Accumulation of selected heavy metals by different genotypes of Salix

The aim of this study was to assess the accumulation of seven heavy metals (lead, cadmium, copper, zinc, chrome, nickel and cobalt). The investigations were conducted on twelve genotypes of willows which grow in the Potasze Forest Division Salicarium. The analysis facilitated quantification of concentrations of selected metals in plants and their comparison in relation to sorption ability of each willow genotype. Simultaneously the studies allowed us to demonstrate essential differentiation of metal size sorption within the species Salix purpurea and in relation to the other genotypes. The results confirm the complexity of factors influencing the efficiency of heavy metal accumulation by willow; they indicate increasing ion absorption in the case of some metals, while the accumulation of other heavy metal ions was limited.     

Morphogenesis of the root cap in adventitious roots of Salix viminalis

The preformed root primordia in stems of Salk viminalis L. consist of undifferentiated cells. Forty-eight hours after activation of the primordia in cuttings a root cap meristem was initiated four to five cell tiers from the surface of the primordia. The cells distal to the meristem divided only in an anticlinal plane, while in the meristem they divided mostly periclinally but sometimes anticlinally. After 72 hours a columella was established and the amyloplasts began to sediment in response to gravity. Shortly after this stage the roots began to bend slightly downward, probably as a geo-tropic response. Six days after activation the root cap consisted of up to 15 tiers of cells. The ultrastucture of the cap cells just prior to emergence was studied in more detail. The plastids in the cells adjoining the root proper were typical proplastids. Distal to this cell tier starch accumulated in the plastids. In the fifth tier the amyloplasts were fully sedimented to the lowermost cell walls. The amount of ER increased with the distance from the initial cells and most of it was located at the distal periclinal cell wall. The nucleus and the vacuoles in the geo-sensitive cells occurred in the space above the sedimented amyloplasts. The cytoplasm was less electron opaque than in the initial cells and the mitochondria had more cristae. In the distal cells of the columella and the lateral root cap secretion of mucilage seemed to have started. Numerous large dictyosomes were associated with large vesicles containing a fibrillar or granular material. The plasmalemma lining the distal periclinal cell wall had separated from the wall. A fibrillar material was present between the plasmalemma and the wall and also in intercellular spaces outside the root cap.     

Ultrastructural features of differentiating protophloem sieve elements in adventitious roots of Salix viminalis

The differentiation of the protophloem in 9- to 14-day-old adventitious roots of Salix viminalis was studied. Ultrastructural observations were mainly made on longitudinal serial sections through an uninterrupted file of 32 differentiating sieve elements. The first cell in the file was located about 50 μm from the apical meristem. At an early stage the nucleus was lobed in outline, and in older cells the nucleoplasm became electron lucent. In the first or second cell from the first mature sieve element the nuclear envelope broke open. The nucleoli decreased gradually in size and disappeared finally. From the 9th cell the plastids contained starch and grew somewhat in size. ER increased in amount and began to form stacks in the 20th cell. These stacks moved to a peripheral position. Callose platelets were first observed on the transverse walls in cell 18. Flattened ER-cisternae covered the sieve pore sites. Gradually the middle lamella was dissolved and the callose aggregations formed cylinders around the pores of the sieve plate. Aggregations of tubular P-protein were present from cell 15. P-protein bodies were also present in parenchyma cells adjoining mature sieve elements. The only cell components remaining in mature sieve elements were plastids, mitochondria, stacked ER, the plasmalemma, remnants of other membranes and bodies consisting of P-protein and of an unidentified granular material. The sieve elements had no ontogenetically related companion cells. At a level where both metaphloem and metaxylem had matured the first formed protophloem sieve elements remained intact.     

Magnesium and iron deficiencies alter Cd accumulation in Salix viminalis L.

