Vertical differences in Phragmites australis culm anatomy along a water depth gradient

We reported on quantitative anatomical comparisons of reed culms grown under a wide range of water depth (from −10 to +230 cm). The study focused mainly on the above water parts of ramets, but also provided an insight into the submerged internodes. Investigated anatomical features were: radial thickness of the internode wall and that of each tissue constituting it (epidermis and subepidermal tissues, aerenchyma channels, cortical sclerenchyma and parenchyma); areas of aerenchyma channels and the summed transversal area of them; areas of the innermost vascular bundles involving the bundle sheath sclerenchyma, phloem and metaxylem vessels; the cross-sectional area of the pith cavity and that of the internode wall. After a 2–3% decrease below the uppermost internode, the thickness of internode tissues, except for cortical sclerenchyma, continuously increased downwards. Quantitative differences between the ramets grown at different water depths were observed only from the lower aerial internodes. Parenchyma taking part considerably in the mechanical stability of culms with its large extension and thick cell walls was more-layered and thicker (with the maximum thicknesses of approximately 1200 and 1000 μm) in ramets from 180–200 than from 20–30 cm water depth. Areas of the innermost vascular bundles increased downwards and decreased after a maximum value appearing in lower internodes of ramets grown in deeper than shallower water. Aerenchyma channels appeared farther up from the water surface in culms in shallower than deeper water: the top ends were found about 50 cm higher in culms from 20–30 than from 180–200 cm water depth. Along the transect at right angles to the shore, the radial thickness and area of aerenchyma channels were higher in ramets grown in medium water depth—the summed transversal area of them was greater than 4 mm² in internodes at the water surface, while plants from the two ends of the water depth gradient had smaller aerenchyma channels (with approximately 1 mm² total areas). In contrast, pith cavity area at the water surface continuously increased – up to 60 mm² – from shallower to deeper water. Therefore, oxygen transport in ramets grown at the open water fringe of stands may occur mostly in the pith cavity.     

Clonal diversity along a water depth gradient in a declining reed stand as detected by three different genetic methods

Isozyme, RAPD (random amplified polymorphic DNA) and microsatellite techniques were applied to determine the genetic diversity of a reed stand declining at the open water fringe for several decades and the accuracy of these methods was evaluated. Sampling was carried out along three transects parallel with and one transect at right angle to the lakeshore. In isoenzyme investigation, esterase and further 11 polymorphic loci of seven isozymes (with a total of 26 alleles) were analysed and 39 clones were distinguished. RAPD analysis involved 11 random decamer primers and, based on 141 repeatably amplifying and polymorphic RAPD fragments, 52 multilocus genotypes were differentiated. In microsatellite investigation, four primer pairs provided 47 polymorphic allelic variants that determined 37 multilocus phenotypes.Besides the differences, all the three methods revealed high number and intermingling (mixed) arrangement of clones at lower water depths, and low clone number and successive structure at the open water fringe of the stands. Only microsatellite analysis revealed different stages of colonization: old successful clone (colonizing the deeper water but almost completely disappearing at the lakeshore edge) and successful but young clone (with large extension at the lakeshore and just reaching the open water edge of the stand) were also distinguished. Finally, the present paper demonstrated that an appropriate data evaluation could make the result of RAPD comparable to that of microsatellite studies.     

Genetic diversity patterns in Phragmites australis at the population,regional and continental scales

Genetic diversity, population structure and interrelationships were investigated in eight populations of the common reed, Phragmites australis, in the Po Plain, Italy, by means of amplified fragments length polymorphisms (AFLPs) and random amplified polymorphic DNAs (RAPDs). Patterns of genetic diversity were analysed in relation to size, age and degree of human impact in the wetlands and compared with that of a distant population in Romania. Genetic distances between Po Plain clones and geographically distant clones were measured to determine the geographical extent of the gene pool.     

