Control of xylem Na+ loading and transport to the shoot in rice and barley as a determinant of differential salinity stress tolerance

Control of xylem Na⁺ loading has often been named as the essential component of salinity tolerance mechanism. However, it is less clear to what extent the difference in this trait may determine differential salinity tolerance between species. In this study, barley (Hordeum vulgare L. cv. CM72) and rice (Oryza sativa L. cv. Dongjin) plants were grown under two levels of salinity. Na⁺ and K⁺ concentrations in the xylem sap, and shoot and root tissues were measured at different time points after stress onset. Salt‐exposed rice plants prevented xylem Na⁺ loading for several days, but failed to control this process in the longer term, ultimately resulting in a massive Na⁺ shoot loading. Barley plants quickly increased xylem Na⁺ concentration and its delivery to the shoot (most likely for the purpose of osmotic adjustment) but were able to reduce this process later on, keeping most of accumulated Na⁺ in the root, thus maintaining non‐toxic shoot Na⁺ level. Rice plants increased shoot K⁺ concentration, while barley plants maintained higher root K⁺ concentration. Control of xylem Na⁺ loading is remarkably different between rice and barley; this difference may differentiate the extent of the salinity tolerance between species. This trait should be investigated in more detail to be used in the breeding programs aimed to improve salinity tolerance in crops.     

Genotypic difference in salinity tolerance in quinoa is determined by differential control of xylem Na loading and stomatal density

Quinoa is regarded as a highly salt tolerant halophyte crop, of great potential for cultivation on saline areas around the world. Fourteen quinoa genotypes of different geographical origin, differing in salinity tolerance, were grown under greenhouse conditions. Salinity treatment started on 10 day old seedlings. Six weeks after the treatment commenced, leaf sap Na and K content and osmolality, stomatal density, chlorophyll fluorescence characteristics, and xylem sap Na and K composition were measured. Responses to salinity differed greatly among the varieties. All cultivars had substantially increased K⁺ concentrations in the leaf sap, but the most tolerant cultivars had lower xylem Na⁺ content at the time of sampling. Most tolerant cultivars had lowest leaf sap osmolality. All varieties reduced stomata density when grown under saline conditions. All varieties clustered into two groups (includers and excluders) depending on their strategy of handling Na⁺ under saline conditions. Under control (non-saline) conditions, a strong positive correlation was observed between salinity tolerance and plants ability to accumulate Na⁺ in the shoot. Increased leaf sap K⁺, controlled Na⁺ loading to the xylem, and reduced stomata density are important physiological traits contributing to genotypic differences in salinity tolerance in quinoa, a halophyte species from Chenopodium family.     

Seasonal plant water uptake patterns in the saline southeast Everglades ecotone

The purpose of this study was to determine the seasonal water use patterns of dominant macrophytes coexisting in the coastal Everglades ecotone. We measured the stable isotope signatures in plant xylem water of Rhizophora mangle, Cladium jamaicense, and Sesuvium portulacastrum during the dry (DS) and wet (WS) seasons in the estuarine ecotone along Taylor River in Everglades National Park, FL, USA. Shallow soilwater and deeper groundwater salinity was also measured to extrapolate the salinity encountered by plants at their rooting zone. Average soil water oxygen isotope ratios (δ ¹⁸O) was enriched (4.8 ± 0.2‰) in the DS relative to the WS (0.0 ± 0.1‰), but groundwater δ ¹⁸O remained constant between seasons (DS: 2.2 ± 0.4‰; WS: 2.1 ± 0.1‰). There was an inversion in interstitial salinity patterns across the soil profile between seasons. In the DS, shallow water was euhaline [i.e., 43 practical salinity units (PSU)] while groundwater was less saline (18 PSU). In the WS, however, shallow water was fresh (i.e., 0 PSU) but groundwater remained brackish (14 PSU). All plants utilized 100% (shallow) freshwater during the WS, but in the DS R. mangle switched to a soil–groundwater mix (δ 55% groundwater) while C. jamaicense and S. portulacastrum continued to use euhaline shallow water. In the DS, based on δ ¹⁸O data, the roots of R. mangle roots were exposed to salinities of 25.4 ± 1.4 PSU, less saline than either C. jamaicense (39.1 ± 2.2 PSU) or S. portulacastrum (38.6 ± 2.5 PSU). Although the salinity tolerance of C. jamaicense is not known, it is unlikely that long-term exposure to high salinity is conducive to the persistence of this freshwater marsh sedge. This study increases our ecological understanding of how water uptake patterns of individual plants can contribute to ecosystem levels changes, not only in the southeast saline Everglades, but also in estuaries in general in response to global sea level rise and human-induced changes in freshwater flows.     

