The Xylem Ureide Assay of Nitrogen Fixation: Sampling Procedures and Sources of Error

Herridge, D. F.; O'Connell, Peter J.; Donnelly, K.A.

doi:10.1093/jxb/39.1.12

Cited by 34 publications

(10 citation statements)

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“…Limitations, Errors Sources of error associated with sampling of stems for extraction of N solutes and of whole shoots for extraction of VES have been examined (9,11,13), and protocols for sampling, treatment of samples, and chemical analysis are now well documented (see also 3,23). However, the ureide technique has restricted application to the measurement of nitrogen fixation in field studies if the relationships between relative ureide-N and P are specific for each combination of plant genotype and strain of B. japonicum.…”

Section: Samplingmentioning

confidence: 99%

“…Errors associated with sampling and subsequent treatment of xylem sap have been identified and sampling protocols defined to minimize errors (13). Comparisons of estimates of nitrogen fixation by field-grown plants based on the ureide and other methods are presented in the companion paper (12) and will be the subject of future reports (DF Herridge, MB Peoples, FJ Bergersen, unpublished data).…”

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confidence: 99%

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Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by ¹⁵N Methods

Herridge¹,

Peoples²

1990

Plant Physiol.

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154

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We report experiments to quantify the relationships between the relative abundance of ureide-N in root-bleeding sap, vacuumextracted sap, and hot water extracts of stems and petioles of nodulated soybean (Glycine max [L.] fixation based on the composition of N solutes in xylem (rootbleeding) sap. These authors and others (9,11,16,21) showed that the relationship between p2 and the relative abundance ofureide-N in xylem sap and extracts of plant parts was highly correlated. Nevertheless, the technique has not been widely adopted as a routine field assay for quantifying nitrogen fixation, but used instead to detect fixation activity or to differentiate treatment effects (22). Reasons for this may include: a general lack of confidence in the calibrations of the technique, because they were done with vegetative plants (16, 21) rather than over a complete growth cycle, or because they relied on the acetylene reduction assay to estimate nitrogen fixation (9, 1 1); doubts about the applicability of a single set of calibration curves to different genotypes of soybean and to different strains of Bradyrhizobium japonicum; uncertainty over sampling protocols and sources of error in the use of the technique; and lack of verification of the technique in field experiments where estimates of nitrogen fixation based on the relative abundance of ureide-N in xylem sap and tissue extracts were compared wtih estimates using other methods, e.g. N difference, 15N.During the past 4 years, we have attempted to resolve these issues. Errors associated with sampling and subsequent treatment of xylem sap have been identified and sampling protocols defined to minimize errors (13). Comparisons of estimates of nitrogen fixation by field-grown plants based on the ureide and other methods are presented in the companion paper (12) and will be the subject of future reports (DF Herridge, MB Peoples, FJ Bergersen, unpublished data). In this paper, we report experiments designed to quantify the relationship between the relative abundance of ureide-N in RBS, VES, and in extracts of the stems (including petioles) of Bragg soybean and Pincr. RBS and VES were sampled at regular intervals throughout the whole growth cycle of soybeans; sampling of stems was terminated during early podfill. Pincr, calculated for each interval of time between successive samplings, was considered a more appropriate measure of nitrogen fixation than P for calibrating the point-of-time ureide technique. Additional experiments examined the effect of plant genotype and strain of B. japonicum on the relationship between composition of xylem N-solutes and nitrogen 2 Abbreviations: P, proportion of plant N derived from nitrogen fixation; Pincr, proportion of plant N in a growth period between successive samplings derived from nitrogen fixation; RBS, root-bleeding xylem sap; VES, vacuum-extracted xylem sap; HUP, uptake hydrogenase. 495www.plantphysiol.org on May 10, 2018 -Published by Downloaded from

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Section: Samplingmentioning

confidence: 99%

mentioning

confidence: 99%

Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by ¹⁵N Methods

Herridge¹,

Peoples²

1990

Plant Physiol.