Genetic diversity and dispersal of Phragmites australis in a small river system

Even though the reed, Phragmites australis, is an extensively studied wetland species, little is known about reproduction and dispersal modes within and among reed populations at the scale of small river systems. Using microsatellite analysis of 189 individuals from three adjacent river catchments in the Czech Republic, we elucidated the role of the river corridors in the dispersal of P. australis. Using Bayesian clustering of individuals, we found that 19% of clusters were distributed only along one river, which implied dispersal by water (or by wind) along river corridors, whereas 38% of clusters were widely distributed and were likely the product of wind long-distance dispersal among rivers. Intensive exchange of propagules among river systems is further demonstrated by only 6% of total variance being attributed to the variance among rivers in the AMOVA-analysis. Spatial autocorrelation analysis revealed a decreasing pattern up to 5–10 km and no clear pattern over longer distances. This gives an evidence for pollen and seed dispersal at short distances (up to 1 km), whereas most likely only seed dispersal at longer distances up to 10 km. We found five multilocus genotypes distributed in two different populations. The distances between populations with the same genotype ranged from 0.5 to 10.8 km. This can be interpreted as long-distance vegetative dispersal.     

Effects of abiotic conditions on Phragmites australis along geographic gradients in Lake Burullus, Egypt

Stand structure and biomass production of Phragmites australis (Cav.) Trin. ex Steud. were analyzed along north-south and east-west transects in the Burullus coastal lagoon (N Egypt, 410 km²) at monthly intervals over a period of 1 year (February 2003 until January 2004). For this purpose, young and old stands were selected at eight different locations in the lagoon. It was found that the north-south transect mainly represented a fertility gradient (207-286 mg l⁻¹ TN, 30-106 mg l⁻¹ TP), while the east-west transect was associated with significantly decreasing salinity (7-4 ppt). All morphological and biomass variables of P. australis were significantly different between young and old stands. On average, the old (7.3 ± 0.2 kg DW m⁻²) accumulated three times more total above-ground biomass than the young stands (2.5 ± 0.1 kg DW m⁻²). Shoot height, diameter and shoot dry weight significantly increased by 25-50% with increasing fertility along the north-south transect. Shoot density significantly decreased from north to south, while it almost doubled in the north sites from 109 ± 6 to 216 ± 7 shoots m⁻² along the west-east transect. In separate stepwise multiple regressions, variation in water quality explained 34-63% of the variation in morphology and total above-ground biomass in the old stands (salinity and water level were most important for biomass, transparency also for height and density) while it explained 16-42% of variation in young stands (mainly transparency).     

Genetic diversity of the endangered and endemic species Psathyrostachys huashanica natural populations using simple sequence repeats (SSRs) markers

Psathyrostachys huashanica Keng., a species endemic to China, is only distributed in Huashan Mountain in Shaanxi Province. It has been listed as “national first-class protected rare species.” In this study, the microsatellites of barley were used to analyze genetic diversity of P. huashanica populations sampled from three valleys (Huangpu, Xian and Huashan Valleys) in Mt. Huashan. A total of 33 alleles of 11 loci were detected from 266 individuals. The observed average number of alleles (A) is 2.75; the effective number of alleles (Ae) is 1.67. The percentage of polymorphic loci (PPB) is 58.33% in Huangpu Valley, 75% in Xian Valley, 83.33% in Huashan Valley, and the total PPB is 83.33%. About 77.6% of (F_ST = 0.324) genetic diversity was observed within the subpopulations. Genetic differentiation within each subpopulations was higher than that among the subpopulations. Mean genetic distance is 0.17 (range: 0.010–0.401). Correlation analysis detected significant correlation between genetic distance and vertical distance of altitude in Huashan Valley. Differentiation mainly occurred between the higher altitude subpopulations and the lower altitude subpopulations, suggesting that altitude might be the major factor that restricted the gene flow between different altitude subpopulations and resulted in differentiation of subpopulations.     

Differences in rhizome aeration of Phragmites australis in a constructed wetland

Waterlogged soils are a challenging environment for plants due to anoxic conditions and enrichment of phytotoxic substances in the rhizosphere. High contents of organic matter in the substrate as present in constructed wetlands (CW) may amplify these effects. The influence of substrate organic matter on the rhizome aeration of Phragmites australis (common reed) was investigated in a CW for sewage treatment on two sites with different organic concentrations: (a) inflow part of the bed with a high organic charge (high-organic site) and (b) outflow zone of the bed with a lower organic charge (low-organic site). Several diurnal oxygen (O₂) courses were recorded inside the rhizomes using micro-optodes. Maximum O₂ percentages in the rhizomes were generally the same at both sites, but minimum O₂ values were lower in the low-organic site than in the high-organic site, leading to higher amplitudes at the low-organic site. The results suggest that oxygen release from the roots is hampered under high-organic compared to low-organic conditions. This observation might be explained by changes in gas conductivity of the roots, but also by alterations in microbial oxygen demand, under different organic burden.     

