Metal ions restore the proteolytic resistance of denatured conglutin gamma, a lupin seed glycoprotein, by promoting its refolding

The susceptibility to trypsin of conglutin gamma, a lupin seed glycoprotein affected by this enzyme only when in a non-native conformation, was used to study the effect of Zn(2+) and other metal ions on the structural dynamics of the protein. When acid-treated trypsin-susceptible conglutin gamma was incubated at neutral pH in the presence of Zn(2+), it became resistant to tryptic attack, contrary to the protein treated in the absence of Zn(2+). The time course of this refolding event has been quantitatively evaluated by SDS-PAGE. Amino acid sequencing of the major polypeptide fragments, produced by trypsin before completion of the refolding process, indicated that only a few cleavable bonds were accessible to the enzyme. This suggested that the presence of metal ions affected the pathway of degradation of the protein, by inducing its folding. Among the other metal ions tested, Ni(2+) also promoted the adoption of a trypsin-resistant conformation of conglutin gamma, whereas Mn(2+) and Ca(2+) had only much lower effects. The relevance of these findings for a deeper understanding of the in vivo degradation of plant food proteins and how it is affected by metal ions are discussed.     

Theoretical speciation of ethylenediamine-N-(o-hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid (o,p-EDDHA) in agronomic conditions

The presence of ethylenediamine-N-(o-hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid (o,p-EDDHA) as the second largest component in commercial EDDHA iron chelates has recently been demonstrated. Here is reported the speciation of o,p-EDDHA by the application of a novel methodology through the determination of the complexing capacity, protonation, and Ca(2+), Mg(2+), Cu(2+), and Fe(3+) stability constants. The pM values and species distribution in solution, hydroponic, and soil conditions were obtained. Due to the para position of one phenol group in o,p-EDDHA, the protonation constants and Ca and Mg stability constants have different values from those of o,o-EDDHA and p,p-EDDHA regioisomers. o,p-EDDHA/Fe(3+) stability constants are higher than those of EDTA/Fe(3+) but lower than those of o,o-EDDHA/Fe(3+). The sequence obtained for pFe is o,o-EDDHA/Fe(3+) >/= o,p-EDDHA/Fe(3+) > EDTA/Fe(3+). o,p-EDDHA/Fe(3+) can be used as an iron chelate in hydroponic conditions. Also, it can be used in soils with limited Cu availability.     

β-Carotene Assay Revisited. Application To Characterize and Quantify Antioxidant and Prooxidant Activities in a Microplate

The β-carotene bleaching assay, a common method for evaluating antioxidant activity, has been widely criticized due to its low reproducibility, problematic quantification, complex reagent preparation, and interference of different factors (temperature, pH, solvents, and metals). In this work we have examined the effects of these factors and developed a highly reproducible procedure for microplate assay, evaluated the critical points of the method, and proposed a kinetic model for quantifying both antioxidant and prooxidant activities. The application of these tools produced very consistent results, which provide robust and meaningful criteria to compare in detail the characteristics of several well-known commercial antioxidants, as well as several predictable prooxidants, and can be easily applied to natural extracts, food samples, and many other type of compounds. As an example, we have tested a set of commercial antioxidants and some typical lipophilic prooxidants. The activity of the tested antioxidants decreased in the following order: ethoxyquin ? α-tocopherol > butylhydroxyanisole > butylhydroxytoluene ? propyl gallate. On the other hand, hemoglobin and Fe(2+), Fe(3+), Co(2+), and Cu(2+) showed a strong prooxidant effect, and the activity was null in Cd(2+), Ni(2+), and Sr(2+), slightly antioxidant in Mg(2+), and strongly antioxidant in Zn(2+) and Mn(2+).     

