Intra-individual heterostyly in Atriplex halimus L. (Amaranthaceae)

Classically, the Atriplex halimus L. female flower is perianthless, has two bracteoles, one ovary and one style. Considering bracteoles, one can distinguish, among the representatives of two Tunisian populations, three phenotypes of female flowers, each of them being distributed in three types according to the style length. Male flowers produce three types of pollen. This is the only known example of intra-individual heterostyly in unisexual flowers. These results question the classical concept of individual. The authors discuss a possible process implying indirectly hybridisation and introgression between A. halimus and A. nummularia.     

On the osmoregulation in Atriplex hymenelytra (Torr.) Wats. (Chenopodiaceae)

Summary Aspects of osmoregulation were studied in leaves of irrigated and nonirrigated plants of Atriplex hymenelytra (Torr.) Wats. (Chenopodiaceae) from their natural habitat in Death Valley, California. Using a set of several data concentrations of inorganic electrolytes (Na⁺, K⁺, Cl^-) and of oxalate in the mesophyll of this salt secreting species were calculated. The osmotic potential resulting from these solutes (under consideration of an empirically estimated osmotic coefficient) is in good agreement with field measurements of the overall osmotic potential in the leaf mesophyll as determined by pressure-volume curves. This indicates that these 4 electrolytes are the main osmotically active solutes. Oxalate is present in comparably high concentrations and is used to achieve ion balance.Organic solutes analyzed include soluble carbohydrates (mono-, di- and oligosaccharides), amino- and organic acids as well as glycinebetaine. Of these, organic- and amino acids (including proline) contribute only little to osmoregulation. Soluble carbohydrates and especially glycinebetaine exhibit concentrations high enough for generating considerable osmotic potentials, at least if these compounds are regarded to be restricted to the cytoplasm acting as compatible solutes.     

Male-specific DNA in the dioecious species Atriplex garrettii (Chenopodiaceae)

The mechanism of sex determination in dioecious species of the genus Atriplex (Chenopodiaceae) has not been determined. This paper reports the discovery of a male-specific DNA fragment in the diploid dioecious species A. garrettii. DNA samples extracted individually from ten male and ten female plants were bulked by sex. Random amplified polymorphic DNA (RAPD) fragments were generated in the two bulks in order to identify markers that were polymorphic between male and female plants. A total of 158 decamer primers were tested. A 2075 base-pair (bp) male-specific DNA fragment generated with the OPAF-14 primer was identified. The fragment was cloned and partially sequenced and 24-mer primers that exclusively amplified this fragment were constructed. When 124 male plants, 126 female plants, and one hermaphroditic plant were tested individually, the male-specific 2075-bp DNA fragment was present in the hermaphrodite and all but one of the male plants, and was absent in all female plants. A smaller DNA fragment (~1800 bp) that was homologous to the 2075-bp fragment was amplified from the single male plant that lacked the 2075-bp fragment. Cytogenetic analysis revealed no apparent heteromorphic sex chromosomes. These observations suggest that sex determination in A. garrettii is genetic, with no evidence of heteromorphic sex chromosomes.     

Taxonomic position and distribution of Atriplex lapponica (Chenopodiaceae)

The arctic, halophytic species Atriplex lapponica Pojark has flowers of two kinds: perfect ones surrounded by a pentamerous perianth and pistillate flowers enclosed in bracteoles. Thus, A. lapponica is closely related to the species of sect. Dichospermum Dumort, not to species of sect. Teutliopsis Dumort. as generally believed.     

Vesiculated Hairs: A Mechanism for Salt Tolerance in Atriplex halimus L

The concentration of salts in the vesiculated hairs of Atriplex halimus L. was measured and was remarkably higher than that of the leaf sap and xylem exudate. In spite of their unusually high salt content, these hairs when immersed seemed unable to absorb water, in apparent contradiction to the previously held hypothesis that vesiculated hairs make it possible for such plants to absorb water from the atmosphere. Although growing the plants under saline conditions increased the salt content of the hairs from 2.3 m Na+K to 11.6 m, salt content of the expressed leaf sap from young leaves did not change significantly. This observation indicates that in A. halimus the vesiculated hairs play a significant role in removing salt from the remainder of the leaf and preventing the accumulation of toxic salts in the parenchyma and vascular tissues. Thus, a nearly constant salt content is maintained in leaf cells other than the hairs.     

