Physiological and biochemical mechanisms of the ornamental Eugenia myrtifolia L. plants for coping with NaCl stress and recovery.

MAIN CONCLUSION: We studied the response of Eugenia myrtifolia L. plants, an ornamental shrub native to tropical and subtropical areas, to salt stress in order to facilitate the use of these plants in Mediterranean areas for landscaping. E. myrtifolia plants implement a series of adaptations to acclimate to salinity, including morphological, physiological and biochemical changes. Furthermore, the post-recovery period seems to be detected by Eugenia plants as a new stress situation. Different physiological and biochemical changes in Eugenia myrtifolia L. plants after being subjected to NaCl stress for up to 30 days (Phase I) and after recovery from salinity (Phase II) were studied. Eugenia plants proved to be tolerant to NaCl concentrations between 44 and 88 mM, displaying a series of adaptative mechanisms to cope with salt-stress, including the accumulation of toxic ions in roots. Plants increased their root/shoot ratio and decreased their leaf area, leaf water potential and stomatal conductance in order to limit water loss. In addition, they displayed different strategies to protect the photosynthetic machinery, including the limited accumulation of toxic ions in leaves, increase in chlorophyll content, changes in chlorophyll fluorescence parameters, leaf anatomy and antioxidant defence mechanisms. Anatomical modifications in leaves, including an increase in palisade parenchyma and intercellular spaces and decrease in spongy parenchyma, served to facilitate CO2 diffusion in a situation of reduced stomatal aperture. Salinity produced oxidative stress in Eugenia plants as evidenced by oxidative stress parameters values and a reduction in APX and ASC levels. Nevertheless, SOD and GSH contents increased. The post-recovery period is detected as a new stress situation, as observed through effects on plant growth and alterations in chlorophyll fluorescence and oxidative stress parameters.

Protective effects of Glomus iranicum var. tenuihypharum on soil and Viburnum tinus plants irrigated with treated wastewater under field conditions.

Currently, irrigation using recycled water is increasing, especially in semiarid environments, but a potential problem of using reclaimed wastewater is its elevated salt levels. The application of arbuscular mycorrhizal fungi (AMF) could be a suitable option to mitigate the negative effects produced by the salinity. In this work, the combined effect of Glomus iranicum var. tenuihypharum and two types of water: Control, C, with EC <0.9 dS m(-1) and reclaimed water (wastewater previously treated in a sewage treatment plant) with EC 4 dS m(-1) during a first saline period (11 weeks) and with EC 6 dS m(-1) during a second saline period (25 weeks), was evaluated for laurustinus (Viburnum tinus) plants under field conditions. This plant is a popular shrub very used for gardening. Chemical properties of soil as well as physiological behavior, leaf nutrition, and esthetic value of plants were evaluated. Due to the high salinity from wastewater at 6 dS m(-1), laurustinus plants decreased their stem water potential values and, to a lesser extent, the stomatal conductance. Also, the visual quality of the plants was diminished. The inoculated AMF satisfactorily colonized the laurustinus roots and enhanced the structure of the soil by increasing the glomalin and carbon contents. Furthermore, G. iranicum var. tenuihypharum inoculation decreased Na and Cl content, stimulated flowering and improved the stem water potential of the plants irrigated with both types of reclaimed water. The AMF also had a positive effect as a consequence of stimulation of plant physiological parameters, such as the stem water potential and stomatal conductance. Effective AMF associations that avoid excessive salinity could provide wastewater reuse options, especially when the plants grow in soils.

Water relations, nutrient content and developmental responses of Euonymus plants irrigated with water of different degrees of salinity and quality.

