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Salinity tolerance of japonica and indica rice (Oryza sativa L.) at the seedling stage.

In order to identify the degree of salinity tolerance of the indica and japonica rice groups, 10 varieties were tested under saline and non-saline conditions. Twelve-day-old seedlings were grown in normal culture solution, then initially salinized at an electrical conductivity (EC) of 6 dS/m for 4 days, and finally salinized at an EC of 12 dS/m for the next 14 days. The growth parameters, and Na and K absorption in the shoot were measured to characterize the tolerance level of the two rice groups. Reduction in all growth parameters of tolerant varieties was significantly lower in indica varieties than in japonica varieties. Tolerant indica varieties were good Na excluders, absorbed high amounts of K, and maintained a low Na/K ratio in the shoot. Tolerant japonica varieties also absorbed less Na but were not as good excluders as indica varieties. Shoot K concentration alone did not show any relationship to salinity tolerance. These results indicate that, for all parameters measured, the tolerance level of indica was higher than that of japonica.

Adaptation, Physiological↗

Metamorphosis in the summer flounder, Paralichthys dentatus: thyroidal status influences salinity tolerance.

Metamorphosis in the summer flounder (Paralichthys dentatus) is controlled by thyroid hormones (TH) and takes place as the larvae move from a salinity of about 35 parts per thousand (ppt) in the ocean to salinity ranging from 0-35 ppt in estuaries. Historically, the role of TH in juvenile and adult teleost osmoregulation has been ambiguous, and it is not known if TH influences larval teleost osmoregulatory development. This study addresses the influence of thyroxine (T4) on the development of tolerance to low (5 ppt) and high salinity (45 and 50 ppt) as determined by salinity tolerance tests. In untreated larvae, tolerance to both low and high salinity was high during early premetamorphosis (early pre-M) and decreased or was very low from late prometamorphosis (late pro-M) through mid-metamorphic climax (mid-MC). Salinity tolerance increased 2-3-fold during late MC when whole-animal T4 levels are highest, and reached maximum tolerance at the juvenile stage. The early induction of metamorphosis by exposing larvae in pre-M to exogenous T4 reduced tolerance to low salinity during early and mid-MC, though tolerance of fish that had developed into juveniles was not impaired. In contrast, T4 increased high salinity tolerance during early and mid-MC, and the juvenile stage. This T4-induced heterochrony in salinity tolerance with regards to developmental stage suggests that the effects of T4 on salinity tolerance may be uncoupled from accelerated metamorphosis. Treatment of larvae with thiourea (TU, an inhibitor of T4 synthesis) inhibited metamorphosis and reduced tolerance to high salinity, but did not affect tolerance to low salinity. Reduced tolerance to high salinity by TU was only partially counteracted by T4 treatment, suggesting that TU also affects hypoosmoregulatory activity by an extrathyroidal mechanism. Our findings suggest that in the summer flounder T4 plays a more important role in the development of hypoosmoregulatory ability than hyperosmoregulatory ability. J. Exp. Zool. 284:414-424, 1999.

Adaptation, Physiological↗

Proteomics reveals elevated levels of PR 10 proteins in saline-tolerant peanut (Arachis hypogaea) calli.

The proteome of a salinity-tolerant Arachis hypogaea L. callus cell line was compared with its sensitive counterpart. Several low molecular weight proteins were detected by two-dimensional electrophoresis as being unique or significantly elevated in the tolerant line. The identities of several of these proteins were established as PR 10 proteins using tandem Mass Spectrometry and are shown to be phosphorylated on the basis of staining with the phosphorylation-specific stain, Pro-Q Diamond. Our results suggest that these differentially phosphorylated PR 10 proteins may play an important role in mediating salinity stress responses.

Amino Acid Sequence↗

Effects of sublethal sodium dichromate exposure in freshwater on the salinity tolerance and serum osmolality of juvenile coho salmon, Oncorhynchus kisutch, in seawater.

To stimulate a potential field situation in which seaward-migrating juvenile coho salmon, Oncorhynchus kisutch, pass through Cr-polluted freshwater (FW) before being exposed to seawater (SW), the effects of sublethal FW exposure to Cr (as sodium dichromate) on salinity tolerance and serum osmolality after transfer to SW were investigated. SW survival was significantly decreased in salmon exposed in FW to 0.23 mg Cr/L for four weeks or to 0.5 mg Cr/L for two weeks and transferred to 20 or 30 parts per thousand (%) salinity, respectively. Serum osmolality, measured after two weeks of FW exposure to 0.0 or 0.5 mg Cr/L and after transfer to 20 or 30% SW, was significantly higher in Cr-exposed fish from one-half to two days after SW transfer but not at the end of the FW exposure period or at seven days after SW transfer. These effects in SW occurred at Cr exposure concentrations which had no effect on survival or serum osmolality in FW. It is suggested that Cr acted non-specifically on osmoregulatory epithelia to cause excessive dehydration in SW, and that measurement of salinity tolerance and serum osmolality may be sensitive and ecologically realistic indicators of sublethal toxicity in aquatic animals.