Antioxidative ability of lactic acid bacteria

Nineteen strains of lactic acid bacteria were investigated for antioxidative activity. These includedLactobacillus acidophilus B, E, N1, 4356, LA-1, and Farr; Lactobacillus bulgaricus 12 278, 448, 449, Lb, 1006, and 11 842; Streptococcus thermophilus 821, MC, 573, 3641, and 19 987; and Bifidobacterium longum B6 and 15 708. Intracellular cell-free extract of all strains demonstrated antioxidative activity with inhibition rates of ascorbate autoxidation in the range of 7-12%. Antioxidative mechanisms including metal ion chelating ability, scavenge of reactive oxygen species, enzyme inhibition, and reducing activity of intracellular cell-free extract of lactic acid bacteria were studied. S. thermophilus 821 had the highest metal ion chelating ability for Fe(2+), and B. longum 15 708 showed the highest Cu(2+) chelating ability among the 19 strains tested. All strains demonstrated reactive oxygen species scavenging ability. L. acidophilus E showed the highest hydroxyl radical scavenging ability, and B. longum B6 had the best hydrogen peroxide scavenging ability. Reducing activity was also found in all strains. Most of the strains tested demonstrated excellent reducing activity. B. longum B6 showed the highest reducing activity among the 19 strains tested. In enzyme inhibition, superoxide dismutase activity was not found in these 19 strains, and the activity of superoxide dismutase was not induced when metal ion Mn(2+), Fe(2+), or Cu(2+)Zn(2+) was present.     

Production, purification, and properties of an endoglucanase produced by the hyphomycete Chalara (Syn. Thielaviopsis) paradoxa CH32

The hyphomycete Chalara (syn. Thielaviopsis) paradoxa produces endoglucanase activity during the late trophophase. The low molecular mass (35 kDa) endoglucanase purified from cultured broths works optimally at 37 degrees C and pH 5.0. The enzyme inactivates at pH below 3.0 and also at temperatures of 50 degrees C or higher, but it is stable at lower temperatures, including refrigeration temperature and freezing. The enzyme is inhibited by detergents, by EDTA, and by the divalent cations Hg(2+) and Ag(2+). It is also inhibited to some extent by 10 mM Zn(2+), Fe(2+), and Mg(2+), but it is stimulated by Mn(2+). Enzyme activity is not affected by reducing agents. In the presence of low concentrations of water miscible organic solvents (20%) endoglucanase activity is inhibited by 7% (for methanol) to 50% (for acetonitrile), and it is totally inhibited at higher solvent concentrations (50%). Enzyme activity is not affected by the water immiscible solvent ethyl acetate. Carboxymethylcellulose is the preferred substrate (K(m(app)) = 8.3 g/L; V(max(app)) = 1.1 microM/min). Hydrolysis of crystalline cellulosic substrates is very limited, but it is greatly enhanced by phosphoric acid swelling. The purified enzyme shows no activity toward disaccharides or aryl-glucosides. Its activity is inhibited by cellobiose.     

Effect of humic substances in nutrient film technique on nutrient uptake

An experiment was conducted to find out how humic substances affected nutrient uptake of plants. The test plants, oregano, thyme, and basil, were grown in nutrient film technique at two pH levels (4.5 and 6.5), in two substrates (peat and perlite), and at three levels of humic substance that was a peat extract (control, low, and high concentration). Nutrient uptake of potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu) were determined by elemental contents in aerial parts of the plant and its weight. Humic substance had no effect on K, Ca, and Mg uptake but lowered the uptake of Fe, Mn, Zn, and Cu for the three test plants, more pronounced with perlite than peat and more at low pH than at high pH. The lowering of the uptake might be caused by complexation of Fe, Mn, Zn, and Cu by the humic substance and the lower availability of these metals in complexed form than as a cation or as EDTA‐chelate in the case of Fe. It is not clear why the effect of the humic substance on micro‐element uptake is larger at low than at high pH. The complexation is expected to be stronger at pH 6.5 than at pH 4.5. At low pH, the high concentration of humic substance caused a low fresh weight of the shoots, perhaps caused by a toxicity of the humic substance at low pH. This was less pronounced at high pH.     

