A New Type of Phytase from Pollen of Typha latifolia L.

A phytase was isolated and partially purified from pollen of cattail, Typha latifolia. Its maximum activity was at pH 8.0 and its Km value was 1.7 × 10^?5 m for phytic acid in the presence of Ca²⁺. Among divalent cations tested only Ca²⁺ affected the activity, increasing it by about 120%, but an excess was inhibitory. The enzyme was specific for phytic acid except for 6% activity for p-nitrophenylphosphate. It seems to be a new type of phytase because it cleaved almost 50% of the total phosphate esters in phytic acid and was product-specific, yielding an inositol triphosphate as a final product.     

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.     

Epifluorescent and histochemical aspects of shoot anatomy of Typha latifolia L., Typha angustifolia L. and Typha glauca Godr

Using epifluorescent and histochemical techniques, we examined anatomical differences in the shoot organs of Typha latifolia, T. angustifolia and T. glauca. The leaf lamina of T. latifolia and T. glauca had enlarged epidermal cells and a thickened cuticle above the subepidermal vascular bundles; that of T. angustifolia lacked these characteristics. Leaf sheaths were similar among the species and all lacked the epidermal thickenings found in the lamina. The fertile stems had typical scattered vascular bundles with a band of fibres that was most prominent in T. glauca. The sterile stems were only 1 cm in length and contained a multiseriate hypodermis and a uniseriate endodermis over part of their length. The rhizomes were similar except for a pronounced band of fibres surrounding the central core in T. angustifolia. The rhizome was also characterized by an outer cortical region with a large multiseriate hypodermis/exodermis and a uniseriate endodermis with Casparian bands, suberin lamellae and secondarily thickened walls.     

Growth strategy of an emergent macrophyte, Typha orientalis Presl, in comparison with Typha latifolia L. and Typha angustifolia L.

The growth strategy of an emergent plant, Typha orientalis Presl, was examined in experimental ponds in comparison with two other Typha species distributed in Japan, Typha latifolia L. and Typha angustifolia L. T. orientalis showed the greatest ability of vegetative reproduction at the expense of growth in height. T. orientalis started to produce new ramets earlier than T. latifolia and T. angustifolia. These results suggest that T. orientalis should be a rather pioneer-like species and would be restricted to disturbed habitats.     

Evapotranspiration by Eichhornia crassipes (Mart.) Solms and Typha latifolia L.

Evapotranspiration (E) by Eichhornia crassipes (Mart.) Solms and Typha latifolia L. growing in 5.77-m² tanks and evaporation (E₀) from control tanks were measured over a 6-month period at Auburn, Alabama (32.5° N latitude). The E/E₀ ratios for E. crassipes and T. latifolia were 1.31–2.52 (mean = 1.75) and 1.05–2.50 (mean = 1.62), respectively. Evidence is presented which demonstrates that E/E₀ values were similar to those which occur in natural populations of the two species. Both plant characteristics and meteorological variables influenced evapotranspiration. Equations for estimating evapotranspiration were E_Ec = (4.19 + (7.32 × 10⁻⁸) S² + (0.00035 × 10⁻³)H²)D R² = 0.92E_Tl = (1.43 + (2.79 × 10⁻¹⁵)S⁴ + 1.44L)D R² = 0.93 where E_Ec and E_Tl are monthly water loss in mm/month for E. crassipes and T. latifolia, respectively; S is the average daily solar radiation in W m⁻² integrated over 24 h for the month; H is plant height in m; L is leaf area index (dimensionless); and D is the number of days in the month.     

Depth distribution of three Typha species, Typha orientalis Presl, Typha angustifolia L. and Typha latifolia L., in an artificial pond

The depth distribution of the aquatic macrophyte Typha orientalis Presl was examined in comparison with two other Typha species: Typha angustifolia L. and Typha latifolia L. Vegetation surveys mapping the depth distributions were conducted at Ushigafuchi Pond, Tokyo, Japan, in autumn 2004 and 2005. All vegetation had been cleared from this artificial pond in spring 2003. In 2004 T. orientalis was distributed in shallow to deep water habitats between T. latifolia (shallow water regions) and T. angustifolia (shallow to deep water regions). However, by 2005 T. orientalis had almost disappeared from the pond. It had been replaced by Leersia japonica Makino at depths of 0–30 cm, by Schoenoplectus validus (Vahl) at depths of 30–60 cm and by T. angustifolia at depths of 60–100 cm. It appears that T. orientalis is not a strong competitor, particularly with taller species, but rather a pioneer species.     

