Genetic diversity of the submerged macrophyte Ceratophyllum demersum depends on habitat hydrology and habitat fragmentation.

The adaptability of plant populations to a changing environment depends on their genetic diversity, which in turn is influenced by the degree of sexual reproduction and gene flow from distant areas. Aquatic macrophytes can reproduce both sexually and asexually, and their reproductive fragments are spread in various ways (e.g. by water). Although these plants are obviously exposed to hydrological changes, the degree of vulnerability may depend on the types of their reproduction and distribution, as well as the hydrological differences of habitats. The aim of this study was to investigate the genetic diversity of the cosmopolitan macrophyte Ceratophyllum demersum in hydrologically different aquatic habitats, i.e. rivers and backwaters separated from the main river bed to a different extent. For this purpose, the first microsatellite primer set was developed for this species. Using 10 developed primer pairs, a high level of genetic variation was explored in C. demersum populations. Overall, more than 80% of the loci were found to be polymorphic, a total of 46 different multilocus genotypes and 18 private alleles were detected in the 63 individuals examined. The results demonstrated that microsatellite polymorphism in this species depends on habitat hydrology. The greatest genetic variability was revealed in populations of rivers, where flowing water provides constant longitudinal connections with distant habitats. The populations of the hydrologically isolated backwaters showed the lowest microsatellite polymorphism, while plants from an oxbow occasionally flooded by the main river had medium genetic diversity. The results highlight that in contrast to species that spread independently of water flow or among hydrologically isolated water bodies, macrophytes with exclusive or dominant hydrochory may be most severely affected by habitat fragmentation, for example due to climate change.

Exploring the trend effects of diffuse anthropogenic pollution in a large river passing through a densely populated area.

The detection of non-point pollution in large rivers requires high-frequency sampling over a longer period of time, which, however presumably provides data with large spatial and temporal variance. Variability may mean that data sets recorded upstream and downstream from a densely populated area overlap, suggesting at first glance that the urban area did not affect water quality. This study presents a simple way to explore trend-like effects of non-point pollution in the Danube based on data that varied strongly in space and time. For one year, biweekly sampling was carried out upstream and downstream from a large city with negligible emission of untreated wastewater and the surrounding settlements, industrial and agricultural areas. Although most of the values of the 34 examined physicochemical characteristics fell within the range of data previously published for the Danube, and the mean values of all parameters indicated unpolluted surface water, different water quality was revealed upstream and downstream from the metropolitan area at each sampling time. Since the physicochemical characteristics causing the separation also differed from time to time, univariate tests and consensus ordination were used to determine which variables changed similarly during most of the examined period. With this evaluation method, several diffuse pollutants of anthropogenic origin contaminating the Danube in the long term were identified, such as nitrogen, phosphorus, sulphate, chloride, potassium and vanadium. The results demonstrated that trend-like effects of non-point pollution can be detected even in a large river, where physicochemical measurements can vary strongly in space and time.

Significant impact of seasonality, verticality and biofilm on element accumulation of aquatic macrophytes.

Submersed macrophytes accumulate large amounts of macro- and trace elements from the environment and, therefore, are frequently used as indicators of water pollution and tools to remove pollutants from contaminated waters. This study provides evidences that the quantity of macro- and trace elements accumulated in the macrophyte Ceratophyllum demersum depends strongly on the seasonality, on the vertical position of the plant material and on the biofilm cover. Element contents of macrophytes with and without biofilm cover and that of vertical plant sections were investigated by an ICP-MS technique in three different habitats, at the beginning and at the end of the vegetation period. Results demonstrated that the element concentrations of Ceratophyllum demersum dropped to one-half and one-eighth by the end of the summer; and the amount of certain elements in the lower part of plants were up to six times higher than in the upper and in plants with well-developed epiphytic microbial community 2-5-fold higher than in plants without biofilm. These results help in phytoremediation practice and in setting up future biomonitoring studies. When it is necessary to calculate the exact amount of elements which can be accumulated by plants in a polluted environment or should be removed from a contaminated water by harvesting macrophytes, it is of high importance to consider the month of the study, the plant parts harvested and the biofilm cover.

