Strain differences in amiloride inhibition of NaCl responses in mice,Mus musculus

Summary The effects of lingual treatment with amiloride, an inhibitor of salt taste responses in several mammalian species, on NaCl responses of the chorda tympani nerve were compared between four inbred strains of mouse (BALB/cCrSlc, DBA/2CrSlc, C57BL/6CrSlc and C3H/HeSlc). In C57BL and C3H mice amiloride significantly suppressed responses of the chorda tympani nerve to NaCl at a concentration 0.1 M or more whereas in BALB and DBA mice the drug did not significantly affect the responses to NaCl at any concentration, suggesting a lack of the amiloride-sensitive receptor component for NaCl in the latter two strains.A two-bottle preference test demonstrated that all strains of mouse usually showed no preference for NaCl at any concentration and avoided NaCl at 0.3 M or more, although some differences were observed in that C57BL and C3H mice showed aversive responses to 0.1 and 0.15 M NaCl, whereas BALB and DBA mice were indifferent to these solutions.The results suggest that there exist prominent differences between mouse strains in the amiloride-sensitive component of their salt receptor systems. However, in mice the taste information derived from the amiloride-sensitive receptor component probably has no remarkable effect on behavioral responses to NaCl except for a possible contribution to decreasing aversion thresholds for NaCl by increasing overall taste information about NaCl.     

Effect of growth conditions on the motility of<Emphasis Type="Italic"> Photorhabdus temperata</Emphasis>

Photorhabdus temperata is a bioluminescent bacterium that lives in mutualistic association with entomopathogenic nematodes of the genus Heterorhabditis. The bacterium exists in two morphologically distinguishable phases (primary and secondary). The swimming behavior of P. temperata was investigated. Both the primary and secondary variants were able to swim in liquid or semisolid media under appropriate conditions. Variation in the oxygen levels had little affect on the chemotaxis and motility of the primary form, but greatly influenced the behavior of the secondary form. Under oxic conditions the secondary form was nonmotile, but motility was induced under anoxic conditions. Several phenotypic traits of the primary form were not expressed under anoxic conditions. The constituents of the growth media affected the motility of both variants. P. temperata required additional NaCl or KCl for optimum motility and chemotaxis. Optimal chemotactic behavior required the presence of bacto-peptone and yeast extract in the swim-migration medium. A mutant that was isolated from the secondary form was able to swim under oxic conditions and possessed an altered salt requirement for motility.     

Developmental changes in chemotactic response and choice of two attractants, sodium acetate and diacetyl, in the nematode Caenorhabditis elegans

The chemotactic behavior of the nematode Caenorhabditis elegans to chemical attractants, water-soluble sodium acetate and odorant diacetyl, was investigated using nematodes at various developmental stages to examine the effects of postembryonic development on chemotactic response and spontaneous locomotion. The chemotactic responses to attractants increased as development progressed, and the largest responses to either 1.0 M sodium acetate or 0.1% diacetyl were seen at the young adult (YA) or day adult (A1) stage, respectively. Responses to the chemicals declined thereafter in-line with increasing age. The chemotaxis indices for attractants correlated with activity of spontaneous locomotion (p<0.01), suggesting that a change in spontaneous locomotion is one of the factors involved with the change in chemotactic responses during development. We also investigated the effect of aging on attractant choice by the simultaneous presentation of 0.6 M sodium acetate and 0.1% diacetyl. In the presence of both attractants, the fraction of larval animals at the sodium acetate location was greater than that at the diacetyl location (p<0.05). The fractions of YA animals that gathered at either location were almost identical, whereas the fraction of adult animals at the diacetyl location was greater than that at the sodium acetate location (p<0.05). The patterns of attractant choice of the long-lived daf-2 mutants and short lifespan mev-1 mutants showed the same tendency as those of wild type nematodes in the presence of both attractants. These results suggest that a change in the neuronal mechanisms controlling attractant choice and preference occurs during developmental progression.     