Metagenome Cloning and Functional Analysis of Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> Antiporter Genes from Keke Salt Lake in China

Na⁺/H⁺ antiporters are ubiquitous membrane proteins and play a central role in cell homeostasis including pH regulation, osmoregulation, and Na⁺/Li⁺ tolerance in bacteria. The microbial communities in extremely hypersaline soil are an important resource for isolating Na⁺/H⁺ antiporter genes. A metagenomic library containing 35,700 clones was constructed by using genomic DNA obtained from the hypersaline soil samples of Keke Salt Lake in Northwest of China. Two Na⁺/H⁺ antiporters, K1-NhaD, and K2-NhaD belonging to NhaD family, were screened and cloned from this metagenome by complementing the triple mutant Escherichia coli strain KNabc (nhaA ⁻ , nhaB ⁻ , chaA ⁻) in medium containing 0.2 M NaCl. K1-NhaD and K2-NhaD have 75.5% identity at the predicted amino acid sequence. K1-NhaD has 78% identity with Na⁺/H⁺ antiporter NhaD from Halomonas elongate at the predicted amino acid sequence. The predicted K1-NhaD is a 53.5 kDa protein (487 amino acids) with 13 transmembrane helices. K2-NhaD has 73% identity with Alkalimonas amylolytica NhaD. The predicted K2-NhaD is a 55 kDa protein (495 amino acids) with 12 transmembrane helices. Both K1-NhaD and K2-NhaD could make the triple mutant E. coli KNabc (nhaA ⁻ , nhaB ⁻ , chaA⁻) grow in the LBK medium containing 0.2–0.6 M Na⁺ or with 0.05–0.4 M Li⁺. Everted membrane vesicles prepared from E. coli KNabc cells carrying K1-NhaD or K2-NhaD exhibited Na⁺/H⁺ and Li⁺/H⁺ antiporter activities which were pH-dependent with the highest activity at pH 9.5. Little K⁺/H⁺ antiporter activity was also detected in vesicles form E. coli KNabc carrying K1-NhaD or K2-NhaD.     

Osmoregulation in juvenile and adult white sturgeon,Acipenser transmontanus

Synopsis Blood samples from cannulated young adult (2.5–15 kg) white sturgeon, acclimated to San Francisco Bay water (24 ppt) had plasma values of 248.8 ± 13.5 mOsm kg⁻¹ H₂O, [Na⁺] = 125 ± 8.0 mEq 1⁻¹, [K⁺] = 2.6 ± 0.8 mEq 1⁻¹ and [CL⁻] = 122 ± 3.0 mEq 1⁻¹. Freshwater acclimated sturgeon had an osmolality of 236 ± 7, [Na⁺] = 131.6 + 4.4, [K⁺] = 2.5 ± 0.7 and [CL⁻] = 110.6 ± 3.6. Freshwater acclimated fish gradually exposed to sea water (increase of 5 ppt h⁻¹) had higher plasma osmolalities than did the bay water acclimated fish. These young adult sturgeon are able to tolerate transfer from fresh water to sea water as well as gradual transfer from sea water to fresh water. Plasma electrolytes in transferred fish are regulated, but tend to differ from long term acclimated fish at the same salinities. There is a gradual increase in the upper salinity tolerance (abrupt transfer) of juvenile white sturgeon with weight: 5–10 ppt for 0.4–0.9 g fish, 10–15 ppt for 0.7–1.8 g fish, and 15 ppt for 4.9–50.0 g fish. The ability of juveniles to regulate plasma osmolality is limited. The young adult fish are able to tolerate higher salinities (35 ppt) than juvenile sturgeon but probably are also characterized by low activity of the necessary ion exchange mechanisms in the gills which permit rapid adjustment of blood electrolytes with graduate change in external salinity.     

