Effect of Salt Stress on Growth,Na+ Accumulation and Proline Metabolism in Potato (Solanum tuberosum) Cultivars

Potato (Solanum tuberosum) is a major crop world-wide and the productivity of currently used cultivars is strongly reduced at high soil salt levels. We compared the response of six potato cultivars to increased root NaCl concentrations. Cuttings were grown hydroponically and treated with 0 mM, 60 mM and 180 mM NaCl for one week. Growth reduction on salt was strongest for the cultivars Mozart and Mona Lisa with a severe senescence response at 180 mM NaCl and Mozart barely survived the treatment. The cultivars Desiree and Russett Burbank were more tolerant showing no senescence after salt treatment. A clear difference in Na⁺ homeostasis was observed between sensitive and tolerant cultivars. The salt sensitive cultivar Mozart combined low Na⁺ levels in root and stem with the highest leaf Na⁺ concentration of all cultivars, resulting in a high Na⁺ shoot distribution index (SDI) for Mozart as compared to Desiree. Overall, a positive correlation between salt tolerance and stem Na⁺ accumulation was found and the SDI for Na⁺ points to a role of stem Na⁺ accumulation in tolerance. In stem tissue, Mozart accumulated more H₂O₂ and less proline compared to the tolerant cultivars. Analysis of the expression of proline biosynthesis genes in Mozart and Desiree showed a clear reduction in proline dehydrogenase (PDH) expression in both cultivars and an increase in pyrroline-5-carboxylate synthetase 1 (P5CS1) gene expression in Desiree, but not in Mozart. Taken together, current day commercial cultivars show promising differences in salt tolerance and the results suggest that mechanisms of tolerance reside in the capacity of Na⁺ accumulation in stem tissue, resulting in reduced Na⁺ transport to the leaves.     

The effect of NaCl on proline accumulation in potato seedlings and calli

The effects of salt stress were studied on the accumulation and metabolism of proline and its correlation with Na⁺ and K⁺ content in shoots and callus tissue of four potato cultivars, viz., Agria, Kennebec (relatively salt tolerant), Diamant and Ajax (relatively salt sensitive). Na⁺ and proline contents increased in all cultivars under salt stress. However, K⁺ and protein contents decreased in response to NaCl treatments. The activities of enzymes involved in proline metabolism, Δ¹-pyrroline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (ProDH) increased and decreased, respectively, in response to elevated NaCl concentrations. The changes of P5CS and ProDH activities in more salt sensitive cultivars (Diamant, Ajax) were more than those in the tolerant ones. Then the stimulation of synthesis in combination with a partially increase of protein proteolysis, a decrease in proline utilization and inhibition of oxidation resulted in high proline contents in seedlings and calli under salt stress. In callus tissue, reduced growth and cell size may be partially responsible for high proline accumulation in response to high NaCl levels. However, although the basic proline contents in the seedlings of more salt tolerant cultivars were higher than the sensitive ones, a clear relationship was not generally observed between accumulation of proline and salt tolerance in potato.     

Comparative salt tolerance analysis between Arabidopsis thaliana and Thellungiella halophila, with special emphasis on K(+)/Na(+) selectivity and proline accumulation

The eco-physiology of salt tolerance, with an emphasis on K⁺ nutrition and proline accumulation, was investigated in the halophyte Thellungiella halophila and in both wild type and eskimo-1 mutant of the glycophyte Arabidopsis thaliana, which differ in their proline accumulation capacity. Plants cultivated in inert sand were challenged for 3 weeks with up to 500 mM NaCl. Low salinity significantly decreased A. thaliana growth, whereas growth restriction was significant only at salt concentrations equal to or exceeding 300 mM NaCl in T. halophila. Na⁺ content generally increased with the amount of salt added in the culture medium in both species, but T. halophila showed an ability to control Na⁺ accumulation in shoots. The analysis of the relationship between water and Na⁺ contents suggested an apoplastic sodium accumulation in both species; this trait was more pronounced in A. thaliana than in T. halophila. The better NaCl tolerance in the latter was associated with a better K⁺ supply, resulting in higher K⁺/Na⁺ ratios. It was also noteworthy that, despite highly accumulating proline, the A. thaliana eskimo-1 mutant was the most salt-sensitive species. Taken together, our findings indicate that salt tolerance may be partly linked to the plants’ ability to control Na⁺ influx and to ensure appropriate K⁺ nutrition, but is not linked to proline accumulation.     