Effect of 5-Aminolevulinic Acid on Development and Salt Tolerance of Potato (Solanum tuberosum L.) Microtubers in vitro

The effects of 5-aminolevulinic acid (ALA), a key precursor in the biosynthesis of porphyrins such as chlorophyll and heme, on development and salt tolerance of microtubers of two potato (Solanum tuberosum L.) cultivars Jingshi-2 and Zihuabai were examined under in vitro conditions. ALA at 0.3–3 mg/l promoted microtuber formation by increasing the average number, diameter, and fresh weight of microtubers especially under 0.5% NaCl stress conditions, but further increase in ALA concentration resulted in a reduction of microtuber yield irrespective of NaCl stress. Under 1.0% NaCl stress conditions, microtuberization was seriously repressed and could not be restored by the addition of ALA. The accumulation of malondialdehyde in the microtubers treated with 30 mg/l ALA increased by 22% compared to the controls (no salinity), while only a 7% increase was observed when the microtubers were exposed to 0.5% NaCl, indicating that ALA functions as a protectant against oxidative damages of membranes. Under 0.5% NaCl stress conditions, the highest activities of peroxidase and polyphenoloxidase were detected in microtubers treated with ALA at 0.3 and 3 mg/l, being by 73% and by 28% greater than those in the untreated controls, respectively. These results demonstrate that ALA at lower concentrations of 0.3–3 mg/l promotes development and growth of potato microtubers in vitro and enhances protective functions against oxidative stresses, but ALA at 30 mg/l and higher concentrations seems to induce oxidative damage probably through formation and accumulation of photooxidative porphyrins.     

3种李属彩叶植物对NaCl胁迫的生理响应

以盆栽4年生紫叶李、紫叶矮樱、黑杆樱李幼苗为试材,设置土壤NaCl含量分别为0.042%(CK)、0.1%、0.2%、0.3%和0.4%的盐胁迫处理,研究3种李属彩叶植物盐害情况及其叶片膜质过氧化程度、抗氧化酶活性、有机渗透调节物质含量的变化,已明确它们的耐盐能力.结果表明:(1)随着土壤中NaCl含量的递增,3种彩叶植物的盐害指数和盐害率不断增大,上述三者盐害指数在50%时土壤NaCl含量分别是0.349%、0.261%和0.327%;(2)它们叶片细胞膜透性和氧自由基产生速率均迅速增加,并以紫叶李各处理的值最低且增加最慢;(3)各植物叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性变化均呈先上升后下降趋势,并均在土壤NaCl含量为0.2%或0.3%时达到峰值;(4)各植物叶片脯氨酸含量不断增加,可溶性糖含量先升高后下降,可溶性蛋白含量则先下降后升高.研究发现,3种李属彩叶植物均能在一定盐胁迫范围内通过自身保护酶系统和渗透调节物质有效降低活性氧的伤害,它们的抗盐性表现为紫叶李>黑杆樱李>紫叶矮樱;脯氨酸和可溶性糖可能是盐胁迫下3种植物的主要渗透调节物质.     

Salicylic Acid Induces Salinity Tolerance in Tomato (Lycopersicon esculentum cv. Roma): Associated Changes in Gas Exchange, Water Relations and Membrane Stabilisation

The present study investigates the role of salicylic acid (SA) in inducing plant tolerance to salinity. The application of 0.1 mM SA to tomato [Lycopersicon esculentum Mill.] plants via root drenching provided protection against 150 mM or 200 mM NaCl stress. SA treated plants had greater survival and relative shoot growth rate compared to untreated plants when exposed to salt stress. At 200 mM salt, shoot growth rates were approximately 4 times higher in SA treated plants than untreated plants. Application of SA increased photosynthetic rates in salt stressed plants and may have contributed to the enhanced survival. Transpiration rates and stomatal conductance were also significantly higher in SA treated plants under saline stress conditions. SA application reduced electrolyte leakage by 44% in 150 mM NaCl and 32% in 200 mM NaCl, compared to untreated plants, indicating possible protection of integrity of the cellular membrane. Beneficial effects of SA in saline conditions include sustaining the photosynthetic/transpiration activity and consequently growth, and may have contributed to the reduction or total avoidance of necrosis. SA, when used in appropriate concentrations, alleviates salinity stress without compromising the plants ability for growth under a favourable environment.     