For 20 weeks, the physiological responses of Euonymus japonica plants to different irrigation sources were studied. Four irrigation treatments were applied at 100 % water holding capacity: control (electrical conductivity (EC) <0.9 dS m(-1)); irrigation water normally used in the area (irrigator's water) IW (EC: 1.7 dS m(-1)); NaCl solution, NaCl (EC: 4 dS m(-1)); and wastewater, WW (EC: 4 dS m(-1)). This was followed by a recovery period of 13 weeks, when all the plants were rewatered with the same amount and quality of irrigation water as the control plants. Despite the differences in the chemical properties of the water used, the plants irrigated with NaCl and WW showed similar alterations in growth and size compared with the control even at the end of the recovery period. Leaf number was affected even when the EC of the irrigation water was of 1.7 dS m(-1) (IW), indicating the salt sensitivity of this parameter. Stomatal conductance (gs) and photosynthesis (Pn), as well as stem water potential (Ψstem), were most affected in plants irrigated with the most saline waters (NaCl and WW). At the end of the experiment the above parameters recovered, while IW plants showed similar values to the control. The higher Na(+) and Cl(+) uptake by NaCl and WW plants led them to show osmotic adjustment throughout the experiment. The highest amount of boron found in WW plants did not affect root growth. Wastewater can be used as a water management strategy for ornamental plant production, as long as the water quality is not too saline, since the negative effect of salt on the aesthetic value of plants need to be taken into consideration.

Morphological performance and seasonal pattern of water relations and gas exchange in Pistacia lentiscus plants subjected to salinity and water deficit.

Soil water deficit and salinity represent a major factor impacting plant survival and agricultural production. The frequency and severity of both abiotic stresses are expected to increase in a context of climate change, especially in arid and semi-arid regions. This work studied the growth pattern, biomass and mineral distribution and the seasonal pattern of water status, photosynthetic rate and stomatal conductance in plant of Pistacia lentiscus grown under different levels of water deficit and salinity. P. lentiscus plants growing under greenhouse conditions were subjected to four irrigation treatments during 11 months: control (C, 1 dS m-1), moderate water deficit (MW, 1dS m-1, 60% of the control), severe water deficit (SW, 1 dS m-1, 40% of the control) and saline (S, 4dS m-1). The results show that Pistacia lentiscus plants were more affected by deficit irrigation than salinity. Deficit irrigation and salinity inhibited plant height, with reductions of 20%, 22% and 35% for S, MW and SW, respectively. Total leaf area was not modified by effect of the treatments, with the result that plant compactness increased in MW. The salt stressed plants only showed lower relative growth rate at the end of the experiment. Plants responded to saline or drought stress by increasing their osmotic adjustment, which was more pronounced under salinity. Saline plants had the highest values in Na+ and Cl- ions and the lowest values for K+/Na+ and Ca2+/Na+ ratios in leaves and stems, which is correlated with a decrease in growth, stomatal conductance, photosynthesis and stem water potential, and can be used as a diagnostic tool to assess plant tolerance to salinity stress. As a measure of plant hydration, relative water content was more sensitive to deficit irrigation than salinity, being a good indicator of water stress. P. lentiscus plants subjected to both deficit irrigation treatments exhibited an increase in their intrinsic water use efficiency, which is an important adaptation for plants growing in environments with water scarcity.

Acute and Rapid Response of Melissa officinalis and Mentha spicata to Saline Reclaimed Water in Terms of Water Relations, Hormones, Amino Acids and Plant Oxylipins.

The use of reclaimed water is considered an efficient tool for agricultural irrigation; however, the high salinity associated to this water could compromise plant quality and yields. Balm and spearmint plants were submitted for 15 days to three irrigation treatments in a controlled chamber: control with EC: 1.2 dS m-1 (control), reclaimed water from secondary effluent (EC: 1.6 dS m-1) (S) and water from secondary effluent with brine (EC: 4.4 dS m-1) (SB). The plant water status, stomatal and hormonal regulation, nutritional response, concentration of amino acids and plant oxidative stress-based markers, as well as growth were evaluated. Both species irrigated with saline reclaimed water reduced leaf water potential and gas exchange in comparison with control plants, following 2 days of exposure to irrigation treatments. Nevertheless, spearmint plants recovered photosynthetic activity from the seventh day onwards, maintaining growth. This was attributed to hormonal changes and a greater accumulation of some amino acids and some plant oxylipins (phytoprostanes) in comparison to balm plants, which contributed to the improvement in the organoleptic and health-promoting properties of spearmint. A longer irrigation period with saline reclaimed water would be necessary to assess whether the quality of both species, especially spearmint, could further improve without compromising their growth.

Effects of nursery preconditioning through mycorrhizal inoculation and drought in Arbutus unedo L. plants.

