Accumulation of myo -Inositol in Actinidia Seedlings Subjected to Salt Stress

Seedlings of kiwifruit (Actinidia deliciosa (A. Chev.) C. F.Liang et A. R. Ferguson vardeliciosa ) and A. arguta (Sieb.et Zucc.) Planch. ex Miq. grown in hydroponic nutrient solutionswith elevated salt (MgSO₄and KCl) concentrations showed visiblesigns of stress at salt concentrations of 50 m M and above.The polyol myo -inositol accumulated in leaf tissue when thesalt was added to 15 m M or more, with increases being similarin the two species. The increase in concentration of myo -inositolwas approximately linear with rising salt. At any given saltconcentration an increase in myo -inositol was linear with timefrom application of salt.myo -Inositol concentrations increasedwithin the first 24 h of salt treatment, and declined againas quickly once the stress was removed. Sucrose also increasedwith salt stress, accumulating only once plants showed physicalsigns of stress. Accumulation of myo -inositol was negativelycorrelated to fructose and glucose. Copyright 1999 Annals ofBotany Company Actinidia arguta, Actinidia deliciosa, kiwifruit, leaf tissue, myo -inositol, salt stress, sucrose.     

Effect of Low Salt Concentrations on Nitrate Reductase and Peroxidase of Sugar Beet Leaves

Nitrate reductase, peroxidase, nitrate and sugar contents ofsugar beet leaves were increased by low NaCl concentrations.The salt was applied in the nutrient solution at concentrationsof 2, 4 and 10 mM and determinations were made at 24, 48, 72and 96 h after salt applications. Nitrate reductase was assayed both in vitro and in vivo. Inthe latter case maximum activity was attained in the first 24h for all salt concentrations after which there was a declineuntil control level was reached. Maximum nitrate content wasobserved at 48 h. It is suggested that in the first hours thesalt stimulated preferentially the flux of nitrate into theinducer nitrate pool. Maximum sugar content occurred in thefirst 24 h. This may be associated with the increase in nitratereductase activity as sugars are a source of reducing powerfor the enzyme and can supply energy and carbon skeletons forthe induction process. The salt treatments also stimulated peroxidase, maximum activitybeing reached at 48 h. Key words: Sugar beet, Sodium chloride, Nitrate reductase, Peroxidase     

The Adaptation of Dunaliella to Widely-Differing Salt Concentrations

Methods are described for adapting pure cell-lines of Dunaliella,a green unicellular alga, to grow at concentrations between0.5 M and 3.5 M NaCl. It is shown that, provided large abruptchanges are avoided, cells of the same cell-line can becomeadapted to grow over this wide range of salt concentration.Once adapted, cultures are able to continue growing at a steadyrate for many generations provided that the salt concentrationremains constant. Viability tests performed after abrupt changesin salt concentration have shown that survival is higher afterdownward than after upward changes (3.5 M to 0.5 M NaCl: 20%;0.5 M to 3.5 M NaCl: 0.01% in D. parva 19/9). When the changein concentration from 0.5 M to 3.5 M NaCl takes place over aperiod of 48 h viability approaches 100%. The time needed for100% survival for a downward change over the same range is only1.5 h. There is no evidence for a genetic difference betweencell-lines adapted to particular concentrations of salt andit is concluded that the so-called ‘halophilic’and ‘halotolerant’ strains are interchangeable.It seems likely that the difference between the two types isa matter of gene expression. Key words: Dunaliella, salt concentration, viability     

The Effect of Salinity Stress upon Protein Synthesis of Germinating Wheat Embryos

Two differently salt-sensitive wheat genotypes were imbibedin 0·4 M NaCl for 72 h or, alternatively, for 48 h andthen transferred to water. Seed germination, fresh weight andprotein synthesis in embryos were determined. The followingdifferences were found in the synthesis of in vivo [³⁵S]methionine-labelledproteins during salt imbibition: (a) a general decrease or disappearanceof polypeptides specific to the radicle emergence phase in thesalt-sensitive genotype; (b) a new synthesis of polypeptideswhich are not found during water imbibition and are common toboth genotypes; (c) a differential synthesis of polypeptidesthat are unique to each cultivar. Upon return to water, salt-inducedproteins ceased to be synthesized while proteins associatedwith an advanced germination phase were actively produced. Theseresults suggest that the expression of 'salt stress' proteinsis related to the adaptation process of seeds to salinity aswell as to the genetic constitution of a selected salt-tolerantgenotype.Copyright 1993, 1999 Academic Press Triticum durum, wheat, embryo, salt stress, protein synthesis     

