Content of Oxalate in Actinidia Deliciosa Plants Grown in Nutrient Solutions with Different Nitrogen Forms

Kiwifruit plants (Actinidia deliciosa cv. Hayward) were grown in Hoagland nutrient solution with calcium nitrate, potassium nitrate, ammonium nitrate or ammonium chloride as the nitrogen source. Plants grown in the solution with nitrate nitrogen displayed a higher oxalate content, greater shoot length and leaf area, and higher content of ascorbic acid and NO₃ ^– ions in the leaves. Plants grown in the solution with ammonium nitrate, and particularly with ammonium chloride, showed low oxalate content, low content of ascorbic acid and NO₃ ^–, high content of Cl^– and Na⁺, low shoot length and leaf area. Oxalate formation appeared to be connected with the assimulation of nitrate, more precisely with nitrate reduction, while ammonium nitrogen assimilation did not induce the synthesis of oxalic acid.     

Oxalic acid and oxalate oxidase enzyme in <Emphasis Type="Italic">Costus pictus</Emphasis> D. Don

The leaves of Costus pictus are sour in taste due to the presence of oxalic acid in the leaves. Different stages of leaves were collected and the samples were designated as stage one, stage two and stage three. It was found that oxalate content and oxalate oxidase activity were maximum in second leaf stage followed by first leaf stage and third leaf stage. Drying causes substantial loss of oxalate content and complete loss of oxalate oxidase activity. With various solvents water recovered more oxalate followed by methanol and ethanol while oxalate oxidase activity was maximum in ethanol followed by methanol and water. The ethanol or methanol extract of second leaf stage of C. pictus can be used for isolating active principles. The oxalate oxidase from C. pictus can be used as a cheap source of oxalate oxidase enzyme which is used in oxalate determination in biological fluids. Moreover, the sensitivity of oxalate determination employing oxalate oxidase from C. pictus will be more as oxalate oxidase in C. pictus has K _m 20 times lesser than the oxalate oxidase enzyme from barley seedling.     

Oxalic acid metabolism and calcium oxalate formation in Lemna minor L.

Abstract Axenic Lemna minor plants, which form numerous calcium oxalate crystals, were exposed to [¹⁴C]-glycolic acid, -glyoxylic acid, -oxalic acid and -ascorbic acid and prepared for microautoradiography by a technique that preserves only insoluble label to determine specifically the pathway leading to oxalic acid used for crystal formation. Label from glycolic, glyoxylic, and oxalic acids was incorporated into crystals. Label from oxalic acid was also found in starch when exposure to label was done in the light but not dark, while plastids specialized for lipid storage were heavily labelled under both conditions. Incorporation of label from glycolic and glyoxylic acids, but not oxalic acid, was inhibited in the presence of the glycolate oxidase inhibitors, αHPMS (2-pyridylhydroxy methanesulphonic acid) and mHBA (methyl 2-hydroxy-3-butynoic acid), and inhibition of labelling was not due to an effect on uptake. These studies show that the glycolate oxidase pathway to oxalic acid is operational in L. minor and that the product is available for crystal formation. Dark-grown plants form almost four times as many crystal cells (idioblasts) as do light-grown plants, indicating crystal formation is not in response to photorespiratory glycolate production. Label from [1-¹⁴C]ascorbic acid was also incorporated into crystals and labelling was inhibited by mHBA, indicating glycolic acid and/or glyoxylic acid are possible intermediates of ascorbic acid catabolism. The effect of nitrogen source on crystal formation was also investigated. Significantly more crystal idioblasts were formed, on a surface area basis, by plants grown on ammonium than by plants grown on nitrate nitrogen. When grown with mixed ammonium and nitrate, an intermediate number of crystal idioblasts were formed.     

