The importance of NaCl concentration in a chemically defined medium for the development of bovine oocytes matured and fertilized in vitro

Bovine oocytes matured and fertilized in vitro were cultured in a chemically defined medium (modified Tyrode's solution) without glucose. When different concentrations of NaCl were added to the medium, the proportions of embryos developed to the >/=8-cell, morula and blastocyst stages 96, 144 and 192 h post insemination, respectively, were significantly higher at 89 to 114 mM than 64 to 76 and 126 to 139 mM NaCl. A high proportion (28%) of blastocyst-stage embryos 192 h post insemination was obtained at 89 mM NaCl. When calculated osmolarity in the medium with 64 mM NaCl was varied by adding D-sorbitol, significantly higher proportions of morula-stage embryos were obtained at 265 to 315 mOsm (27 to 38%) than 215 (9%) and 365 (2%) mOsm, but the development to the blastocyst stage was difficult at any osmolarities (215 to 365 mOsm) tested. In the medium with a fixed osmolarity (315 mOsm) but with different concentrations (64 to 114 mM) of NaCl, there were no differences in the proportions (29 to 33%) of morula-stage embryos among different NaCl concentrations. However, significantly higher proportions of embryos developed to the blastocyst stage at 89 to 101 mM (22 to 23%) than 64 to 76 (0 to 9%) and 114 (11%) mM NaCl. When Cl- concentration in the medium with 64 mM NaCl was adjusted by adding choline chloride, significantly higher proportions of embryos developed to the morula stage at 97 to 122 mM (32 to 40%) than 72 (6%) and 147 (2%) mM Cl-, but few embryos developed to the blastocyst stage at any Cl- concentrations (72 to 147 mM) tested. In the medium with 64 or 114 mM NaCl and each with 2 different Na (+)K (+) ratios, there were no differences in the proportions of morula- and blastocyst-stage embryos between different Na+ K+ ratios (31 and 39 at 64 mM NaCl, and 39 and 47 at 114 mM NaCl) at each NaCl concentration. When glucose was added to the medium with 89 mM NaCl 120 h postinsemination, there were no significant differences in the proportions (40 to 48%) of morula-stage embryos 144 h post insemination among different concentrations (0 to 6.95 mM) of glucose. The proportion (33%) of blastocysts 192 h post insemination at 2.78 mM glucose was significantly higher than the values at 0 (22%), 5.56 (19%) and 6.95 (15%) mM but not different compared with the values at 1.39 (23%) and 4.17 (28%) mM. In conclusion, NaCl concentration in a defined medium is one of the most important factors for the development of bovine embryo to the blastocyst stage, but the development of embryos up to the morula stage is also regulated by osmolarity and/or Cl-concentration.     

The role of salinity in seed maturation of the euhalophyte Suaeda salsa

Controlled conditions were used to investigate how salinity maintains the salt tolerance of seeds and seedlings of the euhalophyte Suaeda salsa. Seeds were harvested from S. salsa plants that had been treated with 1 or 500 mM NaCl for 113 days in a glasshouse. The results showed that high salinity (500 mM NaCl) increased chlorophyll concentration and oxygen production in embryos of maturing seeds. At 500 mM NaCl, the phosphatidylglycerol and sulfoquinovosyldiacylglycerol levels and the digalactosyldiacylglycerol/monogalactosyldiacylglycerol ratio were higher in young seedlings derived from seeds whose source plants were cultured in 500 mM rather than in 1 mM NaCl. When seeds were incubated with 600 mM NaCl, the conductivity and malondialdehyde concentration in the embryos was greater if the source plants had been cultured in 1 mM rather than in 500 mM NaCl. The opposite pattern was evident for seedling survival and shoot weight. In conclusion, salinity during seed maturation may increase the salt tolerance of seeds and seedlings by increasing the oxygen production in the embryos of the maturing seeds and by changing the lipid composition of membranes in the seedlings.     

High salinity induces dehydrin accumulation in Chenopodium quinoa Willd. cv. Hualhuas embryos

Background and Aims

Chenopodium quinoa can grow at altitudes of 3,600–4,000 masl and is adapted to the highly arid conditions typical of the salty soils in the South American Altiplano, with less than 250?mm of annual rain and temperatures below 0°C. The aim of the study was to investigate the effect of salinity on the dehydrin content of mature embryos harvested from salt-stressed Chenopodium quinoa cv. Hualhuas plants grown at 100 to 500?mM NaCl. To date, no studies exist on the dehydrins of seeds from salt-stressed plants, although dehydrins in the root, stems and leaves have been reported as an adaptation to water deficit produced by salinity.

