Physiological responses in two populations of Andropogon glomeratus Walter B.S.P. to short-term salinity

Summary Andropogon glomeratus is a C₄ nonhalophytic grass which exhibits population differentiation for tolerance to short-term salinity exposure. To investigate possible physiological mechanisms whch enable salt-tolerant individuals to survive short-term inundation, gas exchange and water relations parameters were measured before and during a 5-day watering treatment of half-strength synthetic seawater in plants from a tolerant and a non-tolerant population. Photosynthetic recovery was followed for 10 days after the salinity treatment. Photosynthetic CO₂ uptake was substantially inhibited in both populations. Stomatal conductances decreased and intercellular CO₂ concentrations increased, indicating non-stomatal factors were primarily responsible for the decrease in CO₂ uptake. After termination of the salinity treatment photosynthetic capacity increased more rapidly in the tolerant population and reached the pretreatment level after 6 days, whereas the nontolerant population did not recover fully after 10 days. A-Ci curves measured before and after the salinity treatment indicated a decrease in the carboxylation efficiency, and suggested a proportionately greater metabolic inhibition relative to the increase in the stomatal limitation. Osmotic adjustment occurred in a 2-day period in the tolerant population, but there was no change in the osmotic potentials or the water potential at the point of turgor loss in the nontolerant population. Thus short-term salt tolerance in the marsh population is associated with rapid osmotic adjustment and recovcry of photosynthetic capacity shortly after the end of the salinity exposure, rather than maintenance of greater photosynthesis during the salinity treatment.     

RAPDs as an aid to evaluate the genetic integrity of somatic embryogenesis-derived populations of Picea mariana (Mill.) B.S.P.

Summary The usefulness of random amplified polymorphic DNA (RAPD) in assessing the genetic stability of somatic embryogenesis-derived populations of black spruce [Picea mariana (Mill.) B.S.P.] was evaluated. Three arbitrary 11-mer primers were successfully used to amplify DNA from both in-vivo and in-vitro material. Twenty-five embryogenic cell lines, additional zygotic embryos and megagametophytes from three controlled crosses involving four selected genotypes of black spruce were used for the segregation analysis of RAPD variants. Ten markers were genetically characterized and used to evaluate the genetic stability of somatic embryos derived from three embryogenic cell lines (one cell line per cross, 30 somatic embryos per cell line). No variation was detected within clones. The utilization of RAPD markers both for the assessment of genetic stability of clonal materials and to certify genetic stability throughout the process of somatic embryogenesis is discussed.     

Effects of salinity and cutting on the development of Phragmites australis

The effects of increased salinity and cutting the above ground biomass on the growth of Phragmites australis were evaluated by investigating four experimental reed stands grown in outdoor tanks. Two stands were treated with 30 salinity and the other two stands with freshwater; one stand of each treatment was cut to 20 cm during the second growing season. Growth conditions were observed until all the plants were dead at the end of the second year. The number of shoots emerged from the freshwater-treated stand was about 70% higher than that of the saltwater-treated stand. The number of shoots emerged from cut plant stands were markedly lower than uncut stands. The average shoot height was negatively affected by salinity and shoots that emerged after cutting further decreased in height. The average number of leaves on a shoot was not significantly affected by salinity, but reduced by cutting in both treatments. Leaf length, width and the distance between leaves were decreased by both salinity and cutting. In the freshwater-treated uncut stand more than 50% of the shoots formed panicles, but this proportion was reduced to 6% by salinity, to 15% by cutting, and to 0% by the combination of salinity and cutting. This study showed again that salinity reduces the growth of aboveground components. The growth, however, was most severely retarded by cutting combined with salinity, which has many implications for better management of P. australis stands.     

Response to short-term inundation with isoosmotic solutions of seawater and sorbitol in a C4 nonhalophyte: evidence for a salt tolerance mechanism

Summary To examine the importance of Na⁺ and Cl^- to osmotic adjustment in a salt-tolerant ecotype of the C₄ nonhalophyte Andropogon glomeratus, plants were watered with sorbitol, a neutral osmoticum, and synthetic seawater, for five days. Gas exchange measurements were made during the course of the watering treatment and during a recovery period following treatment. Leaf osmotic adjustment occurred only in plants watered with seawater, and was associated with an increase in Na⁺ and Cl^- concentrations. Estimates of the molar concentrations indicated these ions could account for 95% of the leaf osmotic adjustment. Net photosynthetic CO₂ uptake was less effected during the watering treatment, and photosynthetic recovery was greater following the treatment in plants watered with seawater. Photosynthetic inhibition was related primarily to metabolic factors, including a decrease in carboxylation efficiency. A model is presented for a mechanism promoting tolerance to transient seawater inundation in A. glomeratus.     