Purification and Properties of UDP-glucuronate Pyrophosphorylase from Pollen of Typha latifolia Linne

UDP-glucuronate pyrophosphorylase from the pollen of Typha latifoliaLinne was purified about 600-fold by protamine sulfate treatment,ammonium sulfate fractionation, gel filtration, chromatofocusing,affinity chromatography, and isoelectric focusing. The purificationwas carried out using buffer containing 20% sucrose which helpedto prevent enzyme inactivation. This enzyme required equimolarlevels of Mg²⁺ to PP_i or UTP for maximum velocity of enzymecatalysis. Results of experiments on product inhibition andthe initial velocity of the enzyme catalysis reaction suggesteda Theorell-Chance mechanism. ¹ Present address: Japan Spectroscopic Co., 2967-5, Ishikawa-Cho,Hachioji-City, Tokyo 192, Japan. (Received April 5, 1983; Accepted September 26, 1983)     

Accumulation of Free Fatty Acids during Hardening of Chlorella ellipsoidea

Chlorella ellipsoidea Gerneck (IAM C-27) was synchronously grown, and cells at an intermediate stage in the ripening phase of the cell cycle were hardened at 3°C for 48 hours. A nonpolar lipid which increased greatly during hardening was analyzed by gas-liquid chromatography. Palmitic, oleic, linoleic, and linolenic acids were the main components of the lipid. Electron micrographs revealed the appearance of lipid bodies in hardened cells. When formation of free fatty acids and lipid bodies was inhibited with cycloheximide, oligomycin, and 3-(3,4-dichlorophenyl)-1,1-dimethylurea, the development of a high level of hardiness was always inhibited. However, the converse results were not always realized. Cells hardened in the dark in the absence of glucose developed a measurable hardiness in spite of their failure to form free fatty acids. The appearance of lipid bodies was invariably accompanied by the formation of the fatty acids. In pulse-labeling with [¹⁴C]NaHCO₃ for 4 minutes at zero time and at the 12th hour of hardening, initial incorporation rates of ¹⁴C into total lipids of whole cells and the cellular membrane fraction were significantly higher than that into free fatty acids. These results suggest that, although fatty acids are inserted into membrane lipids during hardening, the accumulation of free fatty acids and the appearance of lipid bodies per se are not involved in the development of frost hardiness.     

Dynamics of Typha latifolia L. populations in treatment wetlands in Estonia

The aim of this paper is to evaluate and compare broadleaved cattail (Typha latifolia L.) biomass production and the nitrogen (N) and phosphorus (P) content in phytomass in three treatment wetland systems and to propose suitable areas for treatment wetlands in Estonia for raw material production. The biomass samples (roots/rhizomes, shoots with leaves and spadixes) and litter were collected from 1 m × 1 m plots—15 plots in the Tänassilma semi-natural wetland, 15 plots in the Põltsamaa free water surface (FWS) constructed wetland (CW), and 10 plots in the Häädemeeste FWS CW. The average aboveground biomass of T. latifolia varied from 0.37 to 1.76 kg DW m^?2 in autumn and from 0.33 to 1.38 kg DW m^?2 in winter. The greatest average nitrogen (22,950 mg N kg^?1) concentration was found in spadixes in 2002, and the phosphorus (6500 mg P kg^?1) concentration was measured in roots–rhizomes in 2003. Average standing stock of nitrogen and phosphorus was higher in aboveground than belowground phytomass. In FWS CWs with high hydraulic and nutrient loadings, however, the harvesting of aboveground biomass is not an effective means for the removal of nutrients. Cattail biomass is a valuable insulation material, whereas the fibre from spadixes mixed with clay gives elasticity to clay plasters. According to our estimates, about 5412 km² could be used for Typha cultivation in Estonia.     

Effects of pH on ammonium uptake by Typha latifolia L.

The effects of solution pH on NH₄⁺ uptake kinetics and net H⁺ extrusion by Typha latifolia L. were studied during short-term (days) and long-term (weeks) exposure to pH in the range of pH 3.5–8.0. The NH₄⁺ uptake kinetics were estimated from depletion curves using a modified Michaelis-Menten model. T. latifolia was able to grow in solution culture with NH₄⁺ as the sole N source and to withstand a low medium pH for short periods (days). With prolonged exposure (weeks) to pH 3.5, however, the plants showed severe symptoms of stress and stopped growing. The solution pH affected NH₄⁺ uptake kinetics. The affinity for NH₄⁺, as quantified by the half saturation constant (K_1/2) and C_min (the NH₄⁺ concentration at which uptake ceases), decreased with pH. K_1/2 was increased from 7.1 to 19.2 mmol m^?3 and C_min from 2.0 to 5.7 mmol m^?3 by lowering the pH in steps from 8.0 to 3.5. V_max was, however, largely unaffected by pH (～22 μmol h^?1 g^?1 root dry weight). Under prolonged exposure to constant pH, growth rates were highest at PH 5.0 and 6.5. At pH 8.0 growth was slightly depressed and at pH 3.5 growth completely stopped. NH₄⁺ uptake kinetics were similar at pH 5.0, 6.5 and 8.0 whereas at pH 3.5 NH₄⁺ uptake almost completely stopped. The ratio between net H⁺ extrusion and NH₄⁺ uptake decreased significantly at low pH. The adverse effects of low pH on NH₄⁺ uptake kinetics are probably a consequence of a reduced H⁺-ATPase activity and/or an increased re-entry of H⁺ at low pH, and the associated decrease in the electrochemical gradient across the plasma membranes of the root cells.     