Localization of potential ion transport pathways in vesicular trichome cells of Atriplex halimus L.

The secreting glandular trichomes are recognized as an efficient structure that alleviates salt effects on Atriplex halimus. They are found on buds, young green stems, and leaves. They occupy both the leaf surfaces and give them a whitish color. Their histogenesis and ultrastructure were investigated in the third young leaves. They appear in early stage of plant development and its initiation continuous until just the leaf final development state. Each trichome contains two parts; a stalk which has high electron opacity, embedded in epidermal cells, and bears a second one which is unicellular, called bladder cell and has a low electron density. The bladder cell appears as a huge vacuole and the well-reduced cytoplasm which is pushed close to the wall, contains only a few organelles. Concurrently, the use of silver chloride precipitation technique shows that, in secretion process, salt follows a symplasmatic pathway which is consolidated by the presence of numerous plasmodesmata between the stalk cell(s), and the bladder one and the neighboring mesophyll cells. In addition, according to lanthanum-tracer study, salt can be excreted apoplastically. In fact, the heavy element can be transported via endocytosis vesicles, and by Golgi, endoplasmic reticulum, and lysosome (G.E.R.L.) network toward the storage vacuoles.     

NaCl alleviates polyethylene glycol-induced water stress in the halophyte species Atriplex halimus L

Atriplex halimus L. is a C4 xero-halophyte species well adapted to salt and drought conditions. To collect information on the physiological impact of low salt levels on their water-stress resistance, seedlings were exposed for 6 d to nutrient solution containing either 0% or 15% polyethylene glycol 10,000 (PEG), in the presence or in the absence of 50 mM NaCl. Similar experiments were performed with one PEG-resistant and one PEG-sensitive selected cell line exposed for 50 d to 0% or 15% PEG on standard Linsmaier and Skoog (LS) medium, on LS medium supplemented with 50 mM NaCl, or on Na+-free medium. NaCl mitigated the deleterious impact of PEG on growth of both whole plants and PEG-sensitive cell lines and improved the ability of stressed tissues to perform osmotic adjustment (OA). Water stress reduced CO2 net assimilation rates quantified in the presence of high CO2 and low O2 levels (A), stomatal conductance and transpiration, but NaCl improved water use efficiency of PEG-treated plants through its positive effect on A values, especially in young leaves. PEG increased the internal Na+ concentration. The resistant cell line accumulated higher concentration of Na+ than the PEG-sensitive one. The complete absence of Na+ in the medium endangered the survival of both cell lines exposed to PEG. Although Na+ by itself contributed only for a small part to OA, NaCl induced an increase in proline concentration and stimulated the synthesis of glycinebetaine in response to PEG in photosynthetic tissues. Soluble sugars were the main contributors to OA and increased when tissues were simultaneously exposed to PEG and NaCl compared with PEG alone, suggesting that Na+ may influence sugar synthesis and/or translocation.     

Chloride salinity reduces cadmium accumulation by the Mediterranean halophyte species Atriplex halimus L.

Soil salinity usually increases bioavailability of Cd on heavy metal polluted soils but its impact on Cd absorption and accumulation by plants remains largely unknown. Plants from the halophyte species Atriplex halimus were therefore exposed for 12 and 14 days to nutrient solution containing 50 μM CdCl₂ in the presence of NaCl, KCl or NaNO₃ 50 mM. Most Cd present in solution remained as Cd–EDTA and salinity had no impact on Cd speciation. Chloride salinity (NaCl and KCl) reduced Cd accumulation in shoots and roots while NaNO₃ increased Cd accumulation in leaves. More than 30% of accumulated Cd was found at the leaf surface and accumulated in trichomes but all tested salts decreased the proportion of excreted Cd. Cadmium induced a decrease in the leaf water content. External NaCl and KCl mitigated the deleterious impact of Cd by inducing osmotic adjustment while NaNO₃ and synthesis of protecting compounds such as soluble sugars and glycinebetaine. Free polyamines (putrescine, spermidine and spermine) increased in response to Cd, Cd + NaCl and Cd + KCl while only putrescine increased in response to Cd + NaNO₃. Proline exhibited maximal concentration in the leaves of Cd + NaCl and Cd + KCl-treated plants and was correlated with osmotic adjustment. Our results suggest that chloride salinity improved the resistance of A. halimus to Cd toxicity both by decreasing the absorption of heavy metal and by improving tissular tolerance through an increase in the synthesis of osmoprotective compounds.     