Intrinsic competition between the gregarious parasite,Cotesia glomeratus and the solitary parasite.Cotesia rubecula [Hymenoptera: Braconidae] for their host.Artogeia rapae [Lepidoptera: Pieridae]

The solitary endoparasite,Cotesia rubecula, was superior to the gregarious endoparasite,Cotesia glomeratus, in tests of intrinsic competition for their hostArtogeia rapae. When an egg ofC. rubecula was oviposited in the host prior to those ofC. glomeratus, C. rubecula always won the ensuing competition. When an egg ofC. rubecula was laid in the same host within 48 h after oviposition byC. glomeratus, C. rubecula was successful in more than 90% of the interactions. Even when the egg ofC. rubecula was oviposited in the same host 72 to 96 h after oviposition byC. glomeratus, C. rubecula won the competition approximately 33% of the time. The implications of intrinsic competition are discussed in relation to the establishment ofC. rubecula in an ecosystem containingA. rapae andC. glomeratus.      

Effect of salinity on tissue architecture in expanding wheat leaves

Salinity greatly reduces the leaf cross-sectional area of wheat (Triticum aestivum L.) during its development, which may lead to variation in the architectural properties of growing leaves that would result in a change in leaf physiological functions. Our objective was to characterize the effect of salinity on the spatial distribution of the cross-sectional area and the anatomy of large and small veins of a growing wheat leaf. Spring wheat was grown in a growth chamber in soils with or without 120 mM NaCl. Leaf 4 in both treatments was harvested 2–3 days after its emergence and then cut into five transverse segments. Examination of the transverse sections revealed that salinity significantly reduced the cross-sectional area, width, and radii of both epidermal and mesophyll cells along the leaf axis. Reduction in the cross-sectional area and width occurred mainly at the leaf base, indicating that these reductions occur during the period of leaf initiation. The reduction in cross-sectional area was attributed to a decrease in the size of the vein segments and a reduced number of medium and small veins. The thickness of the leaf was also reduced under the 120 mM NaCl treatment. A greater intercellular air space in the large vein segments under saline conditions was also found. The approximately 35% reduction observed in the number of veins under saline conditions (mainly in the number of small veins) may suggest that salinity reduces the capacity for re-translocation of mineral nutrients and assimilates. The reduced area of protoxylem and metaxylem in midrib and large vein segments in growing tissues may be responsible for lower water deposition into the growth zone under saline conditions.     

Effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee

The effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee were examined in the laboratory. Exposed to 45 different combinations of temperature (10–30 °C) and salinity (0–40) under saturating irradiance, G. instriatum exhibited its maximum growth rate of 0.7 divisions/day at a combination of 25 °C and a salinity of 30. Optimum growth rates (>0.5 divisions/day) were observed at temperatures ranging from 20 to 30 °C and at salinities from 10 to 35. The organism could not grow at ≤10 °C. In addition, G. instriatum burst at a salinity of 0 at all temperatures, but grew at a salinity of 5 at temperatures between 20 and 25 °C. It is noteworthy that G. instriatum is a euryhaline organism that can live under extremely low salinity. Factorial analysis revealed that the contributions of temperature and salinity to its growth of the organism were almost equal. The irradiance at the light compensation point (I₀) was 10.6 μmol/(m² s) and the saturated irradiance for growth (I_s) was 70 μmol/(m² s), which was lower than I_s for several other harmful dinoflagellates (90–110 μmol/(m² s)).     