Thermal Inactivation Properties of Enzymes from Typha latifolia L. Ecotypes

Irreversible thermal denaturation experiments with 3 enzymes from Typha latifolia populations native to distinct thermal climates produced 3 different responses: (1) malate dehydrogenase was much more resistant to high temperature inactivation when obtained from plants native to a hot climate, (2) glutamate-oxaloacetate transaminase was quite resistant to thermal denaturation regardless of origin, and (3) aldolase was rapidly inactivated by heat regardless of origin.     

Root exudation pattern of Typha latifolia L. plants after copper exposure

Aims

Typha latifolia L. is an aquatic plant that has been widely exploited for the aims of phytoremediation. The main reason why we have chosen this plant species for the current study is its capacity to accumulate and detoxify heavy metals. The main topic of the investigation focused on the root uptake of copper (II) nitrate and copper (II) sulfate and the impact of different chemical copper species on the excreted organic acids.

Methods

Oxalic, malic, acetic and lactic acids were determined using capillary electrophoresis; a comparison between the concentration and the time course during 7 days of treatment was performed.

Results

There is a correlation between the total copper (II) sulfate concentration in the roots and the total amount of the excreted organic acids. In addition to that organic acids are involved in the detoxification mechanisms of Typha latifolia for copper (II) nitrate and copper (II) sulfate.

Conclusions

Different from so far investigated plant species the highest amounts of organic acids are excreted from T. latifolia roots not in the first hours after treatment, but up to 7 days later.     

Enzymatic variability in the cattail (Typha angustifolia andT. latifolia,Typhaceae)

The enzymatic spectra ofTypha angustifolia, T. latifolia and their hybrids were examined by disc electrophoresis for three enzymatic systems: esterases, alcohol deshydrogenase, and glutamate deshydrogenase. The intermediate position of the hybrids is confirmed. The study of alcohol deshydrogenase reveales the existence of an intraspecific variability inT. angustifolia between individuals from the Massif Armoricain in France and Dellys in Algeria.     

Release of Free Fatty Acids and Loss of Hill Activity by Aging Spinach Chloroplasts

The free fatty acid content of spinach chloroplasts, isolated at pH 5.8 to 8.0, has been found to vary between 3.1 and 5.5% of the total chloroplast fatty acids. When chloroplasts were incubated at room temperature for 2 hours, the free fatty acids increased by 42% and the Hill activity decreased by 70%. After 2 hours of incubation at 37 degrees , the free fatty acids increased about 3-fold and the Hill activity decreased to almost 0. The addition of crystalline bovine serum albumin largely prevented the loss of Hill activity at room temperature and at 5 degrees , but had little effect during incubation at 37 degrees . Both the release of free fatty acids and the loss of Hill activity were pH dependent. The losses were the least during incubation at pH 5.8 and the greatest during incubation at pH 8.0. The major free fatty acids released at pH 5.8 were saturated, while those released at pH 7.0 or 8.0 were mainly the unsaturated acids, alpha-linolenic acid and hexadecatrienoic acid.     

The Purification and Some Properties of the UDP-Glucose Pyrophosphorylase from Pollen of Typha latifolia Linne

UDP-glucose pyrophosphorylase from the pollen of Typha latifoliaLinné was purified to homogeneity as judged by polyacrylamidegel electrophoresis. Two affinity chromatographies with UDP-glucoseSepharose 6B and glucose-1-phosphate (G-l-P) Sepharose 6B wereused in the purification. The molecular weight of this enzyme was estimated to be about53,000 by gel filtration and 55,000 by SDS-polyacrylamide gelelectrophoresis, evidence that it consists of a single protein.It had a specific activity of 3,400, a molecular activity of180,000 and an equilibrium constant of 2.1 in the directionof UDP-glucose pyrophosphorolysis. Results of experiments on product inhibition and the initialvelocity of the reaction of the enzyme suggested an orderedBi Bi mechanism. (Received July 8, 1982; Accepted November 12, 1982)     

Nitrogen utilization by Typha latifolia L. as affected by temperature and rate of nitrogen application