Analysis of Some Divalent Metal Contents in Tobacco Expressing the Exogenous Soybean Ferritin Gene

Abstract

T2 tobacco lines overexpressing soybean ferritin in the plastids (+TPs) or apoplasm (AFs) under the regulation of CaMV 35S promoter were grown on MS nutrient solution. After 1 month growth, statuses of six major divalent‐metals (Ca, Cu, Fe, Mg, Mn, and Zn) were measured in leaves and roots. Both +TPs and AFs showed enhanced growth (max. 1.7×) in leaves than the control line. The Fe contents in leaves of +TPs and AFs were significantly larger (1.9–2.8×) than that of the control line. The other metal contents in leaves of +TPs and AFs were almost the same as or less than those of the control line. In contrast to the result of leaves, the growth enhancement in roots was not clear in +TPs, but in AFs. Also, some of the non‐ferrous metal contents in roots of +TPs and AFs were dramatically increased compared with those of the control line (Mn, 1.9–10.4×; Zn, 1.6–2.3×), whereas the differences in content of Fe, Cu, Ca, and Mg were insignificant. These results demonstrated that the ferritin overexpression in apoplasm was as effective for inducing Fe accumulation as that in plastid. Under the normal metal‐balanced condition, even if the activation of Fe uptake related enzymes leads to the accumulation of non‐specific accumulation of divalent metal ions in roots, an Fe loading/unloading system and/or an internal translocator in xylem and phloem might specifically deliver Fe to the upper part of plants.     

转大麦烟酰胺合成酶基因提高水稻逆境胁迫耐受性的研究

维持铁等金属离子的动态平衡是保持植物细胞功能正常的先决条件。烟酰胺合成酶基因在单子叶禾本科的缺铁胁迫应答反应中起着关键作用,它的催化产物烟酰胺(NA)是铁及其他二价金属离子在体内吸收和转运的重要载体,且能与Fe2+结合形成Fe2+-NA复合物,从而使植物在酸性土壤中免受铁毒害。利用农杆菌介导法,将大麦烟酰胺合成酶基因NASHOR1转入水稻台北309中,经PCR及PCR-Southern杂交检测,确定目的基因已经整合到水稻基因组中。铁、锰、铜和锌含量的检测结果显示:与非转基因对照植株相比,转基因植株的金属元素含量都明显提高,铜、锌、锰和铁元素含量分别增加了15%、80%、31%和44%,但铁、锰和锌元素增量在株系间差异较大。在干旱胁迫下,转基因植株的超氧化物歧化酶(SOD)活性和脯氨酸含量都高于非转基因对照植株的,暗示大麦烟酰胺合酶基因在一定程度上提高了水稻的耐旱性。     

Purification and characterization of a novel extracellular tripeptidyl peptidase from Rhizopus oligosporus

A novel extracellular tripeptidyl peptidase (TPP) was homogenously purified from the culture supernatant of Rhizopus oligosporus by sequential fast protein liquid chromatography. The purified enzyme was a 136.5 kDa dimer composed of identical subunits. The effects of inhibitors and metal ions indicated that TPP is a metallo- and serine protease. TPP was activated by divalent cations, such as Co(2+) and Mn(2+), and completely inhibited by Cu(2+). Enzyme activity was optimal at pH 7.0 and 45 °C with a specific activity of 281.9 units/mg for the substrate Ala-Ala-Phe-pNA. The purified enzyme catalyzed cleavage of various synthetic tripeptides but not when proline occupied the P1 position. Purified TPP cleaved the pentapeptide Ala-Ala-Phe-Tyr-Tyr and tripeptide Ala-Ala-Phe, confirming the TPP activity of the enzyme.     

