Responses to Water Deficit and Salt Stress in Silver Fir (Abies alba Mill.) Seedlings

Forest ecosystems are frequently exposed to abiotic stress, which adversely affects their growth, resistance and survival. For silver fir (Abies alba), the physiological and biochemical responses to water and salt stress have not been extensively studied. Responses of one-year-old seedlings to a 30-day water stress (withholding irrigation) or salt stress (100, 200 and 300 mM NaCl) treatments were analysed by determining stress-induced changes in growth parameters and different biochemical markers: accumulation of ions, different osmolytes and malondialdehyde (MDA, an oxidative stress biomarker), in the seedlings, and activation of enzymatic and non-enzymatic antioxidant systems. Both salt and water stress caused growth inhibition. The results obtained indicated that the most relevant responses to drought are based on the accumulation of soluble carbohydrates as osmolytes/osmoprotectants. Responses to high salinity, on the other hand, include the active transport of Na+, Cl− and Ca2+ to the needles, the maintenance of relatively high K+/Na+ ratios and the accumulation of proline and soluble sugars for osmotic balance. Interestingly, relatively high Na+ concentrations were measured in the needles of A. alba seedlings at low external salinity, suggesting that Na+ can contribute to osmotic adjustment as a ‘cheap’ osmoticum, and its accumulation may represent a constitutive mechanism of defence against stress. These responses appear to be efficient enough to avoid the generation of high levels of oxidative stress, in agreement with the small increase in MDA contents and the relatively weak activation of the tested antioxidant systems.

The mayfly nymph Austrophlebioides pusillus Harker defies common osmoregulatory assumptions

Osmoregulation is a key physiological function, critical for homeostasis. The basic physiological mechanisms of osmoregulation are thought to be well established. However, through a series of experiments exposing the freshwater mayfly nymph Austrophlebioides pusillus (Ephemeroptera) to increasing salinities, we present research that challenges the extent of current understanding of the relationship between osmoregulation and mortality. A. pusillus had modelled 96 h LC 10 , LC 50 and LC 99 of 2.4, 4.8 and 10 g l −1 added synthetic marine salt (SMS), respectively. They were strong osmoregulators. At aquarium water osmolality of 256 ± 3.12 mmol kg −1 (±s.e.; equivalent to 10 g l −1 added SMS), the haemolymph osmolality of A. pusillus was a much higher 401 ± 4.18 mmol kg −1 (±s.e.). The osmoregulatory capacity of A. pusillus did not break down, even at the salinity corresponding to their LC 99 , thus their mortality at this concentration is due to factors other than increased internal osmotic pressure. No freshwater invertebrate has been previously reported as suffering mortality from rises in salinity that are well below the iso-osmotic point. Recently, studies have reported reduced abundance/richness of Ephemeroptera with slightly elevated salinity. Given that salinization is an increasing global threat to freshwaters, there is an urgent need for studies into the osmophysiology of the Ephemeroptera to determine if their loss at locations with slightly elevated salinity is a direct result of external salinity or other, possibly physiological, causes.

Growth, sodium uptake and antioxidant responses of coastal plants differing in their ecological status under increasing salinity

In the present study, we compared the response to salinity of three plants from Brittany coast with contrasted ecological status: Limonium latifolium (salt marshes), Matricaria maritima (beach tops and sand dunes) and Crambe maritima (fixed dunes). Under controlled glasshouse conditions, the growth of the three plants decreased with increasing external salinity. L. latifolium and C. maritima exhibited the highest and lowest resistance to severe salt stress (400 mM), respectively. M. maritima could be considered as an intermediate species, since it tolerated salinity up to 200 mM. The same observation could be made with sodium absorption and acuumulation in plant tissues, the most tolerant species (L. latifolium being the least Na accumulator. Hydrogen peroxide (H2O2) and malondialdehyde (MDA), commonly produced in conditions of stress, accumulated significantly in salt treated C. maritima and M. maritima while not in the tolerant L. latifolium. The latter used glutathione reductase to maintain constant H2O2 levels under salt stress while peroxidases were very low and ascorbate peroxidase did not respond to salinity stimulation. The medium tolerant halophyte M. maritima used peroxidases to protect from NaCl-induced H2O2, while the sensitive C. maritima failed to detoxify H2O2 despite a sharp increase in catalase activity. Results showed that the three coastal species differ in resistance to salinity. They also suggested that the level of plant resistance to salinity could be attributed to differing mechanisms to manage the accumulation of sodium and cope with the oxidative damages.

