古老铅锌矿区生态型东南景天对锌耐性及超积累特性的研究

 植物长期生长在重金属污染的生境中,逐渐进化成不同的生态型。通过调查中国东南部古老Pb/zn矿和非矿山生境中的植物种群,发现生长在古老Pb／Zn矿的东南景天(Sedum alfredii Hance)是一种新的Zn超积累植物。在自然和控制条件下,古老Pb／Zn矿生态型比非矿山生态型植株的茎粗、叶片大、植株高。在矿山土壤Zn有效含量为105.5～325.4mg·kg-1时,矿山生态型东南景天植株地上部Zn含量为4134～5000mg·kg-1;当营养液中Zn浓度为1223.6μmol时,其Zn含量高达2％。在相同Zn浓度下,矿山生态型地上部Zn含量比非矿山生态型高30倍左右。两种生态型体内Zn分布也不同,古老铅锌矿山生态型的不同器官中Zn含量以茎>叶片>根系,而非矿山生态型则以根系>茎>叶片。古老铅锌矿山生态型地上部积累的Zn占植株总积累量的90％以上,其中叶片和茎分别占41.66％±5.46％和54.75％±5.87％;非矿山生态型各器官中积累的Zn远远低于古老铅锌矿山生态型,各器官中积累的Zn以茎>根系>叶片。本研究表明,这两种生态型东南景天的发现,为今后探讨植物耐高Zn胁迫和超积累Zn的微进化过程提供了非常有价值的材料,也为Zn污染土壤的植物修复提供了一种很有潜力的候选材料。     

Dynamics of zinc uptake and accumulation in the hyperaccumulating and non-hyperaccumulating ecotypes of Sedum alfredii Hance

Sedum alfredii Hance has been identified as a Zn-hyperaccumulating plant species native to China. The characteristics of Zn uptake and accumulation in the hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) of S. alfredii were investigated under nutrient solution and soil culture conditions. The growth of HE was normal up to 1000 μM Zn in nutrient solution, and 1600 mg Zn kg⁻¹ soil in a Zn-amended soil. Growth of the NHE was inhibited at Zn levels ≥250 μM in nutrient solution. Zinc concentrations in the leaves and stems increased with increasing Zn supply levels, peaking at 500 and 250 μM Zn in nutrient solution for the HE and the NHE, respectively, and then gradually decreased or leveled off with further increase in solution Zn. Minimal increases in root Zn were noted at Zn levels up to 50 μM; root Zn sharply increased at higher Zn supply. The maximum Zn concentration in the shoots of the HE reached 20,000 and 29,000 mg kg⁻¹ in the nutrient solution and soil experiments, respectively, approximately 20 times greater than those of the NHE. Root Zn concentrations were higher in the NHE than in the HE when plants were grown at Zn levels ≥50 μM. The time-course of Zn uptake and accumulation exhibited a hyperbolic saturation curve: a rapid linear increase during the first 6 days in the long-term and 60 min in the short-term studies; followed by a slower increase or leveling off with time. More than 80% of Zn accumulated in the shoots of the HE at half time (day 16) of the long-term uptake in 500 μM Zn, and also at half time (120 min) of the short-term uptake in 10 μM ⁶⁵Zn²⁺. These results indicate that Zn uptake and accumulation in the shoots of S. alfredii exhibited a down-regulation by internal Zn accumulated in roots or leaves under both nutrient solution and soil conditions. An altered Zn transport system and increased metal sequestration capacity in the shoot tissues, especially in the stems, may be the factors that allow increased Zn accumulation in the hyperaccumulating ecotype of S. alfredii. Section Editor: F. J. Zhao     

Detection of phytochelatins in the hyperaccumulator Sedum alfredii exposed to cadmium and lead

Phytochelatins (PCs) are known to play an essential role in the heavy metal detoxification of some higher plants and fungi by chelating heavy metals. However, three recent papers reported that no PCs could be detected in the hyperaccumulator Sedum alfredii Hance upon cadmium, lead or zinc treatment, respectively. In this paper, PC synthesis was assayed again in the mine population of S. alfredii with the help of reversed phase high-performance liquid chromatography (HPLC), HPLC-mass spectrometry, and HPLC-tandem mass spectrometry. Our data showed that PC formation could be induced in the leaf, stem and root tissues of S. alfredii upon exposure to 400 microM cadmium, and only in the stem and root when exposed to 700 microM lead. However, no PCs were found in any part of S. alfredii when it was subjected to exposure to 1600 microM zinc.     