盐胁迫对竹柳幼苗生理响应及结构解剖的研究

为了解盐胁迫对竹柳(Salix spp.)幼苗生长的影响,采用土培方法,对NaCl胁迫下半年生竹柳扦插苗的成活率、生理响应和根叶部结构进行研究。结果表明,在0.25%Na Cl胁迫(轻度盐胁迫)下竹柳能正常生长,而在0.5%NaCl(中度、重度盐胁迫)下生长受到抑制,推断竹柳的耐盐阈值是0.5%。随着NaCl浓度的增大,叶片相对含水量、叶绿素a含量、叶绿素总含量和叶绿素a/b均呈下降趋势;但叶绿素b含量、脯氨酸含量和MDA含量呈升高趋势。在轻度盐胁迫下,叶片SOD活性和可溶性蛋白含量均升高,在中、重度盐胁迫下显著下降。从根叶解剖结构来看,叶片、角质层、栅栏组织厚度和根部周皮和直径在轻度盐胁迫时最大,但在中度盐胁迫时叶片栅栏组织细胞长度减小且排列越来越疏松,根部输导组织细胞不正常。这表明竹柳在轻度胁迫时具有一定的耐盐性,但在中高度盐胁迫下生长不良。     

Response of mycorrhizal and nonmycorrhizal Arachis hypogaea to NaCl and acid stress

 The response of peanut to salt (NaCl) and acid (HCl) stress was studied in association with Glomus caledonium, an arbuscular mycorrhizal (AM) fungus. The plants were exposed to salt stress by irrigation on alternate days with 1% or 5% NaCl solutions, or with 0.1 N HCl to induce acid stress. Plant yield almost tripled in mycorrhizal plants compared with nonmycorrhizal control plants. AM inoculation significantly increased plant yield and biomass at 1% NaCl, while at 5% NaCl AM was less effective in alleviating salt stress. Percentage AM colonization was also lowest at 5% NaCl. AM inoculation was found to promote the establishment of peanut plants under acid stress conditions. Accepted: 2 October 1995     

In vitro selection of salinity tolerant variants from triploid bermudagrass (<Emphasis Type="Italic">Cynodon transvaalensis</Emphasis> × <Emphasis Type="Italic">C. dactylon</Emphasis>) and their physiological responses to salt and drought stress

A protocol was established for in vitro selection of salinity tolerant somaclonal variations from suspension cultured calli of triploid bermudagrass cv. TifEagle. To induce somaclonal variations the calli were subcultured for 18 months and were then subject to three-round selections for salt-tolerant calli by placing on solid medium containing 0.3 M NaCl for 10 days followed by a recovery for 2 weeks. The surviving calli were regenerated on regeneration medium containing 0.1 M NaCl. Three somaclonal variant lines (2, 71, and 77) were obtained and analyzed. The selected somaclonal lines showed higher relative growth and less injury than TifEagle under salt stress, indicating that they increased salt tolerance. In addition, they had higher relative water content and lower electrolyte leakage than TifEagle after withholding irrigation, indicating that they also increased drought tolerance. The three somaclonal variant lines had higher proline content than TifEagle under normal growth condition. The line 71 had a higher K⁺/Na⁺ ratio, whereas the lines 2 and 77 had higher CAT activity under control and salt stress conditions, indicating that different mechanisms for salt tolerance might exist in these three lines.     

Effect of Phytohormone Pretreatment on Nitrogen Metabolism in Vigna radiata Under Salt Stress

Application of NaCl (electrical conductivity 4.0 mS cm^–1) resulted in about 52, 50 and 55 % reduction in total nitrogen contents in mung bean [Vigna radiata (L.) Wilczek] leaf, root and nodule, respectively. In nodule, nitrogenase activity was reduced by about 84 % under stress as compared with the control set. Glutamine synthetase activity was reduced by about 31, 16 and 23 %, glutamate oxoglutarate aminotransferase activity was reduced by 78, 57 and 42 % and glutamate dehydrogenase activity was reduced by 9, 8 and 42 % in leaf, root and nodule, respectively, under salt stress. The pretreatment with indole-3-acetic acid, gibberellic acid and kinetin, each ranging from 0.1 to 10 µM, in restoring the metabolic alterations imposed by NaCl salinity was investigated in mung bean. The three phytohormones used were able to overcome to variable extents the adverse effects of stress imposed by NaCl solution.     