Relation of water transport to leaf gas exchange properties in three mangrove species

Mangrove species more tolerant to salinity may function with less efficient water transport, which may be related to more conservative water use. To test the hypothesis, we investigate the gas exchange and hydraulic properties of three mangrove species: Rhizophora mangle L., Laguncularia racemosa Gaert and Avicennia germinans (L.)L. Experiments were performed with adult plants growing naturally in the field under a salinity of 35‰. Gas exchange parameters showed that A. germinans had significantly higher photosynthetic rates, and lower stomatal conductance and transpiration rates, compared to the other two mangroves. In concert with this, instantaneous water use efficiency was significantly high in A. germinans, intermediate in L. racemosa and lowest in R. mangle. The hydraulic parameters of the three mangrove species were in the lowest end of the range reported for tropical trees. However, the three mangrove species exhibited measurable differences in hydraulic parameters related to the control of water requirements for maintenance of carbon gain. L. racemosa and A. germinans showed less efficient water transport at shoot level but were the more efficient species in water use at the leaf level in comparison to R. mangle. Received: 7 April 1999 / Accepted: 25 July 1999     

Occludin expression in goldfish held in ion-poor water

With an emphasis on the tight junction protein occludin, the response of goldfish following abrupt exposure (0–120 h) as well as long-term acclimation (14 and 28 days) to ion-poor water (IPW) was examined. Both abrupt and long-term exposure to IPW lowered serum osmolality, [Na⁺] and [Cl⁻], and elevated serum glucose. After abrupt exposure to IPW, gill tissue exhibited a prompt and sustained decrease in Na⁺–K⁺–ATPase activity, and a transient increase in occludin expression that returned to control levels by 6 h. Following 14 and 28 days in IPW, gill occludin expression was markedly elevated, while Na⁺–K⁺–ATPase activity was only significantly different (elevated) at day 14. Kidney tissue exhibited an elevation in both Na⁺–K⁺–ATPase activity and occludin expression after 28 days; however, in the intestine, occludin expression declined at day 14 but did not differ from FW fish at day 28. These studies demonstrate that goldfish can tolerate abrupt as well as sustained exposure to ion-poor surroundings. Data also suggests that occludin may play an adaptive role in fishes acclimated to ion-poor conditions by contributing to the modulation of epithelial barrier properties in ionoregulatory tissues. Helen Chasiotis and Jennifer C. Effendi contributed equally to this work.     

Seminal plasma features of Prochilodus lineatus and Brycon orbignyanus throughout two consecutives spawning seasons

This study aims to describe seminal plasma characteristics, detect changes during and between two consecutive spawning seasons (SS), and compare plasma features between two important South American fish species. Prochilodus lineatus and Brycon orbignyanus sperm was collected over two (SS1; SS2). Each season was divided into first and second sampling periods (P1; P2). Thus, the four experimental periods were referred to as SS1P1, SS1P2, SS2P1, and SS2P2. Seminal plasma was analyzed for osmolality, pH, and Na⁺, K⁺, and Ca²⁺ concentration. Additionally, sperm concentration, motility rate, and velocities (curvilinear = VCL; straight line = VSL) were determined and correlated with plasma features. In P. lineatus, plasma osmolality was lower in SS1P2, pH was higher in SS2P2, Na⁺ was higher and K⁺ and Ca²⁺ were lower in SS2P1 compared with other experimental periods. Positive correlations were observed between motility and plasma osmolality, motility and Na⁺, and VCL and Na⁺. In B. orbignyanus, plasma osmolality was higher in SS2P1 and SS2P2 and K⁺ concentration was higher in SS1P1 compared with other experimental periods; no correlation was observed. Seminal plasma parameters change during SS; therefore, the composition of a sperm extender and artificial fertilization methods should be adapted to maximize fertilization rates.     