The photosynthetic production of oxalic acid in Oxalis corniculata

The biogenesis of oxalic acid in Oxalis corniculata has beeninvestigated. In O. corniculata the bulk of the oxalic acidis produced by CO₂ fixation both in light and in darkness butthe rate of its photosynthetic formation is much higher thanin darkness. Several other plants some of which are known toaccumulate oxalic acid e.g., Biophytum sensitivum, Averrhoacarambola, Impatiens balsamina, Amorphophallus campanulatusand Colocassia antiquorum also fix ¹⁴CO₂ into oxalic acid photosyntheticallywithin 1 min of exposure to the gas. In O. corniculata ¹⁴C canbe detected in oxalic acid within 5 sec and about half of thetotal ¹⁴C fixed in the 70% ethanol soluble fraction can be locatedin this compound after 5 min. This is accompanied by a declineof radioactivity in two compounds, the chromatographic behaviourand melting points of one of which and its DNP hydrazone aresimilar to those of an authentic sample of glyoxylic acid. Whenglyoxylate 1, 2-¹⁴C is incubated with Oxalis leaf homogenateit is converted to oxalate-¹⁴C. Glycolate is also metabolizedto oxalate. The conversion of both glycolate and glyoxylateare favoured by light. The C₂ compounds acetate and glycinehowever are utilized rather poorly. Sucrose-¹⁴C is also notmetabolized markedly for this purpose. (Received August 20, 1969; )     

Oxalate Accumulation as Regulated by Nitrogen Forms and Its Relationship to Photosynthesis in Rice （Oryza sativa L.）

Four-leaf rice seedlings (Oryza sativa L.), which had been cultivated in Kimura B complete nutrient solution, were treated with two nitrogen forms by replacing the nitrogen element in the complete solution with sole nitrate or ammonium (2.86 mmol/L). Nitrate-N nutrition tended to increase oxalate content in all parts of the plant, including the leaves, stems, roots, and root exudates, whereas ammonium had the opposite effect. Consequently, marked differences in oxalate content were observed between the two treatments throughout the time tested (0-12 d), with maximal differences of approximately 12-fold at 6d after treatment. Photosynthetic/respiratory parameters were examined over time simultaneously with changes in oxalate content. Net photosynthetic rate, chlorophyll fluorescence parameters (i.e. maximal photochemical efficiency (Fv/Fm) and photochemical quantum yields of photosystem (PS)II (ΦPSⅡ)), and respiratory rate were not significantly different between plants treated with the two nitrogen forms, although ammonium-fed plants had apparently higher leaf chlorophyll content than nitrate-fed plants. Leaf glucose content was altered little, but the content of fructose, sucrose, and total soluble sugar was significantly higher in the leaves of ammonium-fed plants than nitrate-fed plants. The results indicate that nitrate/ammonium may serve as efficient regulators of oxalate accumulation owing to regulation of metabolism in rice leaves rather than oxalate downward transfer and root excretion, and that photosynthetic metabolism is not directly correlated with the regulation of oxalate accumulation in rice plants.     

Oxalate Accumulation as Regulated by Nitrogen Forms and Its Relationship to Photosynthesis in Rice (Oryza sativa L.)

Four-leaf rice seedlings (Oryza sativa L.), which had been cultivated in Kimura B complete nutrient solution, were treated with two nitrogen forms by replacing the nitrogen element in the complete solution with sole nitrate or ammonium (2.86 mmol/L). Nitrate-N nutrition tended to increase oxalate content in all parts of the plant, including the leaves, stems, roots, and root exudates, whereas ammonium had the opposite effect. Consequently, marked differences in oxalate content were observed between the two treatments throughout the time tested (0--12 d), with maximal differences of approximately 12-fold at 6 d after treatment. Photosynthetic/respiratory parameters were examined over time simultaneously with changes in oxalate content. Net photosynthetic rate, chlorophyll fluorescence parameters (i.e. maximal photochemical efficiency (Fv/Fm) and photochemical quantum yields of photosystem (PS)Ⅱ (φ PSⅡ)), and respiratory rate were not significantly different between plants treated with the two nitrogen forms, although ammonium-fed plants had apparently higher leaf chlorophyll content than nitrate-fed plants. Leaf glucose content was altered little, but the content of fructose, sucrose, and total soluble sugar was significantly higher in the leaves of ammonium-fed plants than nitrate-fed plants, The results indicate that nitrate/ammonium may serve as efficient regulators of oxalate accumulation owing to regulation of metabolism in rice leaves rather than oxalate downward transfer and root excretion, and that photosynthetic metabolism is not directly correlated with the regulation of oxalate accumulation in rice plants.     