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154

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“…Nitrate-N was determined by the salicylic acid method (3). The relative abundance of ureides was calculated (9) Assay for GS Nodule GS activity was assayed using the ADP-dependent transferase reaction (15) which measures the formation of GHA. One unit of GS activity is defined as 1 ,umol of GHA formed per min.…”

Section: Sample Preparation and Analysis For Relative Abundance Of Urmentioning

confidence: 99%

Nitrogen Fixation of Nodulation Mutants of Soybean as Affected by Nitrate

Wu¹,

Harper²

1990

Plant Physiol.

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It was previously reported that three soybean (Glycine max [L.] Merr.) nodulation mutants (NOD1-3, NOD2-4, and NOD3-7) were partially tolerant to nitrate when nitrate was supplied simultaneously with inoculation at the time of transplanting. The current study evaluated the effect of short-term nitrate treatment on nitrogenase activity (C2H2 reduction per plant and per nodule weight) and on relative abundance of ureides when nitrate application was delayed until plants were 3 weeks old and nodules were fully developed. Nitrogenase activity of the mutants was similar to that of Williams after an initial 3-week growth period, prior to nitrate treatment. Application of 5 millimolar nitrate resulted in greater inhibition of nitrogenase activity in Williams than in the three mutants. NOD1-3 was most tolerant of nitrate among the mutants tested and showed the highest relative abundance of ureides. Although C2H2 reduction activity per plant for NOD1-3 was higher than for Williams in the presence of nitrate, C2H2 reduction activity per gram of nodules was lower for NOD1-3 than for Williams in the presence and absence of nitrate. Compared to Williams, NOD1-3 had higher nodule ureide concentration and had similar glutamine synthetase activity in nodule tissue, indicating its nodules have normal nitrogen assimilation pathways. Nitrate application resulted in ureide accumulation in nodule tissue as well as in all plant parts assayed. Unexpectedly, nitrate treatment also increased the rate of ureide degradative capacity of leaves in both NOD1-3 and Williams. The data confirmed that nitrogenase activity of the selected nodulation mutants was more, but still only partially, tolerant of nitrate compared with the Williams parent.also had higher ureide concentration in its nodule tissue (13), likely due to limited growth of the mutant which may have impaired ureide utilization.Partially nitrate tolerant mutants of soybean (NOD1-3, NOD2-4, and NOD3-7) have also been selected from mutagenized Williams populations, and initial characterization of these mutants has been reported (4). The data indicated many similar characteristics of our mutants to those reported by Carroll et al. (1, 2). However, it is not known whether the nitrogenase activity of our mutants is tolerant to nitrate per se, since in the previous study (4) the effects of nitrate on infection steps and on subsequent nodule function could not be separated due to simultaneous inoculation and nitrate addition. Using "5N analysis it was determined that NOD 1-3 fixed more '5N2 than Williams, when grown on urea, and that all three nodulation mutants symbiotically fixed more N2 than wild type when the mutants were grown on nitrate (1 1). This confirmed the earlier report based on C2H2 reduction assays (4). In the present report, nitrate treatment was delayed until plant roots were well nodulated and capable of relatively high N2 fixation activity. N2 fixation was then assessed by using the in situ C2H2 reduction assay and determining the relative abundance of ureide...

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“…For working on-farm, only two techniques for evaluating BNF inputs are really viable: the ureide abundance technique (Herridge et al, 1988) and the 15 N natural abundance technique (Shearer & Kohl, 1986). The former relies on the fact that many legumes of tropical origin transport fixed nitrogen from nodules to shoot in the form of ureides (allantoin and allantoic acid), but as this is not true for groundnut, one of the legumes used in this study (Peoples et al, 1989), this technique was not used.…”

Section: Introductionmentioning

confidence: 99%

Nitrogen fixation by groundnut and velvet bean and residual benefit to a subsequent maize crop

Okito

Alves

Urquiaga

et al. 2004

Pesq. agropec. bras.