Steady state proline levels in salt-shocked barley leaves

Excised barley (Hordeum vulgare var Larker) leaves were treated with salt solutions or wilted. After the treatment period, the leaves were allowed to recover in a 50 millimolar sucrose and 1 millimolar glutamate solution, and proline, Na⁺, and K⁺ were measured at intervals. Na⁺ and K⁺ concentrations stayed at a constant high level after the salt treatments, and proline increased to a steady state concentration in response. The relationship between the maximum rate of proline accumulation and the Na⁺ concentration reached in each experiment was linear. The final steady state proline concentration reached was also directly proportional to the Na⁺ concentration. For a given Na⁺ concentration in the leaves, the steady state proline level was greater when 410 millimolar NaCl was added to the leaves than when 205 millimolar NaCl was added. These results are consistent with proline acting as a compatible cytoplasmic solute, balancing an accumulation of salts outside of the cytoplasm.

In contrast to the proline levels in salt-shocked leaves, the concentrations in wilted leaves decreased to near control levels within 24 hours of relief of stress.

     

Selection of callus cultures of sugarcane (Saccharum sp.) tolerant to NaCl and their response to salt stress

Stable callus cultures tolerant to NaCl (68 mM) were developed from salt-sensitive sugarcane cultivar CP65-357 by in vitro selection process. The accumulation of both inorganic (Na⁺, Cl⁻ and K⁺) and organic (proline and soluble sugars) solutes was determined in selected and non-selected calli after a NaCl shock in order to evaluate their implication in in vitro salt tolerance of the selected lines. Both salt-tolerant and non-selected calli showed similar relative fresh weight growth in the absence of NaCl. No growth reduction was observed in salt-tolerant calli while a significant reduction about 32% was observed in nonselected ones when both were cultivated on 68 mM NaCl. Accumulation of Na⁺ was similar in both salt-tolerant and non-selected calli in the presence of NaCl. Accumulation of Cl⁻ was lower in NaCl-tolerant than in non-selected calli while proline and soluble sugars were more accumulated in salt-tolerant than in non-selected calli when both were exposed to salt. K⁺ level decreased more severely in non-selected calli than in NaCl-tolerant ones after NaCl shock. The results indicated that K⁺ and Cl⁻ may play a key role in in vitro salt-tolerance in sugarcance cell lines obtained by in vitro selection and that organic solutes could contribute mainly to counteract the negative water potential of the outside medium.     

Dynamics of Na+, K+, and Proline Accumulation in Salttreated Vigna sinensis (L) and Phaseolus aureus (L)

The dynamics of Na⁺, K⁺, and proline accumulation in various organs of non nodulated Vigna sinensis and Phaseolus aureus was followed during their acclimation to two levels of salinities for a period of 35 days and was correlated to the vegetative growth of the two species. The rate of Na⁺ and K⁺ absorption is at a maximum during the first 15 to 20 days of culture. K⁺ absorption is not completely inhibited even at 100 mM NaCl although the endogenous Na⁺ largely surpasses that of K⁺ in certain organs. Low salinity rather accelerates K⁺ absorption in both species. The relative growth rates (RGR) correlate with the rate of Na⁺ and K⁺ accumulation. At low salinity (10 mM NaCl), the RGR of V. sinensis is greater than that of P. aureus. However, at high salinity (100 mM NaCl) the RGR is the same for both species. The growth of the younger parts of the two species is not arrested by salt treatment. Very high accumulation of Na⁺ is avoided in organs with less vacuolated tissues. At no time does the endogenous K : Na ratio in these organs fall below 1.0. Certain organs, especially the roots, hypocotyls, and the lower parts of the stems are capable of storing large quantities of Na⁺. In V. sinensis, the accumulated Na⁺ and K⁺ are evenly distributed among the various organs while in P. aureus they are rather concentrated in the roots. External salinity creates water deficiency in the younger plant parts and as a consequence, proline accumulates especially in the youngest aerial organs - more in P. aureus than in V. sinensis. The accumulation of this amino acid in both the species is dependent on time and correlates directly, not only with the water deficit, but also with the K⁺ contents. In contrast, it does not seem to depend directly on the endogenous Na⁺ content. The relative salt tolerance of the two species and the possible role of K⁺, Na⁺ and proline in the osmotic adjustments of the two species under saline conditions are discussed.     