Accumulation of Phenolic Compounds in the Necrotic Areas of Austrian and Red Pine Needles Due to Salt Spray

Austrian pine (Pinus nigra) and red pine (P resinosa) were testedfor needle damage caused by salt spray. Morphology and histologyof newly grown needles were compared in 3- and 6-year-old plantsgrowing in the greenhouse under controlled conditions. Pottedplants covered with plastic bags were sprayed daily either withdistilled H₂O or saturated salt solution. Needle tips, mid-partsand bases were fixed after 1, 3 and 24 h, and 3, 7 and 30 din glutaraldehyde+0·1 % caffeine for precipitation ofphenolics. Histochemical reactions and electron microscope observationsshowed that the morphological damage is linked to increasedphenolic compound deposition in mesophyll cells, beginning directlyunder the stomata. Disorganization of membranes leads to deathof the mesophyll cells, even before morphological damage isrecognizable as brown spots on the needle surface. Thus thehistochemical reactions and observations of cell structure revealeddamage earlier than macroscopic observations. Salt injury, phenolic compounds, histological damage, ultrastructural damage, salt spray, Pinus nigra, Austrian pine, Pinus resinosa, red pine     

Expression of Genes for Photosynthesis and the Relationship to Salt Tolerance of Alfalfa (Medicago sativa) Cells

The basis for the salt tolerant phenotype of a line of Medicagosativa (alfalfa) cells (HG2-N1) derived by selection from asalt sensitive line (HG2) was studied. The salt tolerant HG2-N1cell line shows eleven fold elevated chlorophyll content overthat of the parent salt sensitive HG2 cell line, with an additionaltwo fold increase in chlorophyll levels when the cells are grownin 1% NaCl. In this study, we demonstrate that the chlorophyllaccumulation and response to salt was associated with largeincreases in the two photosynthesis related mRNAs, rbcL (ribulose-l,5-bis-phosphatecarboxylase [Rubisco] large subunit) and rbcS (Rubisco smallsubunit) and a substantial increase in the activity of the holoenzyme.The salinity-induced increase in catalytically competent Rubiscoprotein in the salt tolerant cell line was highly responsiveto light and correlated with the salt tolerant phenotype. Inaddition, NaCl stimulated rbcL and rbcS mRNA and Rubisco accumulationin dark grown salt tolerant cells, indicating that salt couldsubstitute to some degree for light in stimulating increasesin specific mRNA and protein concentrations. Increased photosyntheticcompetence associated with these increased protein levels wasapparently important in contributing to the salt tolerant phenotypeof HG2-N1, since PS II electron transport inhibitors (DCMU,cyanazine) were found to significantly reduce the growth ofthis cell line in the presence of salt, but not in the absenceof salt. These results suggest that the salt-induced increasein mRNA and protein accumulation involved in photosynthesismay play a significant role in the salt tolerant capabilityof HG2-N1 alfalfa cells. (Received April 2, 1990; Accepted September 10, 1990)     

Experimental Studies on the Stability of the Cortical Microtubule Cytoskeleton in Relation to Polarity and Cell Elongation in the Coenocytic Green Alga, Chaetomorpha moniligera