Methods

Dehydrin-like protein detection was carried out with an antiserum raised against a highly-conserved lysine-rich 15-amino acid sequence known as the K-segment, which is capable of recognizing proteins immunologically related to the dehydrin family.

Results

Dehydrins were analyzed in embryos by both western blot and in situ immunolocalization. Western blot analysis detected at least four dehydrins (55, 50, 34, and 30?kDa) in seeds harvested from quinoa salt-stressed plants treated under a wide range of salinities. The 30?kDa dehydrin increased its accumulation in both 300 and 500?mM NaCl growth conditions as revealed by densitometric analyses. Dehydrin subcellular localization was mostly nuclear at 500?mM of NaCl. A phosphatase treatment of protein extracts caused a mobility shift of the 34 and 30?kDa dehydrin bands suggesting a putative modulation mechanism based on protein phosphorylation.

Conclusions

We propose that these novel observations regarding dehydrin accumulation, subcellular localization and phosphorylation state are related to the high salt stress tolerant phenotype previously reported on this cultivar.     

Excess boron reduces polyphenol oxidase activities in embryo and endosperm of maize seed during germination

The effects of increasing concentrations of boron (0, 0.1, 1, 10 and 20 mM) as boric acid on the rate of germination and polyphenol oxidase activities in embryo and endosperm tissues of maize seeds (Zea mays L. cv. Arifiye) were studied. The germination percentage of maize seeds was not affected by boron concentrations up to 10 mM, and decreased by 20 mM. Distilled water and lower boron concentrations (0.1 and 1 mM) increased polyphenol oxidase activities at the beginning of germination up to 12 h whereas its excess levels (10 and 20 mM) decreased polyphenol oxidase activities in embryos and endosperm during germination. Polyphenol oxidase activities with o-diphenolic substrates (caffeic acid, catechol and dopa) were found to be higher than with a monophenolic substrat (tyrosine) in both embryos and endosperms. Further, caffeic acid oxidizing polyphenol oxidase was found to show more activity in embryos of the seeds germinating in distilled water when compared to other substrates.     

Effect of salinity on seed germination,ion content and photosynthesis of cotyledons in halophytes or xerophyte growing in Central Asia

Aims We investigated the impact of salinity on seed germination, chlorophyll content, chloroplast structure and photosynthesis of the green embryos in desiccated seeds of the xerophyte Haloxylon persicum, xero-halophyte Haloxylon ammodendron and euhalophyte Suaeda physophora.Methods Seeds of H. persicum, H. ammodendron and S. physophora were collected from natural environment in Fukang, Xinjiang province. Pretreatment with 700 mM NaCl was carried out to stimulate the natural 'seed priming'; we analyzed the joint effect of salinity and different species on germination physiology and cotyledonal structure and photosynthetic function changes during germination and recovery stage.Important findings We found that seeds did not suffer ion toxicity for the two halophytes H. ammodendron and S. physophora, as evidenced by the high final germination after ungerminated seeds pretreated with 700 mM NaCl were transferred to distilled water, but the final germination of the xerophyte H. persicum was significantly lower than that of control. The Na + concentration in embryos increased under salinity for all species, while K + concentration decreased by salinity only for H. persicum and H. ammodendron, i.e. the concentration of K + in embryos of H. persicum and H. ammodendron decreased by 36% and 46%, respectively. For all species, whether dry intact seeds or cotyledons of dry seeds imbibed in deionized water and NaCl solution, had high chlorophyll content. Treatment with NaCl also caused chloroplast thylakoids to swell and chlorophyll content to decrease in seeds of H. persicum, but no significant change was observed in the more salt-tolerant species S. physophora and H. ammodendron. Fluorescence measurement showed that 700 mM NaCl decreased the Fv/Fm ratio of cotyledons in seeds for all species, especially for H. persicum and H. ammodendron. Photosynthetic oxygen releasing was detected from the seeds that were moistened with distilled water and 700 mM NaCl for 6 or 24 h and from the seeds that were initially moistened with 700 mM NaCl in darkness for 10 days, then transferred to distilled water for another 6 and 24 h. The results indicated that the chlorophyll in cotyledon of desiccated seed had photosynthetic function in early germination stage, even under high-saline condition. In addition, the photosynthesis of chlorophyll in the embryonic cotyledons of desiccated seeds during germination was similar to that in leaves of young seedlings for all species. In conclusion, the chloroplasts of the two halophytes were more salt resistant compared with the xerophyte H. persicum. The photosynthetic function of chlorophyll in cotyledons of mature seeds may be ecologically important for seedling development in early stage for plants growing in extremely saline or arid environments.     

