Root associated iron oxidizing bacteria increase phosphate nutrition and influence root to shoot partitioning of iron in tolerant plant Typha angustifolia

Aims

Typha angustifolia is a heavy metal tolerant plant that grows in a uranium mine tailings highly contaminated with iron. In this study three iron oxidizing microbes (FeOBs) isolated from Typha rhizoplane were investigated for their role in plant growth promotion (PGP). Their effect on iron nutrition in Typha under iron replete and excess condition was also evaluated.

Methods

The PGP activities of the FeOBs were studied by measuring their influence on plant growth. To investigate the mechanism of growth promotion their ability to solubilize phosphate, and to produce Indole acetic acid and siderophores were studied. The influence of the FeOBs on root to shoot partitioning of iron was tested by measuring total iron content in roots and shoots treated with microbes.

Results

The FeOBs were named as Paenibacillus cookii JGR8, (MTCC12002), Pseudomonas jaduguda JGR2 (LMG25820) and Bacillus megaterium JGR9 (MTCC12001). The siderophore producers, influenced iron accumulation in the plant root. Additionally P. pseudoalcaligenes JGR2 increased shoot iron content overcoming the root- shoot barrier that allows Typha to exclude metals from its shoot. Among the PGP mechanisms tested, ability to solubilize phosphate appeared to be most significant for increasing the plant biomass.

Conclusion

FeOBs that produce siderophore increased iron content in plant and therefore can be of immense biotechnological importance. However Biomass increase was directly correlated with increased phosphate acquisition and not with enhanced iron accumulation in Typha.     

Multiple elements in Typha angustifolia rhizosphere and plants: Wetland versus dryland

In a recent study, researchers found that multiple elements accumulated near the roots of Rumex crispus more under wetland conditions and element uptake was significantly greater in the plants grown under wetland compared to dryland conditions. The study reported here also found that elements accumulated in the root zone (up to 3 mm beyond the rhizoplane) of Typha angustifolia grown under wetland conditions. In comparison to the bulk zone, Be, Cu, Fe, Li, Sr and Zn accumulated more in the root zone of the flooded treatment whereas Ni and Sr accumulated more in the root zone of the non-flooded treatment. On average, T. angustifolia produced 4 times more biomass and element uptake was 2-27 times greater under wetland compared to dryland conditions.     

Interspecific differences in radial oxygen loss from the roots of three <Emphasis Type="Italic">Typha</Emphasis> species

A comparison was made of the radial oxygen loss (ROL) from the roots of three Typha species, Typha latifolia L., Typha orientalis Presl and Typha angustifolia L., which resemble each other in morphology. ROLs were evaluated in the laboratory for seedlings of T. orientalis and T. angustifolia in order to compare them with the ROL value for T. latifolia obtained in our previous study. Measurements were conducted using the highly oxygen-sensitive anthraquinone radical anion as an oxygen indicator, which enabled us to simulate the natural conditions in which the oxygen released from the root is immediately consumed by the soil. Among the three Typha species, the ROL was the highest in T. angustifolia, followed by T. latifolia and T. orientalis. Illumination significantly enhances the ROL of T. orientalis, and this effect was also observed for T. latifolia in our previous study, whereas it did not affect the ROL of T. angustifolia. These results indicate that ROL differs significantly between species, even among members of the same genus that are similar in morphology.     

Typha glauca Godron and its parental plants in Poland: taxonomic characteristics

Natural interspecific hybridization between Typha latifolia L. and Typha angustifolia L. was analyzed by morpho-anatomical and molecular methods to determine whether the hybrid Typha glauca Godr. is present in Poland and to identify the best diagnostic traits for its identification. Eighty-three samples of the Typha species were collected. Nine random amplified polymorphic DNA (RAPD) primers provided 12 fragments specific for T. angustifolia and eight fragments specific for T. latifolia. DNA of all sampled individuals was analyzed with 20 diagnostic RAPD markers. The morpho-anatomical variability of T. glauca F₁ was found to be quite similar to that observed in parental plants. All of the 41 traits examined in the hybrids overlapped with those observed in the parents, however, the hybrids were visibly closer to T. angustifolia than to T. latifolia. The most discriminate characteristics were the length and pedicel width, as well as the epidermal cell thickness located above vascular bundles in leaf blades. Moreover, preliminary observations of seed sculpture showed that the length of testa cells could also be used to identify T. glauca. Clusters and the hybrid index (for molecular and morphological data) were highly coincident and support the hybridization hypothesis.     