Response of root and sediment phosphatase activity to increased nutrients and salinity

Wetlands of northern Belize, distributed along a salinity gradient, are strongly phosphorus limited and dominated largely by three species of emergent macrophytes: Eleocharis cellulosa, Cladium jamaicense, and Typha domingensis. We assessed changes in root and sediment phosphatase activities of each species to simultaneous changes of nutrients (N, P) and salinity in a mesocosm experiment. Phosphorus and nitrogen treatment effects on both root and sediment phosphatase were highly significant for all the species, while salinity significantly affected root phosphatase activity in Cladium only. All species showed a significant negative correlation between root phosphatase activity and increasing tissue P content until a threshold of 0.2% P, 0.15% P and 0.12% P in Eleocharis, Cladium and Typha, respectively. There was also a significant negative correlation between soil available P and root and sediment phosphatases in all species. Activity of root phosphatases of Eleocharis and Typha were positively correlated with root tissue N. Both root and sediment phosphatases of all three species were positively correlated with soil available N. The strongest (positive) correlation was found between phoshatase activites and N/P ratios. The results confirmed that these systems are P-limited and that extracellular phosphatases respond to P enrichment by decreasing their activities. Expression of root phosphatase activity by dry root weight, sediment volume, or whole plant biomass gave very different relative results across nutrient treatments and species, suggesting that root phosphatase activities need to be interpreted in a wider context that considers root density.     

Interaction between CO2 enrichment and salinity stress in the C4 non-halophyte Andropogon glomeratus (Walter) BSP

Abstract Increasing atmospheric CO₂ may result in alleviation of salinity stress in salt-sensitive plants. In order to assess the effect of enriched CO₂ on salinity stress in Andropogon glomeratus, a C₄ non-halophyte found in the higher regions of salt marshes, plants were grown at 350, 500, and 650 cm³ m^?3 CO₂ with 0 or 100 mol m^?3 NaCl watering treatments. Increases in leaf area and biomass with increasing CO₂ were measured in salt-stressed plants, while decreases in these same parameters were measured in non-salt-stressed plants. Tillering increased substantially with increasing CO₂ in salt-stressed plants, resulting in the increased biomass. Six weeks following initiation of treatments, there was no difference in photosynthesis on a leaf area basis with increasing CO₂ in salt-stressed plants, although short-term increases probably occurred. Stomatal conductance decreased with increasing CO₂ in salt-stressed plants, resulting in higher water-use efficiency, and may have improved the diurnal water status of the plants. Concentrations of Na⁺ and Cl^? were higher in salt stressed-plants while the converse was found for K ⁺. There were no differences in leaf ion content between CO₂ treatments in the salt-stressed plants. Decreases in photosynthesis in salt-stressed plants occurred primarily as a result of decreased internal (non-stomatal) conductance.     

Effects of salinity on photosynthesis and respiration of the seagrass Zostera japonica: A comparison of two established populations in North America

Photosynthetic responses were quantified for two Zostera japonica Aschers. and Graebn. populations from the northern and southern limits of distribution exposed to a range of salinities along the Pacific Coast of North America. Plants were collected from Padilla Bay, Washington (northern) and Coos Bay, Oregon, USA (southern) and cultured together in experimental tanks at 3 salinities (5, 20 and 35) under saturating irradiance for 3 weeks. Subsequently, photosynthesis–irradiance (P vs. E curves) relationships for leaf segments from the two populations were assessed using an oxygen electrode system. We found no evidence for diel rhythms in either light saturated photosynthesis (P_max) or dark respiration (R_d). For the Padilla Bay population, P_max ranged from 192 to 390 μmol O₂ g DW⁻¹ h⁻¹; for the Coos Bay population P_max ranged from 226 to 774 μmol O₂ g DW⁻¹ h⁻¹. Photosynthetic maxima of the Coos Bay plants occurred at a salinity of 20, whereas salinity had no effect on the photosynthetic maxima of the Padilla Bay plants. There were significant differences in leaf tissue R_d among salinity treatments but the two populations responded similarly to salinity. North American populations of Z. japonica are best adapted to intermediate salinities, displaying minimum R_d rates, lower compensation irradiance, higher saturation irradiance, and greater P_max rates at a salinity of 20. Additionally, the southern population may be better adapted to southward expansion along the Pacific Coast and changes associated with global climate change.     