Changes in Sugars, Sucrose Synthase Activity and Proteins in Salinity Tolerant Callus and Cell Suspension Cultures of Brassica oleracea L.

Salt tolerant callus and cell suspension cultures of Brassica oleracea L. var. botrytis were obtained by the selection of cells from cultures growing in medium supplemented with 85, 170, and 255 mM NaCl. Salt adapted calli and cell suspensions differed in their RNA and protein concentrations. These concentrations tend to diminish in calli and increase in cell suspensions, both at one or three weeks periods of growth in NaCl. Contents of sucrose and reducing sugars, however, accumulate similarly both in calli and cell suspensions after NaCl treatments. The activity of sucrose synthase was higher in salt adapted cells than in controls. Calli exposed to 255 mM NaCl for six months synthesized a 27 kDa polypeptide, while a 13 kDa polypeptide present in control conditions was absent under salinity. Several high molecular mass polypeptides (> 200 kDa) were visualized in control calli and at moderate salt concentrations, when conditions of the gel were modified.     

Accumulation of ions during seed development under controlled saline conditions of two Suaeda salsa populations is related to their adaptation to saline environments

Controlled conditions were used to investigate the relationship between ion distribution in developing seeds of two Suaeda salsa populations and seed germination and seedling emergence. Seeds were harvested from S. salsa plants that had been treated with 1 or 400 mM NaCl for 122 (saline inland population) or 135 days (intertidal zone population) in a glasshouse. Germination and seedling emergence were evaluated under salinity. In both populations, more ions were accumulated in the pericarps of plants cultured in 400 mM NaCl than in 1 mM NaCl. Pericarps accumulated much higher ion concentrations in the intertidal zone population than in the saline inland population, while the opposite trend occurred for ion accumulation in the embryos. Seeds of plants from the intertidal zone population germinated more rapidly than those from plants of the saline inland population, regardless of the NaCl concentration during seed germination. However, seedling emergence under high salinity was lower with seeds from the intertidal zone population than with seeds from the saline inland population. In conclusion, S. salsa in the intertidal zone employs superior control of ion compartmentalization in the pericarps to tolerate salinity but requires a minimal level of ions in embryos to ensure seedling establishment in highly saline environments. This indicates that euhalophytes require salts during the mature seed stage to maintain seed viability and to ensure seedling emergence and population establishment.     

Protein Profiles of Somatic Embryos and Regenerated Plants from NaCl Selected and Control Cultures of Orchardgrass

The protein profile of cells of control somatic embryos was compared to that of embryos that have become selected and maintained on 200 mM NaCl in order to detect salt inducible proteins. Two proteins (60 and 51.5 kDa) were more abundant in the selected embryos and one protein with molecular mass 18 kDa was unique to the selected embryos. Enhanced content of 27 kDa protein was observed in all somatic embryos indicating its involvement in the embryonal state. Similar pattern of salt inducible proteins in selected somatic embryos and the plantlets regenerated from such embryos was found. This revised version was published online in July 2006 with corrections to the Cover Date.     

Growth and Fatty Acid Composition of Borage (Borago officinalis L.) Leaves and Seeds Cultivated in Saline Medium

The effects of salinity on growth and fatty acid composition of borage (Borago officinalis L.) leaves and seeds grown in hydroponic medium were investigated. Three different levels of NaCl (25, 50, and 75 mM) were applied. The first results showed that salinity significantly reduced plant growth by 56.5 % at 75 mM compared with the control, suppressed seed yield at 50 and 75 mM, and increased lipid peroxidation. Raising NaCl concentrations led to an important decrease in total fatty acid (TFA) content by 77 % at 75 mM NaCl. Moreover, the polyunsaturated fatty acid (PUFA) content decreased, whereas the saturated fatty acids increased with respect to increasing salinity. The 25 mM NaCl level did not modify the fatty acid composition of seeds and their contents.     