Chemical Characteristics of Precipitation at an Atlantic Station

Weekly or half-monthly bulk rainwater samples were collected over 3 yr (1993–1995) at Porspoder, an Atlantic station near Brest, France. They were analyzed for 0.08 M HNO3-leachable trace metals Cd, Cu, Fe, Mn, Ni, Pb and Zn by direct electrothermal atomic absorption spectrometry. Major ions and nitrogen species (NO3- and NH4+) in precipitation were determined by ion chromatography. In coastal precitation, a high correlation was observed for major ions having a seawater origin (Na+, CL- and Mg2+. Zn, Cd, Cu, Ni and Pb were well correlated indicating their anthropogenic origin. In the case of nitrogen species and nssSO42- a high correlation with Zn was observed, taken as an anthropogenic pollution tracer. Major ions and nitrogen species were determined according to wind sector direction showing clearly the impact of the pollution at this station attributed to continental air masses. A relative constancy of pH values in all wind directions was observed. Due to an important oceanic influence, trace metal concentrations and annual deposition densities were significantly lower at Porspoder than at North Sea coastal stations. In the case of nitrogen species, the relatively higher concentration levels probably indicated a significant local agricultural contribution.     

Demetalation of Fe, Mn, and Cu chelates and complexes: application to the NMR analysis of micronutrient fertilizers

The application of nuclear magnetic resonance (NMR) for the quality control of fertilizers based on Fe(3+), Mn(2+), and Cu(2+) chelates and complexes is precluded by the strong paramagnetism of metals. Recently, a method based on the use of ferrocyanide has been described to remove iron from commercial iron chelates based on the o,o-EDDHA [ethylenediamine-N,N'bis(2-hydroxyphenylacetic)acid] chelating agent for their analysis and quantification by NMR. The present work extended that procedure to other paramagnetic ions, manganese and copper, and other chelating, EDTA (ethylenediaminetetraacetic acid), IDHA [N-(1,2-dicarboxyethyl)-d,l-aspartic acid], and complexing agents, gluconate and heptagluconate. Results showed that the removal of the paramagnetic ions was complete, allowing us to obtain (1)H NMR spectra characterized by narrow peaks. The quantification of the ligands by NMR and high-performance liquid chromatography showed that their complete recovery was granted. The NMR analysis enabled detection and quantification of unknown impurities without the need of pure compounds as internal standards.     

Highly effective adsorption of heavy metal ions from aqueous solutions by macroporous xylan-rich hemicelluloses-based hydrogel

Xylan-rich hemicelluloses-based hydrogel was developed as a novel porous bioadsorbent by graft co-polymerization of acrylic acid (AA) and xylan-rich hemicelluloses for adsorption of heavy metal ions (Pd(2+), Cd(2+), and Zn(2+)) from aqueous solutions. The chemical structure, the interaction between the hydrogel and metal ions, and the porous structure of xylan-rich hemicelluloses-g-AA hydrogel were revealed by Fourier transform infrared spectroscopy and scanning electron microscopy. The effects of AA and cross-linker dosage, pH value, contacting time, and initial concentration of metal ion on the adsorption capacity were studied. The adsorption equilibrium time was about 60 min from the adsorption kinetics study. The maximum adsorption capacities of Pd(2+), Cd(2+), and Zn(2+) were 859, 495, and 274 mg/g, respectively. Furthermore, xylan-rich hemicelluloses-g-AA hydrogel also exhibited highly efficient regeneration and metal ion recovery efficiency and can be reused without noticeable loss of adsorption capacity for Pd(2+), Cd(2+), and Zn(2+) after quite a number of repeated adsorption/desorption cycles.     

Reactions of fulvic acid with metal ions

Fulvic acid is a water-soluble humic material that occurs widely in soils and waters and that tends to form water-soluble and water-insoluble complexes with a variety of metal ions, some of which are toxic. This paper presents information on the conditions under which the different types of FA-metal complexes are formed. The solubility in water, separately and after mixing, of FA (2 to 30 mg/100 ml) and eleven metal ions (Fe(III), Al, Cr(III), Pb, Cu, Hg(II), Zn, Ni, Co, Cd and Mn; 1 × 10^?5 moles of each metal ion) was investigated over the pH range 4 to 9. After mixing, the solubility of the components was significantly affected by pH only when less than 20 mg of FA was present. As the systems became richer in FA (22 to 30 mg), most of the metal ions remained in the aqueous phase, likely due to the formation of FA-metal complexes, inhibiting the formation of metal hydroxides. The order in which the eleven metal ions tended to form water-insoluble FA-metal complexes depended on the pH. At pH 6 it was: Fe = Cr = Al > Pb = Cu > Hg > Zn = Ni = Co = Cd = Mn. This order appeared to correlate with the valence, 1st hydrolysis constants and effective hydrated ionic diameters of the metal ions. In general, FA/metals weight ratios of > 2 favored the formation of water-soluble FA-metal complexes; at lower ratios, water-insoluble complexes, which could accumulate in soils and sediments, were formed.     