Evidence exists that Cd and certain nutrient elements, such as Fe and Mg, could share similar mechanisms of plant uptake and accumulation. Here we report that Mg and Fe deficiency in mature plants of Salix viminalis, grown in hydroponic solutions containing 5 µg ml^?1 of Cd, caused a significant increase in Cd accumulation in roots, stems and leaves. Cd (µg g^?1 dry weight) was determined following three treatments: 1) Cd treatment in complete nutrient solution; 2) Cd treatment with Fe deficiency; and 3) Cd treatment with Mg deficiency, yielding, respectively: in young leaves (65.3, 76.1, and 92.2), mature leaves (51.5 to 76.3 and 87.1), upper stems (80.6, 116.8, and 130.6) lower stems (67.2, 119, and 102.3), roots (377.1, 744.8, and 442,5). Our results suggest that Cd utilizes the same uptake and transport pathways as Mg and Fe. Evidence exists that Mg and Fe uptake and translocation could be further facilitated by plants as an adaptive response to deficiency of these elements. Such physiological reaction could additionally stimulate Cd accumulation. Although Cd uptake was mostly confined in roots, high Cd content in aerial plant parts (51.5–130.6 µg g^?1) indicates that the analysed Salix viminalis genotype is suitable for phytoextraction.     

Selective accumulation of heavy metals by microorganisms

Summary An investigation of the removal and recovery of urnnium from aqueous systems using microbial biomass has been described previously (Nakajima et al. 1982). To establish which microorganisms accumulate the most uranium, we extended our investigation of uranium uptake to 83 species of microorganisms, 32 bacteria, 15 yeasts, 16 fungi and 20 actinomycetes. Of these 83 species of microorganisms tested, extremely high uranium-absorbing ability was found in Pseudomonas stutzeri, Neurospora sitophila, Streptomyces albus and Streptomyces viridochromogenes.The selective accumulation of heavy metal ions by various microorganisms has also been examined. Uranyl, mercury and lead ions were readily accumulated by almost all the species of microorganisms tested. Actinomycetes and fungi differ from many bacteria and most yeasts in their selective accumulation of uranium and mercury.In addition to this fundamental research, uranium recovery was investigated in immobilized Streptomyces albus, a microorganism with high uranium-uptake ability. These immobilized cells adsorbed uranium readily and selectively. The immobilized cells recovered uranium almost quantitatively and almost all uranium absorbed was desorbed with 0.1 M Na₂CO₃. The dry weight of the free cells decreased by 50% during 5 adsorption-desorption cycles. However, the dry weight of the immobilized cells decreased by only 2% during 5 cycles. These results showed that microbial cells are more stable after immobilization and can be used repeatedly for the process of uranium adsorption-desorption.     

Effect of heavy metals combined stress on growth and metals accumulation of three Salix species with different cutting position

This study aimed to compare growth performance and heavy metal (HM) accumulation at different cutting positions of Salix species grown in multi-metal culture. Three Salix species stems cut at different positions (apical to basal) were grown hydroponically for four weeks. The plants were then treated for three weeks with 0, 5, 10, and 20 μM Cd, Cu, Pb, and Zn, resulting in total metal concentrations of 0, 20, 40, and 80 μM. The growth parameters and HM content in shoots and initial cutting were measured. Results showed that, compared with S. fragilis, S. matsudana grew more poorly in uncontaminated condition but grew better and accumulated lower metal in shoots under mixed HM treatment. In addition, cuttings from apical parent stem position exhibited poorer growth performance before and after treatment, as well as greater metal content in shoots than base parts under the HM treatment. These results suggest that S. matsudana may undergo a special mechanism to hinder metals in the initial cutting, thus mitigating growth damage. The apical portion also showed poor resistance against the invasion of mixed HMs because of the immature structure. Therefore, in the selection of phytoremediation plants, metal accumulation ability is not proportional to growth performance.     

Endogenous Gibberellins in Elongating Shoots of Clones of Salix dasyclados and Salix viminalis

Elongating shoots of rapidly growing clones of Salix viminalis L. (clone 683-4) and Salix dasyclados Wimm. (clone 908) harvested in early August were analyzed for endogenous gibberellins (GA). Distribution of GA-like activity, determined by Tan-ginbozu dwarf rice microdrop bioassay after reverse phase C₁₈ high performance chromatography, was similar for both species. For S. dasyclados, combined gas chromatography-selected ion monotoring (GC-SIM) yielded identifications of GA₁, GA₈, GA₁₉, GA₂₀, and GA₂₉. Identifications of GA₄ and GA₉ were also made using co-injections of known amounts of [17, 17-²H₂]GAs. By bioassay, the main activity was GA₁₉-like in both species. Gibberellin A₁, GA₁₉, and GA₂₀ concentrations were approximated by GC-SIM using co-injections of known amounts of [17,17-²H₂]GAs. Both bioassay and GC-SIM results indicated very high concentrations of GA₁₉ and GA₂₀ (about 6000 nanograms per kilogram fresh weight shoot tissue using GC-SIM: 800 ng using bioassay), compared to the concentration of GA₁ (about 130 nanograms per kilogram fresh weight using either GC-SIM or bioassay).     

