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Genetic diversity of Typha latifolia (Typhaceae) and the impact of pollutants examined with tandem-repetitive DNA probes.

Genetic diversity at variable-number-tandem-repeat (VNTR) loci was examined in the common cattail, Typha latifolia (Typhaceae), using three synthetic DNA probes composed of tandemly repeated "core" sequences (GACA, GATA, and GCAC). The principal objectives of this investigation were to determine whether: (1) the previously reported almost complete lack of polymorphism at allozyme loci in this species was indicative of a reduced amount of genetic diversity at VNTR loci as well; (2) VNTR markers were informative about possible clonal propagation; and (3) significant differences in genetic structure of sampling sites were associated with differences in environmental levels of pollutants at those sites. Previously, widespread sampling across the eastern United States, surveying across ten allozyme loci, has detected only two genotypes, involving a difference at a single locus, among 104 populations. In this study, the amount of genetic diversity detected at VNTR loci: (1) among ramets (N = 40; 40 genotypes detected) collected at ∼8-km intervals along a 320-km transect; (2) among ramets (N = 220; 117 genotypes detected) from five study sites separated by 50-3000 m; and (3) even among ramets within each study site [N = 44 per site; from 13 to 34 genotypes detected per site (270 m(2))] exceeds that previously found in those more geographically widespread allozyme surveys. Among the 260 ramets analyzed here, the mean number of bands scored per individual was 48.61 (SD = 2.80). Mean genetic similarity among ramets collected along the 320-km transect was 0.91, which was within the range of mean genetic similarity within the five study sites (range: 0.89-0.95). Among the five study sites, 61% of the samples analyzed appeared to be clonal ramets, with up to 12 clones detected for 44 ramets sampled within a site. Clones grew intermingled and ranged up to 39 m in extent. Permutation tests of genetic similarity revealed significant genetic differentiation between each of the five study sites. Consistent with the previous allozyme studies, T. latifolia was characterized by extremely low genetic variation relative to levels of polymorphism detected at VNTR loci in other plant species. Estimated heterozygosity among ramets along the 320-km transect ranged from 0.11 to 0.13, while that within the five study sites ranged from 0.05 to 0.12. Estimates of F(st) (0.32-0.41) also indicated considerable genetic subdivision among these stands. Significantly higher genetic diversity was detected at the two study sites that chemistry and toxicity data indicate to be the most severely impacted by pollutants. Although this correlation does not establish cause and effect, the results of this study indicate that the analysis of genetic diversity at VNTR loci may be a useful tool for monitoring anthropogenic-induced changes in the genetic structure of natural populations of plants.

Journal Article↗

Fate of oxygen losses from Typha domingensis (Typhaceae) and Cladium jamaicense (Cyperaceae) and consequences for root metabolism.

The objective of this work was to determine whether radial oxygen loss (ROL) from roots of Typha domingensis and Cladium jamaicense creates an internal oxygen deficiency or, conversely, indicates adequate internal aeration and leakage of excess oxygen to the rhizosphere. Methylene blue in agar was used to visualize the pattern of ROL from roots, and oxidation of a titanium-citrate solution was used to quantify rates of oxygen leakage. Typha's roots had a higher porosity than Cladium's and responded to flooding treatment by increasing cortical air space, particularly near the root tips. A greater oxygen release, which occurred along the subapical root axis, and an increase in rhizosphere redox potential (E(h)) over time were associated with the well-developed aerenchyma system in Typha. Typha roots, regardless of oxygen release pattern, showed low or undetectable alcohol dehydrogenase (ADH) activity or ethanol concentrations, indicating that ROL did not cause internal deficiencies. Cladium roots also released oxygen, but this loss primarily occurred at the root tips and was accompanied by increased root ADH activity and ethanol concentrations. These results support the hypothesis that oxygen release by Cladium is accompanied by internal deficiencies of oxygen sufficient to stimulate alcoholic fermentation and helps explain Cladium's lesser flood tolerance in comparison with Typha.

Journal Article↗

Monocotyledon fruits and seeds, and an associated palynoflora from Eocene-Oligocene sediments of coastal central Queensland, Australia.