Growth,photosynthesis, and ion distribution in hydroponically cultured <Emphasis Type="Italic">Populus alba</Emphasis> L. cuttings grown under various salinity concentrations

Growth, photosynthesis, and Na⁺, K⁺, and Ca²⁺ distributions were investigated in 2-year-old hydroponically cultured Populus alba L. cuttings exposed to salt stresses (0, 0.85, 8.5, 17, and 85 mM NaCl in experiment 1 and 0, 50, 100, 150, and 200 mM in experiment 2) for 4 weeks in 2/5 Hoagland solution. Salt did not markedly inhibit height growth and diameter increment in 150 and 100 mM NaCl, respectively. The 85 mM NaCl treatment increased the dry weights of roots and total dry weight of plants, while 150 mM NaCl significantly reduced the dry weights of leaves, stems, and total plant weight. The decline in the photosynthetic rate lagged 2 weeks behind that of stomatal conductance in the 50 and 100 mM salt solutions. Different ions exhibited different distributions in different parts of the plant. Most Na⁺ ions were excluded and/or compartmentalized in roots at low and moderate salt stress (≤50 mM). K⁺ content in leaves increased with the increase in the salt concentration in the growth solutions.     

The effects of the interaction between salinity and nitrogen limitation in Agrostis stolonifera L.

Salt-tolerant Agrostis stolonifera ecotypes commonly grow on upper salt marshes, environments regarded as having a limited nitrogen supply. The interaction between salinity and nitrate supply limitation was studied in two ecotypes of A. stolonifera, one isolated from an upper salt marsh and one from an inland habitat. The ion, amino acid, glycine betaine and sugar contents of the two ecotypes were determined over a range of external salt concentrations and levels of nitrate supply. In vivo nitrate reductase activity was also measured. Several low molecular weight nitrogenous compounds accumulated in the salt-stressed plants. Nitrogen supply limitation had a great effect on the way in which the plants responded to salt stress. In particular, the concentrations of the soluble organic nitrogenous compounds were reduced. The results are discussed with respect to the salt marsh environment, and possible models for cytoplasmic osmoregulation are presented.Acknowledgements: One of us (MJH) gratefully acknowledges the receipt of a research studentship from the Science Research Council, U.K. We would also like to thank Mrs E. E. Griffiths for skilled technical assistance, and Dr I. Ahmad for help with the amino acid analysis.     

Patterns of genetic diversity and differentiation in the tsetse fly <Emphasis Type="Italic">Glossina morsitans morsitans</Emphasis> Westwood populations in East and southern Africa

Genetic diversity and differentiation within and among nine G. morsitans morsitans populations from East and southern Africa was assessed by examining variation at seven microsatellite loci and a mitochondrial locus, cytochrome oxidase (COI). Mean COI diversity within populations was 0.63 ± 0.33 and 0.81 taken over all populations. Diversities averaged over microsatellite loci were high (mean number of alleles/locus ≥7.4; mean H _E ≥ 65%) in all populations. Diversities averaged across populations were greater in East Africa (mean number of alleles = 22 ± 2.6; mean h _e = 0.773 ± 0.033) than in southern Africa (mean number of alleles = 18.7 ± 4.0; mean h _e = 0.713 ± 0.072). Differentiation among all populations was highly significant (R _ST = 0.25, F _ST = 0.132). Nei’s G _ij statistics were 0.09 and 0.19 within regions for microsatellites and mitochondria, respectively; between regions, G _ij was 0.14 for microsatellites and 0.23 for mitochondria. G _ST among populations was 0.23 for microsatellite loci and 0.40 for mitochondria. The F, G and R statistics indicate highly restricted gene flow among G. m. morsitans populations separated over geographic scales of 12–917 km.     

Growth and protein profile responses in the halophyte sea aster (Aster tripolium L.) suspension-cultured cells to salinity

We established salt tolerant cell lines, which survived and grew under high salinity conditions with 150 mM (S-150) and 300 mM (S-300) NaCl, to study the effects of salt stress on the proliferation and protein profile of these cells in the halophyte sea aster,Aster tripolium L. These salt-adapted cell lines were produced from leaves and selected by repeated suspension subculture in media containing NaCl every 25 days for five cycles. S-150 cells displayed no inhibition in their growth compared to control cells maintained under non-stressed conditions. S-150 cells exhibited approximately a 15-fold increase in both fresh and dry weight during the 25 days under saline conditions. S-300 cells showed positive growth under severe salt stress, but their dry matter gain was significantly less than that of the S-150 cells, with only a 2.5-fold increase in dry weight. We also detected changes in the protein profile of salt-adapted cells with two specifically induced polypeptides (basic 58.4 and acidic 24.8 kDa) and one enhanced polypeptide (basic 15.1 kDa) in the soluble fraction, and one specifically induced polypeptide (42.0 kDa) in the insoluble fraction.     