The stability and ordered assembly of cytoskeletal microtubules(MTs) and the relationship between cell growth and MT cytoskeletonin the coenocytic green alga, Chaetomorpha moniligera Kjellmanwere examined. The cytoplasm of cylindrical growing cells ofChaetomorpha is covered with dense arrays of longitudinallyarranged cortical MTs which constitute the MT cytoskeleton.Seventy-five percent of MTs of the cytoskeleton disappearedwithin 4 h, with 25% remaining after 20 h following cold treatment.On terminating MT assembly with amiprophos-methyl (APM), thenumber of MTs decreased by 75% within 4 h. The remaining MTsdisappeared gradually within 24 h. The MT cytoskeleton of Chaetomorphawould thus appear to be composed of at least two kinds of MTsdiffering in stability. The MT cytoskeleton returned to normalafter treatment with APM for less than 48 h. However, this didnot occur after treatment with APM for more than 48 h, and theMT arrays became random. Cell elongation ceased completely within24 h after treatment with APM for less than 48 h but was restoredwithin 24 h after removing APM. The restoration of cell elongationwas no longer evident after removaI of APM for more than 48h. The results indicate that assembly of MTs into ordered arraysdepends on cell polarity and that in turn cell elongation isdependent on the polar-dependent arrays of MTs.Copyright 1994,1999 Academic Press Cell polarity, Chaetomorpha moniligera, coenocytic green alga, cold treatment, immunofluorescence, microtubule     

An Experimental Investigation into the Response of Some New Zealand Sand Dune Species to Salt Spray

The tolerance to salt spray of 29 species, mainly from New Zealandsand dunes, was investigated. Plants were grown in water culturein a glasshouse and subjected to overhead salt spraying at intervals.Growth rates in many species were reduced by salt spray buta significant decrease occurred only in six native herbs. However,many species showed sensitivity in leaf necrosis. Tolerant speciesincluded Scirpoides nodosa, Elymus farctus and Desmoschoenusspiralis. Ammophila arenaria, tolerant of spray as an adult,was less so when younger. There was little correlation between tolerance to salt sprayand tolerance to root salinity. Some species were tolerant toboth, e.g. S. nodosa and E. farctus, and some intolerant toboth, e.g. Wahlenbergia congesta. One species, Lupinus arboreus,was glycophytic in respect to root salt but tolerant of aerialsalt. Other species, such as Senecio elegans L. and Austrofestucalittoralis, were intolerant of salt spray but tolerant of mediumroot salinities. For some species salt spray tolerance correlated well with fielddistribution, e.g. D. spiralis and Bromus diandrus. However,some species present in semi-fixed dunes close to the sea havemuch lower tolerance than would be expected from their fieldsituation, e.g. W. congesta. This apparent inconsistency couldbe explained by the ameliorating high rainfall on the West Coast,or protection by ridges. One environmental variable alone, suchas salt spray, could not explain the field distribution formany species. Salt spray, growth rate, live leaf area, New Zealand, dune species, root salinity     

In vitro salt tolerance of cell wall enzymes from halophytes and glycophytes

The halophyte Suaeda maritima grows optimally in high concentrations(40–60% seawater) of salt. In these conditions the concentrationof salt in the apoplast of the leaves is at least 500 mM, aconcentration which severely inhibits the activity of cytoplasmicenzymes of both glycophytes and halophytes. The in vitro salttolerance of a number of cell wall enzymes was assayed in thepresence of a range of concentrations of NaCl. There was nosignificant inhibition of the activity of galactosidase, glucosidase,peroxidase or xyloglucan endo-transglycosylase extracted fromSuaeda maritima by in vitro concentrations of NaCl up to atleast 1 M. In vitro salt tolerance of cell wall enzymes wasnot restricted to the halophyte, similar enzymes from the non-halophilicrelative Kochia tricophylla, and from the glycophytes Vignaradiata and Cicer arietinum, were inhibited little, or not atall, by the same concentrations of salt. Pectin esterase wassomewhat less tolerant, but activity at 500 mM NaCl was stillgreater than at 0 mM NaCl in both Suaeda and Vigna. It is concludedthat these enzymes of the cell wall compartment are much moresalt-tolerant than cytoplasmic enzymes of higher plants. Theresults are discussed in relation to conditions thought to pertainin the apoplast. Key words: Apoplast, cell wall enzymes, halophyte, salt tolerance, Suaeda maritima     

Anatomy, Ultrastructure and Assimilate Concentrations of Roots of Citrus Genotypes Differing in Ability for Salt Exclusion