Fruits of Restionaceae and seeds of Typhaceae are described from a latest Eocene-Oligocene mudstone underlying oil shales in a subsurface sequence near Rockhampton, coastal central Queensland. The Restionaceae fruits are unilocular and encase a single pendulous orthotropous seed with a structured micropylar cap, which may be tannin filled. These are allocated to Restiocarpum gen. nov., which has as its type Restiocarpum latericum sp. nov., and four other taxa described herein; Restiocarpum tesselatum sp. nov., Restiocarpum verrucatum sp. nov., Restiocarpum laeve sp. nov., and Restiocarpum fusiforme sp. nov. Typhaspermum cooksoniae gen. et sp. nov., which accommodates asymmetric claviform seeds, is interpreted as a member of the Typhaceae based on the presence of a lid-like operculum, bitegmic wall, and chalazal cavity.Reconstruction of the source plant community emphasizes similarities to restiad swamps of present day Wallum (swamp heathland) vegetation which is extensively developed along the Queensland coast. Biogeographic implications for the Restionaceae and Typhaceae are discussed.

Journal Article↗

Diversity of lepidopteran stem borers on monocotyledonous plants in eastern Africa and the islands of Madagascar and Zanzibar revisited.

Surveys were completed in Eritrea, Ethiopia, Kenya, Madagascar, Mozambique, Tanzania, Uganda and Zanzibar to assess the lepidopteran stem borer species diversity on wild host plants. A total of 24,674 larvae belonging to 135 species were collected from 75 species of wild host plants belonging to the Poaceae, Cyperaceae and Typhaceae. Amongst them were 44 noctuid species belonging to at least nine genera, 33 crambids, 15 pyralids, 16 Pyraloidea species not yet identified, 25 tortricids and three cossids. The noctuid larvae represented 73.6% of the total number of larvae collected, with 66.3, 3.5 and 3.8% found on Poaceae, Cyperaceae and Typhaceae, respectively. The Crambidae, Pyralidae, Tortricidae and Cossidae represented 19.8, 1.9, 2.5 and 0.1% of the total larvae collected, respectively, with 90.4% of the Crambidae and Pyralidae collected from Poaceae, and 99.7% of the Tortricidae collected from Cyperaceae. The lepidopteran stem borer species diversity in the wild host plants was far more diverse than previously reported.

Africa, Eastern↗

Genetic modification of wetland grasses for phytoremediation.

Wetland grasses and grass-like monocots are very important natural remediators of pollutants. Their genetic improvement is an important task because introduction of key transgenes can dramatically improve their remediation potential. Tissue culture is prerequisite for genetic manipulation, and methods are reported here for in vitro culture and micropropagation of a number of wetland plants of various ecological requirements such as salt marsh, brackish water, riverbanks, and various zones of lakes and ponds, and bogs. The monocots represent numerous genera in various families such as Poaceae, Cyperaceae, Juncaceae, and Typhaceae. The reported species are in various stages of micropropagation and Arundo donax is scaled for mass propagation for selecting elite lines for pytoremediation. Transfer of key genes for mercury phytoremediation into the salt marsh cordgrass (Spartina alterniflora) is also reported here. All but one transgenic lines contained both the organomercurial lyase (merB) and mercuric reductase (merA) sequences showing that co-introduction into Spartina of two genes from separate Agrobacterium strains is possible.

Bacterial Proteins↗

Determination of alkanolamines in cattails (Typha latifolia) utilizing electrospray ionization with selected reaction monitoring and ion-exchange chromatography.

Selected reaction monitoring (SRM) with electrospray ionization was used as a specific detection technique for the analysis of alkanolamines in plant tissue extracts. Ion-exchange chromatography was used as the method of separation. Quantification was based on monitoring the loss of either H2O or 2(H2O) from the protonated molecule [M+H]+. The method provided increased selectivity for all analytes and better detection limits for three of the six analytes investigated compared with an earlier method using selected ion monitoring with liquid chromatography. Instrumental detection limits ranged from 6-300 pg injected for monoethanolamine (MEA), monoisopropanolamine (MIPA), diethanolamine (DEA), methyldiethanolamine (MDEA), diisopropanolamine (DIPA), and triethanolamine (TEA). Method robustness and selectivity were demonstrated by the determination of DIPA and a known transformation product MIPA in over 35 plant extract samples derived from a laboratory study of plant uptake mechanisms.