The influence of a water deficit treatment and mycorrhizal inoculation with Pisolithus tinctorius (Pers.) Coker and Couch on the water relations, gas exchange, and plant growth in Arbutus unedo L. plants was studied in order to evaluate the hardening process during the nursery period. The ability to withstand the adverse conditions after transplantation was also studied. Mycorrhizal and non-mycorrhizal seedlings of A. unedo were pot-grown for 4 months in a greenhouse (nursery period), during which time two irrigation treatments, well watered (100% water holding capacity, leaching 20% of the applied water) and deficit irrigation (50% of the well watered), were applied. Subsequently, the plants were transplanted to the field and well irrigated (transplanting period), after which and until the end of the experiment they received no water (establishment period). At the end of the nursery period, both water deficit and mycorrhizae were seen to have altered the plant morphology. Mycorrhizal plants had lower leaf area and improved leaf color parameters, while the water deficit increased root dry weight and the root/shoot ratio. Mycorrhizal plants had higher leaf water potential values than non-inoculated plants. Mycorrhizae increased stomatal conductance and photosynthesis values, especially in stressed plants. Drought led to an osmotic adjustment and a decrease in the leaf water potential values at turgor loss point in the mycorrhizal plants. Cell wall rigidity, measured as increased bulk modulus of elasticity, was decreased by the mycorrhizae effect. After transplanting, no differences were found in the water relations or gas exchange values between treatments. During the establishment period, the plants that had been exposed to both drought and mycorrhizae showed a better water status (higher leaf water and turgor potential values) and higher gas exchange values. In conclusion, water deficit and mycorrhizal inoculation of A. unedo plants in nursery produced changes in tissue water relations, gas exchange, and growth, related with the acclimation process in the seedlings, which could provide better resistance to drought and stress conditions following planting.

Effect of Pisolithus tinctorious on Physiological and Hormonal Traits in Cistus Plants to Water Deficit: Relationships among Water Status, Photosynthetic Activity and Plant Quality.

Cistus species can form ectomycorrhizae and arbuscular mycorrhizal fungus that can bring benefits when plants are under water stress conditions. However, the application of some ectomycorrhizae on the water uptake under drought through physiological traits and hormonal regulation is less known. The experiment was performed during three months in a growth chamber with Cistus albidus plants in which the combined effect of the ectomycorrhiza Pisolithus tinctorious inoculation and two irrigation treatments (control and water-stressed plants) were applied. Irrigation absence caused significant decrease in aerial growth and tended to decrease soil water potential at the root surface, leading to a decrease in leaf water potential. Under these conditions, the abscisic acid and salicylic acid content increased while the precursor of ethylene decreased. Although the mycorrhization percentages were not high, the inoculation of P. tinctorious improved the water status and slightly cushioned the rise in leaf temperature of water-stressed plants. The ectomycorrhiza decreased the scopoletin values in leaves of plants subjected to deficit irrigation, indicating that inoculated plants had been able to synthesize defense mechanisms. Therefore, Pisolithus tinctorious alleviated some of the harmful effects of water scarcity in Cistus plants, being its use a sustainable option in gardening or restoration projects.

Substrate composition affects the development of water stress and subsequent recovery by inducing physiological changes in Cistus albidus plants.

Organic residues (compost) can be used as growth medium but may contain phytotoxic ions that, combined with a water deficit may alter the behavior of plants. The experiment was carried out in a growth chamber with Cistus albidus in a commercial substrate, C (sphagnum peat, coconut fiber and perlite, 8:7:1) and a mixture of compost substrates, Cp (slurry compost, coconut fiber and perlite, 3:6:1). Plants were grown in pots under well-watered, maintaining values of Ψl around -0.9 MPa (WW) and water-stressed (WS) conditions, where the irrigation was removed until reached values of Ψl around -3.0 MPa (water stress period), after then, water was re-established in all plants (recovery period). Although, the well-watered plants had a leaf water potential (Ψl) around -0.9 MPa, stomatal conductance (gs) was 125 mmol m-2s-1 in the commercial substrate and 30 mmol m-2s-1 in compost. The time taken to reach the threshold value at which water stress occurs was 13 days in the commercial substrate and 53 days in compost. Water-stressed plants in the commercial substrate had significantly lower values of Ψl and gs than well-watered. Plants in compost maintained values of gs similar in both irrigation treatments (WW and WS) and accumulated less biomass than those that grown in commercial. The water stress in compost led an increase in the adaxial epidermis, parenchyma and mesophyll, whereas water stress in commercial the proportions of the different tissues decreased. Higher lipid peroxidation values were found in plants grown in both substrates under water stress. The recovery time of the plants, until manage Ψl values around -0.9 MPa, depended on the type of substrate. The restoration of irrigation in commercial substrate act as a new stress, as reflected in the photochemical mechanisms.