Evaluation of AFLP markers to reveal genetic diversity in Typha

We investigated to what extent DNA-markers can assist species determination in the genus Typha. A set of AFLP markers was used to discriminate samples of the species Typha latifolia and Typha angustifolia collected in Flanders (North Belgium). The T. latifolia samples formed a compact cluster while the T. angustifolia samples were divided into smaller groups. It was not clear whether interspecific hybrids or higher levels of diversity present in the T. angustifolia dataset could account for this. As in previous surveys, using isozyme and VNTR markers, AFLP markers revealed an almost complete lack of genetic variation in Flemish T. latifolia. Despite the low degree of diversity, a significant level of genetic differentiation was found between the T. latifolia samples originating from different river basins. Whether this differentiation has any ecological relevance remains to be investigated. The methodology applied was not able to detect clonal reproduction in T. latifolia. Probably, the low levels of diversity present in this species can account for this, indicating that the usefulness of the methodology applied depends on the level of diversity present in the species studied.     

Potential of Typha angustifolia for phytoremediation of heavy metals from aqueous solution of phenol and melanoidin

Typha angustifolia was evaluated for various heavy metals (Cu, Pb, Ni, Fe, Mn, and Zn) bioremediation potential from aqueous solution containing variable concentrations of phenol (100–800 mg l^?1) and melanoidin (2500–8500 Co–Pt) at 20, 40, and 60 days. The concentration of phenol (200–400 mg l^?1) along with melanoidin 2500 Co–Pt showed optimum for phytoremediation of tested heavy metals, while, higher concentrations of melanoidin (5600–8500 Co–Pt) showed toxic effect on T. angustifolia along with phenol. Phenol and melanoidin showed adverse effect on T. angustifolia of up to 20 days incubation, but this leads to induction of peroxidase and ascorbic acid activity to cope with adverse conditions. Subsequently, as pollutants were decreased along with plant growth, peroxidase and ascorbic acid also declined. However, with reduction of peroxidase, catalase level was increased. The Cu, Zn, and Ni were accumulated at maximum in all tested conditions. The TEM observations of T. angustifolia showed clotted deposition of metals and shrinkage of cell in root, breakdown of spongy and palisade parenchyma of leaves at higher concentration of phenol (100 mg l^?1) and melanoidin (5500 Co–Pt). Thus, this study concluded that T. angustifolia could be a potential phytoremediator for heavy metals from metal, melanoidin, and phenol containing industrial wastewater at optimized condition.     

Effects of copper and mineral nutrition on growth,copper accumulation and mineral element uptake in two Rumex japonicus populations from a copper mine and an uncontaminated field sites

Two Rumex japonicus populations, one from a copper mine and the other from an uncontaminated site, were studied in hydroponic experiments for the plant growth, copper accumulation and mineral nutrient content under excess copper and nutrient deficiency conditions. The tolerance indices of the contaminated population were significantly higher than that of the uncontaminated population, indicating the evolution of Cu resistance in the former. At control and low Cu treatment, there was no difference in Cu accumulation in roots between the two populations. At high Cu (100 μM) treatment, however, the contaminated population accumulated less Cu in roots than the uncontaminated one, suggesting the root exclusion mechanism existing in the former. The contaminated population was also more tolerant to general nutrient deficiency than the uncontaminated one. The results indicated that the contaminated population had evolved not only Cu-tolerance but also tolerance to low nutrient supply. Under Cu stress, the contaminated population had less change in nutrient composition than the uncontaminated one. The similar result was observed in general nutrient deficiency experiment. The results indicated that the mineral composition homeostasis under the stresses was important in metal tolerance and colonizing the Cu-enriched soils for the Cu-tolerant population. At high Cu (100 μM) treatment and general nutrient deficiency treatment, the contaminated population accumulated significantly lower copper and higher phosphorus in both roots and shoots than the uncontaminated one. This was not the case for other mineral nutrients Ca, Mg and Fe except for root Mg and root Fe at Cu treatment. The result suggested that the high Ca-metabolism in R. japonicus was uncorrelated with high Cu-tolerance and that P might play an important role in governing Cu bioaccumulation.     