Re-evaluation of the role of ureides in the xylem transport of nitrogen in Arachis species

There are conflicting reports in the literature of the possible role of the ureides, allantoin and allantoic acid, in the nitrogen economy of Arachis species. Therefore, xylem sap composition in food peanut ( Arachis hypogaea L.), and two forage peanuts ( A. pintoi L. and A. glabrata Benth.) has been studied in detail. Xylem saps were collected from peanuts grown under different nutritional regimes and environmental conditions in the glasshouse and field in Australia, Malaysia and Indonesia, and the N-containing solutes analysed. The relative amounts and concentrations of ureides in these peanut exudates were compared with those of soybean ( Glycine max [L.] Merr.) – a species known to export ureides in its xylem stream as the major product of N₂ fixation.
Xylem concentrations of ureides in soybean were high in N₂-fixing plants (2.9 to 3.7 μmol ml⁻¹), representing 60 to 88% of xylem solute nitrogen, but it contributed only 9% (0.7 μmol ml⁻¹) if plants were unnodulated and supplied nitrate. In all species of peanut, concentrations of ureides measured in xylem sap were generally much smaller (0.02 to 0.37 μmol ml⁻¹; 1–7% of xylem nitrogen) and were unaffected by peanut species or cultivar, rhizobial strain, plant size, growth rate, or stage of development, and were not related to N₂ fixation (less than 0.1% of currently fixed nitrogen exported as ureides) or the assimilation of nitrate. Apparently high levels of ureides in sap from some field-grown plants were shown to be due to interference with the ureide colorimetric assay by some contaminating compound rather than represent increased ureides per se.     

Effects of root temperature on leaf gas exchange and xylem sap abscisic acid concentrations in six Cucurbitaceae species

Roots of six Cucurbitaceae species were exposed to low (14 °C), middle (24 °C), and high (34 °C) temperatures while aerial parts of plants were maintained at ambient temperatures between 23 and 33 °C. The highest dry mass (DM), photon-saturated rate of net photosynthesis (P _Nsat), and stomatal conductance (g _s) were found at 14 °C in figleaf gourd and turban squash plants, at 24 °C in cucumber and melon plants, while bitter melon and wax gourd plants had lower DM, P _Nsat, and g _s at 14 °C than at 24 or 34 °C. Sub-or supra-optimum root temperatures did not induce photoinhibition but induced slight changes in the quantum efficiency of photosystem 2, PS2 (Φ_PS2) and photochemical quenching (q_p). Meanwhile, xylem sap abscisic acid (ABA) concentration followed a contrasting change pattern to that of g _s. Thus the change in P _Nsat was mainly due to the change in g _s and roots played an important role in the regulation of stomatal behaviour by delivering increased amount of ABA to shoots at sub-or supra-optimum root temperatures.     

Phosphorus-31 nuclear magnetic resonance studies of intracellular pH,phosphate compartmentation and phosphate transport in yeasts

³¹P NMR spectra were obtained from suspensions of Candida utilis, Saccharomyces cerevisiae and Zygosaccharomyces bailii grown aerobically on glucose. Direct introduction of substrate into the cell suspension, without interruption of the measurements, revealed rapid changes in pH upon addition of the energy source. All ³¹P NMR spectra of the yeasts studied indicated the presence of two major intracellular inorganic phosphate pools at different pH environments. The pool at the higher pH was assigned to cytoplasmic phosphate from its response to glucose addition and iodoacetate inhibition of glycolysis. After addition of substrate the pH in the compartment containing the second phosphate pool decreased. A parallel response was observed for a significant fraction of the terminal and penultimate phosphates of the polyphosphate observed by ³¹P NMR. This suggested that the inorganic phosphate fraction at the lower pH and the polyphosphates originated from the same intracellular compartment, most probably the vacuole. In this vacuolar compartment, pH is sensitive to metabolic conditions. In the presence of energy source a pH gradient as large as 0.8 to 1.5 units could be generated across the vacuolar membrane. Under certain conditions net transport of inorganic phosphate across the vacuolar membrane was observed during glycolysis: to the cytoplasm when the cytoplasmic phosphate concentration had become very low due to sugar phosphorylation, and into the vacuole when the former concentration had become high again after glucose exhaustion.Non-Standard Abbreviations NMR nuclear magnetic resonance - ppm parts per million - PP polyphosphate - P_i,c cytoplasmic inorganic phosphate - P_i,v vacuolar inorganic phosphate - pH_in,c cytoplasmic pH - pH_in,v vacuolar pH - FCCP carbonyl p-trifluoromethoxyphenylhydrazone     