Seedlings of Rangpur lime (Citrus reticulata var. austera hybrid?)and Etrog citron (C. medica) were treated in water culture with0, 25, 50 or 100 mol m^–3 NaCl, and in sand culture with0 or 100 mol m^–3 NaCl. Leaf chloride analyses indicatedthat Etrog citron accumulated the most chloride at all levelsof salinity. The structure, ultrastructure and concentrationsof chloride and reserve assimilates of the primary root up to50 mm back from the tip were compared between genotypes andbetween salt treatments. There were no differences in root anatomy in the absence ofsalt between the two genotypes. The hypodermal cells developedlignified and suberized walls which blocked the plasmodesmataand resulted in degeneration of the cell contents. Frequentthin-walled passage cells in the hypodermis had living contentsand may represent major sites of ion uptake into the symplasm,which was connected with the stele via plasmodesmatal connectionswith and between the cortical cells. The primary endodermalcells had lignified casparian strips and plasmodesmata in othercell wall areas. These connections were blocked by secondarysuberization of all except the endodermal passage cells oppositethe protoxylem arcs. Suberization of the hypodermis and endodermis and the appearanceof granular deposits in the vacuole occurred closer to the roottip of both genotypes after treatment with 100 mol m^–3NaCl. Levels of starch and triglyceride in 10 mm serial segmentswere similar between roots of control and salt-treated plantsand increased with distance back from the root tip. Chlorideconcentrations increased with salt treatment but values (ona tissue water basis) were similar between genotypes and betweenthe apical and proximal ends of the root. Reducing sugar concentrationsdecreased with salt treatment to a similar extent in roots ofboth genotypes. Key words: Citrus, Roots, Salinity     

Ion Absorption and Allocation of Carbon Resources in Excised Pea Roots Grown in Liquid Medium in Absence or Presence of NaCl

Freshly excised pea roots (Pisum sativum cv. Alaska) when transferredto growth medium (130 mOsm) or growth medium containing salt( 370 mOsm) suffer an initial osmotic shock and lose water.Contol roots tended to accumulate potassium, particularly inthe apical zone, while those exposed to NaCl accumulated mainlysodium, potassium accumulation being depressed. Exposure tosalinity for 6 d caused increases in root protein, cellulose,uronic acid and lignin content per cell. In roots supplied with¹⁴C-glucose for 24 h immediately after excision there was littledifference in uptake of glucose and in its use in respirationbetween control and salt treated roots. However, there werenoticeable differences in incorporation of labelled carbon intoseveral cell fractions, and particularly into the cellulosefraction in the upper parts of the root. When roots were grownfor six days in culture before being supplied with [¹⁴C]glucose,uptake per root was greater in the 120 mM NaCl treatment, andthe fraction diverted to respiration was decreased by salinity.On a per cell basis incorporation into soluble starch, uronicacid and cellulose fractions was increased in the salt treatedroots. The data obtained are in accord with the previous findingsand are suggestive of increased synthesis of cell wall materials.No conclusion could be drawn as to whether the changes describedare of adaptive value. Pisum sativum L. cv. Alaska, root culture, salinity, osmoregulation, cell wall     

Salt Stress Induced Nuclear and DNA Degradation in Meristematic Cells of Barley Roots

Root growth of barley (Hordeum vulgare L., cv. Akashinriki)was inhibited by 200 raM NaCl, when 1 mM CaCl₂ was present inthe hydroponic culture solution. Increasing the CaCl₂ up to10 mM partially prevented this inhibition. However, inhibitionalso occurred with 100 mM NaCl in the presence of 0.1 mM CaCl₂.The nuclei of meristematic cells in roots in which growth hadbeen inhibited by salt stress were studied after staining withDAPI (4',6-diamino-2-phenylindol). Nuclear deformation of thecells occurred with 12 h of salt stress with 500 mM NaCl, andwas followed by degradation. The nuclear degradation was alsoobserved when the roots were exposed to more than 300 mM NaClfor 24 h. Biochemical analysis revealed that nuclear degradationwas accompanied by apoptosis-like DNA fragmentation. The intracellularmechanisms of nuclear degradation in cells after salt stressare discussed. ¹Emertius professor, Okayama University.     