Structure and development of adventitious roots in Salix caprea and S. caprea × S. viminalis

Formation and structure of adventitious roots in cuttings of two clones of S. caprea and a hybrid betwen S. caprea and 5. viminalis was studied with light microscopy (LM) and transmission electron microscopy (TEM). The hybrid contained large preformed root primordia, which in cuttings placed in water developed into roots in only three days. One of the S. caprea clones contained minute preformed root primordia, which developed into roots in about eight days. Treatment with indolebutyric acid (IBA) increased the percentage of rooted cuttings and the number of roots formed. Roots emerging from IBA-treated cuttings contained both mature protophloem and protoxylem, while in roots of untreated cuttings only some sieve elements of the protophloem were mature. In the other 5. caprea clone no preformed root primordia were detected, but after treatment with IBA roots appeared in about two weeks. The cambium in treated stems produced a large number of cells, most of which differentiated into xylem. Root primordia were initiated in the newly produced tissue external to the cambium. The roots contained both mature protophloem and protoxylem at emergence. A few roots emerged also from extensive callus tissues formed external to the basal end of the cuttings. Cell enlargement and cell divisions in various parts of the base of the cuttings caused disruption of the peripheral tissues, which made the cuttings susceptible to infection by microorganisms.     

The Effect of Cooling on Translocation in Salix viminalis

Radioactive phosphate was applied externally to a small areaof the bark of the main stem below the point of insertion ofa leafy branch. When photosynthesis and hence translocationwas proceeding, phosphate was carried down the stem and wasmonitored externally by two Geiger counters spaced 10 cm apartfurther down the stem. Sudden cooling to 0 °C of the leaves,or the stem between the leaves and the point of applicationof the isotope, caused a rapid and seemingly simultaneous responseby the two Geiger counters. It is concluded that cooling atone point causes an instantaneous reduction of velocity alongthe whole pathway. If cooling is continued for several hoursthe rate of translocation of phosphate recovers. The evidencesuggests that this recovery is due to a compensatory increasein concentration of radioactive phosphate in the translocationstream rather than a recovery of velocity.     

Immobilization of selected heavy metals in sewage sludge by natural zeolites

This contribution presents the possibility of application of natural sorbent (Transcarpathian clinoptylolite (KL)) for immobilization of selected heavy metals in the sewage sludge. The influence of ion-exchange parameters (e.g. time, amount of zeolite) were discussed. Process of immobilization was performed using a static method (Batch). It was found that best possible conditions for immobilization of heavy metal ions were as follows: zeolite fraction 0.7–1.0 mm, 5 h of shaking, zeolite/sewage sludge ratio 2/98.     

Genetic mapping of sex-linked markers in Salix viminalis L

A total of 88 selective primer combinations were screened using bulked males and females sampled from four families of Salix viminalis. A total of more than 1000 polymorphic fragments was obtained, of which only four cosegregated with sex. These four sex-linked markers were subsequently scored in individuals that were used for bulked sample preparation in additional individuals of the same families, and in individuals in other families. A pair of primers that amplified the sex-linked fragments was constructed from one of the sex-linked amplified fragment length polymorphism (AFLP) fragments. In hybridization of Southern blot filters with the sex-linked DNA fragments, the band was present in females and absent in males, but the opposite pattern of band segregation (a band found in males and no band in females) was never observed in either the AFLP or RFLP experiments. Two of the sex-linked markers were placed on a linkage map. They both map at the same location in a linkage group comprising other markers not segregating with sex. Our data suggest that a single locus governs the sex determination and that nonrecombining sex chromosomes are absent in S. viminalis. A close association was found between skewed sex ratio and segregation distortion at this locus.     