Different sensitivity of Phragmites australis and Glyceria maxima to high availability of ammonium-N

The ability to cope with NH₄⁺-N was studied in the littoral helophytes Phragmites australis and Glyceria maxima, species commonly occupying fertile habitats rich in NH₄⁺ and often used in artificial wetlands. In the present study, Glyceria growth rate was reduced by 16% at 179 μM NH₄⁺-N, and the biomass production was reduced by 47% at 3700 μM NH₄⁺-N compared to NO₃⁻-N. Similar responses were not found in Phragmites. The amounts (mg g⁻¹ dry wt) of starch and total non-structural carbohydrates (TNC) in rhizomes were significantly lower in NH₄⁺ (8.9; 12.2 starch; 20.1; 41.9 TNC) compared to NO₃⁻ treated plants (28.0; 15.6 starch; 58.5; 56.3 TNC) in Phragmites and Glyceria, respectively. In addition, Glyceria showed lower amounts (mg g⁻¹ dry wt) of soluble sugars, TNC, K⁺, and Mg²⁺ in roots under NH₄⁺ (5.6; 14.3; 20.6; 1.9) compared to NO₃⁻ nutrition (11.6; 19.9; 37.9; 2.9, for soluble sugars, TNC, K⁺, and Mg²⁺, respectively), while root internal levels of NH₄⁺ and Ca²⁺ (0.29; 4.6 mg g⁻¹ dry wt, mean of both treatments) were only slightly affected. In Phragmites, no changes in soluble sugars, TNC, Ca²⁺, K⁺, and Mg²⁺ contents of roots (7.3; 14.9; 5.1; 17.3; 2.6 mg g⁻¹ dry wt, means of both treatments) were found in response to treatments. The results, therefore, indicate a more pronounced tolerance towards high NH₄⁺ supply in Phragmites compared to Glyceria, although the former may be susceptible to starch exhaustion in NH₄⁺-N nutrition. In contrast, Glyceria's ability to colonize fertile habitats rich in NH₄⁺ is probably related to the avoidance strategy due to shallow rooting or to the previously described ability to cope with high NH₄⁺ levels when P availability is high and NO₃⁻ is also provided.     

Competitive effects of Phragmites australis on the endangered artesian spring endemic Eriocaulon carsonii

We investigated the relative importance of above- and below-ground competition by reeds (Phragmites australis (Cav.) Trin. ex Steud) on the growth rate of Eriocaulon carsonii F.Muell. subsp. carsonii, an endangered plant threatened by reeds on artesian springs in Australia. Soil-filled buckets containing E. carsonii were frequently watered to simulate artesian spring conditions and subject to three treatments: (1) no Phragmites (control), (2) Phragmites (ABG), and (3) Phragmites with shoots tied back (BG). After thirteen months, Phragmites mean below-ground biomasses had increased to c. 3 kg m⁻² and mean above-ground biomasses to c. 1 kg m⁻². After the same period, mean root biomass of E. carsonii plants was significantly lower in buckets subject to both Phragmites treatments compared with control plants, as was E. carsonii foliage area. Comparison of the two Phragmites treatments indicated that below-ground competition was the primary cause of this reduced growth in E. carsonii. The vulnerability of E. carsonii to competitive exclusion by P. australis is in part due to the highly synchronized phenologies of the two species.     