Salt tolerance and survival thresholds for two species of Antarctic soil nematodes

We evaluated the response of the Antarctic soil nematodes Scottnema lindsayae and Plectus antarcticus to various salts (NaCl, MgSO₄, KNO₃ and NaCl + MgSO₄) and salt concentrations in prepared salt solutions ranging from 0.1 to 3 M, and in saturation paste extracts of soils collected from multiple locations where nematode abundance varied from zero to numerous, and where electrical conductivity ranged from 108 to >12,000 μS/cm. Nematode salt tolerance was salt specific; both nematode species survived in low-experimental concentrations of NaCl and MgSO₄, and neither species survived in KNO₃ solutions of any concentration. There was no survival of nematodes in the saturation paste extracts of highly saline soils (4,100 μS/cm), while survival was over 80–97% in less saline soils (1,945 μS/cm). A 1:1 dilution of these highly saline saturation paste extracts increased S. lindsayae survival to 80%, while survival of P. antarcticus was not observed until dilutions of greater than 200%. The results complement previous studies demonstrating niche partitioning of S. lindsayae and P. antarcticus across salinity gradients and strengthen interpretations of the physiological mechanisms underlying previously reported spatial correlation between soil salinity and nematodes abundance in the Antarctic Dry Valleys.     

Taste responses of Drosophila melanogaster

The amounts of sugar solution consumed by Drosophila melanogaster flies were determined. Starved and desiccated flies of a wild type strain (QA) consume 7?9 × 10^?2 λ of a 0.3 M sucrose solution per fly during the first hour and less later. They consume more of the 0.3 M sucrose solution than of the more diluted and the more concentrated solutions. In preference-aversion tests the flies discriminated between water and various sugar solutions, and between different sugar concentrations. Contrary to other fly species these flies did not prefer 0.05 M fructose over 0.05 M glucose. 0.3–0.5 M NaCl added to 0.1 M sucrose turned a preference over 0.01 sucrose into an aversion. A mutant, Lot-94, selected for its increased consumption of a 1 M NaCl solution was found to consume more of all test solutions. The amount of NaCl that had to be added to 0.1 M sucrose to turn the preference over 0.01 M sucrose by the mutant flies into aversion was not different from that found for the wild type flies.     

The insulin/PI 3-kinase pathway regulates salt chemotaxis learning in Caenorhabditis elegans

The insulin-like signaling pathway is known to regulate fat metabolism, dauer formation, and longevity in Caenorhabditis elegans. Here, we report that this pathway is also involved in salt chemotaxis learning, in which animals previously exposed to a chemoattractive salt under starvation conditions start to show salt avoidance behavior. Mutants of ins-1, daf-2, age-1, pdk-1, and akt-1, which encode the homologs of insulin, insulin/IGF-I receptor, PI 3-kinase, phosphoinositide-dependent kinase, and Akt/PKB, respectively, show severe defects in salt chemotaxis learning. daf-2 and age-1 act in the ASER salt-sensing neuron, and the activity level of the DAF-2/AGE-1 pathway in this neuron determines the extent and orientation of salt chemotaxis. On the other hand, ins-1 acts in AIA interneurons, which receive direct synaptic inputs from sensory neurons and also send synaptic outputs to ASER. These results suggest that INS-1 secreted from AIA interneurons provides feedback to ASER to generate plasticity of chemotaxis.     