Germination responses of Diplotaxis harra to temperature and salinity

Diplotaxis harra (Forssk.) Boiss, an annual herb in the family of Brassicaceae, is widely distributed in many sandy and gypseous areas in southern Tunisia. Laboratory experiments were carried out to assess the effects of temperature and salinity on seed germination and recovery responses after seed transfer to distilled water. The germination responses of the seeds in complete darkness were determined over a wide range of temperatures (5, 10, 15, 20, 25 and 30 °C) and salinities (0, 50, 100, 150 and 200 mM NaCl). Germination was inhibited by either an increase or decrease in temperature from the optimal temperature (15 °C). Highest germination percentages were obtained under non-saline conditions and an increase in NaCl concentrations progressively inhibited seed germination. Rate of germination decreased with an increase in salinity at all temperatures but comparatively higher rates were obtained at 15 °C. Salt stress decreased both the percentage and the rate of germination. An interaction between salinity and temperature yielded no germination at 200 mM NaCl. Seeds were transferred from salt solution to distilled water after 20 days, and those from low salinities recovered at all temperatures. At NaCl concentration of 200 mM, the recovery of germination was completely inhibited.     

Mechanisms of response to salinity in halotolerant microalgae

Summary A limited number of organic solutes are used by microalgae to adjust their internal osmotic pressure in response to changing external salinities. Glycerol and proline are used by the most extremely halotolerant algae. Only glycerol allows growth at salinities approaching saturation. In addition to organic osmoregulatory solutes, inorganic ions also play an important role in osmoregulation. The ability of microalgae to maintain intracellular ions at levels compatible with metabolic functions may set upper limits for their salt tolerance. Requirements for NaCl in the external medium for nutrient transport may define the lower salinity limits for growth observed for some euryhaline algae.Osmotic upshocks generally cause severe temporary inhibition of photosynthesis in euryhaline microalgae. Extensive osmotic downshocks have little effect on photosynthesis in microalgae with strong cell walls, while wall-less species appear to be more sensitive. Rapid glycerol synthesis takes place in response to increased external salinity inChlamydomonas pulsatilla both in light and dark. Starch supplies carbon for glycerol synthesis in the dark and also during the initial periods of inhibition of photosynthesis in the light. Turnover of osmoregulatory solutes such as glycerol and isofloridoside may be an important aspect of the osmoregulatory mechanism.At salinities beyond the growth limit for the green flagellateChlamydomonas pulsatilla, resting spores are formed that enable this alga to survive extreme salinities.     

Screening of rice (Oryza sativa L.) genotypes for physiological characters contributing to salinity resistance,and their relationship to overall performance

Summary Phenotypic resistance of salinity is expressed as the ability to survive and grow in a salinised medium. Some subjective measure of overall performance has normally been used in plant breeding programmes aimed at increasing salinity resistance, not only to evaluate progeny, but to select parents. Salinity resistance has, at least implicitly, been treated as a single trait. Physiological studies of rice suggest that a range of characteristics (such as low shoot sodium concentration, compartmentation of salt in older rather than younger leaves, tolerance to salt within leaves and plant vigour) would increase the ability of the plant to cope with salinity. We describe the screening of a large number of rice genotypes for overall performance (using an objective measure based on survival) and for the aforementioned physiological traits. There was wide variation in all the characters studied, but only vigour was strongly correlated with survival. Shoot sodium concentration, which a priori is expected to be important, accounted for only a small proportion of the variability in the survival of salinity. Tissue tolerance (the cellular component of resistance reflecting the ability to compartmentalise salt within leaves) revealed a fivefold range between genotypes in the tolerance of their leaves to salt, but this was not correlated positively with survival. On the basis of such (lack of) correlation, these traits would be rejected in normal plant breeding practice, but we discuss the fallacies involved in attempting correlation between individual traits and the overall performance of a salt-sensitive species in saline conditions. We conclude that whilst overall performance (survival) can be used to evaluate the salt resistance of a genotype, it is not the basis on which parents should be selected to construct a complex character through breeding. It was the norm for varieties which had one good characteristic affecting salt resistance to be unexceptional or poor in the others. This constitutes experimental evidence that the potential for salt resistance present in the rice genome has not been realised in genotypes currently extant. The results are discussed in relation to the use of physiological traits in plant breeding, with particular reference to environmental stresses that do not affect a significant part of a species' ecological range.     