Salt stress responses of a halophytic grass <Emphasis Type="Italic">Aeluropus lagopoides</Emphasis> and subsequent recovery

To investigate the salt tolerance mechanisms, Aeluropus lagopoides as a halophytic plant was used. Plants were treated with 0, 150, 450, 600, and 750 mM NaCl and harvested at 0, 4, 8, and 10 days after treatment and 1 day and 1 week after recovery. Optimal growth, measured as fresh and dry weights, occurred at 150 mM NaCl, but it was suppressed by 450, 600, and 750 mM NaCl. Recovery significantly increased fresh and dry weights only in 750 mM NaCl-treated plants. Water content was decreased after NaCl treatment and increased after recovery. Na⁺ and proline contents and activity of superoxide dismutase (SOD) were increased after NaCl treatment and decreased after recovery in all treated plants. In contrast, K⁺ content and ascorbate peroxidase activity decreased after NaCl treatment and increased after recovery in all treated plants. Catalase (CAT) was activated only in 750 mM NaCl-treated plants. Total content of soluble protein was slightly changed after NaCl treatment. It was concluded that proline accumulation for osmotic adjustment, SOD activation for O₂^·− scavenging, and CAT activation at the higher level of salt stress to detoxify produced H₂O₂ were main A. lagopoides strategies under salt stress. A. lagopoides salt tolerance was not based on the restriction of Na⁺ uptake.     

Physiological and biochemical parameters for evaluation and clustering of rice cultivars differing in salt tolerance at seedling stage

Salinity tolerance levels and physiological changes were evaluated for twelve rice cultivars, including four white rice and eight black glutinous rice cultivars, during their seedling stage in response to salinity stress at 100 mM NaCl. All the rice cultivars evaluated showed an apparent decrease in growth characteristics and chlorophyll accumulation under salinity stress. By contrast an increase in proline, hydrogen peroxide, peroxidase (POX) activity and anthocyanins were observed for all cultivars. The K⁺/Na⁺ ratios evaluated for all rice cultivars were noted to be highly correlated with the salinity scores thus indicating that the K⁺/Na⁺ ratio serves as a reliable indicator of salt stress tolerance in rice. Principal component analysis (PCA) based on physiological salt tolerance indexes could clearly distinguish rice cultivars into 4 salt tolerance clusters. Noteworthy, in comparison to the salt-sensitive ones, rice cultivars that possessed higher degrees of salt tolerance displayed more enhanced activity of catalase (CAT), a smaller increase in anthocyanin, hydrogen peroxide and proline content but a smaller drop in the K⁺/Na⁺ ratio and chlorophyll accumulation.     

Evaluation of salt tolerance at the seedling stage in rice genotypes by growth performance, ion accumulation, proline and chlorophyll content

Aims

The objectives of this study were to evaluate salt tolerance level of rice genotypes using the well-established screening criteria; the salt injury score, survival percentage and ratio between Na⁺ and K⁺, as well as the contents of proline and chlorophyll, and to identify the relationship between salt tolerance and physiological characters.

Methods

One hundred and six rice genotypes were grown in hydroponic solutions subjected to salt stress and evaluated for salt tolerance ability and the physiological parameters. Multivariate cluster analysis was performed based on salinity tolerance scores (ST scores; score 1 being the most tolerant, score 9 the most sensitive), survival percentage and Na⁺/K⁺ ratio.

Results

ST scores based on salt injury symptoms were negatively correlated with survival percentage and chlorophyll concentration in the stressed seedlings but positively correlated with Na⁺/K⁺ ratio and proline content. Rice genotypes were classified into five salt tolerance groups: tolerant (T), moderately tolerant (MT), moderately sensitive (MS), sensitive (S) and highly sensitive (HS). The means of ST scores were significantly different among the five tolerance groups indicating that the ST score was the most reliable index for identifying salt tolerance. The means of Na⁺/K⁺ ratio and proline content in stressed seedlings were distinctively different between the extreme T and HS groups, but the means among the intermediate groups (MT, MS and S) were not significantly different. Chlorophyll content, on the other hand, was not related to the levels of salt tolerance.