Intercontinental dispersal of Typha angustifolia and T. latifolia between Europe and North America has implications for Typha invasions

The full effects of biological invasions may be underestimated in many areas because of cryptogenic species, which are those that can be identified as neither native nor introduced. In North America, the cattails Typha latifolia, T. angustifolia, and their hybrid T. × glauca are increasingly aggressive invaders of wetlands. There is a widespread belief that T. latifolia is native to North America and T. angustifolia was introduced from Europe, although there has so far been little empirical support for the latter claim. We used microsatellite data and chloroplast DNA sequences to compare T. latifolia and T. angustifolia genotypes from eastern North America and Europe. In both species, our data revealed a high level of genetic similarity between North American and European populations that is indicative of relatively recent intercontinental dispersal. More specifically, the most likely scenario suggested by Approximate Bayesian Computation was an introduction of T. angustifolia from Europe to North America. We discuss the potential importance of our findings in the context of hybridization, novel genomes, and increasingly invasive behaviour in North American Typha spp.     

Lignified and nonlignified fiber cables in the lacunae of <Emphasis Type="Italic">Typha angustifolia</Emphasis>

The leaves of Typha are noteworthy in terms of their mechanical properties. We determined the mechanical properties of the fiber cables within the leaf. We found that in vegetative plants, the lignified fiber cables isolated from the leaf sheath and nonlignified fiber cables isolated from the leaf blade of Typha angustifolia differ in their diameter, swelling capacity, Young’s modulus, tensile strength, and break load. These differing properties are related to their contributions to stability in the two regions of the leaf.     

Genetic relationship and genetic diversity among Typha taxa from East Asia based on AFLP markers

The genetic relationship and diversity among four Typha taxa in East Asia were evaluated using amplified fragment length polymorphism (AFLP) markers. Three AFLP selective primer combinations generated a total of 707 amplification products, of which 704 (99.6%) were polymorphic. The unweighted pair-group method with arithmetic mean (UPGMA) dendrogram and principal component analysis (PCA) plot confirmed the taxonomic status of four separate species. East Asian Typha taxa separated into two groups: the first with Typha angustifolia and the second with T. orientalis, T. laxmanni, and T. latifolia with a high bootstrap value for UPGMA (93%) and a low first score for PCA (25%). The two clusters corresponded with two sections based on the bracteoles in the female flower: section Bracteolatae and section Ebracteolatae. T. angustifolia showed the highest genetic diversity among the four Typha taxa (percentage of polymorphic loci [PPL] = 71%, H_o = 0.157), whereas T. latifolia had the lowest genetic diversity (PPL = 40%, H_o = 0.117). Genetic diversity was related to the presence of the gap between male and female inflorescences. A positive correlation between genetic distance and geographic distance was clearly found in the two species with continuous inflorescences (T. latifolia and T. orientalis). This positive correlation was not observed in the other species with discontinuous spikes (T. angustifolia and T. laxmanni).     