The effect of industrial pollution on Zinc,Cadmium and Copper concentration in the xylem rings of Scot's pine (Pinus sylvestris L.) and in the soil

Summary The concentrations of heavy metals, including zinc, cadmium, and copper were studied in the xylem rings of Pinus sylvestris trees and in the soil of three heavily polluted sites and one non-polluted site. There was a high correlation between the concentration of heavy metals in the soil and recently produced xylem rings. However, the initiation of industrial pollution did not abruptly increase heavy metal concentrations in the xylem rings. There was a high correlation between the zinc content of trees growing near a zinc smelter and the values obtained by other investigators for pine trees growing on the same site.     

Evidence for operation of the direct zinc ligand exchange mechanism for trafficking, transport, and reactivity of zinc in mammalian cells

In addition to its critical role in normal cell function, growth, and metabolism, zinc is implicated as a major factor in the development and progression of many pathological conditions and diseases. Despite this importance of zinc, many important factors, processes, and mechanisms of the physiology, biochemistry, and molecular biology of zinc remain unknown. Especially important is the unresolved issue regarding the mechanism and process of the trafficking, transport, and reactivity of zinc in cells; especially in mammalian cells. This presentation focuses on the concept that, due to the existence of a negligible pool of free Zn²⁺ ions in the mammalian cell environment, the trafficking, transport and reactivity of zinc occurs via a direct exchange of zinc from donor Zn-ligands to acceptor ligands. This Zn exchange process occurs without the requirement for production of free Zn²⁺ ions. The direct evidence from mammalian cell studies is presented in support of the operation of the direct Zn-ligand exchange mechanism. The paper also provides important information and conditions that should be considered and employed in the conduct of studies regarding the role and effects of zinc in biological/biomedical research; and in its clinical interpretation and application.     

Supercooling of xylem ray parenchyma cells in tropical and subtropical hardwood species

 The freezing behavior of xylem ray parenchyma cells in several woody species, Ficus elastica, F. microcarpa, Mangifera indica, Hibiscus Rosa-sinensis, and Schefflera arboricola, that are native to non-frost tropical and subtropical zones, was investigated by differential thermal analysis (DTA), cryo-scanning electron microscopy (cryo-SEM) and freeze-replica electron microscopy. Although profiles after DTA did not exhibit clear evidence of supercooling in the xylem ray parenchyma cells, electron microscopy revealed that the majority of xylem ray parenchyma cells in all of the woody species examined were supercooled to around –10°C upon freezing temperatures and were not frozen extracellularly. It seems likely that DTA failed to reveal the low temperature exotherm (LTE), that is produced by breakdown of supercooling in the xylem ray parenchyma cells as a consequence of the overlap between the high temperature exotherm and the LTE in each case. The xylem ray parenchyma cells in these woody species were very sensitive to dehydration, and supercooling had, to some extent, a protective effect against freezing injury. It is suggested that the capacity for supercooling of xylem ray parenchyma cells of tropical and subtropical woody species might be the result of inherent structural characteristics, such as rigid cell walls and compact xylem tissues, rather than the result of positive adaptation to freezing temperatures. The present and previous results together indicate that the responses of xylem ray parenchyma cells in a wide variety of hardwood species to freezing temperatures can be explained as a continuum, the specifics of which depend upon the temperatures of the growing conditions. Received: 24 January 1997 / Accepted: 13 May 1997     

Root xylem transport of amino acids in the root hemiparasitic shrub Olax phyllanthi (Labill) R.Br. (Olacaceaea) and its multiple hosts