Accumulation of Oxalate in Tissues of Sugar-beet, and the Effect of Nitrogen Supply

In field-grown sugar-beet concentration of insoluble oxalatewas low in roots and high (about 12 per cent of ethanol insolublematerial) in leaves, and for a particular leaf the concentrationincreased continuously during its life. Of the insoluble oxalate,15–30 per cent was present as the magnesium salt and theremainder as the calcium salt. Oxalate contents of plants grownin culture solutions with nitrate as nitrogen source were similarto those of plants grown in soil, but when nitrogen was suppliedas ammonium sulphate or ammonium nitrate both soluble and insolubleoxalate were low. Plants grown in soil with regular additionsof ammonium sulphate or ammonium nitrate also had very low concentrationsof soluble oxalate although insoluble oxalate was only slightlylower than with nitrate nitrogen. Disks of root or leaf tissuewashed for several days in distilled water lost insoluble oxalatebut when washed in tap water insoluble oxalate increased morethan twofold. Addition of calcium and nitrate to the distilledwater caused an increase of insoluble oxalate, while additionof potassium caused a decrease. Use of ¹⁴C labelled oxalateand washing experiments showed that oxalate can be metabolizedby tissue disks and so is not necessarily a final product ofmetabolism. The accumulation of oxalate appears to be connectedwith the assimilation of nitrate and the preservation of thecation-anion balance of the plant.     

Genetic Variability in Taro, Colocasia esculenta (L.) Schott (Araceae)

An examination of two seedling populations indicates that geneticvariability can be obtained in taro by means of sexual propagation. Colocasia esculenta, dasheen, taro, alkaloids, anthocyanins, calcium oxalate, chlorophyll, genetic variability, nitrogen content, protein levels, in taro seedlings     

不同因子对荞麦中草酸含量的影响

用不同化合物从根部喂养麦幼苗,测定其根叶中草酸含量的变化。结果表明：异柠檬酸、抗坏血酸及其前体物均可不同程度地降低荞麦根叶中草酸含量;而乙醇酸与乙醛酸则显著提高其草酸含量,表明荞麦叶片草酸合成主要来自乙醇酸途径,而非来自抗坏血酸等途径。水培条件下,以铵态氮或尿素等作唯一氮源时,荞麦中草酸含量远低于以硝态氮培养的;将谷氨酸或丝氨酸加到含硝态氮培养液中也能显著降低其草酸含量,不同氮素影响荞麦草酸含量可能与乙醇酸途径有关。     

Further experiments on the acid grassSetaria sphacelata

Summary Field trials carried out in 1963 and 1964 have confirmed the ammonia-acidity relationship in the grassSetaria sphacelata. The addition of nitrogenous fertilisers enhanced ammonia accumulation in the grass but not titratable acidity. Sand- and water-culture experiments confirmed the essentially acidic nature ofS. sphacelata and the presence of a relatively highly dissociated acid which was found to be oxalic acid. In these experiments ammonia accumulated inS. sphacelata even when nitrogen was supplied as nitrate alone. Darkening the plants for seven days during growth in the field led to a marked accumulation of ammonia inS. sphacelata particularly when covered during early growth.From the evidence obtained it is concluded thatS. sphacelata is essentially an acid grass and that ammonia accumulation is a consequence of this. The mechanism proposed is that part of the ammonia in the plant, whether derived from direct uptake, nitrate reduction or decomposition (following darkening) of organic nitrogenous compounds, is present as ammonium oxalate. Since plant protoplasts are relatively impermeable to ions ammonium accumulates in the grass. The subsequent decline in concentration in the plant, with increasing maturity, is ascribed to the increase in structural carbohydrates and lignin (a dilution effect) and possible slow utilisation of the ammonium.     