Conclusions

In addition to the commonly used Na⁺/K⁺ ratio, proline content is suggested to be another useful criterion to differentiate salt-tolerant from salt-sensitive rice. This study also identified several Thai improved and local cultivars with the level of salt tolerance and physiological characters comparable to Pokkali, the standard salt-tolerant donor and may be utilized as alternative sources of salt tolerance alleles.     

Enhanced tolerance to salinity following cellular acclimation to increasing NaCl levels in Medicago truncatula

Salinity is a major abiotic stress that limits plant productivity. Plants respond to salinity by switching on a coordinated set of physiological and molecular responses that can result in acclimation. Medicago truncatula is an important model legume species, thus understanding salt stress responses and acclimation in this species is of both fundamental and applied interest. The aim of this work was to test whether acclimation could enhance NaCl tolerance in calli of M. truncatula. A new protocol is described incorporating multi-step up acclimation over 0–350 mM exogenous NaCl. By the end of the experiment, calli were tolerant to 150 mM and competent for embryogenesis at 100 mM NaCl. Positive and negative linear relationships between Na⁺ and K⁺ uptake and exogenous NaCl concentration intercepted at 160 mM suggesting a Na⁺/K⁺ homeostasis. Proline level peaked at 100/150 mM whilst highest osmolarity and lowest water content occurred at 250/350 mM NaCl. The concentration of water soluble sugars was positively related to 0–250 mM NaCl whilst callus growth and embryogenesis occurred regardless of endoreduplication. Expression of genes linked to growth (WEE1), in vitro embryogenesis (SERK), salt tolerance (SOS1), proline synthesis (P5CS) and ploidy level (CCS52 and WEE1) peaked at 100/150 mM NaCl. Hence, these genes and various physiological traits except sugar levels, served as useful markers of NaCl tolerance. To our knowledge, this is the first report of a multi-step acclimation conferring tolerance to 150 mM NaCl in leaf-derived calli of M. truncatula.     

The expression of salt tolerance from Triticum tauschii in hexaploid wheat

Summary Accessions of Triticum tauschii (Coss.) Schmal. (D genome donor to hexaploid wheat) vary in salt tolerance and in the rate that Na⁺ accumulates in leaves. The aim of this study was to determine whether these differences in salt tolerance and leaf Na⁺ concentration would be expressed in hexaploid wheat. Synthetic hexaploids were produced from five T. tauschii accessions varying in salt tolerance and two salt-sensitive T. turgidum cultivars. The degree of salt tolerance of the hexaploids was evaluated as the grain yield per plant in 150 mol m^-3 NaCl relative to grain yield in 1 mol m^-3 NaCl (control). Sodium concentration in leaf 5 was measured after the leaf was fully expanded. The salt tolerance of the genotypes correlated negatively with the concentration of Na⁺ in leaf 5. The salt tolerance of the synthetic hexaploids was greater than the tetraploid parents primarily due to the maintenance of kernel weight under saline conditions. Synthetic hexaploids varied in salt tolerance with the source of their D genome which demonstrates that genes for salt tolerance from the diploid are expressed at the hexaploid level.     

In vitro responses of grape rootstocks to NaCl

An investigation was undertaken to adjudge the in vitro salt tolerance and biochemical changes due to NaCl on four grape rootstocks (Dogridge, SO4, H-144 and 3309C). The in vitro two-node micro-cuttings from the established cultures were sub-cultured on rooting medium comprising Murashige and Skoog (MS) basal medium supplemented with 0.5 μM indolebutyric acid (IBA), 200 mg dm⁻³ activated charcoal and different NaCl concentrations ranging from 0 to 125 mM. The surviving and proliferated cultures were further sub-cultured four times on respective media. Dogridge and H-144 tolerated 125 and 100 mM NaCl, respectively, while SO4 and 3309C survived only up to 75 mM NaCl. Contents of proteins, proline, K⁺ and Na⁺ in tissue increased in all genotypes due to NaCl supplementation, while contents of chlorophyll and total soluble sugars declined. Higher K⁺/Na⁺ ratio was registered in Dogridge and H-144 than in SO4 and 3309C. The relative NaCl tolerance for different grape rootstocks under study could be ranked as Dogridge > H-144 > SO4 and 3309C.     