Interactive effects of aluminum and chromium stresses on the uptake of nutrients and the metals in barley

Aluminum (Al) and chromium (Cr) stresses often occur simultaneously in agricultural soils, and pose a great damage to crop growth, yield formation and product safety. In the current study, the influence of combined Al and Cr stresses on plant biomass, metal and nutrient contents was determined in comparison with that of Al or Cr stress alone. A hydroponic experiment was conducted to investigate the effect of pH, Al and Cr in the medium solution on the uptake of mineral elements as well as Al and Cr in the two barley genotypes differing in Al tolerance. Aluminum sensitive genotype Shang 70-119 had significantly higher Cr and Al contents in plants than Al-tolerant genotype Gebeina. Barley roots had much higher Al and Cr contents than above-ground plant parts. Chromium contents were much higher in the solution with pH 4.0 than in that with pH 6.5. Aluminum stress reduced phosphorus (P), calcium (Ca), magnesium (Mg), sulfur (S), copper (Cu), manganese (Mn), zinc (Zn) and boron (B) contents in roots and restrained potassium (K) and iron (Fe) from being translocated into shoots and leaves. Chromium stress resulted in reduced P, K, Mg, S, Fe, Zn and Mn contents in roots at pH 6.5 and P, K, Ca, Mg, S, Zn and Mn contents at pH 4.0. Translocation of all nutrients from roots to upper parts of plants was inhibited except Ca in pH 6.5 with Cr addition. Lower contents of all nutrients were observed at pH 4.0 as compared to pH 6.5. Combined stress of Cr and Al, on the whole, caused further reduction in mineral content in all plant parts of the two barley genotypes as compared to Al or Cr stress alone. Moreover, the reduction was more pronounced in Al sensitive genotype Shang 70-119.     

Characterization of iron,manganese, and copper synthetic hydroxyapatites by electron paramagnetic resonance spectroscopy

The incorporation of micronutrients (e.g., Fe, Mn, Cu) into synthetic hydroxyapatite (SHA) is proposed for slow release of these nutrients to crops in NASA's Advanced Life Support (ALS) program for long-duration space missions. Separate Fe3+ (Fe-SHA), Mn2+ (Mn-SHA), and Cu2+ (Cu-SHA) containing SHA materials were synthesized by a precipitation method. Electron paramagnetic resonance (EPR) spectroscopy was used to determine the location of Fe3+, Mn2+, and Cu2+ ions in the SHA structure and to identify other Fe(3+)-, Mn(2+)-, and Cu(2+)-containing phases that formed during precipitation. The EPR parameters for Fe3+ (g=4.20 and 8.93) and for Mn2+ (g=2.01, A=9.4 mT, D=39.0 mT and E=10.5 mT) indicated that Fe3+ and Mn2+ possessed rhombic ion crystal fields within the SHA structure. The Cu2+ EPR parameters (g(z)=2.488, A(z)=5.2 mT) indicated that Cu2+ was coordinated to more than six oxygens. The rhombic environments of Fe3+ and Mn2+ along with the unique Cu2+ environment suggested that these metals substituted for the 7 or 9 coordinate Ca2+ in SHA. The EPR analyses also detected poorly crystalline metal oxyhydroxides or metal-phosphates associated with SHA. The Fe-, Mn-, and Cu-SHA materials are potential slow release sources of Fe, Mn, and Cu for ALS and terrestrial cropping systems.     