Clonal diversity and structure related to habitat of the marine angiosperm Thalassia testudinum along the Atlantic coast of Mexico

The clonal structure of the tropical seagrass Thalassia testudinum was studied at 16 sites along the Mexican Atlantic coast, situated in back-reef, shallow coastal and lagoon habitats. Thalassia testudinum was highly clonal, with an overall average clonal richness (R) of 0.55. The largest genet found in this study extended throughout the sampling area (∼230 m), with an estimated max age almost reaching 600 years. Lagoons with higher nutrient availability reflected by nutrient content of leaves (mean leaf C:N ratio 11.4) and lower hydrodynamic regimes reflected by the percentage of fine sediments (on average 23%), sustained larger genets of T. testudinum (mean of the largest genets over populations was 167.3 m) than the shallow coastal areas (C:N 12.3, 6.2% fine sediment, mean largest genet 10.3 m) and the more oligotrophic back-reefs (C:N 16.3, 2.7% fine sediment, mean largest genet 6.5 m). Population genetic analysis showed different levels of clonality, genotypic diversity and spatial genetic relatedness for this seagrass per habitat, with the lagoons presenting much lower levels of clonal diversity than the other two habitats.     

Growth-mortality trade-offs along a depth gradient in Cancer borealis

Intertidal and shallow subtidal ecosystems experience steep environmental gradients over short distances. Individual foraging rate, predation risk, and physiologic stress vary along these gradients, resulting in growth-mortality trade-offs with depth. In the summer, Cancer borealis commonly forage in the shallow subtidal in the Gulf of Maine. C. borealis are the favored invertebrate prey of the Herring Gull and the Great Black-backed Gull, which consume 25%-50% of available C. borealis (those in < 1 m water) during each daytime low tide. We investigated three possible explanations for the presence of C. borealis in the risky gull-predation zone. First, we tested whether predation risk in the gull-predation zone was matched at deeper depths by subtidal predators; we found predation risk decreases with depth. Second, we tested whether water temperatures were warmer in the gull-predation zone and whether these warmer temperatures resulted in increased growth rates. We found that, while waters were warmer in the gull-predation zone, crabs grew at similar rates above and below the thermocline when fed similar diets. Finally, we tested for differences in food availability with depth and whether these differences influenced C. borealis growth rates. Our results suggest a growth-mortality trade-off, where increased food availability provides sufficient growth benefit to outweigh the risk of foraging at shallower depths.     

Detection of genetic diversity in closely related bread wheat using microsatellite markers

Wheat microsatellites (WMS) were used to estimate the extent of genetic diversity among 40 wheat cultivars and lines, including mainly European elite material. The 23 WMS used were located on 15 different chromosomes, and revealed a total of 142 alleles. The number of alleles ranged from 3 to 16, with an average of 6.2 alleles per WMS. The average dinucleotide repeat number ranged from 13 to 41. The correlation coefficient between the number of alleles and the average number of repeats was only slight (r _s = 0.55). Based on percentage difference a dendrogram is presented, calculated by the WMS-derived data. All but two of the wheat cultivars and lines could be distinguished. Some of the resulting groups are strongly related to the pedigrees of the appropriate cultivars. Values for co-ancestry (f) of 179 pairs of cultivars related by their pedigrees (f0.1) averaged 0.29. Genetic similarity (GS) based on WMS of the same pairs averaged 0.44. The rank correlation for these pairs was slight, with r _s = 0.55, but highly significant (P<0.001). The results suggest that a relatively small number of microsatellites can be used for the estimation of genetic diversity and cultivar identification in elite material of hexaploid bread wheat.     

Treatment of industrial wastewater with two-stage constructed wetlands planted with Typha latifolia and Phragmites australis

Industrial wastewater treatment comprises several processes to fulfill the discharge permits or to enable the reuse of wastewater. For tannery wastewater, constructed wetlands (CWs) may be an interesting treatment option. Two-stage series of horizontal subsurface flow CWs with Phragmites australis (UP series) and Typha latifolia (UT series) provided high removal of organics from tannery wastewater, up to 88% of biochemical oxygen demand (BOD₅) (from an inlet of 420 to 1000 mg L⁻¹) and 92% of chemical oxygen demand (COD) (from an inlet of 808 to 2449 mg L⁻¹), and of other contaminants, such as nitrogen, operating at hydraulic retention times of 2, 5 and 7 days. No significant (P < 0.05) differences in performance were found between both the series. Overall mass removals of up to 1294 kg COD ha⁻¹ d⁻¹ and 529 kg BOD₅ ha⁻¹ d⁻¹ were achieved for a loading ranging from 242 to 1925 kg COD ha⁻¹ d⁻¹ and from 126 to 900 kg BOD₅ ha⁻¹ d⁻¹. Plants were resilient to the conditions imposed, however P. australis exceeded T. latifolia in terms of propagation.     

