Qualitative and Quantitative Differences in Osmolytes Accumulation and Antioxidant Activities in Response to Water Deficit in Four Mediterranean Limonium Species.

Limonium is a genus represented in the Iberian Peninsula by numerous halophytic species that are affected in nature by salinity, and often by prolonged drought episodes. Responses to water deficit have been studied in four Mediterranean Limonium species, previously investigated regarding salt tolerance mechanisms. The levels of biochemical markers, associated with specific responses-photosynthetic pigments, mono- and divalent ions, osmolytes, antioxidant compounds and enzymes-were determined in the control and water-stressed plants, and correlated with their relative degree of stress-induced growth inhibition. All the tested Limonium taxa are relatively resistant to drought on the basis of both the constitutive presence of high leaf ion levels that contribute to osmotic adjustment, and the stress-induced accumulation of osmolytes and increased activity of antioxidant enzymes, albeit with different qualitative and quantitative induction patterns. Limonium santapolense activated the strongest responses and clearly differed from Limonium virgatum, Limonium girardianum, and Limonium narbonense, as indicated by cluster and principal component analysis (PCA) analyses in agreement with its drier natural habitat, and compared to that of the other plants. Somewhat surprisingly, however, L. santapolense was the species most affected by water deficit in growth inhibition terms, which suggests the existence of additional mechanisms of defense operating in the field that cannot be mimicked in greenhouses.