High chromosome numbers in Mojavean and Sonoran desert Atriplex canescens (Chenopodiaceae)

Cytological, flavonoid, and morphological data are provided for several varieties of the shrubby species Atriplex canescens (Pursh) Nutt. (x = 9) (fourwing saltbush) in the Mojavean and Sonoran deserts of southwestern United States and northern Mexico. These include var. linearis (S. Wats.) Munz (2x); var. angustifolia (Torr.) S. Wats. (2x, 4x); var. occidentalis (Torr. & Frem.) Welsh & Stutz (4x, 6x), the common variety; var. laciniata Parish (12x); and var. macilenta Jeps. (12x). Atriplex canescens var. grandidentatum Stutz & Sanderson (20x) is newly described. An autoploid origin from 12x var. laciniata is suggested for the 14x and 20x polyploids, through unreduced gametes. Founder populations of odd-ploid products arising during such a sequence of events could probably have returned to even-ploidy through genetic segregation and the rapid elimination of aneuploids. Morphological characters suggest an origin for 12x var. laciniata by interspecific hybridization of var. occidentalis with A. polycarpa (Torr.) S. Wats.     

Loss of genetic variation in geographically marginal populations of Atriplex tatarica (Chenopodiaceae)

BACKGROUND AND AIMS: Genetic variability was estimated for Atriplex tatarica from 25 populations in the Czech Republic. Since its north-western range margin is in central Europe, a relationship between marginality and low within-population genetic diversity was tested in accordance with the Central-Marginal Model. METHODS: Population genetic diversity was expressed by assessing patterns of variation at 13 putatively neutral allozyme loci (comprising 30 putative alleles) within and between 25 natural populations of A. tatarica along a north-west-south-east transect in the Czech Republic. KEY RESULTS: Atriplex tatarica is a species of human-made habitats with a mixed mating system and wide geographic distribution. Overall, A. tatarica displayed moderate levels of genetic diversity in comparison with other herbaceous plants. The percentage of loci that were polymorphic was 47.1%, with average values of 1.55, 0.151 and 0.155 for the average number of alleles per polymorphic locus (A), observed heterozygosity (Ho) and expected heterozygosity (He), respectively. There was only weak evidence of inbreeding within populations (FIS=0.031) and significant population differentiation (FST=0.214). Analysis of the data provides no evidence for isolation-by-distance for the whole study area. However, Mantel tests were highly significant for the marginal Bohemian region and non-significant for the central Moravian region. While northern populations of A. tatarica showed significantly lower allelic richness (A=1.462) than populations from the southern part of the study area (A=1.615), they did not differ in observed heterozygosity (Ho), gene diversity (HS), inbreeding within populations (FIS) or population differentiation (FST), despite generally lower values of particular genetic measurements in the marginal region. CONCLUSIONS: Genetic diversity, with the exception of allelic richness, was not significantly lower at the margins of the species' range. This, therefore, provides only weak support for the predictions of the Central-Marginal Model.     