Purification and characterization of the sea urchin embryo hatching enzyme

The sea urchin hatching enzyme provides an interesting model for the control of gene expression during early development. In order to study its properties and developmental regulation, the hatching enzyme of the species Paracentrotus lividus has been purified. The fertilization envelopes of the embryos were digested before hatching by a crude culture supernatant previously made. The enzyme was then solubilized by 1 M NaCl and 0.5% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate and purified by hydrophobic chromatography on Procion-agarose. A 470-fold increase in specific activity was obtained. The kinetic parameters of the proteolytic activity using dimethylcasein as substrate are: Km = 120 micrograms x ml-1, Vm = 200 mumol x min-1 x mg-1, and kcat = 180 s-1 at 500 mM NaCl, 10 mM CaCl2, pH 8.0, at 35 degrees C. The purified enzyme is highly active on fertilization envelopes: at 20 degrees C and 500 mM NaCl, 10 mM CaCl2, pH 8.0, 100 ng of enzyme completely denudes embryos in about 20 min under standard conditions. The molecular mass of the enzyme was estimated as 57 kDa by gel filtration, 51 kDa by gel electrophoresis, and 52 kDa by amino acid analysis. The hatching enzyme was shown to be a glycoprotein which autolyzes to a 30-kDa inactive form. Antibodies raised against the 51- or 30-kDa forms reacted with both these forms. Immunoblotting experiments showed that the hatching supernatants contain important amounts of the autolyzed species.     

Effects of Dormancy Regulating Chemicals on Innate and Salinity Induced Dormancy in the Invasive <Emphasis Type="Italic">Prosopis juliflora</Emphasis> (Sw.) DC. Shrub

The effect of different types and concentrations of dormancy regulating chemicals (DRCs) on innate and induced dormancy was evaluated under optimal germination conditions in the invasive Prosopis juliflora shrub. Lower concentrations of gibberellic acid (0.3 mM) and kinetin (0.05 mM) were more effective in enhancing germination % and rate at higher concentrations of NaCl, but the reverse was true for thiourea. None of the DRCs alleviated innate dormancy of P. juliflora. Germination % and rate decreased as salinity increased. Percent final germination of non-treated seeds was significantly reduced at 500 mM NaCl and virtually inhibited in 600 mM NaCl. Germination reduction in 500 mM NaCl was not alleviated by any of the DRCs, but inhibition induced at 600 mM NaCl was partially alleviated by all the DRCs. Gibberellic acid had a significantly greater effect than kinetin in alleviating germination inhibition. For restoration of saline soil through the use of P. juliflora, results suggest using DRCs, particularly gibberellic acid and thiourea, as a preseeding treatment can overcome the problem of reduced germination.     

Esterase and lipase activity in Jatropha curcas L. seeds

Two new esterases (JEA and JEB) and a lipase (JL) were extracted from the seeds of Jatropha curas L. Lipase activity was only found during germination of the seeds and increased to a maximum after 4 days of germination. All enzymes were found to be most active in the alkaline range at around pH 8 and the purified (fractionated precipitation with ethanol and gel filtration) esterases were very stable at high temperatures. The molecular weight (SDS-PAGE) of both esterases was determined to be 21.6-23.5 kDa (JEA) and 30.2 kDa (JEB) and the isoelectric point was 5.7-6.1 for esterase JEA and 9.0 for esterase JEB. Most ions caused a negative influence on the activity of both esterases. Using p-nitrophenyl butyrate as a substrate JEA showed a K(m) of 0.02 mM and a v(max) of 0.26 micromol mg(-1) min(-1). Under the same conditions JEB showed a K(m) of 0.07 mM and a v(max) of 0.24 micromol mg(-1) min(-1). Both esterases hydrolyzed tributyrin, nitrophenyl esters up to a chain length of =C4 and naphtylesters up to a chain length =C6. In transesterification reactions, JL was found to be most active at very low water activities (0.2) and in high water activities, the lipase hydrolysed triglycerides into conversions above 80%. The lipase hydrolysed both short chain and long chain triglycerides at about the same rate but was inactive on alpha-methylbenzyl acetate. JL is a potentially useful biocatalyst in the hydrolysis of triglycerides in organic solvents.     