Amino Alcohols↗

Isolation and characterization of polymeric galloyl-ester-degrading bacteria from a tannery discharge place.

The culturable bacteria colonizing the rhizosphere of plants growing in the area of discharge of a tannery effluent were characterized. Relative proportions of aerobic, denitrifying, and sulfate-reducing bacteria were determined in the rhizosphere of Typha latifolia, Canna indica, and Phragmites australis. Aerobic bacteria were observed to be the most abundant group in the rhizosphere, and plant type did not seem to influence the abundance of the bacterial types analyzed. To isolate bacteria able to degrade polyphenols used in the tannery industry, enrichments were conducted under different conditions. Bacterial cultures were enriched with individual polyphenols (tannins Tara, Quebracho, or Mimosa) or with an undefined mixture of tannins present in the tannery effluent as carbon source. Cultures enriched with the effluent or Tara tannin were able to degrade tannic acid. Six bacterial isolates purified from these mixed cultures were able to use tannic acid as a sole carbon source in axenic culture. On the basis of 16S ribosomal DNA sequence analysis, these isolates were closely related to organisms belonging to the taxa Serratia, Stenotrophomonas maltophilia, Klebsiella oxytoca, Herbaspirillum chlorophenolicum, and Pseudomonas putida.

Bacteria↗

Periphytic photosynthetic stimulation of extracellular enzyme activity in aquatic microbial communities associated with decaying typha litter.

We examined the effect of light on extracellular enzyme activities of periphytic/endogenous microbial assemblages associated with decomposing litter of an emergent macrophyte Typha angustifolia within a small inland wetland in southeastern Michigan. Standing-dead Typha leaf litter was collected, placed into floating wire mesh litter baskets, and submerged in a wetland pool. Enzyme saturation assays were conducted on three occasions following litter submergence (days 9, 28, and 44) to generate saturation curves for the individual enzymes tested and to examine potential differences in enzyme saturation kinetics during microbial colonization and development. Experimental light manipulations were conducted on two occasions during microbial development (days 10 and 29). Short-term (30 min) light exposure significantly increased extracellular beta-glucosidase activity of litter-associated microbial communities. Activities of beta-xylosidase and leucine-aminopeptidase were not stimulated, and stimulation of phosphatase activity was variable. The exact mechanism for increased enzyme activity remains unknown, but it may have been increased pH arising from periphytic algal photosynthesis. These results suggest that extracellular enzyme activity in microbial communities colonizing natural organic substrata may be influenced by light/photosynthesis, as has previously been demonstrated for periphyton communities grown on artificial, inert substrata. Thus, light/photosynthetic mediated stimulation of extracellular enzyme activities may be a common occurrence in microbial communities associated with natural decaying plant litter in wetlands and might engender diurnal patterns in other microbial decay processes (e.g., production, organic matter decomposition, and mineralization).

Bacteria↗

Applicability of a septic tank/engineered wetland coupled system in the treatment and recycling of wastewater from a small community.

A septic tank (ST)/engineered wetland coupled system used to treat and recycle wastewater from a small community in Dar es Salaam, Tanzania was monitored to assess its performance. The engineered wetland system (EWS) had two parallel units each with two serial beds packed with different sizes of media and vegetated differently. The larger-sized medium bed was upstream and was planted with Phragmites (reeds) and the smaller-sized medium bed was downstream and was planted with Typha (cattails). The ST/EWS coupled system was able to remove ammonia by an average of 60%, nitrate by 71%, sulfate by 55%, chemical oxygen demand by 91%, and fecal coliform as well as total coliform by almost 100%. The effluent from the ST/EWS coupled system is used for irrigation. Notably, users of the recycled irrigation water do not harbor any negative feelings about it. This study demonstrates that it is possible to treat and recycle domestic wastewater using ST/ EWS coupled systems. The study also brings attention to the fact that an ST/EWS coupled system has operation and maintenance (O&M) needs that must be fulfilled for its effectiveness and acceptability. These include removal of unwanted weeds, harvesting of wetland plants when the EWS becomes unappealingly bushy, and routine repair.