某冶炼厂周围8种植物对重金属的吸收与富集作用

采用野外采样系统分析法,研究了沈阳冶炼厂的8种植物对重金属(Pb、Zn、Cu和Cd)的吸收与富集作用以及可能的耐性机制.结果表明,植物对重金属的吸收和富集,因植物种类、部位及重金属种类而不同.茼麻(Abutilon theophrasti)对Pb的吸收和富集能力较强,小白酒花(Conyza canadensis)、三裂叶豚草(Ambrosia trifida)、酸模叶蓼(Polygonum lapathifolium)、茼麻、龙葵(Solanum nigrum)、绿珠藜(Chenopodium acuminatum)和菊芋(Helianthus tuberosus)对Zn的吸收和富集效果较好,绿珠藜和茼麻对Cu的吸收和富集能力较强,龙葵、绿珠藜、茼麻、酸模叶蓼和小白酒花对Cd的吸收和富集能力较强.这些植物向地上部转移某些重金属的能力很强,转移系数大于1,可用于植物提取方式的污染土壤修复.其他转移系数小于1的植物,适合于重金属污染土壤的植物稳定.     

Growth, membrane potential and endogenous ion currents of willow (Salix viminalis) roots are all affected by abscisic acid and spermine

Growth measurements of hormone-treated roots from willow cuttings were combined with electrophysiological recordings to study hormone-induced changes in membrane potential and in endogenous ion currents. The mean growth rate of roots was 10 ± 2 μm min^?1 in regular nutrient solution. It increased to 13 ± 2 μm min⁺¹ after application of spermine and decreased to 0.07 ± 0.01 μm min^?1 after treatment with abscisic acid (ABA). Transient depolarizations were elicited in root cortex cells by spermine, while ABA caused a transient hyperpolarization. All changes in membrane potential were accompanied by transient responses of the endogenous current. These responses suggest that first anions, then cations leave the root during spermine-induced depolarizations. From the changes of the endogenous current an apparent efflux of anions (presumably Cl^?) and cations (presumably K⁺) of 200 to 700 pmol cm^?2 per depolarization was calculated. To further investigate a possible relation between endogenous ion currents, growth and the growth regulators ABA and spermine, long-lasting extracellular vibrating-probe measurements were performed. Control roots showed an inward current of about 1.5 μA cm^?2 at the apical elongation zone and an outward current with a maximum density of 1.3 μA cm^?2 at the central and basal elongation zone. The addition of ABA and spermine (final concentration 0.1 mM) to the bathing medium affected the endogenous current in opposite ways: ABA caused a reduction of inward and outward current, while spermine stimulated both. Since protons are a major component of the endogenous current, and sucrose can be taken up by root cells from the apoplast via symport with H⁺, a role of the endogenous current in growth regulation is indicated.     

P-protein and inclusion bodies in root protophloem of Salix viminalis

The formation of P-protein in the protophloem of 9- to 14-day-old adventitious roots of Salix viminalis was studied. In immature sieve elements a finely granular material was present. This was considered to be nascent P-protein. Small aggregations of tubular P-protein were observed 17 cells from the first "cleared" sieve element. In older cells the bodies were up to 7 μm long. Nondispersed and disaggregating P-protein bodies were present in mature sieve elements. P-protein bodies were also observed in parenchyma cells adjoining mature sieve elements. In addition, inclusion bodies of unknown origin are described. They had a granular content and were most often found in mature sieve elements.     

Blockade of heavy metals accumulation in Chlorella vulgaris cells by 24-epibrassinolide

This study was conducted on the influence of 24-epibrassinolide (24-epiBL) mixed with varying concentrations of heavy metals (copper, lead, cadmium, zinc) upon the growth and accumulation of these heavy metals in the cell of the alga Chlorella vulgaris Beijerinck (Chlorophyceae). Heavy metals at the concentration of 10^–3 M, alone or mixed with 24-epiBL, showed a lethal effect on C. vulgaris. At metal concentrations of 10^–6–10^–4 M, a combination with 24-epiBL appeared to have a stronger stimulatory effect on a number of cells than a single metal (a stronger inhibitory effect). 24-EpiBL at the concentration of 10^–8 M in combination with heavy metals (in the range 10^–6–10^–4 M) blocked metal accumulation in algal cells. 24-EpiBL has an anti-stress effect on C. vulgaris contaminated by heavy metals. The inhibitory effect on metal accumulation of 24-epiBL mixed with different heavy metals was arranged in the following order: zinc > cadmium > lead > copper. This process is correlated with the stimulation of growth of C. vulgaris. The stimulatory effect of 24-epiBL mixed with heavy metals leading to an increased pH in the medium (5.28–6.20) was significantly higher than the impact due to the increased acidity in the medium due to metals alone (pH 3.10–5.85). Lower pH increased the toxicity of heavy metals in C. vulgaris cells.     