Effects of excess aluminum on mineral uptake in mycorrhizal sorghum

Soil acidity is often associated with toxic aluminum (Al), and mineral uptake usually decreases in plants grown with excess Al. This study was conducted to evaluate the effects of Al (0, 35, 70, and 105 μM) on Al, phsophorus (P), sulfur (S), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn,) and copper (Cu) uptake in shoots and roots of sorghum [Sorghum bicolor (L.) Moench, cv. SC283] colonized with the vesicular‐arbuscular mycorrhizal (VAM) fungi isolates Glomus intraradices UT143–2 (UT143) and Glomus etunicatum UT316A‐2 (UT316) and grown in sand (pH 4.8). Mycorrhizal (+VAM) plants had higher shoot and root dry matter (DM) than nonmycorrhizal (‐VAM) plants. The VAM treatment had significant effects on shoot concentrations of P, K, Ca, Fe, Mn, and Zn; shoot contents of P, S, K, Ca, Mg, Fe, Mn, Zn, and Cu; root concentrations of P, S, K, Ca, Mn, Zn, and Cu; and root contents of Al, P, S, K, Ca, Mg, Fe, Mn, Zn, and Cu. The VAM effects on nutrient concentrations and contents and DM generally followed the sequence of UT316 > UT143 > ‐VAM. The VAM isolate UT143 particularly enhanced Zn uptake, and both VAM isolates enhanced uptake of P and Cu in shoots and roots, and various other nutrients in shoots or roots.     

RELATIONSHIPS AMONG METALS IN THE SOLID PHASE OF SOILS AND IN WILD PLANTS

The relationships between plant and soil systems were investigated using multivariate statistical methods and relative ionic impulsions. Soil samples were taken from three locations and wild plant species consisted of:Poa, Pteridium aquilinum, Diplotaxis, Plantago lanceolata andTrifolium repens. The content of Mg, Ca, Na, K, Mn, Fe, Pb, Co, Ni, Cu, Zn and Cr in soils and plants was determined. A five-step chemical fractionation procedure (speciation) was applied to soil samples. Total metal contents were determined and amounts extracted with HCl, EDTA and DTPA were measured. Pb, Co, Ni, Cu, Fe, Zn, Cd and Cr in soils (considered as microconstituents for plants) show a greater number of statistically significant relationships with plant contents than those shown by macroconstituents (Mg, Ca, Na, K and Mn). On the other hand, only Zn and Pb extracted with EDTA and DTPA seem to be related to metal contents in wild plants. When using relative ionic impulsions, any soil fraction (obtained from the fractionation procedure, including the soluble fraction, which is the sum of all the fractions except the residual) is suitable for the study of soil-plant system, suggesting a global balance among all the fractions. Our results suggested an active assimilation for K and a passive uptake for Na, Ca and Mn. Furthermore, Fe seemed to favour the active assimilation of the other microelements.     

营养物质及金属离子对DLL-E4菌降解对硝基苯酚的影响

研究了酵母膏、葡萄糖、蛋白胨、土壤浸液及Ca2 、Mg2 、Fe2 、Fe3 、Al3 、Mn2 、Co2 、Zn2 、Li 、Cu 、Cu2 、Ba2 、Ni2 等 13种金属离子对甲基对硫磷降解菌DLL E4降解对硝基苯酚的影响。结果表明 :适量添加酵母膏、葡萄糖和蛋白胨都能有效提高菌株对对硝基苯酚的降解 ,土壤浸液没有影响 ;金属离子中 ,除Li 和 0 1mmolL-1Fe3 外均对DLL E4降解对硝基苯酚的性状有一定的影响 ,其中Ca2 、Mg2 、Mn2 影响不大 ,Fe3 、Fe2 、Al3 、Ba2 、Zn2 高浓度时影响较大 ,Co2 、Cu2 、Cu 、Ni2 对DLL E4降解对硝基苯酚有较大的影响     