Na and k accumulation and salt tolerance of Atriplex canescens (Chenopodiaceae) genotypes

Sixteen accessions of the xerohalophyte, Atriplex canescens (Pursh.) Nutt., differing in tendency to accumulate Na or K in leaf tissues, were compared for salt tolerance in a greenhouse study. Plants were grown along a salinity gradient from 72 to 2017 mol/m³ NaCl measured in the root zone. Growth rates (RGR) were negatively affected by salinity for all accessions. Initial leaf levels of Na (measured before exposing plants to saline solutions) were positively correlated with subsequent RGR's of accessions on the salinity gradient (r = 0.60 - 0.88, P < 0.05 across salinity levels), whereas initial leaf K levels were negatively correlated (r = -0.68 to -0.85, P < 0.01 across salinity levels). Varieties linearis (S. Wats.) Munz and grandidentatum Stutz & Sanderson had greater tendency for Na accumulation, lower tendency for K, and higher growth rates on saline solutions than var. occidentalis (Torr. & Frem.) Welsh & Stutz accessions. Within var. occidentalis accessions, RGRs were negatively correlated with initial leaf levels of K but not Na. Postexposure leaf Na and K levels were not strongly correlated with RGR's. All accessions responded to salinity by increasing their uptake of Na, which is the primary mechanism of osmotic adjustment to salinity in this species. It is suggested that differences in tendency to accumulate Na or K among A. canescens genotypes are related to their specialization for saline or xeric habitats, respectively.     

Genetic structure of Atriplex halimus populations in the Mediterranean Basin

BACKGROUND AND AIMS: The saltbush Atriplex halimus is a chenopodiaceous plant well adapted to dry saline habitats and widely distributed in the Mediterranean Basin. A study was carried out to analyse the genetic diversity of A. halimus at the level of the Mediterranean Basin. METHODS: To assess the intra- and interpopulational variation of A. halimus a total of 51 populations and six plants per populations was analysed with the RAPD-PCR technique. For the study of the phylogeny of the populations, 21 samples of A. halimus and seven samples of other species of Atriplex were analysed by the sequencing of the ITS (internal transcribed spacer) region of the ribosomal DNA. KEY RESULTS: The AMOVA analysis of the RAPD results showed that populations were divided into two discrete genetic groups, as the variation among groups accounted for 54.36 % of the total variance of the collection. At the same time, the intrapopulational diversity was high, as 301 out of 306 plants analysed constituted an individual RAPD haplotype. The sequencing of the ITS region also showed a significant separation of the two genetic groups, with a genetic distance of 0.023 nucleotide substitutions per site. Using A. breweri, A. canescens, A. glauca and A. prostrata as outgroups in the phylogenetic analysis, A. breweri and A. canescens are the species closest to A. halimus from this group, while A. prostrata is the most distant. CONCLUSIONS: The present work indicates that two genetic groups of A. halimus can be distinguished after analysing the genetic diversity of 51 populations from ten countries in the Mediterranean Basin.     

Competition between Salicornia europaea and Atriplex prostrata (Chenopodiaceae) along an experimental salinity gradient

Salicornia europaea L. is a halophyte that often occupies the lowestand most saline (>3.5% total salt) areas of salt marshes. Atriplexprostrata Boucher is less salt tolerant than S. europaea and oftengrows in a less saline (<2.0% total salts) zone adjacent to S. europaea. The purpose of this experiment was to determine thecompetitive outcome when these two species are grown at differentsalinities to ascertain the extent salinity and competition affect plantzonation. Plants were grown in a de Wit replacement series at 85, 170,and 340 mM NaCl in half-strength Hoagland's no. 2 nutrient solution fortwo months. There was a significant effect of salt concentration,competition, and their interaction on biomass production of S. europaea plants. However, only salt concentration significantly affectedbiomass production of A. prostrata plants. Results of thisexperiment confirmed the results of other studies that demonstrated thatthe more salt tolerant species were less competitive at lower salinities. Atriplex prostrata was the better competitor at 85 mM NaCl, whereasS. europaea was the better competitor at 340 mM NaClbecause growth of A. prostrata was inhibited. At 170 mMNaCl, A. prostrata biomass production decreased more than S. europaea biomass in mixed culture.     