Osmotic adjustment,water relations and growth attributes of the xero-halophyte <Emphasis Type="Italic">Reaumuria vermiculata</Emphasis> L. (Tamaricaceae) in response to salt stress

Reaumuria vermiculata (L.), a perennial dwarf shrub in the family of Tamaricaceae, is a salt-secreting xero-halophyte found widely in arid areas of Tunisia. In the present study, physiological attributes of R. vermiculata were investigated under salt stress. Four-month-old plants were subjected to various salinity levels (0, 100, 200, 300, 400 or 600 mM NaCl) for 30 days under greenhouse conditions. Results showed that plants grew optimally when treated with standard nutrient solution without NaCl supply. However, increasing osmolality of nutrient solutions caused a significant reduction in biomass production and relative growth rate. This reduction was more pronounced in roots than in shoots. In addition, this species was able to maintain its shoot water content at 30% of the control even when subjected to the highest salt level, whereas root water content seemed to be unaffected by salt. Shoot water potential declined significantly as osmotic potential of watering solutions was lowered and the more negative values were reached at 600 mM NaCl (−3.4 MPa). Concentrations of Na⁺ and Cl⁻ in the shoots of R. vermiculata were markedly increased with increasing osmolality of nutrient solutions, whereas concentration of K⁺ was not affected by NaCl supply. Salt excretion is an efficient mechanism of Na⁺ exclusion from the shoots of this species exhibiting high K⁺/Na⁺ selectivity ratio over a wide range of NaCl salinity. Proline accumulation in shoots was significantly increased with increase in salt level and may play a role in osmoregulation.     

Elevated Na+/K+-ATPase responses and its potential role in triggering ion reabsorption in kidneys for homeostasis of marine euryhaline milkfish (Chanos chanos) when acclimated to hypotonic fresh water

The milkfish (Chanos chanos) is an economic species in Southeast Asia. In Taiwan, the milkfish are commercially cultured in environments of various salinities. Na⁺/K⁺-ATPase (NKA) is a key enzyme for fish iono- and osmoregulation. When compared with gills, NKA and its potential role were less examined by different approaches in the other osmoregulatory organs (e.g., kidney) of euryhaline teleosts. The objective of this study was to investigate the correlation between osmoregulatory plasticity and renal NKA in this euryhaline species. Muscle water contents (MWC), plasma, and urine osmolality, kidney histology, as well as distribution, expression (mRNA and protein), and specific activity of renal NKA were examined in juvenile milkfish acclimated to fresh water (FW), seawater (SW 35‰), and hypersaline water (HSW 60‰) for at least two weeks before experiments. MWC showed no significant difference among all groups. Plasma osmolality was maintained within the range of physiological homeostasis in milkfish acclimated to different salinities, while, urine osmolality of FW-acclimated fish was evidently lower than SW- and HSW-acclimated individuals. The renal tubules were identified by staining with periodic acid Schiff’s reagent and hematoxylin. Moreover, immunohistochemical staining showed that NKA was distributed in the epithelial cells of proximal tubules, distal tubules, and collecting tubules, but not in glomeruli, of milkfish exposed to different ambient salinities. The highest abundance of relative NKA α subunit mRNA was found in FW-acclimated milkfish rather than SW- and HSW-acclimated individuals. Furthermore, relative protein amounts of renal NKA α and β subunits as well as NKA-specific activity were also found to be higher in the FW group than SW and the HSW groups. This study integrated diverse levels (i.e., histological distribution, gene, protein, and specific activity) of renal NKA expression and illustrated the potential role of NKA in triggering ion reabsorption in kidneys of the marine euryhaline milkfish when acclimated to a hypotonic FW environment.     