Evaluation of salinity tolerance in sorghum (<Emphasis Type="Italic">Sorghum bicolor</Emphasis> L.) using ion accumulation,proline and peroxidase criteria

Salt tolerance of sorghum varieties in terms of fresh weight, ion accumulations, proline content and peroxidase activity was analyzed in this study. Three sorghum varieties, Payam, Kimia, and Jambo, differing in salt tolerance, were grown in a greenhouse-hydroponic culture with a complete nutrition solution to which 0, 50, 100, 150 and 200 mM NaCl was added. Plant roots and leaves were harvested at 15 and 30 days after treatment and subjected to analysis. Clear decline in K⁺ and Ca²⁺ concentrations and increase in Na⁺ and proline contents were observed in the root and leaf tissues at each NaCl concentration in all varieties during the NaCl treatment. The Ca²⁺ concentration in leaves was higher than in roots, and had the following order in the tested cultivars: Jambo, Kimia, and Payam. Total peroxidase activity increased under salinity stress and it was proportional with the salt concentration. Payam had the largest decrease (46.95%) in fresh weight caused by NaCl, while Jambo had the lowest decrease, 28.63%. Linear regression analysis revealed significant relationships between the estimated factors and fresh weight. The profiles of isoperoxidases were modified under stress conditions. Two isoforms, A1 and A2, were detected in all three varieties with different intensities. Under NaCl stress, isoperoxidases were strongly expressed and a third isoform, A3, was specifically found in variety Jambo suggesting that A3 is implicated in salt adaptation of this variety.     

Physiological and proteomic analysis of salinity tolerance of the halotolerant cyanobacterium <Emphasis Type="Italic">Anabaena</Emphasis> sp

The halotolerant cyanobacterium Anabaena sp was grown under NaCl concentration of 0, 170 and 515 mM and physiological and proteomic analysis was performed. At 515 mM NaCl the cyanobacterium showed reduced photosynthetic activities and significant increase in soluble sugar content, proline and SOD activity. On the other hand Anabaena sp grown at 170 mM NaCl showed optimal growth, photosynthetic activities and comparatively low soluble sugar content, proline accumulation and SOD activity. The intracellular Na⁺ content of the cells increased both at 170 and 515 mM NaCl. In contrast, the K⁺ content of the cyanobacterium Anabaena sp remained stable in response to growth at identical concentration of NaCl. While cells grown at 170 mM NaCl showed highest intracellular K⁺/Na⁺ ratio, salinity level of 515 mM NaCl resulted in reduced ratio of K⁺/Na⁺. Proteomic analysis revealed 50 salt-responsive proteins in the cyanobacterium Anabaena sp under salt treatment compared with control. Ten protein spots were subjected to MALDI-TOF–MS/MS analysis and the identified proteins are involved in photosynthesis, protein folding, cell organization and energy metabolism. Differential expression of proteins related to photosynthesis, energy metabolism was observed in Anabaena sp grown at 170 mM NaCl. At 170 mM NaCl increased expression of photosynthesis related proteins and effective osmotic adjustment through increased antioxidant enzymes and modulation of intracellular ions contributed to better salinity tolerance and optimal growth. On the contrary, increased intracellular Na⁺ content coupled with down regulation of photosynthetic and energy related proteins resulted in reduced growth at 515 mM NaCl. Therefore reduced growth at 515 mM NaCl could be due to accumulation of Na⁺ ions and requirement to maintain higher organic osmolytes and antioxidants which is energy intensive. The results thus show that the basis of salt tolerance is different when the halotolerant cyanobacterium Anabaena sp is grown under low and high salinity levels.     