The amino acid compositions of the root xylem saps of Olax phyllanthi and a range of its common hosts were examined in native coastal heath in Western Australia and in pot cultures of Olax reliant on single hosts. When hosts specializing in the xylem transport of one major solute (asparagine, glutamine, histidine, arginine or proline) were exploited, the endophytic tissue of haustoria and the xylem sap of Olax showed much lower proportions of this than of other solutes, suggesting pronounced metabolic transformation prior to xylem loading by the parasite. However, the xylem sap of Olax did partly reflect the compositions of its hosts; for example, djenkolic acid and pipecolic acid were present when Olax was parasitic on species of Acacia, and levels of citrulline and aspartic acid were higher than normal when it exploited hosts transporting large amounts of these compounds. Back-flow of S-ethenyl cysteine, a novel amino acid specific to Olax, was observed to another root hemiparasite (Exocarpos sparteus) in native habitat and to certain non-parasitic hosts in water-stressed pot cultures. Haustoria exhibited high levels of glutamine synthetase but showed appreciable in vivo nitrate reductase activity only when on hosts with high xylem levels of nitrate.     

Sequential depolarization of root cortical and stelar cells induced by an acute salt shock - implications for Na(+) and K(+) transport into xylem vessels

Early events in NaCl-induced root ion and water transport were investigated in maize (Zea mays L) roots using a range of microelectrode and imaging techniques. Addition of 100 mm NaCl to the bath resulted in an exponential drop in root xylem pressure, rapid depolarization of trans-root potential and a transient drop in xylem K(+) activity (A(K+) ) within ～1 min after stress onset. At this time, no detectable amounts of Na(+) were released into the xylem vessels. The observed drop in A(K+) was unexpected, given the fact that application of the physiologically relevant concentrations of Na(+) to isolated stele has caused rapid plasma membrane depolarization and a subsequent K(+) efflux from the stelar tissues. This controversy was explained by the difference in kinetics of NaCl-induced depolarization between cortical and stelar cells. As root cortical cells are first to be depolarized and lose K(+) to the environment, this is associated with some K(+) shift from the stelar symplast to the cortex, resulting in K(+) being transiently removed from the xylem. Once Na(+) is loaded into the xylem (between 1 and 5 min of root exposure to NaCl), stelar cells become more depolarized, and a gradual recovery in A(K+) occurs.     

Photosynthesis, water use efficiency and stable carbon isotope composition are associated with anatomical properties of leaf and xylem in six poplar species

Although fast‐growing Populus species consume a large amount of water for biomass production, there are considerable variations in water use efficiency (WUE) across different poplar species. To compare differences in growth, WUE and anatomical properties of leaf and xylem and to examine the relationship between photosynthesis/WUE and anatomical properties of leaf and xylem, cuttings of six poplar species were grown in a botanical garden. The growth performance, photosynthesis, intrinsic WUE (WUE_i), stable carbon isotope composition (δ¹³C) and anatomical properties of leaf and xylem were analysed in these poplar plants. Significant differences were found in growth, photosynthesis, WUE_i and anatomical properties among the examined species. Populus cathayana was the clone with the fastest growth and the lowest WUE_i/δ¹³C, whereas P. × euramericana had a considerable growth increment and the highest WUE_i/δ¹³C. Among the analysed poplar species, the highest total stomatal density in P. cathayana was correlated with its highest stomatal conductance (g_s) and lowest WUE_i/δ¹³C. Moreover, significant correlations were observed between WUE_i and abaxial stomatal density and stem vessel lumen area. These data suggest that photosynthesis, WUE_i and δ¹³C are associated with leaf and xylem anatomy and there are tradeoffs between growth and WUE_i. It is anticipated that some poplar species, e.g. P. × euramericana, are better candidates for water‐limited regions and others, e.g. P. cathayana, may be better for water‐abundant areas.     