SALT TOLERANCE WITHIN A TYPHA POPULATION

Mc Millan , Calvin . (U. Texas, Austin.) Salt tolerance within a Typha population. Amer. Jour. Bot. 46(7): 521–526. Illus. 1959.—Typha in a disturbed salt flat near Lincoln, Nebraska, provided material for an examination of population dynamics. Within the population, clones of T. angustifolia L. tended to occupy the drier sites and those of T. latifolia L. occupied the sites of greater moisture probability. Clones of intermediate morphological characteristics were distributed with both T. angustifolia and T. latifolia. Rhizomes taken from the clones were grown in various NaCl solutions in the greenhouse. Results indicated greatest salt tolerance by T. angustifolia and least salt tolerance by T. latifolia. The intermediate, probably hybrid, clones were intermediate in salt tolerance. Seeds of the 3 clone-types germinated over the same range of salt concentration. The seeds of all 3 types withstood 4 months submergence in a 2% salt solution and germinated upon being returned to tap water. In the salt flat habitat, the clones of T. latifolia were not vigorous during the years 1956–1957 and many died or were reduced considerably in area of occupancy. The clones of T. angustifolia remained vigorous and flowered over the same period. The intermediate clones were vigorous and increased their coverage, primarily in areas that were occupied prior to 1956 by T. latifolia. The spatial adjustments within the population probably resulted from the selective action of increased salt concentration accompanying the drier conditions of 1956 and 1957.     

Effects of Cadmium,Chromium and Lead on Growth,Metal Uptake and Antioxidative Capacity in <Emphasis Type="Italic">Typha angustifolia</Emphasis>

This study investigates the modulation of antioxidant defence system of Typha angustifolia after 30 days exposure of 1 mM chromium (Cr), cadmium (Cd), or lead (Pb). T. angustifolia showed high tolerance to heavy metal toxicity with no visual toxic symptom when exposed to metal stress, and Cd/Pb addition also increased plant height and biomass especially in Pb treatment. Along with increased Cr, Cd, and Pb uptake in metal treatments, there was enhanced uptake of plant nutrients including Ca and Fe, and Zn in Pb treatment. A significant increase in malondialdehyde (MDA) content and superoxide dismutase (SOD) and peroxidase (POD) activities were recorded in plants subjected to Cr, Cd, or Pb stress. Furthermore, Pb stress also improved catalase (CAT), ascorbate peroxidase (APX), and glutathione peroxidase (GPX) activities; whereas Cr stress depressed APX and GPX. The results indicate that enzymatic antioxidants and Ca/Fe uptake were important for heavy metal detoxification in T. angustifolia, stimulated antioxidative enzymes, and Ca, Fe, and Zn uptake could partially explain its hyper-Pb tolerance.     

Pigment formation by callus of Lavandula angustifolia

Callus cultures of Lavandula angustifolia accumulated and secreted the (Z,E)-2-(3,4-dihydroxyphenyl)ethenyl ester of 3-(3,4-dihydroxyphenyl)-2-propenoic acid and its (E,E)-isomer under a wide range of culture conditions. The secreted compounds formed intensely blue pigments by chelation with Fe(su2+)in the media. These unusual enol esters could not be detected in the parent plant but the (Z,E)-isomer occurred in shoots and foliage of Plectranthus caninus.     

Remediation of AMD Contaminated Soil by Two Types of Reeds

Acid mine drainage (AMD) adversely impacts many regions in the world. The interactions among citric acid (CA), rhizosphere bacteria and metal uptake in different types of Phragmites australis cultured in spiked AMD contaminated soil were investigated. Compared with non-contaminated reeds cultured under the same conditions, wild reeds harvested from a contaminated site accumulated more metals into tissues. Rhizosphere iron oxidizing bacteria (Fe(II)OB) enhanced the development of Fe plaque but had no significant impact on the formation of Mn and Al plaque on the root surface of either reeds. Plaque may restrain the accumulation of Fe and Mn into tissues of reeds. CA inhibited the growth of Fe(II)OB, reduced the formation of metal plaque and significantly elevated metal accumulations into both underground and aboveground biomass of reeds. The concentrations of Fe, Al and Mn were higher in belowground organs than aboveground tissues. The roots contained 0.28 ± 0.01 mg/g Mn, 3.09 ± 0.51 mg/g Al, 94.47 ± 5.75 mg/g Fe, while the stems accumulated 0.19 ± 0.01 mg/g Mn, 1.34 ± 0.02 mg/g Al, 10.32 ± 0.60 mg/g Fe in wild reeds cultured in soil added with 33,616 ppm CA. Further field investigations may be required to study the effect of CA to enhance phytoremediation of metals from real AMD contaminated sites.     