Concurrent removal and accumulation of heavy metals by the three aquatic macrophytes

Under the present investigation effectiveness of three aquatic macrophytes Pistia stratiotes L. (water lettuce), Spirodela polyrrhiza W. Koch (duckweed) and Eichhornia crassipes were tested for the removal of five heavy metals (Fe, Zn, Cu, Cr and Cd). These plants were grown at three different concentrations (1.0, 2.0 and 5.0mgl(-1)) of metals in laboratory experiment. Result revealed high removal (>90%) of different metals during 15 days experiment. Highest removal was observed on 12th day of experiment, thereafter it decreased. Results revealed E. crassipes as the most efficient for the removal of selected heavy metals followed by P. stratiotes and S. polyrrhiza. Results from analysis confirmed the accumulation of different metals within the plant and a corresponding decrease of metals in the water. Significant correlations between metal concentration in final water and macrophytes were obtained. Plants have accumulated heavy metals in its body without the production of any toxicity or reduction in growth. Selected plants shown a wide range of tolerance to all of the selected metals and therefore can be used for large scale removal of heavy metals from waste water.     

Rooting Substances in Water Extracts of Castanea sativa and Salix viminalis

The rooting activity of water extracts of cuttings of chestnut (Castanea sativa Mill.) and willow (Salix viminalis L.) was studied throughout the year by the bean rooting test. The chestnut extract nullified the rooting activity of 10^?5M IAA, but on the whole it did not modify the rooting pattern of the bean cuttings. The willow extract increased rooting and showed a rhythm in the content of endogenous growth substances. IAA was found at the time when the extract showed the strongest rooting activity. The extract from willow contained two synergistic root-promoting fractions with the R_f's of 0.0–0.2 and 0.6–0.8 in isopropanol: ammonia: water. The latter one promoted rooting without added IAA. In this fraction a high amount of catechol was detected. No clear correlation was found between the ability to induce roots and the total phenol content in the two species.     

Nitrate Supply and the Biophysics of Leaf Growth in Salix viminalis

The influence of nitrogen on leaf area development and the biophysicsof leaf growth was studied using clonal plants of the shrubwillow, Salix viminalis grown with either optimal (High N) orsub-optimal (Low N) supplies of nitrate. Leaf growth rate andfinal leaf size were reduced in the sub-optimal treatment andthe data suggest that in young rapidly growing leaves, thiswas primarily due to changes in cell wall properties, sincecell wall extensibility (% plasticity) was reduced in the LowN plants. The biophysical regulation of leaf cell expansion also differedwith nitrogen treatment as leaves aged. In the High N leaves,leaf cell turgor pressure (P) increased with age whilst in theLow N leaves P declined with age, again suggesting that foryoung leaves, cell wall plasticity limited expansion in theLow N plants. Measurements of cell wall properties showed thatcell wall elasticity (%E) was not influenced by nitrogen treatmentand remained constant regardless of leaf age. Key words: Salix, cell wall extensibility, nitrogen nutrition, biophysics of leaf growth     

Toxicity of 2,3,5,6-tetrachlorophenol to willow trees (Salix viminalis)

Chlorinated phenols have been intensively investigated from an eco-toxicological point of view, however almost nothing is known about toxicity of tetrachlorophenol (TeCP) to higher terrestrial plants. This article applied the willow tree acute toxicity test to study the toxicity of 2,3,5,6-TeCP to willows Salix viminalis (S. viminalis) at neutral and acidic conditions (roughly pH 7 and 4) with inhibition of transpiration as toxic endpoint. At neutral pH the EC₅₀ was >10 mg L^?1 while the EC₅₀ at acidic conditions was 0.32 ± 0.17 mg L^?1, clearly indicating that toxicity is exerted by the non-ionic chemical fraction. Standard tests running at neutral pH are therefore not capturing the full toxicity of weak acids and bases.