Effects of Salinity and Temperature on Seed Germination in a Japanese Endangered Halophyte Triglochin maritimum (Juncaginaceae)

Triglochin maritimum (Juncaginaceae). Germination tests were carried out at three salinity levels (0,200,400 mM NaCl in which seeds were exposed to increasing- or decreasing temperatures. Effects of moist-chilling pretreatment (stratification) in 0,200 and 400 mM NaCl on seed germination at 0,200 and 400 mM NaCl, respectively, were also examined. Under the highest salinity condition (400 mM NaCl), no germination was observed. The seeds germinated very well (88%) in fresh-water after 5-month moist-chilling pretreatment. Longer moist-chilling pretreatment resulted in higher germination percentages. Moist chilling pretreatment in 0,200 and 400 mM NaCl for 5 month enhanced germination percentages to 10% (in 400 mM NaCl) –88% (in 0 mM NaCl). Received 10 December 1998/ Accepted in revised form 8 October 1999     

Effect of salinity on seed germination and seedling growth of the halophyte Suaeda japonica Makino

Seed germination and seedling growth of the annual halophyte species Suaeda japonica Makino were investigated in response to variable salinity of sediment pore water. The germination percentage of S. japonica’s soft brown seeds, which are dominant among dimorphic seeds, decreased with an increase in salinity, although germination was still observed at 1200‐mM NaCl concentration. The germination percentage and germination speed observed in April were higher than those observed in December when treated with sediment water with 400–1200 mM of NaCl concentrations. These data suggest that S. japonica seedlings could be established on sediments that experience high temperatures. Germination recovery of S. japonica seeds transferred from 600‐mM NaCl containing sediment (seawater equivalent) was lowest among 0–1200‐mM NaCl treatments, implying the low tolerance of seawater conditions of S. japonica seeds. Seeds germinated in 900‐ to 1200‐mM NaCl medium showed poor growth, but survived, in hypersaline conditions, and exhibited improvement in growth upon transfer to lower salinity.     

Alleviation of Salinity Stress During Germination in Brassica Juncea by Pre-sowing Chilling Treatments to Seeds

Influence of pre-sowing chilling treatments to seeds on seed germination and accumulation of osmotics in Indian mustard [Brassica juncea (L.) Czernj & Cosson] under NaCl stress was investigated. Germination was 100 % in water soaked non-chilled (control) seeds upto 100 mM NaCl, whereas it was upto 200 mM NaCl in chilling treated seeds. Pre-sowing chilling treatments to seeds for 5, 10 and 15 d also enhanced the dry mass of 6-d-old seedlings, and concentrations of saccharides and amino acids. The alleviation of NaCl stress by pre-sowing chilling treatments was also associated with decrease in Na⁺ and proline accumulation and a slight increase in K⁺ and Ca²⁺ contents of seedlings. This revised version was published online in June 2006 with corrections to the Cover Date.     

Changes in nitrogen content and protein profiles following <Emphasis Type="Italic">in vitro</Emphasis> selection of NaCl resistant mung bean and tomato

Mung bean and tomato were in vitro selected on media containing 0, 25, 50, 100 and 150 mM NaCl. Two types of media (hormone supplemented media, CB and hormone free media, MS) were used for mung bean using cotyledon explants whereas two types of explants (cotyledons and shoot apices) were used for tomato on MS media. Total-N, protein content, nitrite reductase (NiR) activity and protein protein profiles were checked in selected plants and compared to original non selected ones. NaCl at low concentrations slightly increased total-N in shoots and roots of in vitro selected mung bean and tomato whereas higher concentrations induced significant reductions. Similar increases in protein content were detected at lower concentrations with no significant effects thereover. On the contrary, NaCl gradually inhibited NiR activity. Similar responses of total-N, protein and NiR activity, but with greater magnitudes, were detected in original plants. In addition, NaCl significantly reduced dry weights of shoots and roots of either in vitro selected or, in particular, original intact plants. Moreover, electrophoresis (SDS-PAGE) of protein from shoots of either in vitro selected or intact plants showed that NaCl induced new protein bands while some others were concomitantly disappeared. The induction of one or more of the 86.4, 79, 77.6, 77 and 71.5 kDa bands following in vitro selection and/or the disappearance of the 86 kDa band from intact plants seemed necessary for mung bean resistance. Also, the presence of 86.2 kDa band and/or the loss of the 85.8 and 57.5 kDa bands might be included in tomato resistance. Of these induced bands in mung bean selected on CB media, only two bands were detected in plants selected on MS media. In tomato, two bands lost following selection from cotyledons but only one band lost following selection from shoot apices. These changes in protein pattern therefore might serve as adaptive regulators for resistance to NaCl.     