The long-term resistance mechanisms, critical irrigation threshold and relief capacity shown by Eugenia myrtifolia plants in response to saline reclaimed water.

Salts present in irrigation water are serious problems for commercial horticulture, particularly in semi-arid regions. Reclaimed water (RW) typically contains, among others elements, high levels of salts, boron and heavy metal. Phytotoxic ion accumulation in the substrate has been linked to different electric conductivities of the treatments. Based on these premises, we studied the long-term effect of three reclaimed water treatments with different saline concentrations on Eugenia myrtifolia plants. We also looked at the ability of these plants to recover when no drainage was applied. The RW with the highest electric conductivity (RW3, EC = 6.96 dS m-1) provoked a number of responses to salinity in these plants, including: 1) accumulation and extrusion of phytotoxic ions in roots; 2) a decrease in the shoot/root ratio, leaf area, number of leaves; 3) a decrease in root hydraulic conductivity, leaf water potential, the relative water content of leaves, leaf stomatal conductance, the leaf photosynthetic rate, water-use efficiency and accumulated evapotranspiration in order to limit water loss; and 4) changes in the antioxidant defence mechanisms. These different responses induced oxidative stress, which can explain the damage caused in the membranes, leading to the death of RW3 plants during the relief period. The behaviour observed in RW2 plants was slightly better compared with RW3 plants, although at the end of the experiment about 55% of the RW2 plants also died, however RW containing low salinity level (RW1, EC = 2.97 dS m-1) can be effective for plant irrigation.

Changes in growth, physiological parameters and the hormonal status of Myrtus communis L. plants irrigated with water with different chemical compositions.

Myrtus communis, an important Mediterranean ornamental shrub, was used to study the effect of irrigation water with different chemical compositions in the plant response. A treatment with NaCl was used to establish the plant resistance to high salinity at long term. Plants were subjected to four irrigation treatments with drainage for three months: Control (0.8 dS m(-1)); two treatments using reclaimed water (RWs): RW1 (2.0 dS m(-1)) and RW2 (5.0 dS m(-1)); and NaCl (10.0 dS m(-1)). High levels of electric conductivity of RWs not affected plant growth, while NaCl decreased leaf dry weight. Coinciding with the accumulation of Na(+) and Cl(-) in the roots, soil water potential decreased, which hinders the mobilization of water to the leaves, decreasing leaf water potential. The osmotic adjustment in the NaCl treatment was due to Na(+) and Cl(-) ions, although the proline could contribute as an Osmo compatible solute, increasing the turgor plants. Also changes in cell walls rigidity minimize the negative effects on the water balance; however, a higher lipid peroxidation was observed in these plants. Stomatal closure was associated with a decrease in K(+) and an increase in abscisic acid. NaCl produced an increase in salicylic acid and did not affect jasmonic acid contents at the end of the experiment. Similar behavior in soil and leaf water potentials, although less pronounced than in NaCl, was shown in RW2 plants. The abscisic acid increased in the RW2 with respect to the control and a decrease in stomatal conductance was observed at the end of the experiment. Plants irrigated with RW1 behaved similarly to the control.

Influence of arbuscular mycorrhizal fungi and treated wastewater on water relations and leaf structure alterations of Viburnum tinus L. plants during both saline and recovery periods.