Tolerance of extreme salinity in two stem-succulent halophytes (Tecticornia species)

Communities of Tecticornia on the margins of ephemeral salt lakes in Australia often exhibit species zonation, such as at Hannan Lake (Western Australia) where Tecticornia indica subsp. bidens (Nees) K.A.Sheph. and Paul G.Wilson occupies the less saline dune habitat on lake margins and Tecticornia pergranulata (J.M.Black) K.A.Sheph. and Paul G.Wilson subsp. pergranulata occupies both the dunes and the more saline and moist lake playa. Here we tested the hypothesis that these two species differ in tolerance to extreme salinity. Plants were grown in drained sand cultures with treatments of 10–2000 mM NaCl for 85 days. Both species were highly salt tolerant, maintaining growth at treatments of up to 2000 mM NaCl, although the death of two replicates of T. indica at 2000 mM NaCl suggests this salinity is close to the species tolerance limit. Both Tecticornia species maintained a favourable gradient in tissue water potential via osmotic adjustment as external salinity increased, also with reduced tissue water content at very high external salinity. Regulated accumulation of Na+ and Cl–, maintenance of net K+ to Na+ selectivity, high tissue concentrations of glycinebetaine and presumed cellular solute compartmentation, would have contributed to salt tolerance. The growth rate of T. pergranulata was 11–29% higher than T. indica suggesting, in addition to these moderate differences in salinity tolerance, other factors are likely to contribute to species zonation at salt lakes. The higher water use efficiency of the C4 T. indica compared with the C3 T. pergranulata may provide an advantage in the drier dune habitat on salt lake margins. An additional experiment confirmed the hypothesis that survival of T. pergranulata seedlings is enhanced by the duration of reduced salinity after germination, as would occur following significant rainfall, as older seedlings maintained higher growth rates during subsequent increases in salinity.

Synthesis, Release, and Recapture of Compatible Solute Proline by Osmotically Stressed Bacillus subtilis Cells

ABSTRACTBacillus subtilissynthesizes large amounts of the compatible solute proline as a cellular defense against high osmolarity to ensure a physiologically appropriate level of hydration of the cytoplasm and turgor. It also imports proline for this purpose via the osmotically inducible OpuE transport system. Unexpectedly, anopuEmutant was at a strong growth disadvantage in high-salinity minimal media lacking proline. Appreciable amounts of proline were detected in the culture supernatant of theopuEmutant strain, and they rose concomitantly with increases in the external salinity. We found that the intracellular proline pool of severely salinity-stressed cells of theopuEmutant was considerably lower than that of itsopuE+parent strain. This loss of proline into the medium and the resulting decrease in the intracellular proline content provide a rational explanation for the observed salt-sensitive growth phenotype of cells lacking OpuE. None of the known MscL- and MscS-type mechanosensitive channels ofB. subtilisparticipated in the release of proline under permanently imposed high-salinity growth conditions. The data reported here show that the OpuE transporter not only possesses the previously reported role for the scavenging of exogenously provided proline as an osmoprotectant but also functions as a physiologically highly important recapturing device for proline that is synthesizedde novoand subsequently released by salt-stressedB. subtiliscells. The wider implications of our findings for the retention of compatible solutes by osmotically challenged microorganisms and the roles of uptake systems for compatible solutes are considered.