Cell Cycle Changes in the Shoot Apex of Silene coeli-rosa During the Second and Third Days of Floral Induction

The cell cycle in Silene coeli-rosa shoot apices was measuredto test whether or not early components of the floral stimulus,produced during the 2nd and 3rd long days (LD) of an inductiveLD treatment, resulted in an increase in the duration of G₂phase in constant 20–24 h cell cycles. Plants were grownat 20°C in short days (SD) of 8 h light and 16 h darknessfor 28 d (day 0). Starting on day 0, plants were given SD or3 LD each comprising an identical 8 h day and 16 h photo-extension,or 3 dark-interrupted (d.i.) non-inductive LD, interrupted at1700 h of each day with 1 h of darkness. The cell cycle (percentagelabelled mitoses method) and changes in cell number were determinedin the shoot apical meristem. During days 1–2 of the SDtreatment, the cell cycle and mean cell generation time (MCGT)was 18 and 32 h, respectively, giving a growth fraction of 56%.During days 2–3, the cell cycle and MCGT shortened to15 and 23 h, respectively (growth fraction = 65%). During days1–2 of the LD and d.i. LD treatments, cell cycles andMCGTs were 9–10 and 27–29 h, respectively, resultingin smaller growth fractions (about 33%). Thus, shortened cellcycles and altered growth fractions occurred regardless of whetheror not the treatment was inductive. The LD treatment resultedin a marked shortening of G₁ and, to a lesser extent, S-phase,whilst G₂ remained constant. These changes were consistent withincreases in the proportion of cells in G₂ during the photoextensionof each LD which were suppressed during the comparable periodsof the d.i. LD treatment. The latter treatment resulted in eachphase occupying virtually identical proportions of the cellcycle as in the SD treatment. Thus, the unique cell cycle responsesto the initial part of the inductive LD treatment were increasesin the proportion of cells in G₂ coupled with G₁ and G₂ beingof similar duration. Cell cycle, mean cell generation time, shoot apex, Silene coeli-rosa     

The Mineral Ion Relations of Mangroves I. Root Cell Compartments in a Salt Excluder and a Salt Secreter Species at Low Salinities

The efflux kinetics of ²²Na and ³⁶Cl from 48 h pre-loaded Rhizophoramangle and Avicennia germinans mangrove seedling roots grownin low salinity conditions have been documented. Estimates ofthe percent isotope distributed throughout the whole plant,the uptake rates, the amounts calculated to be present in theroot cell compartments, and the rate constants and half-timesof efflux were made. Little to no ²²Na or ³⁶Cl was noted asbeing distributed to leaf tissue in the red mangrove, Rhizophoramangle, substantiating its characterization as a non-secreting,salt excluding species. Significantly greater net uptake ratesfor Na and Cl were observed for the black mangrove, Avicenniagerminans, a non-excluding, salt-secreting species. The typicalintracellular compartmentation found in higher plant root tissueswas noted to be present in these mangroves and the efflux kineticsclearly indicated that Na and Cl were handled differentiallyby these compartments and their membranes. Rhizophora manglerevealed greater apparent contents of Na than Cl in both thevacuolar and cytoplasmic phases (5.4 and 7.7 fold greater, respectively).The inverse was found for Avicennia germinans, with the apparentvacuolar compartment amount for Cl being 4.6 times greater thanthe Na amount and the cytoplasmic Cl amount was 1.5 times morethan Na contents. The rate constants of efflux showed a patternof significantly greater Na than Cl efflux from each respectivered mangrove compartment, while the black mangroves had a patternof greater efflux rates of Cl than Na in each compartment. Thehalf-times of efflux for Na from the black mangrove cytoplasmiccompartment were 3.5 times longer, while vacuolar half-timesof efflux were near unity for Na and Cl. The results are discussedin terms of Scholander's model of salt secreting and salt excludingspecies. (Received March 26, 1984; Accepted June 26, 1984)     