Long-term competitive displacement of Typha latifolia by Typha angustifolia in a eutrophic lake

This study follows the outcome of long-term competition between a broad-leaved and a narrowleaved Typha species, T. latifolia and T. angustifolia respectively, in a eutrophic lake. The lake was bordered by a zone of T. latifolia, at one location interrupted by a T. angustifolia stand. Distributional changes of the T. angustifolia stand and the adjacent zone of T. latifolia were measured on aerial photographs (13 years) and along ground-level transects (6 years). A second stand of T. angustifolia was established with transplanted ramets within a formerly homogeneous zone of T. latifolia, and displacement between the two species was measured along ground-level transects after 6 years. Differences between the species in shoot performance were investigated to help explain the relative competitive abilities of the two Typha species. T. angustifolia expanded at the expense of T. latifolia at all water depths where both species occurred, except in very shallow water. Expansion rates suggest that T. angustifolia was not affected by the presence of T. latifolia in water depths exceeding 0.25 m. The Typha species were significantly negatively associated according to rank correlations of shoot densities, and changes of shoot densities, along the transects. These results suggest that T. angustifolia is competitively superior to T. latifolia, contradicting earlier studies. The higher competitive ability of T. angustifolia is consistent with its having taller shoots and a higher standing crop in early summer. Further, shoot height distributions indicated a closer integration of shoot emergence during spring in T. angustifolia than in T. latifolia. A high leaf area/shoot weight ratio suggest that T. latifolia may instead be relatively fast-growing, achieving competitive superiority over narrower-leaved Typha species during a transient period after simultaneous seedling establishment.     

Fibre cables in the lacunae of Typha leaves contribute to a tensegrity structure

Background and Aims

Cables composed of long, non-lignified fibre cells enclosed in a cover of much shorter thin-walled, crystal-containing cells traverse the air chambers (lacunae) in leaves of the taller species of Typha. The non-lignified fibre cables are anchored in diaphragms composed of stellate cells of aerenchyma tissue that segment the long air chambers into smaller compartments. Although the fibre cables are easily observed and can be pulled free from the porous-to-air diaphragms, their structure and function have been ignored or misinterpreted.

Methods

Leaves of various species of Typha were dissected and fibre cables were pulled free and observed with a microscope using bright-field and polarizing optics. Maximal tensile strength of freshly removed cables was measured by hanging weights from fibre cables, and Instron analysis was used to produce curves of load versus extension until cables broke.

Key Results and Conclusions

Polarized light microscopy revealed that the cellulose microfibrils that make up the walls of the cable fibres are oriented parallel to the long axis of the fibres. This orientation ensures that the fibre cables are mechanically stiff and strong under tension. Accordingly, the measured stiffness and tensile strength of the fibre cables were in the gigapascal range. In combination with the dorsal and ventral leaf surfaces and partitions that contain lignified fibre bundles and vascular strands that are strong in compression, the very fine fibre cables that are strong under tension form a tensegrity structure. The tensegrity structure creates multiple load paths through which stresses are redistributed throughout the 1–3 m tall upright leaves of Typha angustifolia, T. latifolia, T. × glauca, T. domingensis and T. shuttleworthii. The length of the fibre cables relative to the length of the leaf blades is reduced in the last-formed leaves of flowering individuals. Fibre cables are absent in the shorter leaves of Typha minima and, if present, only extend for a few centimetres from the sheath into the leaf blade of Typha laxmannii. The advantage of the structure of the Typha leaf blade, which enables stiffness to give way to flexibility under windy conditions, is discussed for both vegetative and flowering plants.     

Gas chromatography mass spectrometry based profiling of alkyl coumarates and ferulates in two species of cattail (Typha domingensis P., and Typha latifolia L.)