Monitoring of Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase mRNA expression in the cinnamon clownfish, <Emphasis Type="Italic">Amphiprion melanopus</Emphasis>, exposed to an osmotic stress environment: profiles on the effects of exogenous hormone

We examined changes in the expression of Na⁺/K⁺-ATPase mRNA in the gills of the cinnamon clownfish using quantitative real-time PCR in an osmotically changing environment [seawater (35 psu; practical salinity unit, 1 psu ≈ 1‰) → brackish water (17.5 psu) and brackish water with prolactin]. The expression of Na⁺/K⁺-ATPase mRNA in gills was increased after the transfer to brackish water, and the expression was repressed by prolactin treatment. Also, activities of gill Na⁺/K⁺-ATPase and plasma cortisol levels increased after the transfer to brackish water and were repressed in brackish water with prolactin treatment. Na⁺/K⁺-ATPase-immunoreactive cells were almost consistently observed in the gill filaments, but absent from the lamella epithelia. The plasma osmolality level decreased in brackish water, but the level of this parameter increased in brackish water with prolactin treatment during salinity change. These results suggest that the Na⁺/K⁺-ATPase gene plays an important role in osmoregulation in gills, and prolactin improves the hyperosmoregulatory ability of cinnamon clownfish in a brackish water (hypoosmotic) environment.     

Long-Term Growth and Succession in Restored and Natural Mangrove Forests in Southwestern Florida

We compared colonization, growth and succession from 1989 to 2000 in a restored mangrove site and in gap and closed canopy sites in a natural mangrove forest. The restored site was created in 1982 and planted with Rhizophora mangle (≈2 m⁻²) propagules. By 1989, Laguncularia racemosa, with densities up to 12.9 tree m⁻², was a dominant in all plots, although densities were greater at edge plots relative to inner plots, and near open water (west plots) relative to further inland (east plots), and in tall mangrove plots relative to scrub plots. Rhizophora mangle (1989 tree densities about 2 m⁻²) was a codominant in inner and scrub plots, while Avicennia germinans had the lowest densities (<1 tree m⁻²) in all plots. From 1989 to 2000 L. racemosa experienced reduced recruitment and apparent density-dependent mortality of canopy individuals in plots with high initial densities. Scrub plots experienced high rates of colonization by R. mangle and L. racemosa, rapid growth in height of all species (1989–1996), followed by a dieoff of L. racemosa in later years (1997–2000) as the canopy came to resemble that of tall mangrove plots. Colonization and growth rates were lower in gap and closed canopy regions of the natural forest relative to rates in the restored site. After 11 years, densities of L. racemosa were 10–20× lower and R. mangle slightly less in the gap relative to densities in tall mangrove plots in the restored site at the same age. Although the restored stand had converged with the natural forest by 2000 in terms of some factors such as species richness, vegetation cover, litterfall, and light penetration, trees were still much smaller and stem densities much higher. Full development of mature structure and ecological function will likely require decades more development.     

Influence of external salinity on the osmolality of xylem sap,leaf tissue and leaf gland secretion of the mangrove<Emphasis Type="Italic"> Laguncularia racemosa</Emphasis> (L.) Gaertn