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-Chemical fertilisers are rarely avaiable to poor farmers, for whom the nitrogen (N) is often the most limiting element for cereal grain production. The objective of this study was to quantify the contribution of biological nitrogen fixation (BNF) to groundnut (Arachis hypogaea) and velvet bean (Mucuna pruriens) crops using the 15 N natural abundance (δ 15 N) technique and to determine their residual effect and that of a natural fallow, on growth and N accumulation by two rustic maize varieties. The contribution of BNF calculated from δ 15 N data was 40.9, 59.6 and 30.9 kg ha -1 , for groundnut, velvet bean and the natural fallow, respectively. The only legume grain harvested was from the groundnut, which yielded approximately 1.000 kg ha -1 . The subsequent maize varieties ("Sol de Manhã" and "Caiana Sobralha") yielded between 1.958 and 2.971 kg ha -1 , and were higher after velvet bean for both maize varieties and "Sol da Manhã" groundnut, followed by "Caiana" after groundnut and, finally, the natural fallow. For a small-holder producer the most attractive system is the groundnut followed by maize, as, in this treatment, both groundnut and maize grain harvest are possible. However, a simple N balance calculation indicated that the groundnut-maize sequence would, in the long term, deplete soil N reserves, while the velvet bean-maize sequence would lead to a build up of soil nitrogen. Index terms: Arachis hypogaea, Mucuna pruriens, Zea mays,15 N natural abundance technique, green manures, N balance, small-holder producers. Fixação de nitrogênio por amendoim e mucuna e benefício residual para uma cultura de milhoResumo -Fertilizantes químicos raramente estão disponíveis aos agricultores com poucos recursos econômi-cos, e assim o N é, freqüentemente, um elemento mais limitante para a produção de grãos. O objetivo deste trabalho foi quantificar a contribuição da fixação biológica de nitrogênio (FBN) às culturas de amendoim (Arachis hypogaea) e mucuna (Mucuna pruriens), por meio da técnica de abundância natural de 15 N e determinar o efeito residual das leguminosas e do pousio sobre o crescimento e acumulação de N em duas variedades de milho. A contribuição da FBN calculada a partir dos dados de δ 15 N, foi de 40,9, 59,6 e 30,9 kg ha -1 , respectivamente, para amendoim, mucuna e o pousio. A única leguminosa de grão colhida foi amendoim que produziu aproximadamente 1,000 kg ha -1 . A produtividade da cultura de milho (variedades Caiana e Sol da Manhã) variou de 1,958 a 2,971 kg ha -1 de grãos, sendo mais alta após mucuna em ambas as variedades de milho e após amendoim na Sol da Manhã, seguida por Caiana após amendoim e por último o pousio normal. Para pequenos agricultores o sistema mais atrativo é o que inclui amendoim seguido pelo milho, uma vez que duas colheitas de grãos são possíveis. Contudo, o cálculo do balanço de N indicou que a seqüência amendoim-milho diminuiria, a longoprazo, a reserva de N do solo, enquanto a seqüência mucuna-milho ajudaria a aumentar a reserva de N do solo.Termos para indexação...

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The Xylem Ureide Assay of Nitrogen Fixation: Sampling Procedures and Sources of Error

Cited by 34 publications

References 14 publications

Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by ¹⁵N Methods

Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by ¹⁵N Methods

Nitrogen Fixation of Nodulation Mutants of Soybean as Affected by Nitrate

Nitrogen fixation by groundnut and velvet bean and residual benefit to a subsequent maize crop

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The Xylem Ureide Assay of Nitrogen Fixation: Sampling Procedures and Sources of Error

Cited by 34 publications

References 14 publications

Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by 15N Methods

Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by 15N Methods

Nitrogen Fixation of Nodulation Mutants of Soybean as Affected by Nitrate

Nitrogen fixation by groundnut and velvet bean and residual benefit to a subsequent maize crop

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Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by ¹⁵N Methods

Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by ¹⁵N Methods