Several long-chain n-alkyl coumarates and ferulates were identified in cattails (Typha domingensis and Typha latifolia) from the Florida Everglades. Characterization of these compounds was achieved based on the interpretation of mass spectra obtained by GCMS as their trimethylsilyl ether derivatives, comparison with published mass spectra and available standards. Both n-alkyl p-coumarates and n-alkyl ferulates were identified in roots and leaves of both Typha species, featuring unique distribution patterns and differences between leaf and root biomass. For both Typha species, roots have higher concentrations and a much greater diversity of n-alkyl p-coumarates and ferulates but with different side chain carbon numbers ranging from C₁₄ to C₂₈. Typha domingensis leaves only contained n-alkyl ferulates with traces of n-alkyl p-coumarates, while both types of compounds were present in Typha latifolia leaf material. These chemicals were not found in the other dominant wetland vegetation, which suggests their potential for application as phytochemical tracers of fresh cattail-derived organic matter in the Everglades ecosystem.     

Comparative Analysis of Subcellular Distribution of Heavy Metals in Organs of the Bivalve Mollusks <Emphasis Type="Italic">Crenomytilus grayanus</Emphasis> and <Emphasis Type="Italic">Modiolus modiolus</Emphasis> in a Continuously Polluted Environment

The distribution of Zn, Fe, Ni, Cu, Mn, Cd, and Pb in subcellular fractions, and of Cd, Zn, and Cu in cytoplasm proteins of the kidney and digestive gland of the mussels Crenomytilus grayanus and Modiolus modiolus, sampled from contaminated and conditionally clean areas, was studied. It was found that, in a contaminated environment, the organs of mussels were more highly enriched with metals. It was shown that essential trace elements were accumulated mostly in the cytosol of organs of both molluscan species from contaminated areas, whereas in the background areas the trace elements were associated mostly with membrane structures in Gray's mussel, C. grayanus, and with the cytosol in M. modiolus, the northern horse mussel. The lead was bound mostly to membrane structures in organs of both mussel species at all stations. The method of gel chromatography enabled us to isolate metallothionein-like proteins from the kidney of the northern horse mussel sampled in contaminated areas, whereas their concentration in the kidney of Gray's mussels was lower than the limiting error of the method. It is supposed that in the kidney of Gray's mussel the synthesis of metallothionein-like proteins was quenched by the integrated effect of the accumulated metals.     

Molecular genetic data reveal hybridization between Typha angustifolia and Typha latifolia across a broad spatial scale in eastern North America

A recent increase in the abundance of cattails (Typha spp.) in North American wetlands has been anecdotally linked with hybridization between Typha latifolia and Typha angustifolia. In this study, we used molecular genetic markers (microsatellites) to investigate whether the hybrid lineage (Typha × glauca) is restricted to The Great Lakes region, or exists across a much broader spatial scale. We also investigated the possibility of backcrossing and genetic introgression in natural populations. Parental species could be distinguished from one another based on the distribution of alleles at six microsatellite loci. Species identification based on genetic data corresponded well with species identifications based on leaf width, a key morphological trait that can distinguish the two parental species. We found that hybrids occur in Ontario, Quebec, New Brunswick, and Nova Scotia, but we did not detect hybrids in Maine. F₁s are more abundant than backcrossed or intercrossed hybrids, although we also found evidence of backcrossing, particularly in Ontario. This indicates that hybrids are fertile, and are therefore potential conduits of gene flow between the parental species. Further work is needed to determine whether T. × glauca is particularly successful in the Great Lakes region relative to other areas in which the two parental species co-exist, and to assess whether introgression may lead to increased invasiveness in the species complex.     

Synergistic impact of water level fluctuation and invasion of Glyceria on Typha in a freshwater marsh of Lake Ontario

The effects of multiple stressors on the native Typha marsh community (mainly Typha latifolia) were examined using historical records of water levels, human census population, and field vegetation maps. Percent cover of the major plant species was estimated in a GIS, and the percent cover of Typha was related to changes in water level, human population growth, and percent cover of exotic Glyceria maxima and invasive Phragmites australis. Water level fluctuation was the major natural disturbance and it alone accounted for 88% of the variation in Typha. After partitioning out the effect of water level, both human population growth and the presence of exotic species were still significantly related to the decline of native Typha. We suggest that multiple stressors interact with each other to influence changes in native Typha community and cause greater detrimental impact. An important implication of our results is that projected water level decline due to climate change may not necessarily favor the restoration of a desirable native marsh because of the presence of other disturbances such as exotic and invasive species and altered nutrient regime.