This study assessed if mature leaves of Laguncularia racemosa were able to demonstrate salt secretion, and if the magnitude of secretion was a function of soil salinity. Thus, salinity influence on the osmolality of leaf tissue, xylem sap and leaf secretion was assessed in field and glasshouse experiments. As salinity increased, solutes were accumulated in sufficient quantity to decrease osmotic potential over the whole range of water potential. In the field, xylem osmolality (mol m^–3) increased with salinity from 32.4±2.9 at 17 to 38.2±0.6 at 28. Similarly, in the glasshouse, xylem sap osmolality (mol m^–3) increased from 33.4±1.8 (15) to 40.6±1.5 (30). Changes in Na⁺ concentration explained about 51–58% of increase in xylem osmolality. Rates of secretion (mmol m^–2 day^–1) in the field increased from 0.80±0.12 (17) to 1.16±0.14 (28), and in the glasshouse the secretion increased from 0.73±0.07 (15) to 1.25±0.07 (30). The Na⁺ accounted for 40–53% of total secretion. This study presented evidence of the capability of mature leaves of L. racemosa to secrete salt for the first time, and that the rates of secretion were enhanced as soil salinity increased.     

Ability of multicellular salt glands in <Emphasis Type="Italic">Tamarix</Emphasis> species to secrete Na<Superscript>+</Superscript> and K<Superscript>+</Superscript> selectively

The present study aimed to determine the mechanism of cation-selective secretion by multicellular salt glands. Using a hydroponic culture system, the secretion and accumulation of Na⁺ and K⁺ in Tamarix ramosissima and T. laxa under different salt stresses (NaCl, KCl and NaCl+KCl) were studied. Additionally, the effects of salt gland inhibitors (orthovanadate, Ba²⁺, ouabain, tetraethylammonium (TEA) and verapamil) on Na⁺ and K⁺ secretion and accumulation were examined. Treatment with NaCl (at 0–200 mmol L⁻¹ levels) significantly increased Na⁺ secretion, whereas KCl treatment (at 0–200 mmol L⁻¹ levels) significantly increased K⁺ secretion. The ratio of secretion to accumulation of Na⁺ was higher than that of K⁺. The changes in Na⁺ and K⁺ secretion differed after adding different ions into the single-salt solutions. Addition of NaCl to the KCl solution (at 100 mmol L⁻¹ level, respectively) led to a significant decrease in K⁺ secretion rate, whereas addition of KCl to the NaCl solution (at 100 mmol L⁻¹ level, respectively) had little impact on the Na⁺ secretion rate. These results indicated that Na+ secretion in Tamarix was highly selective. In addition, Na⁺ secretion was significantly inhibited by orthovanadate, ouabain, TEA and verapamil, and K⁺ secretion was significantly inhibited by ouabain, TEA and verapamil. The different impacts of orthovanadate on Na⁺ and K⁺ secretion might be the primary cause for the different Na⁺ and K⁺ secretion abilities of multicellular salt glands in Tamarix.     

Relationship between cation accumulation and water content of salt-tolerant grasses and a sedge

Abstract Salt-tolerant grasses and a sedge were grown at three salinities in a controlled-environment greenhouse. They were measured for growth rate, ash content, water content and cations. Fourteen species from the genera Sporobolus, Aeluropus, Leptochloa, Paspalum, Puccinellia, Hordeum, Elymus, Distichlis and Spartina survived up to the highest salt treatment (540 mol m^?3 NaCl). These were designated halophytes. Eleven species from the genera Triticum, Phragmites, Dactylotenium, Cynodon, Polypogon, Panicum, Jovea and Heleocharis only survived up to 180 mol m^?3 NaCl and were designated salt-tolerant glycophytes. All species except Distichlis palmeri grew fastest on the non-saline control treatment. All species tended to have higher Na⁺ contents and lower K⁺ and water contents on saline treatments compared to control plants. Halophytes differed from glycophytes in having statistically significant lower water contents on the non-saline treatment, and lower ash contents and Na:K ratios on 180 mol m^?3. However, the range of values among species was greater than the differences between halophytes and glycophytes. All species appeared to use Na⁺ accumulation and loss of water as the main means of osmotic adjustment. Three halophytic species were grown for a longer period of time to check the above results. The osmolality of the cell sap was measured directly by the vapour pressure method and compared to calculated values based on Na⁺, K⁺ and water contents (and assuming a balancing anion such as Cl^?). Na⁺ and K⁺ alone could account for greater than 75% of the osmotic potential at all salinities. Hence, the accumulation of organic solutes did not appear to be an important factor in the osmotic adjustment of these species. The results support the conclusion that grasses coordinate Na⁺ uptake and water loss to maintain a constant osmotic potential gradient between the shoot tissues and the external solution. The results were compared to a previous study with dicotyledonous halophytes at the same location.     