In vitro screening of mulberry (<Emphasis Type="Italic">Morus</Emphasis> spp.) for salinity tolerance

An efficient in vitro screening method has been developed for mulberry (Morus spp. ) to screen salinity-tolerant genotypes from a large population. Axillary buds from field-grown plants were cultured on MS medium containing five different concentrations (0.0%, 0.25%, 0.5%, 0.75% and 1.00%) of sodium chloride (NaCl) in order to study the shoot growth pattern. Rooting was also tested at four different concentrations of NaCl (0.0%, 0.1%, 0.2% and 0.3.%). NaCl has been found to inhibit the growth and development of mulberry shoots and roots in vitro. The survivability of the axillary buds of the genotypes tested was reduced from 83.7% for the controls to 6.1% in 1.0% NaCl. The average number of roots developed by the genotypes ranged from 11.9 (controls) to 0.2 (0.3% NaCl). Out of the 63 genotypes tested, only seven—Rotundiloba, English black, Kolitha-3, Berhampore-A, Kajli, BC₂59 and C776—developed roots in 0.3% NaCl. Root growth was also reduced drastically from 1.8 cm for the controls to 0.1 cm in 0.3% NaCl. To test the reproducibility of the results in soil, five tolerant and two susceptible genotypes, identified in this in vitro study, were selected and tested under ex vitro conditions. The significant correlation coefficients obtained between the performances of these genotypes under both types of cultural conditions revealed that in vitro screening of mulberry through axillary bud culture is an easy and efficient method to identify salt-adapted genotypes within a limited space and time period.Abbreviations BA 6-Benzylaminopurine - dS m^-1 Deci-Siemens per meter - EC Electric conductivity - NAA -Naphthaleneacetic acidCommunicated by G.C. Phillips     

Effects of NaCl on physiology and leaf ultrastructure in the halophyte Kalidium foliatum

We studied the effects of salt exposure on the growth and physiology of the xerohalophyte Kalidium foliatum. Plants were grown for 21 days under greenhouse conditions in the presence of between 0 and 500 mM NaCl. Optimum root activity and chlorophyll content were observed at 200 mM and 300 mM NaCl, respectively. Superoxide production increased with increasing NaCl concentration throughout the studied range. These results indicate that moderate salinity has a stimulating effect on the growth of K. foliatum. NaCl also induced leaf ultra‐structural changes. The chloroplasts and cell nuclei all displayed an elliptic shape between 0 and 300 mM NaCl. However, they appeared to be swollen between 400 and 500 mM NaCl. The mitochondria were unaffected by salinity, and all organelles remained intact under NaCl stress. These results provide insights into the mechanism of salt tolerance in K. foliatum.     

Seed germination of the halophyteSuaeda japonica under salt stress

Suaeda japonica Makino belonging to the family Chenopodiaceae, is a halophyte and grows at the shore of Ariake sea in Japan. This plant presumably possesses high salt resistant nature, thus, we examined the mechanisms of seed germination under salt stress. The seeds maintained 80% germination rates on the medium containing 0.7 M NaCl. Germination rates varied depending on salt type; the germination rates under NaCl or KCI exhibited relatively lower values than ones under sodium gluconate or potassium gluconate. This different responses for salts seemed to be as a result of the presence of Cl⁻ ions. Although very high levels of betaine (compatible solute), were kept in the seedlings grown under no salt stress, the contents gradually increased as concentration of NaCl increased. Betaine is a factor present in plants that works to alleviate the effects of excessive soil salts. It is synthesized in leaves from betaine aldehyde, and this process is catabolized by betaine aldehyde dehydrogenase (BADH). When the seedlings were cultivated on the medium without NaCl, relatively high level of BADH activity was found. The activity increased 5-fold in the seedlings grown under 0.5 M NaCl stress. Increases in betaine content and BADH activity were found during seed germination. InS. japonica, the salt stress promoted BADH activity, subsequently endogenous betaine contents were increased, and increased betaine seemed to secure seed germination under salt stress.     