Assessment of genetic diversity in Calamus thwaitesii BECC. (Arecaceae) using RAPD markers

Calamus thwaitesii Becc. is a potentially useful rattan found in the Western Ghats of India and Sri Lanka. The wild stock of this rattan species is greatly diminished due to overexploitation for the furniture industry and increasingly rare. Genetic diversity was estimated in 80 samples representing eight populations from the Western Ghats and Sri Lanka using Random Amplified Polymorphic DNA (RAPD) markers. RAPDs generated a total of 120 markers with 10 decamer primers, of which 85% were found to be polymorphic. The percentage of polymorphic loci varied from 40.00 to 60.83 and genetic distance between populations ranged from 0.0332 to 0.2777. Among the analysed populations, Goa was found to be genetically superior followed by Achenkovil, Sinharaja and Talakkaveri. Majority of the genetic diversity was distributed within populations (70.79%) and only (29.21%) among populations. Genetic relationships estimated by the unweighted pair-group method with arithmetic averaging (UPGMA) cluster analysis and principal co-ordinate analysis failed to separate Indian and Sri Lankan populations geographically into two distinct groups.     

Chloroplast sequences reveal a diversity gradient in the Mediterranean Ruppia cirrhosa species complex

Two subcosmopolitan species Ruppia maritima and Ruppia cirrhosa are recognized throughout Europe, whereas Ruppia drepanensis is endemic to SW Europe. We aimed at characterizing the geographic structure of the chloroplast DNA haplotype diversity of 56 Ruppia populations across the European part of the Mediterranean. On the basis of five cpDNA markers (total length of 2300 bp) 16 haplotypes were obtained for 1546 individuals. Three populations of R. maritima showed a single haplotype and differed in five insertions/deletions and 16 substitutions from a highly variable R. cirrhosa species complex, which included R. drepanensis. The haplotype diversity of this species complex was much higher in the western Mediterranean basin than in the eastern basin. Analysis of molecular variance (AMOVA) showed significant differentiation of R. cirrhosa between the two basins and also within the western Mediterranean thereby suggesting the latter as an important centre of Ruppia diversity. An isolation-by-distance (IBD) pattern was stronger between the West-East basin populations than within basins. A PCO analysis of the western basin populations indicated a diversity gradient with those of Sardinia as polymorph intermediates. The low diversity within the eastern basin suggests that the observed gradient could be hypothesized as a historical dispersal of only a limited number of haplotypes from west to east.     

Roots of the wetland plants Typha latifolia and Phragmites australis are inhabited by methanotrophic bacteria in biofilms

Wetland plants create partly aerobic conditions in the rhizosphere by releasing oxygen to the waterlogged substrate. The present study was conducted to characterise the arrangement of rhizobacteria, especially those active in methane oxidation, in root-associated biofilms of wetland plants. Root cross sections sampled from Typha latifolia L. (broadleafed cattail) and Phragmites australis (Cav.) Trin. ex Steud. (common reed) were scanned using light and electron microscopy. Methane-oxidising bacteria were identified and quantified by immunological labelling of the α-subunit of the methanol dehydrogenase (α-MDH; encoded in mxaF). On roots of both species there was a diverse subset of bacteria arranged in a microbial biofilm around the roots’ exodermis. Similar bacterial densities in the root-associated biofilm were detected in more basal regions and closer to the root tip. Many microbes carried notable internal membrane systems that are characteristic of methanotrophic bacteria. This morpho-anatomical characterisation was confirmed by immunogold labelling with α-MDH antibodies. Quantification of labelled bacteria revealed that 34–43% of the biofilm bacteria were potentially capable of methane turnover. These findings confirm the presence of methane-oxidising bacteria in the root-associated biofilms of the two common macrophytes T. latifolia and P. australis. This implies that the methanotrophs participate essentially in the microbial processes related to oxygen-releasing roots of wetland plants.     