Rectal water absorption in seawater-adapted Japanese eel Anguilla japonica

Marine teleosts drink large amounts of seawater to compensate for continuous osmotic water loss. We investigated a possible significant role of the rectum in water absorption in seawater-adapted eel. In rectal sacs filled with balanced salt solution (BSS) and incubated in isotonic BSS, water absorption was greater in seawater-adapted eel than in freshwater eel. Since rectal fluid osmolality was slightly lower than plasma osmolality in seawater-adapted eel, effects of rectal fluid osmolality on water absorption were examined in rectal sacs filled with artificial rectal fluid with different osmolality. Rectal water absorption was greater at lower rectal fluid osmolality, suggesting that an osmotic gradient between the blood and rectal fluid drives the water movement. Ouabain, a specific inhibitor of Na⁺/K⁺-ATPase, inhibited water absorption in rectal sacs, indicating that an osmotic gradient favorable to rectal water absorption was created by ion uptake driven by Na⁺/K⁺-ATPase. Expression levels of aquaporin 1 (AQP1), a water-selective channel, were significantly higher in the rectum than in the anterior and posterior intestines. Immunoreaction for Na⁺/K⁺-ATPase was detected in the mucosal epithelial cells in the rectum with more intense staining in the basal half than in the apical half, whereas AQP1 was located in the apical membrane of Na⁺/K⁺-ATPase-immunoreactive epithelial cells. The rectum is spatially separated from the posterior intestine by a valve structure and from the anus by a sphincter. Such structures allow the rectum to swell as intestinal fluid flows into it, and a concomitant increase in hydrostatic pressure may provide an additional force for rectal water absorption. Our findings indicate that the rectum contributes greatly to high efficiency of intestinal water absorption by simultaneous absorption of ions and water.     

Transport of Na<Superscript>+</Superscript> and K<Superscript>+</Superscript> by an antiporter-related subunit from the <Emphasis Type="Italic">Escherichia coli </Emphasis>NADH dehydrogenase I produced in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

The NADH dehydrogenase I from Escherichia coli is a bacterial homolog of the mitochondrial complex I which translocates Na⁺ rather than H⁺. To elucidate the mechanism of Na⁺ transport, the C-terminally truncated NuoL subunit (NuoL_N) which is related to Na⁺/H⁺ antiporters was expressed as a protein A fusion protein (ProtA–NuoL_N) in the yeast Saccharomyces cerevisiae which lacks an endogenous complex I. The fusion protein inserted into membranes from the endoplasmatic reticulum (ER), as confirmed by differential centrifugation and Western analysis. Membrane vesicles containing ProtA–NuoL_N catalyzed the uptake of Na⁺ and K⁺ at rates which were significantly higher than uptake by the control vesicles under identical conditions, demonstrating that ProtA–NuoL_N translocated Na⁺ and K⁺ independently from other complex I subunits. Na⁺ transport by ProtA–NuoL_N was inhibited by EIPA (5-(N-ethyl-N-isopropyl)-amiloride) which specifically reacts with Na⁺/H⁺ antiporters. The cation selectivity and function of the NuoL subunit as a transporter module of the NADH dehydrogenase complex is discussed. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The SIDECAR POLLEN gene encodes a microspore-specific LOB/AS2 domain protein required for the correct timing and orientation of asymmetric cell division

