Comparison of<i>In Vivo</i>and<i>In Vitro</i>assays for nitrate reductase activity determination in soybean tissues

Hatam, Mir

doi:10.1080/00380768.1978.10433110

Soil Science and Plant Nutrition

1978

DOI: 10.1080/00380768.1978.10433110

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Comparison ofIn VivoandIn Vitroassays for nitrate reductase activity determination in soybean tissues

Mir Hatam

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1980

1982

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“…They also showed that cumulative NRA of soybean leaf discs, stems plus cotyledons, and root tips calculated on a whole plant basis agreed with actual reduced-N accumulation. In a more recent study, Hatam (8) showed soybean leaf disc and root tip NRA to be comparable on a fresh weight basis. However, leaflet NRA was much lower than is normally reported (7,14,18).…”

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Nitrate Reduction by Roots of Soybean (Glycine max [L.] Merr.) Seedlings

Crafts‐Brandner

Harper²

1982

Plant Physiol.

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Studies were conducted with 9 to 12 day-old soybean (Glycine max IL.I Merr. cv. Williams) seedlings to determine the contribution of roots to whole plant N03-reduction. Using an in vivo -NO3-nitrate reductase (NR) assay (no exogenous N03-added to incubation medium) developed for roots, the roots accounted for approximately 30% of whole plant nitrate reductase activity (NRA) of plants grown on 15 mM N03-.Nitrogen analyses of xylem exudate showed that 53 to 66% of the total-N was as reduced-N, depending on the time of day of exudate collection. These observations supported enzyme data that suggested roots were contributing significantly to whole plant N03-reduction. In short-term feeding studies using 15N-NO3-significant and increasing atom percent "5N excess was found in the reduced-N fraction of xylem exudate at 1.5 and 3 hours after feeding, respectively, which verified that roots were capable of reducing N03-.Estimated reduced-N accumulation by plants based on in vivo -NO3-NR assays of all plant parts substantially over-estimated actual reduced-N accumulation by the plants. Thus, the in vivo NR assay cannot be used to accurately estimate reduced-N accumulation but still serves as a useful assay for relative differences in treatment conditions. Soybean plants grown under midwest field conditions have been estimated to obtain from 25 to 60% of their total-N from symbiotic N2 fixation (22 (9) reported considerable fresh weight NRA in soybean root tips relative to leaf discs. They also showed that cumulative NRA of soybean leaf discs, stems plus cotyledons, and root tips calculated on a whole plant basis agreed with actual reduced-N accumulation. In a more recent study, Hatam (8) showed soybean leaf disc and root tip NRA to be comparable on a fresh weight basis. However, leaflet NRA was much lower than is normally reported (7,14, 18). The in vivo incubation medium used by Hatam and Hume (9) contained NADH which is known to interfere with the colorimetric assay of nitrite (20) and may have contributed to the low NRA values for leaves. In the more recent study, Hatam (8) used the in vivo NR assay method described by Jaworski (10) and yet the values reported for leaves were still quite low.A knowledge of root contribution to NO3-reduction is needed to understand N03-metabolism of the whole plant. Thus, the objectives of this study were to (a) investigate the contribution of roots to whole plant NO3-reduction based on in vivo -NO3-NR assays and xylem exudate analyses, (b) verify with '5N-NO3-the reduction of and (c)

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Nitrate Reduction by Roots of Soybean (Glycine max [L.] Merr.) Seedlings

Crafts‐Brandner

Harper²

1982

Plant Physiol.

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Seasonal and diurnal variations in nitrate reductase activity of soybean (Glycine max (L.) Merr.)

Hatam

1980

Plant Soil

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Comparison ofIn VivoandIn Vitroassays for nitrate reductase activity determination in soybean tissues

Cited by 2 publications

References 13 publications

Nitrate Reduction by Roots of Soybean (Glycine max [L.] Merr.) Seedlings

Nitrate Reduction by Roots of Soybean (Glycine max [L.] Merr.) Seedlings

Seasonal and diurnal variations in nitrate reductase activity of soybean (Glycine max (L.) Merr.)

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