不同氮素形态培养下荞麦叶片中草酸积累的变化

用1/5浓度Hoagland(pH6.0)营养液培养荞麦幼苗3d后,取其中一部分继续用此营养液(硝态氮);另一部分用硫酸氨和氯化钙取代硝态氮(氨态氮)的营养液,均培养至荞麦第一片真叶完全展开。结果表明,以氨态氮为唯一氮源培养荞麦时,植株叶片中草酸含量显著下降。进一步研究表明,氨态氮培养下荞麦根中及根分泌草酸的速率也显著下降,结果排除了叶片中草酸含量的下降是由于叶片中草酸向其根系转运或是因为根分泌草酸速率的差异造成的,而可能与其草酸代谢改变有关。氨态氮培养下叶片中与草酸代谢相关的有机酸含量以及相关酶活性也显著下降,这可能意味着荞麦叶片草酸形成积累可能与相关有机酸代谢有关。     

氮素形态及配比对夏播菘蓝生长及活性成分含量的影响

以来自山西的菘蓝(Isatis indigotica Fort.)为实验对象,采用盆栽法研究铵态氮(NH4+-N)、硝态氮(NO3--N)和酰胺态氮〔CO(NH2)2〕的不同配比对夏播菘蓝生长,叶和根中的可溶性蛋白质及总氮含量,根中多糖含量,叶中叶绿素相对含量,以及叶中靛玉红和靛蓝、根中(R,S)-告依春的含量和积累量的影响.结果表明:各施氮处理组的单株叶干质量均高于对照(不施用氮素)组,但单株根干质量或高于或低于对照组,其中,T4〔n(铵态氮):n(硝态氮):n(酰胺态氮)=25:75:0〕处理组的单株叶和根干质量均最大,且总体上显著高于对照组及其他施氮处理组(P<005);而施氮处理组的根冠比均显著低于对照组.各施氮处理组叶中的可溶性蛋白质含量与对照均无显著差异,但各施氮处理组根中的可溶性蛋白质含量、叶和根中的总氮含量以及叶中的叶绿素相对含量总体上显著高于对照组,而根中的多糖含量或高于或低于对照组,其中,T6〔n(铵态氮):n(硝态氮):n(酰胺态氮)=0:75:25〕处理组根中的多糖含量和叶中的叶绿素相对含量均最高,T3〔n(铵态氮):n(硝态氮):n(酰胺态氮)=50:50:0〕处理组叶和根中的可溶性蛋白质含量均较高.各施氮处理组叶中靛玉红含量总体上显著高于对照组,多数施氮处理组叶中靛蓝含量则显著低于对照组,但各施氮处理组的单株叶中靛蓝和靛玉红积累量总体上高于对照组;其中,T2〔n(铵态氮):n(硝态氮):n(酰胺态氮)=75:25:0〕处理组叶中靛玉红含量及其单株积累量均最高,T6处理组叶中靛蓝含量最高,而单株叶中靛蓝积累量则以T3处理组最高.各施氮处理组根中(R,S)-告依春含量总体上显著低于对照组,其中,以T1〔n(铵态氮):n(硝态氮):n(酰胺态氮)=100:0:0〕处理组根中(R,S)-告依春含量最高,T4处理组单株根中(R,S)-告依春积累量最高.综合分析结果表明:按不同配比施用不同形态氮素,夏播菘蓝的生长及活性成分含量有明显差异,因此,若以收获叶为目的,结合叶中靛玉红含量,建议施用铵态氮和硝态氮物质的量比为75:25的复合氮肥;若以收获根为目的,结合根中(R,S)-告依春含量,建议施用铵态氮和硝态氮物质的量比为25:75的复合氮肥.     