Animals↗

Effect of chronic Taura syndrome virus infection on salinity tolerance of Litopenaeus vannamei.

Taura syndrome virus (TSV) is one of the most important shrimp viruses affecting farmed shrimp worldwide. After an acute phase during which the likelihood of mortality is elevated, infected shrimp enter a chronic phase during which shrimp appear to resume normal behavior and display no gross signs of infection. This study was designed to determine if chronically TSV-infected shrimp Litopenaeus vannamei are compromised by the infection. Specifically we investigated whether chronically infected shrimp could tolerate a drop in salinity as strongly as uninfected shrimp. The study consisted of 3 trials that compared survival of uninfected and chronically TSV-infected L. vannamei after drops in salinity from 24 ppt to salinities varying from 18 to 0 ppt. Logistic regression detected a significant effect of TSV infection on survival of chronically infected shrimp (p < 0.05). Salinity drops from 24 ppt to 3 and 6 ppt resulted in statistically different survivals (p < 0.05). Survival rates were similar among groups for salinity drops to greater than 6 ppt or less than 3 ppt. Salinities at which 50% of the shrimp died (LC50) were 3.06 ppt for the uninfected and 6.65 ppt for the chronically infected groups. Moreover, histopathological analysis of chronically infected shrimp that were moribund or recently dead showed no signs of having reverted to the acute stage of the disease. These results suggest that chronically infected shrimp are not able to tolerate a salinity drop as strongly as uninfected shrimp.

Adaptation, Physiological↗

Amino Acid Substitutions in the Na+/K+-ATPase May Contribute to Salinity Tolerance in Insects.

Environmental salinity levels vary naturally across terrestrial ecosystems but can be heightened locally by coastal proximity and desertification as well as human activities such as road salt application and agriculture. Since salt is essential for many physiological processes in insects, rising environmental sodium concentrations may drive behavioral changes, where insects select environments and food sources with suitable sodium levels, or evolutionary changes in constitutive or plastic physiological mechanisms to process salt, potentially altering ecological dynamics and species interactions.Numerous hematophagous (blood feeding) insects such as the yellow-fever mosquito Aedes aeqypti are known to be able to breed in relatively saline environments. Among phytophagous (plant feeding) insects, grasshoppers can be important herbivores in arid and coastal salt-affected regions, whereas the monarch butterfly (Danaus plexippus) appears to perform relatively well on milkweed host plants growing in roadsides influenced by salt runoff. Several of these insects share a common trait: amino acid substitutions in the first extracellular loop of the Na+/K+-ATPase (NKA), a sodium pump crucial for maintaining ion balance. For the monarch these substitutions confer resistance to toxic cardenolides from milkweeds, but it is unclear whether NKA substitutions may influence salt tolerance.Here, we investigate whether the NKA substitutions found in these insects may contribute to salt tolerance using gene-edited Drosophila melanogaster mutant strains as models. We show that flies with substitution Q111L (found in Aedes mosquitoes) or a combination of Q111L and A119S (found in grasshoppers) exhibited greater salt tolerance, whereas flies carrying the combination of substitutions found in the monarch (Q111V, A119S, and N122H) did not.Our results suggest that the monarch may rely on alternate mechanisms for salt tolerance and that its NKA substitutions are important primarily for cardenolide resistance. However, substitution Q111L and the combination of Q111L and A119S may be relevant for salt tolerance in a variety of insects. Uncovering mechanisms of salt tolerance enhances our understanding of species distributions, ecological interactions, and evolutionary physiology in response to changing environmental salinity levels.

Journal Article↗

Salinity tolerance and structure of external and internal gills in tadpoles of the crab-eating frog, Rana cancrivora.