Cloning and characterization of calreticulin and its association with salinity stress in P. trituberculatus

Calreticulin (CRT) is a highly conserved and multifunctional endoplasmic reticulum (ER) chaperone protein and plays important roles in salinity stress response. Portunus trituberculatus is a commercially important fishery species, and water salinity conditions influence its commercial farming significantly. In order to research the function of calreticulin under salinity stress, the full-length cDNA sequence of calreticulin from P. trituberculatus (PtCRT) was firstly cloned and characterized. The complete cDNA sequence of PtCRT is 1676 bp with 1218 bp open reading frame (ORF), encoding a polypeptide of 405 amino acids. Multiple sequence alignments showed that the deduced acid amino sequences of PtCRT shared the highest homology to CRT of Fenneropenaeus chinensis (89 %). Fluorescent quantitative real-time PCR analysis indicated that PtCRT was expressed in all detected tissues and showed the highest expression level in hepatopancreas. In addition, salinity challenge significantly influenced the expression level of PtCRT in gill. Six single nucleotide polymorphisms (SNPs) were detected in cDNA sequence of PtCRT, and one SNP was associated with the salt tolerant trait. All results indicated that PtCRT plays an important role in mediating the salinity adaption of P. trituberculatus.     

Characterization of a microbial community capable of reducing perchlorate and nitrate in high salinity

Denitrifying up-flow packed-bed bioreactors fed with perchlorate and nitrate allowed for the examination of the impact of a variety of salt conditions (up to 10% w/v NaCl) on the complete perchlorate and nitrate removal capacity of the reactor using activated sludge taken from a municipal wastewater treatment plant. Based on the evaluation of the microbial community in the bioreactor by cloning analysis, Clostridium sp. and a Rhodocyclaceae bacteria were identified as the dominant clones. This suggests that they may be tolerant to high salt and can reduce both nitrate and perchlorate in such conditions.     

Effect of salinity on the distribution, growth, and toxicity of Karenia spp.

The first recorded bloom of Karenia spp., resulting in brevetoxin in oysters, in the low salinity waters of the Northern Gulf of Mexico (NGOMEX) occurred in November 1996. It raised questions about the salinity tolerance of Karenia spp., previously considered unlikely to occur at salinities <24 psu, and the likelihood that the bloom would reoccur in the NGOMEX. Salinity was investigated as a factor controlling Karenia spp. abundance in the field, using data from the NGOMEX 1996 bloom and Florida coastal waters from 1954 to 2004, and growth and toxin production in cultures of Karenia brevis (Davis) G. Hansen and Moestrup. During the NGOMEX bloom, Karenia spp. occurred much more frequently at low salinities than in Florida coastal waters over the last 50 years. The data suggest that the NGOMEX bloom started on the NW Florida Shelf, an area with a higher frequency of Karenia spp. at low salinities than the rest of Florida, and was transported by an unusual westward surface current caused by Tropical Storm Josephine. The minimum salinity at which growth occurred in culture ranged between 17.5 and 20 psu, but the optimal salinity ranged between low values of 20 or 25 and high values of 37.5–45 psu, depending on the clone. The effect of salinity on toxin production in one clone of K. brevis was complex, but at all salinities brevetoxin levels were highest during the stationary growth phase, suggesting that aging, high density blooms may pose the greatest public health threat. The results demonstrate that Karenia spp. can be a public health threat in low salinity areas, but the risk in the NGOMEX is relatively low. No bloom has occurred since the 1996 event, which was probably associated with a special set of conditions: a bloom along the Florida Panhandle and a tropical storm with a track that set up a westward current.