Agriculture↗

Use of microbial community to evaluate performance of a wetland system in treating Pb/Zn mine drainage.

The performance of a wetland system in treating lead (Pb)/zinc (Zn) mine drainage was evaluated by using the polyurethane foam unit (PFU) microbial community (method), which has been adopted by China as a standardized procedure for monitoring water quality. The wetland system consisted of four cells with three dominant plants: Typha latifolia, Phragmites australis and Paspalum distichum. Physicochemical characteristics [pH, EC, content of total suspended solid (TSS) and metals (Pb, Zn, Cd, and Cu)] and PFU microbial community in water samples had been investigated from seven sampling sites. The results indicated that the concentrations of Pb, Zn, Cd, Cu, and TSS in the mine drainage were gradually reduced from the inlet to the outlet of the wetland system and 99%, 98%, 75%, 83%, and 68% of these metals and TSS respectively, had been reduced in concentration after the drainage passed through the wetland system. A total of 105 protozoan species were identified, the number of protozoa species and the diversity index (DI) gradually increased, while the heterotrophic index (HI) gradually decreased from the inlet to the outlet of the wetland system. The results indicated that DI, HI, and total number species of protozoa could be used as biological indicators indicating the improvement of water quality.

Animals↗

Agrobacterium-mediated transformation of the wetland monocot Typha latifolia L. (Broadleaf cattail).

An Agrobacterium-mediated model transformation system was standardized for the wetland monocot Typha latifolia L. to achieve the long-term objective of introducing candidate genes for phytoremediation. Two binary plasmid vectors, pCAMBIA1301/EHA105 and pTOK233/LBA4404, both containing the gus (beta-glucuronidase) and hptII (hygromycin phosphotransferase II) genes, were used for transformation. Fifty-day-old 5 mg/l picloram-derived calli were cocultivated and selected on medium containing 20 mg/l or 40 mg/l hygromycin. Resistant calli were regenerated on medium supplemented with 5 mg/l 6-benzylaminopurine, with or without 20 mg/l or 40 mg/l hygromycin and with or without charcoal (10 g/l). Transient GUS activity in explants ranged between 28% and 36%. Hygromycin-resistant calli, selected after 3 months, showed stable GUS expression. A total of 46 plants were regenerated and established in the greenhouse; 13 showed stable GUS expression. Cocultivation of dark culture-derived calli, directly selected on regeneration medium containing 20 mg/l hygromycin and rooted on medium with 20 mg/l hygromycin was the best protocol. The addition of charcoal did not have any effect on regeneration. PCR and Southern analyses of transgenic calli and transgenic plants confirmed the presence of the introduced genes. In conclusion, T. latifolia could be genetically transformed by Agrobacterium tumefaciens.

Blotting, Southern↗

Decolorization of basic, direct and reactive dyes by pre-treated narrow-leaved cattail (Typha angustifolia Linn.).

The efficiency of basic, direct and reactive dye removal from water by narrow-leaved cattail (NLC) powder treated with distilled water (DW-NLC), 37% formaldehyde+0.2 N sulfuric acid (FH-NLC), or 0.1 N sodium hydroxide (NaOH-NLC) at various pH levels (3, 5, 7, and 9) was tested. Desorption of the adsorbed dyes was also investigated. The type of NLC treatment and pH of the dye solution had little effect on removal of basic dyes, and efficiencies ranged from 97% to 99% over the range of pH used. Over a wide range of pH levels, all types of treated cattail powder had negative charges and probably attracted the basic dyes possessing positive charges. Efficiency of removal by the three NLC treatments ranged from 37% to 42% for direct dyes and from 22% to 54% for direct dyes at pH 7. The pH of the dye solution had substantial effects on the efficiency of removal in direct and reactive dyes. Dye removal was highest at pH 3, with 99% for a direct dye (Sirius Red Violet RL) and 96% for a reactive dye (Basilen Red M-5B). There was mutual attraction between negatively charged direct dye molecules and positively charged molecules on the surface of the FH-treated cattail. In tests of desorption of dyes from cattail in distilled water, the desorption percentage for FH-NLC after adsorbing basic, direct and reactive dyes was 6%, 10% and 35%, respectively, which indicated a chemisorption mechanism for basic and direct dyes and some physiosorption for reactive dyes.