Salinity tolerance and histology of gills were studied in Rana cancrivora larvae. The tadpoles at the external gill stages (W stages 21-22) were able to survive in media containing up to 40% seawater, but died in water of higher salinity. Their external gills appear to have no critical role in adaptation to seawater. However, advanced tadpoles with internal gills (T-K stages I-XVIII) were able to tolerate 50% or higher seawater. In the internal gills, there are numerous mitochondria-rich cells (MR cells) scattered on the ventral and lateral epithelia of the gill arches and the gill tufts in both freshwater- and seawater-acclimated tadpoles. In freshwater-acclimated tadpoles there are three types of MR cell: (1) microplicated, (2) microvillous, and (3) apically vacuolated. In tadpoles acclimated to dilute seawater, the ratio of type-1 to type-2 cells is lower, although all three types of MR cell are present. In 60%-seawater-acclimated tadpoles, a few MR cells with a lumen and concave cytoplasm at the apical membrane (type 4) are present. The changes in MR cell morphology under ambient conditions of low or high salinity may reflect alterations in the physiological roles of the gills with regard to transport of ions.

Acclimatization↗

Genotype-by-environment interaction for salinity tolerance in the freshwater-invading copepod Eurytemora affinis.

This study examined the extent of phenotypic plasticity for salinity tolerance and genetic variation in plasticity in the invasive copepod Eurytemora affinis. Euryemora affinis is a species complex inhabiting brackish to hypersaline environments but has invaded freshwater lakes and reservoirs within the past century. Reaction norm experiments were performed on a relatively euryhaline population collected from a brackish lake with fluctuating salinity. Life history traits (hatching rate, survival, and development time) were measured for 20 full-sib clutches that were split and reared at four salinities (fresh, 5, 10, and 27 practical salinity units [PSU]). On average, higher salinities (10 and 27 PSU) were more favorable for larval growth, yielding greater survival and faster development rate. Clutches differed significantly in their response to salinity, with a significant genotype-by-environment interaction for development time. In addition, genetic (clutch) effects were evident in response to low salinity, given that survival in fresh (lake) water was significantly positively correlated with survival at 5 PSU for individual clutches. Clutches raised in fresh water could not survive beyond metamorphosis, suggesting that acclimation to fresh water could not occur in a single generation. Results suggest the importance of natural selection during freshwater invasion events, given the inability of plasticity to generate a freshwater phenotype, and the presence of genetic variation for plasticity upon which natural selection could act.

Acclimatization↗

In vitro screening of mulberry ( Morus spp.) for salinity tolerance.

An efficient in vitro screening method has been developed for mulberry ( Morus spp. ) to screen salinity-tolerant genotypes from a large population. Axillary buds from field-grown plants were cultured on MS medium containing five different concentrations (0.0%, 0.25%, 0.5%, 0.75% and 1.00%) of sodium chloride (NaCl) in order to study the shoot growth pattern. Rooting was also tested at four different concentrations of NaCl (0.0%, 0.1%, 0.2% and 0.3.%). NaCl has been found to inhibit the growth and development of mulberry shoots and roots in vitro. The survivability of the axillary buds of the genotypes tested was reduced from 83.7% for the controls to 6.1% in 1.0% NaCl. The average number of roots developed by the genotypes ranged from 11.9 (controls) to 0.2 (0.3% NaCl). Out of the 63 genotypes tested, only seven--Rotundiloba, English black, Kolitha-3, Berhampore-A, Kajli, BC(2)59 and C776--developed roots in 0.3% NaCl. Root growth was also reduced drastically from 1.8 cm for the controls to 0.1 cm in 0.3% NaCl. To test the reproducibility of the results in soil, five tolerant and two susceptible genotypes, identified in this in vitro study, were selected and tested under ex vitro conditions. The significant correlation coefficients obtained between the performances of these genotypes under both types of cultural conditions revealed that in vitro screening of mulberry through axillary bud culture is an easy and efficient method to identify salt-adapted genotypes within a limited space and time period.

Adaptation, Physiological↗

The physiology of salinity tolerance in larvae of two species of Culex mosquitoes: the role of compatible solutes.

We investigated the physiological basis for differences in salinity tolerance ranges in mosquito larvae of the genus Culex. We examined the response of larvae of C. quinquefasciatus, a freshwater obligate, and C. tarsalis, a euryhaline osmoconformer, following transfer from fresh water to 34% sea water. Hemolymph Na(+) and Cl(-) levels increased similarly in both species, indicating that ion regulation does not differ under these conditions. C. quinquefasciatus responded to increased environmental salinity with increased hemolymph levels of serine, but suffered a significant reduction in levels of trehalose. C. tarsalis responded to increased environmental salinity with increased hemolymph levels of both proline and trehalose. When C. tarsalis larvae were held in 64% sea water, which C. quinquefasciatus larvae cannot tolerate, hemolymph proline and trehalose were accumulated approximately 50-fold and twofold, respectively, relative to freshwater values. We found that proline serves as both an intra- and extracellular compatible solute in C. tarsalis, the first such circumstance documented in an animal in response to increased environmental salinity. Analyses of the acute responses of the two species to an increase in salinity (from 30% to 50% sea water) indicate that larvae of C. tarsalis are able to volume-regulate via drinking and to attenuate increases in hemolymph NaCl concentration using unknown mechanisms during large, rapid increases in salinity.