Copper uptake and translocation in chicory (Cichorium intybus L. cv Grasslands Puna) and tomato (Lycopersicon esculentum Mill. cv Rondy) plants grown in NFT system. II. The role of nicotianamine and histidine in xylem sap copper transport

The effect of rooting media Cu concentration (0.05–20 mg Cu L^-1) on amino acid concentrations and copper speciation in the xylem sap of chicory and tomato plants was measured using 6 week old plants grown in a nutrient film technique system (NFT). Irrespective of the Cu concentration in the nutrient solutions, more than 99.68% and 99.74% of total Cu in tomato and chicory xylem sap was in a bound form. When exposed to high Cu concentrations in the rooting media, amino acid concentrations in the sap increased. Relative to other amino acids, the concentrations of glutamine (Gln), histidine (His), asparagine (Asn), valine (Val), nicotianamine (NA) and proline (Pro) in tomato xylem saps, and His, γ-aminobutyric acid (Gaba), glutamic acid (Glu), leucine (Leu), NA and phenylalanine (Phe) in chicory xylem saps showed the greatest increases. The data indicate that induced synthesis of some free amino acids as a specific and proportional response to Cu treatment. For a single complexation amino acid, the solution Cu²⁺concentration vs pH titration curve for NA at 0.06–0.07 mM was most similar, closely followed by His at 0.5–0.6 mM, to the solution Cu²⁺concentration behaviour in both tomato and chicory xylem sap. It is concluded that increased Cu concentrations in the rooting media induced selective synthesis of certain amino acid which include NA, His, Asn and Gln which have high stability constants with Cu. NA and His have the highest binding constants for Cu and the concentrations of NA and His in chicory and tomato xylem saps can account for all the bound Cu carried in the sap. This revised version was published online in June 2006 with corrections to the Cover Date.     

Characterization of Separated Cell Types from the Developing Rat Cerebellum: Transport of Glutamate and Aspartate by Preparations Enriched in Purkinje Cells, Granule Neurones, and Astrocytes

Preparations of structurally preserved cerebellar perikarya (cells) were found to express high-affinity transport systems for glutamate but not for certain putative transmitter substances (including monoamines, glycine and taurine) and non-transmitter amino acids. The characteristics of the high-affinity glutamate transport system were similar to those of other preparations of brain tissue: [³H]glutamate uptake by the cells was Na⁺-dependent and was inhibited competetively by other acidic amino acids. The rank order of apparent affinities of the carrier for acidic amino acids was L-aspartate > L-glutamate > D-aspartate ? D-glutamate (the affinity for D-glutamate being over two orders of magnitude lower than for the other three amino acids). Comparison of high-affinity [³H]glutamate uptake in preparations enriched in different cell types showed that although the affinities are similar (2-4 fiM), the rate is outstandingly high in astrocytes (V_max 18 nmol/min per mg protein). Significantly, uptake into the putatively glutamatergic granule cells was very low. These observations were supported by autoradiographic findings which showed that the predominant sites of [3H]glutamate uptake in cerebellar cultures enriched in interneurones are the astrocytes. Furthermore, the V_max in cultures enriched in astrocytes was as high as that in separated astrocytes. Thus, it seems that the principal cell type involved in acidic amino acid uptake in the cerebellum is the astrocyte, and this must be taken into consideration when high-affinity uptake is used as a marker for glutamatergic transmitter systems. Furthermore, the selective cellular distribution of glutamate transport sites, together with the uneven distribution of enzymes related to glutamate metabolism observed previously, indicates that a metabolic interaction takes place between the different cell types, supporting the current hypothesis on metabolic compartmentation in the brain.     

Evaluation of role of the peptide transport system in absorption of dipeptides in the rat small intestine in chronic experiments <Emphasis Type="Italic">in vivo</Emphasis>