Bioreactors↗

Application of response surface methodology for optimization of acidogenesis of cattail by rumen cultures.

Acidogenesis of cattail using rumen cultures was carried out to produce volatile fatty acids (VFA) in this study. The influences of pH and substrate concentration on cattail degradation, VFA yield and microbial growth were investigated by using response surface methodology (RSM). Experimental results showed that a low substrate concentration and pH of 6.9 were optimal for acidogenesis of cattail. The highest cattail degradation efficiency, VFA yield and microbial yield were 75.9%, 0.41 g/g VS and 0.110 g/g VS, respectively. Further experiments confirmed that the main VFA in the acidogenesis of cattail were acetate, propionate and butyrate, while i-butyrate, valerate and i-valerate were also produced at low levels. The results suggested that acidogenesis using rumen cultures is a promising method for cattail disposal.

Animals↗

In situ determination of slurry nutrient content by electrical conductivity.

Land application of animal slurries has both agronomic and environmental implications. It can be supported by the quantification of available nutrients in the field. A prototype device for indirect measurement of the nutrient content of slurry based on electrical conductivity (EC) was calibrated on manure samples collected from farms with different livestock typologies. The resulting correlations between EC and nutrient contents of slurries from laboratory analyses have shown good agreement (r(2) from 0.73 to 0.95) with total and ammoniacal nitrogen and, with some exception, Potassium, but failed to demonstrate any significant relationship with total phosphorous. The mean errors obtained using the device in field conditions for nitrogen content were always lower or equal to 10%, while the standard deviations were 12-13% for pig and calf slurries, and 20-21% for dairy cow slurry. The results obtained suggest that the equipment, provided the regression line used to convert EC readings to nutrient contents is related to the livestock typology under observation, can provide good support to practical slurry spreading, even though it does not reach an accuracy comparable to laboratory methods and does not give reliable information on phosphorus.

Animals↗

Accumulation of heavy metals in Typha angustifolia (L.) and Potamogeton pectinatus (L.) living in Sultan Marsh (Kayseri, Turkey).

Concentrations of heavy metals (Cd, Pb, Cr, Ni, Zn and Cu) were measured in bottom sediments, water and Typha angustifolia and Potamogeton pectinatus in Sultan Marsh. Sultan Marsh is one of the largest and most important wetlands in Turkey, Middle East and Europe, embodying saline and fresh water ecosystems and providing a shelter for 426 bird species. The organs of T. angustifolia have a larger quantity of the measured elements than the P. pectinatus. Considerably higher contents of Cd were found rather than in helophytes (P. pectinatus) in submerged plant (= emergent, T. angustifolia) species. The percentage of Cd in plant tissues points to a certain degree of water pollution in Sultan Mash. Analyses of water, bottom sediments and plant samples indicated that the Marsh were polluted with Pb, Cd, and partly with Cu and Zn. All sampling sites in the study area basin are generally more or less polluted when compared with the control values. Strong positive correlation was found between concentrations of Pb in water and in plants. Ni and Pb were accumulated by plants at a higher rate from bottom sediments than from water. Leaves of T. angustifolia accumulated less heavy metal than the corresponding roots. There was a significant relationship between Cd concentration in samples of plants and water pH value. It has been found that the tissues of T. angustifolia accumulate more heavy metals than the tissues of P. pectinatus. Therefore, all plants can be used as a biological indicator while determining environmental pressures; however, T. angustifolia is proved more appropriate for such studies.

Environmental Monitoring↗

Methyl parathion toxicity to and removal efficiency by Typha latifolia in water and artificial sediments.