Increase of ascorbic acid content and nutritional quality in spinach leaves during physiological acclimation to low temperature

The effect of acclimation to 10 °C on the leaf content of ascorbic and oxalic acids, was investigated in spinach (Spinacia oleracea L.). At 10 °C the content of ascorbic acid in leaves increased and after 7 days it was about 41% higher than in plants remaining under a 25 °C/20 °C day/night temperature regime. In contrast, the content of oxalate, remained unchanged. Transfer to 10 °C increased the ascorbic but not the oxalic acid content of the leaf intercellular washing fluid (IWF). Oxalate oxidase (OXO EC 1.2.3.4) activity was not detected in extracts of leaf blades. Therefore, oxalic acid degradation via OXO was not involved in the control of its content. Our results show that low temperature acclimation increases nutritional quality of spinach leaves via a physiological rise of ascorbic acid that does not feed-forward on the content of oxalic acid.     

The effect of nitrogen fertilization and rust fungus infection, singly and combined, on the leaf chemical composition of Rumex obtusifolius

1. The chrysomelid beetle Gastrophysa viridula occurs on Rumex obtusifolius growing in a range of nutrient conditions and also on plants infected with the foliar fungus Uromyces rumicis . In a controlled environment, we investigated the effect of fertilizing plants with eight nitrate and four ammonium concentrations, with or without infection, on leaf nutritional quality.
2. Increasing nitrate fertilization increased leaf oxalate, total nitrogen and nitrate concentrations and water content, and decreased total non-structural carbohydrate (NSC) concentrations. Increasing ammonium fertilization increased leaf nitrogen concentration and water content, decreased nitrate and NSC concentrations, and had no effect on oxalate concentrations.
3. Infection produced a mainly additive effect to fertilization, increasing NSC and oxalate, and decreasing nitrate and nitrogen concentration in whole plants fed nitrate, and increasing nitrate and NSC in whole plants fed ammonium.
4. Young leaves on infected plants remained uninfected and had greater nitrogen and NSC concentrations, and lower oxalate and nitrate concentrations, than infected leaves on the same plant.
5. These results are discussed in relation to changes in C:N and NSC:organic nitrogen ratios, the effect of nitrate and oxalate, and the known feeding and oviposition preferences of the beetle. The results suggest that there is an optimum nitrogen fertilization level for G. viridula development.     

氮素水平对花生氮素代谢及相关酶活性的影响

 在大田高产条件下研究了氮素水平对花生(Arachis hypogaea)可溶性蛋白质、游离氨基酸含量及氮代谢相关酶活性的影响, 结果表明, 适当提高氮素水平既能增加花生各器官中可溶性蛋白质和游离氨基酸的含量, 又能提高硝酸还原酶、谷氨酰胺合成酶和谷氨酸脱氢酶等氮素同化酶的活性, 使其达到同步增加; 氮素水平过高虽能提高硝酸还原酶和籽仁蛋白质含量, 但谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH)的活性下降; N素施肥水平不改变花生植株各器官中可溶性蛋白质、游离氨基酸含量以及硝酸还原酶(NR)、谷氨酰胺合成酶、谷氨酸脱氢酶活性的变化趋势, 但适量施N (A2和A3处理)使花生各营养器官中GS、GDH活性提高; 氮素水平对花生各叶片和籽仁中GS、GDH活性的高低影响较大, 但对茎和根中GDH活性大小的影响较小。     

Growth Analysis of Contrasting Cultivars of Zea mays L. at Different Rates of Nitrogen Supply