Seasonal dynamics of mineral nutrients and carbohydrates by walnut tree leaves

The dry weight accumulation per leaf as well as the concentration per gram of dry weight and the accumulation of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu) were determined in walnut tree leaves (Juglans regia L.) during a complete life cycle. Additionally, the dynamics of plant nutrient concentration in leaf petiole sap and carbohydrate accumulation in leaves were studied in relation to the main life cycle events of the walnut tree. Total N, P, K, Cu, and Zn concentrations decreased, whereas that of Ca, Mg, and Mn increased during the season. Iron concentration fluctuated around a mean value. Total N, P, K, Mg, and Cu concentrations detected in younger mature leaves were at the sufficient level, whereas Ca, Fe, Mn, and Zn concentrations were at higher levels as compared to those previously reported. All the detected nutrient accumulations increased abruptly during leaf ontogeny and leaf maturation until a maximum level was attained in the younger mature leaves. Similarly, sucrose, glucose, and fructose accumulation were observed at the same period. The rates of total N, P, Cu, and Zn accumulation were lower than the rates of the observed dry matter accumulation and nutrient concentration dilution. Potassium and Mn accumulation rates were almost equal, whereas those for Ca and Mg were higher as compared to the dry matter accumulation rate. The fast embryo growing phase resulted in a considerable decrease in dry weight, total N, P, K, Cu, Zn, and carbohydrate accumulation, and to a lesser degree in Ca, Mg, and Mn accumulation. Nutrient accumulation reduction in leaves by the influence of the growing fruits were estimated to be: total N 52%, K 48%, P 29.5%, Mg 16.3%, Ca 15%, Fe 51.2%, Cu 55.2%, Zn 37.3%, and Mn 5.4% of the maximum nutrient value of the younger mature leaves. Old leaves preserved nutrients before leaf fall as follows: total N 25.4%, P 45%, K 31%, Ca 74.8%, Mg 76.5%, Mn 89.2%, Fe and Zn 50%, and Cu 37%. Nutrient remobilization from the senescing old leaves before leaf fall were: total N 22.6%, P 25.5%, K 21%, Ca 10.2%, Mg 7%, Fe 3.2%, Mn 5.4%, Cu 8%, and Zn 13.3% of the maximum value in the younger mature leaves. In early spring, the absorption rates of N, P, and Ca were low while those of Mg, Fe, Mn, Cu, and Zn were high. During the fast growing pollen phase, the N, P, Fe, Mn, Cu, and Zn concentrations were reduced. Calcium concentration is supposed to be more affected by the rate of transpiration rather than during the growing of embryo. Calcium and Mg concentrations in the sap were negatively correlated. The detected K concentration level in the sap was as high as 33 to 50 times that of soluble N, 12 to 21 times to that of P, 5 times to that of Ca, and 10 to 20 times to that of Mg. The first maximum of starch accumulation in mature leaves was observed during the slow growing embryo phase and a second one after fruit ripening. Old senescing leaves showed an extensive carbohydrate depletion before leaf fall.     

（Mn＋Zn）-LS螯合微肥的研制及肥效实验

以废碱锰电池和木质素为原料,采用＂焙烧-酸解-螯合＂工艺制成（Mn＋Zn）-LS螯合微肥,通过红外光谱分析改性前后木质素的结构变化,并利用盆栽试验验证其肥效。红外分析结果表明,微肥的红外光谱图在峰形、峰位和强度上都发生了较大的变化,木质素结构单元上引入了新的基团（-SO3H、-SO-O-）和金属离子的络合导致了芳环吸收峰的变化,而且其中的络合也与芳环有关。盆栽试验结果表明,螯合微肥施用效果较好、增产明显,具有一定的抗病能力和抗倒伏性,且施用量范围广、易控制,适用范围广;在本试验中,Mn以2mg·kg-1土的施用量为最佳,鲜重增加46.3%,干重增加28.5%;同时盆栽后可以改善土壤pH和有效Zn、Mn含量,为后期利用提供肥力保证。