Effects of NaCl and Mycorrhizal Fungi on Antioxidative Enzymes in Soybean

The effects of different concentrations of NaCl on the activities of antioxidative enzymes in the shoots and roots of soybean (Glycine max [L.] Merr cv. Pershing) inoculated or not with an arbuscular mycorrhizal fungus, Glomus etunicatum Becker & Gerdemann, were studied. Furthermore, the effect of salt acclimated mycorrhizal fungi on the antioxidative enzymes in soybean plants grown under salt stress (100 mM NaCl) was investigated. Activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were increased in the shoots of both mycorrhizal (M) and nonmycorrhizal (NM) plants grown under NaCl salinity. Salinity increased SOD activity in the roots of M and NM plants, but had no effect on CAT and polyphenol oxidase activities in the roots. M plants had greater SOD, POD and ascorbate peroxidase activity under salinity. Under salt stress, soybean plants inoculated with salt pre-treated mycorrhizal fungi showed increased SOD and POD activity in shoots, relative to those inoculated with the non pre-treated fungi.     

Increased salinity stress tolerance of Nicotiana tabacum L. in vitro plants with the addition of xyloglucan oligosaccharides to the culture medium

Xyloglucan oligosaccharides (XGOs), derived from the hydrolysis of plant cell wall xyloglucan, are a novel class of biostimulants that exert positive effects on plant growth and morphology and can enhance plant stress tolerance. The aim of this study was to determine the influence of the application of exogenous Tamarindus indica L. cell wall-derived XGOs on Nicotiana tabacum L. tolerance to salt stress by evaluating morphology, physiological, and metabolic changes. N. tabacum plants were grown in agar-gelled media for 2 mo under salt stress with 100 mM of sodium chloride (NaCl) ± 0.1 μM XGOs. The germination percentage (GP), number of leaves (NL), foliar area (FA), primary root length (PRL), and density of lateral roots (DLR) were measured. In addition, unaffected 21-d-old N. tabacum plants were treated with a salt shock (100 mM NaCl) ± 0.1 μM XGOs. Proline, total chlorophyll, and total carbonyl levels, in addition to lipid peroxidation degree and activities of four enzymes related to oxidative stress, were quantified. The results indicated that XGOs significantly improved N. tabacum plants development after exposure to salt stress. XGOs caused a significant increase in NL and PRL, promoted lateral root formation, and produced an increase in proline and total chlorophyll contents, while reducing protein oxidation and lipid peroxidation. Although the XGOs modulated the activity of the enzymes analyzed, they were not statistically different from the salt control. It was concluded that XGOs may act as metabolic inducers that trigger the physiological responses that counteract the negative effects of oxidative stress under saline conditions.

     

盐胁迫对中国柽柳幼苗生理特性的影响

以中国柽柳(Tamarix chinensis Lour.)插穗为研究材料,在装有不同土壤盐分梯度(0.4%、0.8%、1.2%、1.6%、2.0%、2.4%)的盆钵中进行扦插试验,测定柽柳扦插成活率、叶绿素含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性及丙二醛(MDA)含量等指标,研究分析柽柳扦插成活率及幼苗生理特性对盐分胁迫的响应特征。结果表明:(1)柽柳扦插成活率随盐胁迫增强逐渐降低,适合扦插繁殖的土壤含盐量低于0.8%;土壤含盐量超过0.8%后,扦插成活率太低,不适合柽柳进行扦插繁殖。(2)柽柳可通过提高叶绿素含量来适应盐胁迫,随着盐胁迫增强,柽柳扦插幼苗叶片中叶绿素含量先升高后降低,但过高的盐胁迫会破坏叶绿素的合成。(3)柽柳扦插幼苗叶片SOD和POD活性随盐胁迫增强先升高后降低,通过提高SOD和POD活性来清除细胞内多余的活性氧自由基,但活性氧自由基积累过多时,膜脂过氧化作用会破坏细胞膜的稳定性,导致抗氧化酶活性降低。(4)柽柳扦插幼苗叶片MDA含量随盐胁迫的增强先升高后降低。在土壤含盐量0.4%—1.2%范围内,MDA含量虽高于CK,但各盐分处理之间差异不显著。在含盐量为1.6%时,叶片细胞受到膜脂过氧化作用加强,MDA含量显著升高,但含盐量为2.0%时,MDA含量降低。