The effects of nitrogen supply on the growth and nitrogen contentsof four cultivars of Zea mays L. of different origins were examinedunder water-culture conditions at the seedling stage. Seedlingsof cultivars CNIA12, LG11, Tusa Finn, and UNPHU XC301 were grownunder three different relative addition rates of nitrogen. Growthparameters were determined by means of functional growth analysisconducted on 10 to 19 d-old seedlings. No differences in relativegrowth rates were found among cultivars when nitrogen supplywas high. However, at the lowest rate of nitrogen supply, TusaFina and LG11 showed lower relative growth rates than CNIA12and UNPHU XC301, where relative growth rates were sustainedeven at the lowest rate of nitrogen supply, due to a higherunit leaf rate. The higher unit leaf rate of these two cultivarscorresponds directly to higher leaf and plant nitrogen contents.High positive correlations were found between plant nitrogencontents and both relative growth rate and unit leaf rate. Theresults suggest a potential for selection of genotypes withimproved performance under conditions where high rates of nitrogen-fertilizerapplication are too costly or not desirable.Copyright 1994,1999 Academic Press Maize, Zea mays, growth analysis, maize, nitrogen nutrition, nitrogen content, relative addition rates, relative growth rates, unit leaf rate     

Evidence for a cytoplasmic pathway of oxalate biosynthesis in Aspergillus niger

Oxalate accumulation of up to 8 g/liter was induced in Aspergillus niger by shifting the pH from 6 to 8. This required the presence of Pi and a nitrogen source and was inhibited by the protein synthesis inhibitor cycloheximide. Exogenously added 14CO2 was not incorporated into oxalate, but was incorporated into acetate and malate, thus indicating the biosynthesis of oxalate by hydrolytic cleavage of oxaloacetate. Inhibition of mitochondrial citrate metabolism by fluorocitrate did not significantly decrease the oxalate yield. The putative enzyme that was responsible for this was oxaloacetate hydrolase (EC 3.7.1.1), which was induced de novo during the pH shift. Subcellular fractionation of oxalic acid-forming mycelia of A. niger showed that this enzyme is located in the cytoplasm of A. niger. The results are consistent with a cytoplasmic pathway of oxalate formation which does not involve the tricarboxylic acid cycle.     

Simultaneous detection of 35S- and 32P-labeled proteins on electrophoretic gels

A new method for the determination of oxalic acid in urine, which does not require isolation of oxalic acid, was developed by derivatizing oxalic acid and separating and quantitating the product by automated liquid chromatography. Oxalic acid in urine was reacted with o-phenylenediamine to form the strongly uv-absorbing compound 2,3-dihydroxyquinoxaline. Isolation and quantitation of this derivative were accomplished using a reverse-phase C₈ column, 5% methanol in 0.1 m ammonium acetate buffer (pH 6.6) as eluant, and absorption at 314 nm. The method was linear from 1 to 151 μg oxalic acid/ml of sample and the conversion of oxalic acid to the dihydroxyquinoxaline over this concentration range was 94.9%. The precision of duplicates averaged ±1.1%. Analyses of urine before and after treatment with oxalate decarboxylase were employed to differentiate actual urinary oxalic acid from oxalogenic compounds. Under the conditions employed, no urine was found to contain inhibitors of oxalate decarboxylase. No significant contribution to the method was found in a study of 19 potentially interfering urinary constituents. Levels of oxalic acid found in 27 urine samples from patients by this method averaged 71% of levels found using an earlier colorimetric method.     

Evidence for a cytoplasmic pathway of oxalate biosynthesis in Aspergillus niger.

Oxalate accumulation of up to 8 g/liter was induced in Aspergillus niger by shifting the pH from 6 to 8. This required the presence of Pi and a nitrogen source and was inhibited by the protein synthesis inhibitor cycloheximide. Exogenously added 14CO2 was not incorporated into oxalate, but was incorporated into acetate and malate, thus indicating the biosynthesis of oxalate by hydrolytic cleavage of oxaloacetate. Inhibition of mitochondrial citrate metabolism by fluorocitrate did not significantly decrease the oxalate yield. The putative enzyme that was responsible for this was oxaloacetate hydrolase (EC 3.7.1.1), which was induced de novo during the pH shift. Subcellular fractionation of oxalic acid-forming mycelia of A. niger showed that this enzyme is located in the cytoplasm of A. niger. The results are consistent with a cytoplasmic pathway of oxalate formation which does not involve the tricarboxylic acid cycle.