Salt Tolerance in the Halophyte Suaeda maritima L. Dum. Growth, Ion and Water Relations and Gas Exchange in Response to Altered Salinity

Clipson, N. J. W. 1987. Salt tolerance in the halophyte Suaedamaritima L. Dum. Growth, ion and water relations and gas exchangein response to altered salinity.—J. exp. Bot. 38: 1996–2004. Shoot and root fresh and dry weights and shoot sodium, chlorideand potassium contents were measured and shoot relative growthrates calculated in seedlings of Suaeda maritima over a periodof 11 d following a raising of culture solution salinity from0 to 200 mol m₃– NaCl. Growth, growth rates and sodiumand chloride contents, as compared to plants growing in theabsence of salt were increased whilst potassium contents declined.Shoot sodium accumulation rate and the rate of transport ofsodium from root to shoot, osmotic potential, and rates of photosynthesisand transpiration were also measured for up to 72 h after transferof plants originally growing at 0 and 200 mol₃– NaCl to200 and 400 mol m₃– NaCl respectively. Ion uptake andtransport rates were maximal 6-12 h after transfer and thendeclined to new steady-state levels within 48 h; osmotic potentialswere lowered over a 72 h period on average by approximately1·0 MPa; and after 9 h photosynthetic and transpirationrates were reduced by about 20percnt; and 30% respectively.Results are discussed in terms of the ability of halophytesto adjust to fluctuating salinity and to salt tolerance mechanismsin general. Key words: Suaeda maritima, salinity, gas exchange, growth, ion and water relations     

Plasma membrane isolation from freshwater and salt-tolerant species of Chara: antibody cross-reactions and phosphohydrolase activities

Plasma membranes were isolated using the aqueous polymer two-phasepartition method from the algae Chara corallina and Chara longifolia,algae which differ in their ability to grow in saline environments.Enrichment of plasma membrane and depletion of tonoplast relativeto the microsomal fraction was monitored using phosphohydrolaseassays and crossreactions to antibodies raised against higherplant transporters. Antibodies to the vacuolar ATPase and pyrophosphatasecross-reacted with epitopes in the microsomal fraction, butshowed little affinity for the plasma membrane fraction. Pyrophosphataseactivity also declined in the plasma membrane fraction relativeto the microsomal fraction. The V-type H⁺ -ATPase activity,sensitive to nitrate or bafilomycin, was low in both fractions,though the cross-reaction to the antibody was reduced in theplasma membrane fraction. By contrast, the antibody recognitionof a P-type H⁺-ATPase amino acid sequence from Arabidopsis didnot occur strongly in the anticipated 90–100 kDa range.While there was enhanced recognition of a polypeptide at around140 kDa in the plasma membrane fraction, salt treatment of Charalongifolia resulted in plasma membrane fractions with reducedamounts of this epitope, but no change in vanadate-sensitiveATPase activity, suggesting that it does not represent the onlyP-type ATPase. Microsomal membranes from saltadapted C. longifoliahave higher reactivity with the antibody to the tonoplast ATPase. Key words: Chara, plasma membrane, salt tolerance, ATPase     

Effect of a Light-break on the Growth and Development of Epilobium hirsutum and Lythrum salicaria in Short Photoperiods

Plants of Epilobium hirsutumand Lythrum salicaria were grownin light for 9 h per day with the following adjustments of lightand dark periods. (a) Photoperiod of 9 h and an uninterrupteddark period of 15 h, (b) photoperiod of 8 h and 1 h of lightof the same intensity in the middle of the dark period. Plants of both the species receiving the 9 h continuous photoperiodremained dwarfed in habit. In the 8 + 1 h light-break treatment,Epilobium showed no response but Lythrum responded by a completedevelopmental shift from a short-day to a long-day form. Leafexpansion, stem elongation, dry matter production and flowerinitiation were promoted. In Epilobium, the pattern of total dry weight distribution toleaves, stem and roots remained similar in the two treatmentsbut in Lythrum light-break treatment caused more dry matterto be diverted to stems at the expense of leaves. These changesin morphogenetic pattern of differentiation were accompaniedby physiological changes as the chlorophyll content and therespiration rates were also higher.