Adaptation, Physiological↗

Larval salinity tolerances of the sibling species of Anopheles farauti.

Experiments conducted with laboratory colonies of the sibling species of Anopheles farauti showed larvae of An. farauti No. 1 had a higher salinity tolerance than larvae of An. farauti No. 2 and An. farauti No. 3. The salinity response of field-collected larvae of An. farauti No. 1 from Cowley Beach, Queensland, Australia was similar to that of larvae from two colonies of this species which originated from Papua New Guinea. These results indicate that An. farauti No. 1 is the species which is likely to be found breeding in brackish water whereas the other species may be restricted to freshwater habitats. Laboratory experiments conducted with the colonies and with specimens collected from three localities in northern Queensland indicated that a simple test, based on exposure of first-instar larvae to sea water for 1 hr, should enable identification of An. farauti No. 1 in the field.

Animals↗

Salinity tolerance of stable fly (Diptera: Muscidae).

Effects of salinity on the survival, growth, and development of stable fly, Stomoxys calcitrans (L.), were investigated in the laboratory. Larvae failed to develop to pupation when reared in media containing a salinity of 40 parts per thousand (ppt) sodium chloride (NaCl). Maximum salinity supporting larval development equaled the salinity of seawater (34 ppt); the larval LC90 was 24.2 ppt. Deleterious effects of high salinity decreased as larvae matured. Six-day-old larvae reared at a salinity of 34 ppt weighed 79% less than controls, compared with a 36% difference in 9-d-old larvae; by pupation, the difference was only 24%. Salinity did not influence the duration of larval, pupal, or adult stages. Survival of pupae was unimpaired despite a slight increase in number of pupal deformities, and normal adults emerged. Eggs were highly tolerant to saline. They hatched at salinity concentrations lethal to larvae; greater than 50% hatch occurred even when eggs were maintained at 80 ppt NaCl. Sensitivity of larvae to salinities close to that of seawater might be important for control of stable flies inhabiting marine areas.

Animals↗

Grafting between tobacco plants to enhance salinity tolerance.

We analysed the technique of grafting as a tool to increase salt-stress resistance in tobacco plants. With this aim, we performed two experiments. First, we selected, from among 6 commercial tobacco cultivars (cv. BB-162, cv. H-20, cv. Jarandilla, cv. ZB-3, cv. Havana II and cv. Havana 307) those most tolerant and sensitive to salinity, studying the response of certain nutritional and biochemical indicators of resistance in these plants. In the second experiment, we analysed the response to salinity in grafted tobacco plants using the rootstock of the most tolerant plants, and the scion of the most sensitive ones. In addition, these plants were subjected to salinity to test the viability and efficiency of this grafting technique, assessing the production of foliar biomass and the different quality parameters in this crop. In the first experiment, we found that the most tolerant tobacco cultivars were cv. BB-162 and cv. H-20, which were characterized by reduced uptake and foliar accumulation of Na(+) and Cl(-), together with greater synthesis of sucrose and proline, thereby reducing lipid peroxidation and thus oxidative damage, reflected in higher foliar biomass with respect to the other cultivars studied (primarily cv. Jarandilla, defined as the most salt-sensitive). In the second, we demonstrated that the grafting of salt-sensitive tobacco scions to salt-tolerant rootstocks improves the production and quality of tobacco leaves under conditions of saline stress. Our results show that the rootstocks cv. BB-162 and cv. H-20 best induced salt resistance in tobacco cv. Jarandilla, registering the lowest foliar concentrations of Na(+) and Cl(+), the lowest lipid peroxidation, and the highest proline and sugar concentrations. Overall, this is reflected in better biomass production and quality of the aerial part of the plant.

Adaptation, Physiological↗

Ontogeny of osmoregulation and salinity tolerance in a mangrove crab, Sesarma curacaoense (Decapoda: Grapsidae).