An increase in pectin methyl esterase activity accompanies dormancy breakage and germination of yellow cedar seeds

Pectin methyl esterase (PME) (EC 3.1.1.11) catalyzes the hydrolysis of methylester groups of cell wall pectins. We investigated the role of this enzyme in dormancy termination and germination of yellow cedar (Chamaecyparis nootkatensis [D. Don] Spach) seeds. PME activity was not detected in dormant seeds of yellow cedar but was induced and gradually increased during moist chilling; high activity coincided with dormancy breakage and germination. PME activity was positively correlated to the degree of dormancy breakage of yellow cedar seeds. The enzyme produced in different seed parts and in seeds at different times during moist chilling, germination, and early post-germinative growth consisted of two isoforms, both basic with isoelectric points of 8.7 and 8.9 and the same molecular mass of 62 kD. The pH optimum for the enzyme was between 7.4 and 8.4. In intact yellow cedar seeds, activities of the two basic isoforms of PME that were induced in embryos and in megagametophytes following dormancy breakage were significantly suppressed by abscisic acid. Gibberellic acid had a stimulatory effect on the activities of these isoforms in embryos and megagametophytes of intact seeds at the germinative stage. We hypothesize that PME plays a role in weakening of the megagametophyte, allowing radicle emergence and the completion of germination.     

Selection in Vitro for NaCI-tolerance in Vitis rupestris Scheele

Suspension cultures were made from the vegetative tissues ofanthers of the grape rootstock Vitis rupestris Scheele cv. RupestrisSt George. Cell lines and somatic embryos were isolated whichgrew in agitated liquid media containing up to 150 mM NaCl.Exposure to NaCl (50–100 mM) promoted outgrowth of secondarysomatic embryos but inhibited development of plantlets. Withinsomatic embryos the cotyledon and hypocotyl tissues were moretolerant of NaCl than root tissues. Elongation of the radiclein embryos in the presence of NaCl (50 mM) was associated withnecrosis and death of root tissue but hypocotyls and cotyledonsremained viable and continued to produce callus and secondarysomatic embryos. The usefulness of the cell culture approachin selecting for salt tolerance is questioned.     

Changes in molecular species of triacylglycerols in developing cotton seeds under salt stress

Cotton plants (Gossypium hirsutum L. cv. S4) were grown and irrigated with increasing salt concentrations: 0, 50, 100 and 150 mM NaCl. Lipids from developing seeds, at stages of 3, 4, 5 and 6 weeks after anthesis, were analysed. High salt dose (150 mM NaCl) affected triacylglycerol levels drastically at the end of ripening. HPLC analysis of triacylglycerols in control seeds showed ten molecular species. Palmitodilinolein (PLL) is the major fraction, representing about 25% of total triacylglycerols. The levels of trilinolein (LLL), palmitolinoleo-olein (PLO) and oleodilinolein (OLL) molecules varied between 13 and 16% of the total. The other molecular species: triolein (OOO), dioleopalmitin (POO) and dipalmito-olein (POP) are minor and do not exceed 5%. Moreover, the composition of triacylglycerol molecular species was almost constant during developing stages of control seeds. However, amounts of triacylglycerol species LLL, OLL and palmitodilinolein (PLL), as expressed on a dry-matter basis (mg/g), decreased severely under the highest NaCl concentration, while contents of triacylglycerol species OOO, POO, POP were unchanged. These findings confirm our previous results, which indicated that the amount of linoleic acid in cotton seeds was reduced by salt stress and that the amount of oleic acid remained unchanged.     

Alleviation of Seed Dormancy in the Desert ForbZygophyllum simplexL. from Pakistan

Zygophyllum simplexL. is a succulent annual that grows on thecoastal and inland saline flats around Karachi, Pakistan. Theseeds are moderately salt tolerant during germination. GerminationofZygophyllum simplexseeds under various salinity, proline,betaine, GA and kinetin treatments was determined. Proline (0.1and 1 mM) and betaine (0.1 and 1 mM) alleviated the innate dormancyof seeds, and germination reached 60–70% compared to 12%in the control set. At low salinity compatible osmotica alleviatedsome effects of salinity, but at higher NaCl concentrationsboth proline and betaine were ineffective. Gibberellic acid(0.3 and 3 mM) and kinetin (0.05 and 0.5 mM) substantially alleviatedboth innate as well as salinity-induced seed dormancy. At highersalinity (125 mM), low concentrations of kinetin (0.05 mM) andhigh concentrations of GA (3 mM) were more effective. GA completelyalleviated the effect of salinity at all concentrations used. Betaine; desert; dormancy; forb; GA; germination; halophyte; kinetin; proline; seeds; Zygophyllum simplex