Histological study of some Echium vulgare, Pulmonaria officinalis and Symphytum officinale populations.

Plants living in different ecological habitats can show significant variability in their histological and phytochemical characters. The main histological features of various populations of three medicinal plants from the Boraginaceae family were studied. Stems, petioles and leaves were investigated by light microscopy in vertical and transverse sections. The outline of the epidermal cells, as well as the shape and cell number of trichomes was studied in leaf surface casts. Differences were measured among the populations of Echium vulgare in the width and height of epidermis cells in the stem, petiole and leaf, as well as in the size of palisade cells in the leaves. Among the populations of Pulmonaria officinalis significant differences were found in the length of trichomes and in the slightly or strongly wavy outline of epidermal radial cell walls. Populations of Symphytum officinale showed variance in the height of epidermal cells in leaves and stems, length of palisade cells and number of intercellular spaces in leaves, and the size of the central cavity in the stem. Boraginaceae bristles were found to be longer in plants in windy/shady habitats as opposed to sunny habitats, both in the leaves and stems ofP. officinalis and S. officinale, which might be connected to varying levels of exposure to wind. Longer epidermal cells were detected in the leaves and stems of both E. vulgare and S. officinale plants living in shady habitats, compared with shorter cells in sunny habitats. Leaf mesophyll cells were shorter in shady habitats as opposed to longer cells in sunny habitats, both in E. vulgare and S. officinale. This combination of histological characters may contribute to the plant's adaptation to various amounts of sunshine. The reported data prove the polymorphism of the studied taxa, as well as their ability to adapt to various ecological circumstances.

Deglycosylation by small intestinal epithelial cell beta-glucosidases is a critical step in the absorption and metabolism of dietary flavonoid glycosides in humans.

BACKGROUND: Pharmacokinetic studies have shown that the small intestine is the major site of absorption for many flavonoid glucosides. Flavonoids are generally present as glycosylated forms in plants and foods, but there is increasing evidence that the forms reaching the systemic circulation are glucuronidated, sulphated and methylated derivatives. Hence, first-pass metabolism (small intestine-liver) appears to involve a critical deglycosylation step for which the mechanisms are not known. AIMS: To explore the hypothesis that deglycosylation is a prerequisite to absorption and metabolism of dietary flavonoid glycosides, to identify the enzymes responsible, and relate their specificities with absorption kinetics. METHODS: Flavonoid glycoside hydrolysing enzymes were isolated from samples of human small intestine and liver using chromatographic techniques. The proteins were characterised with respect to the cellular fraction with which they were associated, molecular weight, specificity for various substrates, and cross-reactions with antibodies. Cellular models were used to mimic the small intestine. RESULTS: Protein extracts from human jejunal mucosa were highly efficient in hydrolysing flavonoid glycosides, consistent with an enterocyte-mediated deglycosylation process. Considerable inter-individual variation was observed [e. g. range, mean and standard deviation for rate of hydrolysis of quercetin-3-glucoside (n = 10) were 6.7-456, 96, and 134 nmol min(-1) (mg protein)(-1), respectively]. Two beta-glucosidases with activity towards flavonoid glycosides were isolated from human small intestine mucosa: lactase-phlorizin hydrolase (LPH; localised to the apical membrane of small intestinal epithelial cells) and cytosolic beta-glucosidase (CBG), indicating a role of human LPH and CBG from small intestine in flavonoid absorption and metabolism. Hydrolysis of flavonoid glycosides was only detected in cultured cells exhibiting beta-glucosidase activity. CONCLUSIONS: The absorption of dietary flavonoid glycosides in humans involves a critical deglycosylation step that is mediated by epithelial beta-glucosidases (LPH and CBG). The significant variation in beta-glucosidase activity between individuals may be a factor determining variation in flavonoid bioavailability.