Nowadays, irrigation with low quality water is becoming an alternative to satisfy the needs of crops. However, some plant species have to deal with high salinity of reclaimed water, by adapting their physiological behaviour during both saline and recovery periods and developing morphological changes in their leaves. The application of arbuscular mycorrhizal fungi (AMF) could also be a suitable option to mitigate the negative effects of this kind of water, although the effectiveness of plant-AMF association is influenced by many factors. In this work, during forty weeks, the combined effect of Glomus iranicum var. tenuihypharum and two types of water: control, C, EC<0.9 dS m(-1) and reclaimed water, RW (with EC: 4 dS m(-1) during a first saline period and EC: 6 dS m(-1) during a second saline period) was evaluated for laurustinus plants (Viburnum tinus L.) transplanted in soil. This was followed by a recovery period of eight weeks, when all the plants were irrigated in the control irrigation conditions. Seasonal and daily changes in stem water potential (Ψstem), stomatal conductance (gs), photosynthesis (Pn) and leaf internal CO2 concentration (Ci) of laurustinus plants were evaluated. Leaf structure alterations, nutrient imbalance, height and leaf hydraulic conductivity (Kleaf) were also determined. Due to the high difficulty of absorbing water from the soil, RW plants showed a high volumetric water content (θv) in soil. The stem water potential and the stomatal conductance (gs) values were reduced in RW plants throughout the second saline period. These decreases were also found during the day. Leaf Ca(2+)/Na(+) and K(+)/Na(+) ratios diminished in RW plants respect to the C plants due to the Na(+) accumulation, although height and chlorophyll content values did not show statistical differences. Leaves from RW plants showed a significantly thicker mesophyll than Control leaves as a consequence of high EC. The area of palisade parenchyma (PP) increased while the area of spongy parenchyma (SP) decreased in RW leaves with respect to the C leaves. These structural changes could be considered as a strategy to maximize photosynthesis potential in saline conditions. Mycorrhizal inoculation improved the water status of both C and RW plants by increasing their Ψstem and gs values. As regards leaf structure, AMF showed an opposite effect to salinity for PP and SP. At the end of the recovery period, hardly any statistical differences of physiological parameters were found between treatments, although a tendency to improve them was observed in inoculated plants. In any case, the leaf structural changes and the great reduction in Kleaf observed at Ψleaf below -1.5 MPa would constitute an important mechanism for laurustinus plants to reduce the water loses produced by salinity.

NaCl-induced physiological and biochemical adaptative mechanisms in the ornamental Myrtus communis L. plants.

Physiological and biochemical changes in Myrtus communis L. plants after being subjected to different solutions of NaCl (44, and 88 mM) for up to 30 days (Phase I) and after recovery from the salinity period (Phase II) were studied. Myrtle plants showed salinity tolerance by displaying a series of adaptative mechanisms to cope with salt-stress, including controlled ion homeostasis, the increase in root/shoot ratio, the reduction of water potentials and stomatal conductance to limit water loss. In addition, they displayed different strategies to protect the photosynthetic machinery, including limiting toxic ion accumulation in leaves, increase in chlorophyll content, and changes in chlorophyll fluorescence parameters, leaf anatomy and increases in catalase activity. Anatomical modifications in leaves, including a decrease in spongy parenchyma and increased intercellular spaces, allow CO2 diffusion in a situation of reduced stomatal aperture. In spite of all these changes, salinity produced oxidative stress in myrtle plants as monitored by increases in oxidative stress parameter values. The post-recovery period is perceived as a new stress situation, as observed through effects on plant growth and alterations in non-photochemical quenching parameters and lipid peroxidation values.

Effects of irrigation and air humidity preconditioning on water relations, growth and survival of Rosmarinus officinalis plants during and after transplanting.

The effect of different irrigation and air humidity conditioning treatments on the morphological and physiological responses of Rosmarinus officinalis in nursery conditions was investigated in order to evaluate the degree of hardening resulting from these conditions. Rosmarinus officinalis seedlings were pot-grown during 4 months in two greenhouses (nursery period), in which two irrigation treatments were used (control and deficit). In one of these greenhouses, air humidity was controlled using a dehumidifying system (low humidity), in the other greenhouse the air conditions were not artificially modified (control humidity). After the nursery period, the plants of all treatments were transplanted and well watered (100% water holding capacity for 1 month, transplanting period). After this period, they received no water (establishment period). At the end of the nursery period it was seen that deficit irrigation had altered the morphology of the R. officinalis plants by reducing plant height, stem diameter, leaf area, total dry weight, and root length, while humidity influenced the parameters related with plant water relations. Low air humidity and deficit irrigation-induced tissue dehydration and lower stomatal conductance values (gs). The plants subjected to deficit irrigation developed leaf osmotic adjustment, which was maintained during the transplanting period. At that time, the plants that had been exposed to deficit irrigation and low humidity showed efficient stomatal regulation (lower gs values). After transplanting and during the establishment period, these plants showed a better water status (higher psil and gs values). Their post-planting survival rate improved as a result of acclimation processes.