Functional plasticity of mitochondrion-rich cells in the skin of euryhaline medaka larvae (Oryzias latipes) subjected to salinity changes

A noninvasive technique, the scanning ion-selective electrode technique (SIET) was applied to measure Na+ and Cl− transport by the yolk-sac skin and individual mitochondrion-rich cells (MRCs) in intact medaka larvae ( Oryzias latipes ). In seawater (SW)-acclimated larvae, significant outward Na+ and Cl− gradients were measured at the yolk-sac surface, indicating secretions of Na+ and Cl− from the yolk-sac skin. With Na+ pump immunostaining and microscopic observation, two groups of MRCs were identified on the yolk-sac skin of SW-larvae. These were single MRCs (s-MRCs), which do not have an accompanying accessory cell (AC), and multicellular complex MRCs (mc-MRCs), which usually consist of an MRC and an accompanying AC. The percentage of mc-MRC was ∼60% in 30 parts per thousand of SW, and it decreased with the decrease of external salinity. By serial SIET probing over the surface of the MRCs and adjacent keratinocytes (KCs), significant outward fluxes of Na+ and Cl− were detected at the apical opening (membrane) of mc-MRCs, whereas only outward Cl− flux, but not Na+ flux, was detected at s-MRCs. Treatment with 100 μM ouabain or bumetanide effectively blocked the Na+ and Cl− secretion. Following freshwater (FW) to SW transfer, Na+ and Cl− secretions by the yolk-sac skin were fully developed in 5 h and 2 h, respectively. In contrast, both Na+ and Cl− secretions downregulated rapidly after SW to FW transfer. Sequential probing at individual MRCs found that Na+ and Cl− secretions declined dramatically after SW to FW transfer and Na+/Cl− uptake was detected at the same s-MRCs and mc-MRCs after 5 h. This study provides evidence demonstrating that ACs are required for Na+ excretion and MRCs possess a functional plasticity in changing from a Na+/Cl−-secreting cell to a Na+/Cl−-absorbing cell.

Synthesis of the Compatible Solute Ectoine in Virgibacillus pantothenticus Is Triggered by High Salinity and Low Growth Temperature

ABSTRACT The quantification of the intracellular concentration of ectoine in Virgibacillus pantothenticus revealed that the production of this compatible solute is triggered either by an increase in the external salinity or by a reduction in the growth temperature. This finding reflects increased transcription of the ectoine biosynthetic operon (ectABC) under both environmental conditions.

Productivity and Mineral Nutrition of Limonium Species Irrigated with Saline Wastewaters

To explore the possibility that saline wastewaters may be used to grow commercially acceptable floriculture crops, a study was initiated to determine the effects of salinity on two statice cultivars. Limonium perezii (Stapf) F. T. Hubb. `Blue Seas' and L. sinuatum (L.) Mill `American Beauty' were grown in greenhouse sand cultures irrigated with waters prepared to simulate saline drainage waters typically present in the western San Joaquin Valley (SJV) of California. Seven salinity treatments were imposed on 3-week-old seedlings. Electrical conductivities of the irrigation waters (EC) were 2.5 (control), 7, 11, 15, 20, 25, and 30 dS·m–1. Vegetative shoots were sampled for biomass production and ion analysis ten weeks after application of stress. Flower stem numbers, length, and weight were determined at harvest. Stem length of L. perezii was significantly reduced when irrigation water salinity exceeded a threshold of 2.5 dS·m–1. Salt tolerance threshold based on stem length for L. sinuatum was 7 dS m-1. The species exhibited significant differences in shoot-ion relations which appear to be related to differences in salt tolerance. Sodium, K+, Mg2+, and total-P were more strongly accumulated in the leaves of L. sinuatum than L. perezii. Both species accumulated K+ in preference to Na+, but selectivity for K+ over Na+ was significantly higher in L. sinuatum than in the more salt-sensitive L. perezii. Chloride concentration in L. sinuatum leaves increased significantly as salinity increased, whereas the 20-fold increase in substrate-Cl had no effect on leaf-Cl in L. perezii. Both Limonium species completed their life cycles at salt concentrations exceeding 30 dS·m–1, a character associated with halophytic plants. Maximum growth of each species, however, occurred under relatively low salt stress, and steadily declined as external salinity increased. Based on this crop productivity response, L. perezii should be rated as sensitive and L sinuatum as moderately tolerant.