Seed banks of Phragmites australis-dominated brackish wetlands: Relationships to seed viability, inundation, and land cover

In tidal wetlands of the eastern United States, buried seeds of the non-native haplotype of Phragmites australis may be a source of propagules for re-establishment after eradication efforts but factors controlling the development and expression of seed banks in non-native Phragmites stands have not been examined. We sampled surface soil at four Chesapeake Bay brackish tidal wetlands dominated by the non-native (European) haplotype M of Phragmites and used the seedling emergence method to quantity species of seedlings emerging under flooded and non-flooded soil conditions. Within each subestuary, one site was dominated by Phragmites that produced viable seeds (high viability) and the other by Phragmites that did not (low viability). We also described standing vegetation in plots, measured soil salinity, analyzed soil characteristics, and described surrounding land cover. Based on number of emerging seedlings, we found that 284 and 698 Phragmites seeds m⁻² occurred at the two high-viability sites, which was significantly higher than seed densities at the low-viability sites (10 seeds m⁻²), and greater than densities reported elsewhere. We also found that emergence of Phragmites seedlings from soil samples was prevented by continuous flooding of 3.5 cm of standing water, suggesting that colonization of deep water areas is due to vegetative clonal expansion from Phragmites in adjacent higher elevations. The density of Phragmites seeds was not related to soil salinity or abundance of other species in the seed bank or vegetation, but instead was positively related to greater wave energy disturbance (much longer fetch and more open water) and lower area of wetlands nearby. The seed bank was more species-rich (15-22 species observed) than standing vegetation (3-15 species) at all sites, meaning that the dominance of Phragmites in vegetation does not prevent the development of a diverse seed bank and implying that a species-rich community may establish rapidly following control efforts. Based on these results and our findings in related studies, we postulate that wave energy disturbance generates repeated opportunities for seedling recruitment by Phragmites, which creates stands of Phragmites with higher genotypic diversity. In turn, genetically diverse stands favor greater cross-pollination and production of viable seed. These findings suggest that, in North America, targeting control efforts on non-native Phragmites patches in areas of higher exposure to wave energy may be more effective in reducing source populations than efforts in more protected locations.     

Combined effect of NaCl-salinity and hypoxia on growth,photosynthesis, water relations and solute accumulation in Phragmites australis plants

Aim of the present study was to investigate the effects of two key environmental factors of estuarine ecosystems, salinity and hypoxia, on the physiological attributes in reed plants (Phragmites australis (Cav.) Trin. ex Steudel). Growth, leaf gas exchange, water (and ion) relations, and osmotic adjustment were determined in hydroponically grown plants exposed to hypoxia at varying NaCl-salinity concentrations (0, 50, 100, and 200 mM). Plants grew well under hypoxia treatment with standard nutrient solution without added salt and at NaCl concentrations up to 100 mM. Reed plants were able to produce and allocate phytomass to all their organs even at the highest salt level (200 mM NaCl). In plants subjected to hypoxia at various water potentials no clear relationships were found between growth and photosynthetic parameters except for g_s, whereas growth displayed a highly significant correlation with plant–water relations. A and g_s of reed plants treated with hypoxia at varying water potential of nutrient solutions were positively correlated and the former variable also had a strong positive relationship with E. Leaf Ψ_w and Ψ_π followed a similar trend and declined significantly as water potential of watering solutions was lowered. Highly significant positive correlations were identified between leaf Ψ_w and photosynthetic parameters. At all NaCl concentrations, the increase in total inorganic ions resulted from increased Na⁺ and Cl⁻ while K⁺, Ca²⁺, and Mg²⁺ concentrations decreased with increasing osmolality of nutrient solutions. Common reed has an efficient mechanism of Na⁺ exclusion from the leaves and exhibited a high leaf K⁺/Na⁺ selectivity ratio over a wide range of salinities under hypoxia treatment. In Phragmites australis grown in 200 mM NaCl, K⁺ contributed 17% toΨ_π, whereas Na⁺ and Cl⁻ accounted for only 11% and 6%, respectively. At the same NaCl concentration, the estimated contribution of proline to Ψ_π was less than 0.2%. Changes in leaf turgor occurred with a combined effect of salinity and hypoxia, suggesting that reed plants could adjust their water status sufficiently.