The grapsid crab Sesarma curacaoense is believed to represent the closest saltwater relative to the ancestor which gave rise to an adaptive radiation of endemic freshwater and terrestrial species on the island of Jamaica. Living in mangrove swamps with variable salinity conditions and showing semiterrestrial behaviour, S. curacaoense exhibits ecological adaptations to non-marine conditions. In laboratory experiments, we studied the salinity tolerance during development from hatching to the end of the first juvenile stage. Successful development through metamorphosis occurred in the full salinity range tested (15-32 per thousand), although mortality was significantly enhanced and development delayed at 15 per thousand. In another series of experiments, we studied the ontogeny of the capability for osmoregulation, which is considered as the physiological basis of osmotic stress tolerance. Our results show that S. curacaoense is from hatching a fairly strong hyperosmoregulator in dilute media. This capability increased gradually from hatching throughout the larval and juvenile development. In seawater (32 per thousand) and at an enhanced salt concentration (44 per thousand), the zoeal stages remained hyperosmoconformers. The capability for hypoosmoregulation in concentrated media appeared first in the megalopa stage and increased thereafter. Adult crabs were observed to be strong hyper-hypo-osmoregulators in a salinity range from at least 1 per thousand to 44 per thousand. The unusually early appearance of strong regulatory capabilities, particularly in dilute media, is interpreted as a physiological preadaptation that should have facilitated the evolutionary process of adaptive radiation in non-marine environments on Jamaica.

Journal Article↗

Engineering drought and salinity tolerance in plants: lessons from genome-wide expression profiling in Arabidopsis.

World food security is increasingly dependent on continuous crop improvement and, in particular, the development of crops with increased drought and salinity tolerance. The completed genomic sequence of the model plant Arabidopsis thaliana and the development of whole-genome microarrays, together with increasing repositories of publicly available data and data analysis tools, have opened new avenues to genome-wide systemic analysis of plant stress responses. Here we outline examples of how this full-genome expression profiling can contribute to our understanding of complex stress responses and the identification and evaluation of novel transgenes that could hold the key to the development of commercially viable and sustainable crop plants.

Arabidopsis↗

Screening and characterization of koji molds producing saline-tolerant protease.

Three mold strains isolated from soil in the Taipei area of Taiwan were compared with a commercial strain of Aspergillus oryzae for their proteolytic activities in an 18% NaCl aqueous solution system. Among these strains, the one subsequently identified and designated as Aspergillus sp. FC-10 produced protease with superior saline tolerance. In aflatoxin tests, this strain did not generate detectable aflatoxin after growing on steamed grain polished rice substrate for 24 days. Two types of extracellular proteases were preliminary fractionated by column chromatography on DEAE Sepharose CL-6B. Proteolytic activity of the nonadsorbed protease (P-I) was reduced to 9.4% in the 18% NaCl solution compared to its original activity determined in the buffer solution. However, the adsorbed protease (P-II) was particularly salt tolerant and stable, with 50% proteolytic activity retained throughout the 6-h stability test in 18% NaCl solution.

Aflatoxins↗

Reciprocal expression of gill Na+/K+-ATPase alpha-subunit isoforms alpha1a and alpha1b during seawater acclimation of three salmonid fishes that vary in their salinity tolerance.

The upregulation of gill Na+/K+-ATPase activity is considered critical for the successful acclimation of salmonid fishes to seawater. The present study examines the mRNA expression of two recently discovered alpha-subunit isoforms of Na+/K+-ATPase (alpha1a and alpha1b) in gill during the seawater acclimation of three species of anadromous salmonids, which vary in their salinity tolerance. Levels of these Na+/K+-ATPase isoforms were compared with Na+/K+-ATPase activity and protein abundance and related to the seawater tolerance of each species. Atlantic salmon (Salmo salar) quickly regulated plasma Na+, Cl- and osmolality levels within 10 days of seawater exposure, whereas rainbow trout (Oncorhynchus mykiss) and Arctic char (Salvelinus alpinus) struggled to ionoregulate, and experienced greater perturbations in plasma ion levels for a longer period of time. In all three species, mRNA levels for the alpha1a isoform quickly decreased following seawater exposure whereas alpha1b levels increased significantly. All three species displayed similar increases in gill Na+/K+-ATPase activity during seawater acclimation, with levels rising after 10 and 30 days. Freshwater Atlantic salmon gill Na+/K+-ATPase activity and protein content was threefold higher than those of Arctic char and rainbow trout, which may explain their superior seawater tolerance. The role of the alpha1b isoform may be of particular importance during seawater acclimation of salmonid fishes. The reciprocal expression of Na+/K+-ATPase isoforms alpha1a and alpha1b during seawater acclimation suggests they may have different roles in the gills of freshwater and marine fishes; ion uptake in freshwater fish and ion secretion in marine fishes.

Acclimatization↗