NaCl胁迫对流苏幼苗生长、钠钾离子分布及渗透调节物质的影响

以2年生的流苏播种苗为材料,采用不同浓度(50、100、200、300 mmol·L^-1)NaCl溶液进行胁迫处理,研究盐胁迫对流苏的生长、Na^+和K^+分布格局、渗透调节物质的影响,以明确其耐盐阈值。结果表明:(1)随着NaCl胁迫浓度的增加,流苏幼苗生长量逐渐降低,盐害指数升高、存活率下降;幼苗耐盐阈值为98.693 mmol·L^-1(0.577%W/V)。(2)随着NaCl胁迫浓度的增加,流苏幼苗各器官中的Na^+含量持续增加,并在浓度为50 mmol·L^-1时表现为根>叶>茎,在其余各处理组表现为叶>根>茎;幼苗根、叶中的K^+含量表现为先增后减的变化趋势,茎中K^+含量总体表现为下降趋势,且器官中K^+含量表现为根>叶>茎;幼苗根部到茎部和茎部到叶部的离子选择性运输能力、各器官中的K^+/Na^+比值均呈下降趋势。(3)随着NaCl浓度的增加,流苏幼苗叶片可溶性糖、可溶性蛋白含量总体呈上升趋势,其脯氨酸含量呈先上升后下降的趋势。研究发现,流苏幼苗根系可通过对Na^+的吸收和累积来阻止其向地上部运输进而避免盐害发生;叶片和茎中通过提高对K^+的选择性吸收和累积,从而增大K^+/Na^+比值以减缓盐分对其生理代谢的伤害。     

Influence of exogenous application of glycinebetaine on antioxidative system and growth of salt-stressed soybean seedlings (Glycine max L.)

Glycinebetaine is one of the most competitive compounds which play an important role in salt stress in plants. In this study, the enhanced salt tolerance in soybean (Glycine max L.) by exogenous application of glycinebetaine was evaluated. To improve salt tolerance at the seedling stage, GB was applied in four different concentrations (0, 5, 25 and 50 mM) as a pre-sowing seed treatment. Salinity stress in the form of a final concentration of 150 mM sodium chloride (NaCl) over a 15 day period drastically affected the plants as indicated by increased proline, MDA and Na⁺ content of soybean plants. In contrast, supplementation with 50 mM GB improved growth of soybean plants under NaCl as evidenced by a decrease in proline, MDA and Na⁺ content of soybean plants. Further analysis showed that treatments with GB, resulted in increasing of CAT and SOD activity of soybean seedlings in salt stress. We propose that the role of GB in increasing tolerance to salinity stress in soybean may result from either its antioxidant capacity by direct scavenging of H₂O₂ or its role in activating CAT activity which is mandatory in scavenging H₂O₂.     

The effects of NaCl on growth,water relations,osmolytes and ion content in <Emphasis Type="Italic">Kochia prostrata</Emphasis>

The effects of NaCl (0, 50, 100, 150 and 200 mM) on growth, water relations, glycinebetaine, free proline, ion contents, stomata number and size of Kochia prostrata (L.) Schard were determined. Shoot and root fresh and dry matter, root and shoot length, relative growth rate, net assimilation rate, relative water content, water use efficiency, soluble sugars and glycinebetaine contents were not changed at low NaCl concentrations, but they were significantly decreased at 200 mM NaCl. The K⁺, Mg²⁺ and Ca²⁺ contents, water potential, chlorophyll a+b and carotenoides contents, and stomata number and size were reduced already at low concentrations of NaCl. In contrast, the Na⁺, Cl^– and proline contents increased several times with increasing NaCl concentration. Kochia prostrata is a salt tolerant species, the optimal growth of this plant occurred up to 150 mM NaCl. The mechanisms of salt tolerance in the plant may be balance among ion accumulation and production of glycinebetaine, proline, soluble sugars for maintenance of pressure potential.     