Hydrolysis and absorption of glycylglycine and glycyl-L-leucine as well as absorption of glycine and leucine were studied in chronic experiments on rats with their isolated small intestine loop. Values of the “true” kinetic constants (with taking into account effect of the preepithelial layer) were determined to be as follows: (1) K _t = 46.7 ± 4.0 and 2.15 ± 0.59 mM, J _max = 0.74 ± 0.15 and 0.16 ± 0.03 μmol min^?1 cm^?1 (for transport of free glycine and leucine, respectively); (2) K _t = 4.4 ± 0.6 and 4.8 ± 0.9 mM, J _max = 0.24 ± 0.02 and 0.23 ± 0.02 μmol min^?1 cm^?1 (for transport of glycylglycine and glycyl-L-leucine, respectively); (3) K _M = 5.4 ± 1.0 and 38.2 ± 4.4 mM, V _max = 0.09 ± 0.02 and 0.24 ± 0.07 μmol min^?1 cm^?1 (for membrane hydrolysis of these dipeptides, respectively). According to our calculations, in the wide range of the initial glycylglycine concentrations (2.5–40 mM) a part of the peptide component in its total absorption accounts for 0.77–0.80. In the case of glycyl-L-leucine a part of the peptide component in the total glycine absorption decreases from 0.89 to 0.84, while in the total leucine absorption—from 0.86 to 0.71, the initial dipeptide concentration rising from 5 to 40 mM. The obtained results show that the peptide component prevails in absorption of the studied dipeptides in the rat small intestine, but its role is much lesser than what many authors believe. In the case of glycyl-L-leucine, the peptide component can achieve saturation in the range of high substrate concentrations, its part decreasing essentially to become compared with absorption of free amino acids formed as a result of the dipeptide membrane hydrolysis.     

The effect of Brevicoryne brassicae on leaf area, dry matter distribution and amino acids of the Brussels sprout plant

Twenty-six direct or derived criteria were assessed, eight wk after infestation, of the effect of an initial inoculum of 0, 5, 25 or 50 Brevicoryne brassicae on the growth and dry matter distribution of Brussels sprout plants. Twenty-four of these criteria showed significant (P > 0.05) effects of aphid infestation, though increasing aphid infestation above 5/plant had little further effect. Total fresh and dry weight (as well as of the components — leaves, stems and roots) was reduced by 25–730 with infestation, particularly so the root. The number of leaves as well as individual leaf area was reduced, and leaves on infested plants were less efficient assimilators. This could be explained by the drain on assimilates caused by the aphids, the leaf curling and the reduced density of stomata. The latter was also considered to account partly for the increased moisture content of infested plants. There was a general reduction in the concentration of leaf amino acids.     

Reducing protein content in the diet of growing goats: implications for nitrogen balance,intestinal nutrient digestion and absorption,and rumen microbiota

Reducing dietary CP content is an effective approach to reduce animal nitrogen excretion and save protein feed resources. However, it is not clear how reducing dietary CP content affects the nutrient digestion and absorption in the gut of ruminants, therefore it is difficult to accurately determine how much reduction in dietary CP content is appropriate. This study was conducted to investigate the effects of reduced dietary CP content on N balance, intestinal nutrient digestion and absorption, and rumen microbiota in growing goats. To determine N balance, 18 growing wether goats (25.0 ± 0.5 kg) were randomly assigned to one of three diets: 13.0% (control), 11.5% and 10.0% CP. Another 18 growing wether goats (25.0 ± 0.5 kg) were surgically fitted with ruminal, proximate duodenal, and terminal ileal fistulae and were randomly assigned to one of the three diets to investigate intestinal amino acid (AA) absorption and rumen microbiota. The results showed that fecal and urinary N excretion of goats fed diets containing 11.5% and 10.0% CP were lower than those of goats fed the control diet (P < 0.05). When compared with goats fed the control diet, N retention was decreased and apparent N digestibility in the entire gastrointestinal tract was increased in goats fed the 10% CP diet (P < 0.05). When compared with goats fed the control diet, the duodenal flow of lysine, tryptophan and phenylalanine was decreased in goats fed the 11.5% CP diet (P < 0.05) and that of lysine, methionine, tryptophan, phenylalanine, leucine, glutamic acid, tyrosine, essential AAs (EAAs) and total AAs (TAAs) was decreased in goats fed the 10.0% CP diet (P < 0.05). When compared with goats fed the control diet, the apparent absorption of TAAs in the small intestine was increased in goats fed the 11.5% CP diet (P < 0.05) and that of isoleucine, serine, cysteine, EAAs, non-essential AAs, and TAAs in the small intestine was increased in goats fed the 10.0% CP diet (P < 0.05). When compared with goats fed the control diet, the relative richness of Bacteroidetes and Fibrobacteres was increased and that of Proteobacteria and Synergistetes was decreased in the rumen of goats fed a diet with 10.0% CP. In conclusion, reducing dietary CP content reduced N excretion and increased nutrient utilization by improving rumen fermentation, enhancing nutrient digestion and absorption, and altering rumen microbiota in growing goats.