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.

Responses to Salt Stress of the Interspecific Hybrid Solanum insanum × Solanum melongena and Its Parental Species.

Soil salinity is becoming one of the most critical problems for agriculture in the current climate change scenario. Growth parameters, such as plant height, root length and fresh weight, and several biochemical stress markers (chlorophylls, total flavonoids and proline), have been determined in young plants of Solanum melongena, its wild relative Solanum insanum, and their interspecific hybrid, grown in the presence of 200 and 400 mM of NaCl, and in adult plants in the long-term presence of 80 mM of NaCl, in order to assess their responses to salt stress. Cultivated eggplant showed a relatively high salt tolerance, compared to most common crops, primarily based on the control of ion transport and osmolyte biosynthesis. S. insanum exhibited some specific responses, such as the salt-induced increase in leaf K+ contents (653.8 µmol g-1 dry weight) compared to S. melongena (403 µmol g-1 dry weight) at 400 mM of NaCl. Although there were no substantial differences in growth in the presence of salt, biochemical evidence of a better response to salt stress of the wild relative was detected, such as a higher proline content. The hybrid showed higher tolerance than either of the parents with better growth parameters, such as plant height increment (7.3 cm) and fresh weight (240.4% root fresh weight and 113.3% shoot fresh weight) at intermediate levels of salt stress. For most biochemical variables, the hybrid showed an intermediate behaviour between the two parent species, but for proline it was closer to S. insanum (ca. 2200 µmol g-1 dry weight at 200 mM NaCl). These results show the possibility of developing new salt tolerance varieties in eggplant by introducing genes from S. insanum.

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.

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.

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 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.

Comparative analysis of the responses to water stress in eggplant (Solanum melongena) cultivars.

Little information is available on the physiological and biochemical responses to water stress in eggplant (Solanum melongena). We evaluated four genetically diverse eggplant varieties (MEL3-MEL6) under control and water stress conditions. Measurements were taken for plant growth, tissue water content, levels of chlorophylls a and b, carotenoids, proline, malondialdehyde, total phenolics, total flavonoids, superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) activities. For most traits, the water stress treatment had a greater contribution than the variety effect to the total sums of squares in an ANOVA analysis, except for total flavonoids, SOD, APX, and GR. The water stress treatment had a strong effect on plant growth and tissue water content. In general, water stress reduced the three photosynthetic pigments, increased proline, malondialdehyde, total phenolics, and total flavonoids, although some varietal differences were observed. Different patterns were also detected in the activities of the four enzymes evaluated, but few differences were observed for individual varieties between the control and water stress treatments. Many significant phenotypic correlations were observed among the traits studied, but only eight environmental correlations were detected. A PCA analysis distinctly separated individuals according to the treatment, and revealed a clearer separation of varieties under water stress than under control conditions, pointing to varietal differences in the responses to stress. Our results suggest that proline could be used as a marker for drought stress tolerance in this species. The information obtained provides new insight on the physiological and biochemical responses of eggplant to drought stress.