Control of ion loading into the xylem has been repeatedly named as a crucial factor determining plant salt tolerance. In this study we further investigate this issue by applying a range of biophysical [the microelectrode ion flux measurement (MIFE) technique for non‐invasive ion flux measurements, the patch clamp technique, membrane potential measurements] and physiological (xylem sap and tissue nutrient analysis, photosynthetic characteristics, stomatal conductance) techniques to barley varieties contrasting in their salt tolerance. We report that restricting Na⁺ loading into the xylem is not essential for conferring salinity tolerance in barley, with tolerant varieties showing xylem Na⁺ concentrations at least as high as those of sensitive ones. At the same time, tolerant genotypes are capable of maintaining higher xylem K⁺/Na⁺ ratios and efficiently sequester the accumulated Na⁺ in leaves. The former is achieved by more efficient loading of K⁺ into the xylem. We argue that the observed increases in xylem K⁺ and Na⁺ concentrations in tolerant genotypes are required for efficient osmotic adjustment, needed to support leaf expansion growth. We also provide evidence that K⁺‐permeable voltage‐sensitive channels are involved in xylem loading and operate in a feedback manner to maintain a constant K⁺/Na⁺ ratio in the xylem sap.     

Leaf Longevity,Construction, and Maintenance Costs of Three Mangrove Species Under Field Conditions

This study assessed the effect of leaf age on construction cost (CC) in the mangrove species Avicennia germinans, Laguncularia racemosa, and Rhizophora mangle growing in their natural habitat. Leaf osmolality values were species-specific, the highest in A. germinans (1 693 mmol kg^–1) and the lowest in L. racemosa (1 270 mmol kg^–1). In the three species, contents of chlorophyll (a+b) (Chl_a+b) and nitrogen (N) per unit of leaf area were maximal in adult leaves and tended to decline with age. Leaf mass to leaf area ratio (LMA) and ash content increased during leaf ageing. Similarly, as leaves aged, a significant increase in leaf construction cost per leaf area (CC_a) was observed, while per leaf mass (CC_m) it remained almost constant, suggesting a sustained production of leaf compounds as leaves became older. CC was positively correlated with LMA and heat of combustion (Hc) per leaf area, suggesting differences among species in the quantity and composition of expensive compounds. Leaf half lifetime (t_0.5) showed contrasting values in the three mangrove species (60, 111, and 160 d in L. racemosa, R. mangle, and A. germinans, respectively). Overall, L. racemosa was the species with less expensive leaves to construct while leaves of A. germinans and R. mangle had the highest CC_m and CC_a, respectively. Leaf longevity was positively correlated with the ratio between CC and maximum photosynthetic rate (P _max), clearly showing the existence of a balance between leaf costs and benefits.     

Sodium transport in basil

In order to diversify the production of plants with pharmacological interest, it is important to understand the mechanisms involved in their tolerance to environmental constraints, such as salinity. Basil (Ocimum basilicum), known for its therapeutic uses, has been claimed to be salt tolerant, but physiological aspects of this behavior remain unknown. Since salt tolerance is known to be associated with several characteristics concerning Na⁺ transport to leaves, we studied this function in hydroponically grown basil. We analyzed the response of 30-day-old seedlings to 25–50 mM NaCl applied for 15 days. Growth was poorly affected, indicating that these concentrations corresponded to the tolerated salinity range. Leaves accumulated Na⁺ at relatively high concentration, without dehydrating. Potassium concentration in leaf tissues was maintained close to control level, indicating that K⁺ was 15- to 25-fold preferred over Na⁺ for ion transport and deposition. Collection of xylem sap on detopped plants revealed that this preference was only 10-fold for ion introduction into root xylem sap. Short-term (24 h) changes in Na⁺ distribution between organs after stem (steam) girdling suggested that Na⁺ downward recirculation by phloem occurred. Although modest, this transport might have augmented K⁺ selectivity of ion deposition in leaves.