Regulation of metabolomics in Atriplex halimus growth under salt and drought stress

Forty-day-old Atriplex halimus seedlings were treated with either NaCl (50, 300 and 550?mM) for the subsequent 30?days or PEG for the following 3, 6 and 10?days. Shoot fresh and dry weights were significantly increased by 50?mM NaCl; nevertheless, the other concentrations had no effect. However, the growth was reduced by drought only after 10?days. Meanwhile, Na⁺ was accumulated in treated plants; the magnitude of accumulation was highest with high NaCl concentration or PEG for 10?days. The metabolite profiles showed discrimination particularly up-regulation of the amino acids proline, valine, isoleucine, and methionine. Moreover, the macro analysis revealed that NaCl- and PEG-treated plants shared 10?% of the metabolites in the positive mode, however, 87?% were unique to NaCl and 46?% were unique to PEG whereas in the negative mode, 8?% were in share while 90 or 53?% were restricted to NaCl or PEG, respectively. Additionally, sucrose in particular was significantly increased up to threefold and fivefold by 300 and 550?mM NaCl, respectively and up to 2.5-fold by drought for 10?days, nevertheless, the other sugar fractions remained largely unchanged. Also, proline was significantly increased by only the high NaCl concentrations and the long-term drought, nonetheless, the other treatments led, if any, to decreases. These results conclude that NaCl affects the metabolite profiles more than PEG and these metabolites might contribute to osmotic adjustments to act as osmoprotectants rather than osmolytes. These changes of metabolomics might function in many resistance and stress responses.     

Genetic structure of experimental populations and reproductive fitness in a heterocarpic plant Atriplex tatarica (Chenopodiaceae)

Atriplex tatarica is a heterocarpic species of disturbed habitats. Seeds of Atriplex tatarica do not germinate immediately after shedding, but may remain in a dormant but viable state indefinitely. We investigated whether there were genetic and fitness differences between plants derived from seeds of the different fruit types germinated in different temperatures and salinities. Seeds that germinated in optimal and suboptimal conditions differed significantly in their genetic composition due, in part, to their source population. Seeds that germinated in the suboptimal conditions produced more homozygous plants. Plants that were primarily heterozygous were generated from nondormant fruit types as well as from fruits that germinated in the optimal conditions. Moreover, there was a positive correlation between the degree of heterozygosity and plant fitness measured as the mass of the stem and reproductive structures. In conclusion, the genetic variation of natural populations may be at least partly due to the ability of particular seed genotypes to germinate in the specific environmental conditions of a particular locality. In some circumstances, the process of differential germination may select not only for genetic variability but also for higher fitness if heterozygosity-fitness correlations are present.     

Phenotypic and genetic variability of three natural populations of Atriplex halimus

Three natural populations of Atriplex halimus, located in three different climatic contexts, were studied using leaf characteristics (leaf area, leaf length-to-maximum-width ratio, average width of the leaves and leave-to-branch ratio) and isoenzymatic markers. The study showed the existence of a highly significant phenotypical variability. This variability is all the more significant, as populations are geographically distant and located in different climates. The gradual character of this morphological variability, along a climatic gradient, indicates that it is almost a clinical differentiation. Clones obtained from semi-woody cuttings taken on the level of each population and placed in a common parcel have maintained the same leaf characteristics as the population sources, suggesting the genetic origin of this variability. The study of four isoenzymatic systems confirms the existence of this variability. Thus, the percentage of polymorphic loci (P), the expected heterozygosity (He) and the mean number of alleles per locus (A) are of 77.52%, 0.319, and 1.99, respectively. The genetic diversity index (Fst) obtained is 0.089.     

The agro-ecological suitability of Atriplex nummularia and A. halimus for biomass production in Argentine saline drylands

The choice of the best species to cultivate in semi-arid and arid climates is of fundamental importance, and is determined by many factors, including temperature and rainfall, soil type, water availability for irrigation and crop purposes. Soil or water salinity represents one of the major causes of crop stress. Species of the genus Atriplex are characterized by high biomass productivity, high tolerance to drought and salinity, and high efficiency in use of solar radiation and water. Based on a search of the international literature, the authors outline an agro-climatic zoning model to determine potential production areas in Argentina for Atriplex halimus and Atriplex numularia. Using the agroclimatic limits presented in this work, this model may be applied to any part of the world. When superimposed on the saline areas map, the agroclimatic map shows the suitability of agro-ecological zoning for both species for energy purposes on land unsuitable for food production. This innovative study was based on the implementation of a geographic information system that can be updated by further incorporation of complementary information, with consequent improvement of the original database.