Greenhouse and growth chamber studies were conducted to evaluate growth and N utilization by Typha latifolia L. in flooded organic soil under varying temperatures and rates of N additions. Elevation of temperature from 10 to 25°C increased shoot biomass yields by 275%. Root biomass yields were lowest at 10°C and increased linearly as a function of temperature. Shoot/root ratios were low (0.72–0.82) at lower temperatures (10–15°C) and ratios increased by about three times at higher temperatures (20–30°C). Biomass yields were increased by addition of N fertilizers, while the shoot/root ratios were directly related to plant-available N present in the soil.Fertilizer ¹⁵N uptake (expressed as % of applied N) by the whole plant was 5.3% at 10°C, 37.5% at 20°C and at 30°C decreased to 20.8%. Fertilizer N accumulation in shoots was 2.1–29.8% of applied N, while roots accumulated 3.2–7.7%. Under greenhouse conditions, N uptake by T. latifolia was found to increase with increased rate of N application. Fertilizer N uptake by both shoots and roots was in the range of 61–77%. Plants cultured in growth chambers were affected by low light conditions resulting in poor growth and low fertilizer ¹⁵N uptake, as compared to plants grown under greenhouse conditions. Added fertilizer N was the major source of N during the early part of the growing season, while soil organic N was the major and perhaps the sole source of N during the latter part of the growing season.     

Effects of soil moisture regimes on photosynthesis and growth in cattail (Typha latifolia)

Both waterlogging and water deficiency are major environmental factors affecting plant growth and functioning in many wetland and floodplain ecosystems across North America. Wetland plants possess various characteristics that enable them to survive and function in the intermittently flooded wetland environments, while their sensitivity to drought has received less attention. The present study quantified the photosynthetic and growth responses of cattail (Typha latifolia), an important species of freshwater wetlands, to a wide range of soil moisture regimes. In addition, changes in the efficiency of photosynthetic apparatus following initiation of the treatments were investigated. Under greenhouse conditions, seedlings were subjected to four soil moisture regimes: (1) drained (control), (2) continuous flooding, (3) periodic flooding, and (4) periodic drought. Results indicated that dark fluorescence yield was increased in response to periodic drought, while it showed decreases under continuous flooding. Net photosynthesis and stomatal conductance were enhanced by continuous flooding and periodic flooding. In contrast, these parameters exhibited reduction under periodic drought. In addition, leaf chlorophyll content was adversely affected by periodic drought. Recovery of net photosynthesis was noted, along with enhanced height growth, in both continuously and periodically flooded plants. Meanwhile, continuous flooding enhanced biomass production while periodic drought led to biomass reduction. Periodic drought also contributed to substantial reduction in root growth compared with shoot growth. Therefore, the combined photosynthetic performance and growth responses of cattail are likely to contribute to the ability of this species to thrive in flooded condition but be susceptive to periodic drought.     

Shoot regeneration and plant acclimatization of the wetland monocot Cattail (Typha latifolia)

A regeneration system for broadleaf cattail (Typha latifolia), a common freshwater wetland monocot, was established. Callus was induced from 3 day-old seedlings on Murashige and Skoog medium supplemented with picloram or 2,4-dichlorophenoxyacetic acid (2,4-D). Picloram induced more callus growth than 2,4-D. Callus proliferated predominantly from the root-shoot junction and sporadically from the roots. Upon transfer to N⁶-benzyladenine-supplemented medium, callus regenerated shoots. The mode of regeneration was organogenesis, with shoots arising from organized areas of callus. The regenerated shoots rooted in vitro and, on transfer to pots containing a commercial potting mix, were established in a greenhouse. The plants grew vigorously and produced a high root mass. No albinos and no morphological aberrations were observed in the regenerates. Received: 18 March 1998/ Revision received: 12 June 1998/ Accepted: 14 July 1998     

Effect of Sucrose on Phase Behavior of Membranes in Intact Pollen of Typha latifolia L., as Measured with Fourier Transform Infrared Spectroscopy

In previous studies, we have shown that the temperature dependent vibrational frequency of the CH₂ stretch in hydrocarbons in intact pollen grains can be recorded with Fourier transform infrared spectroscopy and used to measure phase transition temperatures (T_m) in these hydrocarbons. Circumstantial evidence was provided that the major contribution to the signal seen in these samples was from membrane phospholipids, and that sucrose in the dry pollen grains reduced T_m of those phospholipids. In the current study, we clarify why a major constituent of the pollen grains, neutral lipids contained in discrete lipid droplets, does not contribute significantly to the signal. Further, we have isolated membranes from the pollen and show that T_m in the isolated membranes rises from −6°C in the hydrated membranes to 58°C when the membranes are dried without the addition of sucrose. However, when the isolated membranes are dried in the presence of increasing amounts of sucrose, T_m fell steadily, reaching a minimal value of 31°C, a figure in good agreement with that seen in the intact pollen grains. The amount of sucrose required to depress T_m maximally in these membranes is also apparently in agreement with that found in the intact pollen, suggesting that sucrose depresses T_m in the pollen.