Ion fluxes and abscisic Acid-induced proline accumulation in barley leaf segments

The increase in proline induced by ABA, a process stimulated by NaCl or KCl in barley leaves, did not occur when Na⁺ (or K⁺) was present in the external medium as the gluconate salt, namely with an anion unable to permeate the plasma membrane. However, proline increase was restored, to different extents, by the addition of various chloride salts but not by ammonium chloride. Moreover, it was shown that the stimulation of the process by NaCl (or KCl) was variously affected by the presence of different salts; all the ammonium salts (10 millimolar NH₄⁺ concentration) inhibited this stimulation almost completely. Inhibition by NH₄⁺ was accompanied by a decreased Na⁺ influx (−40%). Also, in the case of Na-gluconate, Na⁺ uptake was reduced and the addition of Cl⁻ as the calcium or magnesium salt (but not as ammonium salt) restored both the ion influxes and the increase in proline typical of NaCl treatments. Both 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene (DIDS), an anion transport inhibitor, and tetraethylammonium chloride (TEA), a K⁺ channels-blocking agent, caused, as well as with a reduction of ion influxes, an inhibition of the proline accumulation. The inhibition was practically total with 1 millimolar DIDS and about 80% with 20 millimolar TEA. A possible role of ion influxes in the process leading to the increase in proline induced by ABA is proposed.     

Evaluation of salinity tolerance in seedlings of sugar beet (Beta vulgaris L.) cultivars using proline, soluble sugars and cation accumulation criteria

Salinity tolerance of sugar beet (Beta vulgaris L.) cultivars in terms of growth, proline and soluble sugars concentrations, and Na⁺/K⁺ and Na⁺/Ca²⁺ ratios were analyzed in this study. Three-week-old seedlings of three sugar beet cultivars, ‘Gantang7’, ‘SD13829’, and ‘ST21916’, differing in salinity tolerance, were treated with 0, 50, 100, and 200 mM NaCl. Plant shoots and roots were harvested at 7 days after treatment and subjected to analysis. Low concentration of NaCl (50 mM) enhanced fresh and dry weights of shoot and root in ‘Gantang7’, whereas high one (200 mM) reduced growth in all cultivars and the less reduction was observed in ‘ST21916’. Shoot proline was strongly induced by salinity stress in both ‘Gantang7’ and ‘SD13829’, while it remained unchanged in ‘ST21916’. The addition of 50 mM NaCl significantly increased shoot soluble sugars concentrations in ‘Gantang7’ while it had no significant effects in the other two cultivars. ‘Gantang7’ also showed a higher level of root soluble sugars concentration as compared to the other two cultivars. At 50 mM NaCl, the lower shoot Na⁺ concentration, and the higher shoot K⁺ and root Ca²⁺ concentration in ‘Gantang7’ resulted in the lower shoot Na⁺/K⁺ and root Na⁺/Ca²⁺ ratio. However, ‘SD13829’ maintained a lower Na⁺/K⁺ ratio in both shoot and root when subjected to 200 mM NaCl treatment. According to comprehensive evaluation on salinity tolerance, it is clear that ‘Gantang7’ is more tolerant to salinity than the other two cultivars. Therefore, it is suggested that ‘Gantang7’ should be more suitable for cultivating in the arid and semi-arid irrigated regions.     

Growth, proline accumulation and ion content in sodium chloride-stressed callus of date palm

Summary Growth and physiological responses of date palm. Phoenix dactylifera L. cv. Barhee, callus to salinity stress were examined. Callus induced from shoot tips of offshoots was cultured on Murashige and Skoog medium supplemented with NaCl at concentrations ranging from 0 to 225 mM, in consective increments of 25 mM. Data obtained after 6 wk of exposure to salt have shown a significant increase in callus proliferation in response to 25 mM NaCl the lowest level tested, beyond which callus weight decreased. At 125 mM NaCl and higher, callus growth was nearly completely inhibited. Physiological studies on callus exposed to salt stress have shown an increase in proline accumulation in response to increased salinity. Proline accumulation was correlated to callus growth inhibition. Furthermore, increasing the concentration of NaCl in the culture medium generally resulted in a steady increase in Na⁺ and reduction in K⁺ concentrations. However, at 25 mM NaCl, the only level at which callus growth was significantly enhanced, an increase in K⁺ content was noted, in comparison to the NaCl free control. In response to increasing external NaCl level, the Na⁺/K⁺ ratio increased The Na⁺/K⁺ ratio was positively correlated to proline accumulation and hence callus growth inhibition. This study provides, an understanding of the response of date palm callus to salinity, which is important for future studies aimed at developing strategies for selecting and characterizing somaclonal variants tolerant to salt stress.