Methyl parathion (MeP) is a very hazardous pesticide freely used in agriculture in Mexico. This pesticide and others, arriving through different processes, exert significant effects on water quality with serious consequences for environmental and human health. This study evaluates the removal efficiency of common cattail Typha latifolia L. on MeP in water and artificial sediments. The effects of the pesticide on this macrophyte after 10 days of exposure were determined using a concentration range of 0-200 mg l(-1), 198.1+/-1.79 g average biomass, pH 7.0, 18-22 degrees C temperature and natural daylight/darkness periods, using chlorophyll production as a biomarker. Removal kinetics were conducted under similar conditions on days 0, 3, 7, 9, 11 and 14 of exposure, using 6 mg l(-1) in each system. Pesticide concentration, chlorophyll content and glutathione S-transferase (GST) activity were quantified. Results show a high removal efficiency of cattails on MeP in water and sediments relative to controls. An increase in GST activity and a decline in chlorophyll content in the test systems were not significantly different relative to controls. Cattails may thus be a good candidate for development of a phytoremediation system for MeP-contaminated water and artificial sediments.

Biodegradation, Environmental↗

Assessment of application-rate dependent effects of a long-term fire retardant chemical (Fire Trol 934) on Typha domingensis germination.

Although long-term fire retardants (LTR) gain increasingly acceptance as effective tools for wildfire management, recent studies indicate their potential harmfulness in the environment, especially in aquatic ecosystems. This study comprises a first laboratory impact assessment of LTR action in temporal Mediterranean wetlands, using seed germination of Typha domingensis as indicator of impact. Our aim was to identify application rates upon which seed germination could be significantly affected. We tested for low (1 l m(-2)) and high (3 l m(-2)) application rates of Fire Trol 934 which are recommended by the manufacturers as a function of fuel characteristics. In addition, we simulated the impact of a higher application rate of 5 l m(-2) because inhomogeneous dispersal of the LTR during fire control and prevention operations can result in locally elevated applications. Results of a microcosm experiment indicate that application rates of 1 or 3 l m(-2) can impact Typha germination rates in the short-term via indirect LTR-mediated effects on water quality, which suppressed necessary cues for germination. However, a subsequent experiment with Petri dishes, using seeds isolated from the LTR treated sediments did not show significantly different germination rates between the control and the treatments with application rates of 1 or 3 l m(-2). This suggests that retardant pre-application germination success could be recovered in nature once the retardant is eliminated. By contrast, seeds almost completely failed to germinate in the microcosm experiment and the subsequent Petri dish essay when an application rate of 5 l m(-2) was used. This suggests a critical level upon which Typha seed germination may be perpetually limited. Research should be extended to other plant species to provide fire managers with guidelines for environmentally safe use of LTR in the Mediterranean region.

Dose-Response Relationship, Drug↗

Mosquito larvae density and pollutant removal in tropical wetland treatment systems in Honduras.

Constructed wetlands offer a low-cost wastewater treatment option for tropical developing countries. The vast majority of published treatment wetland research has been conducted in temperate regions. Because the function of treatment wetlands is related to the environmental conditions, more research specific to the tropics should be completed. A six-cell free water surface (FWS) wetland mesocosm was constructed in Santa Rosa de Copán, Honduras that received input from an open-sewer/wastewater-impacted stream. Three cells were planted with Typha domingensis Pers., and three cells were left unplanted. Both planted and unplanted wetlands were constructed with three different surface areas to concurrently study different hydraulic retention times (HRTs) and hydraulic loading rates (HLRs). Results from 6 months of operation showed improved water quality and mosquito larvae populations affected by their specific environment. Five-day biochemical oxygen demand (BOD) removal appeared to be proportional to HRT and HLR, and BOD concentration in the effluent was higher in unplanted cells than in planted cells (P<0.05). BOD removal approached 60% for greater than 3.5 days HRT. Total phosphorus (P) removal and coliform bacteria removal were found to be correlated with pollutant load, and P removal was found to be correlated with HRT but not HLR. A second municipal wastewater FWS treatment wetland in Copán Ruinas, Honduras that had been established for 15 years, was also evaluated. BOD removal in the Copán Ruinas system was determined to be 93% with 2.6 day HRT. These pollutant removal rates are higher than would be expected in temperate regions. Mosquito larvae density was correlated with depth of the wetland. Mosquito larvae densities were higher in cells that were operating below design depth of 0.2 m (P<0.005). Results indicate that it is possible to design FWS wetlands in the tropics for effective BOD removal and reduced mosquito larvae development.

Animals↗