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Inducible UDP-glucose dehydrogenase from French bean (Phaseolus vulgaris L.) locates to vascular tissue and has alcohol dehydrogenase activity.

ABSTRACT: UDP-glucose dehydrogenase is responsible for channelling UDP-glucose into the pool of UDP-sugars utilized in the synthesis of wall matrix polysaccharides and glycoproteins. It has been purified to homogeneity from suspension-cultured cells of French bean by a combination of hydrophobic-interaction chromatography, gel filtration and dye-ligand chromatography. The enzyme had a subunit of Mr 40,000. Km values were measured for UDP-glucose as 5.5 +/- 1.4 mM and for NAD+ as 20 +/- 3 microM. It was subject to inhibition by UDP-xylose. UDP-glucose dehydrogenase activity co-purified with alcohol dehydrogenase activity from suspension-cultured cells, elicitor-treated cells and elongating hypocotyls, even when many additional chromatographic steps were employed subsequently. The protein from each source was resolved into virtually identical patterns of isoforms on two-dimensional isoelectric focusing/PAGE. However, a combination of peptide mapping and sequence analysis, gel analysis using activity staining and kinetic analysis suggests that both activities are a function of the same protein. An antibody was raised and used to immunolocalize UDP-glucose dehydrogenase to developing xylem and phloem of French bean hypocotyl. Together with data published previously, these results are consistent with an important role in the regulation of carbon flux into wall matrix polysaccharides.

SUBMITTER: Robertson D

PROVIDER: S-EPMC1216900 | biostudies-other | 1996 Jan

REPOSITORIES: biostudies-other

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Project description:Biological nitrogen fixation is the most important eco-friendly approach to nitrogenous fertilizer management in the rhizosphere. Rhizobium is considered the most important symbiotic N-fixing microorganism. Native strains of Rhizobium perform better than the non-native strains by getting ambient conditions for growth and proliferation. Native strains enhance the soil fertility and productivity of pulses. The study was carried out in three phases, i.e., pot experiment, field experiment, and farmers' field demonstrations. In a pot experiment, two isolated rhizobia were inoculated to seeds of French bean (Phaseolus vulgaris) and applied with and without lime to evaluate crop growth, photosynthetic activity, and nodule characteristics of the target crop. In the field, strains were inoculated to seeds of French bean, which received different combinations of inputs- inorganic fertilizers, lime, and boron- to study the influence of native stains on crop productivity and agronomic efficiency. In comparison to non-limed packaging, the amounts of chlorophyll a, chlorophyll b, total chlorophyll, and chlorophyll a:b were, respectively, 13% to 30%, 1% to 15%, 10% to 27%, and 1% to 20% greater in limed packages. In limed packages compared to non-limed packages, the root length, biomass, density, and growth rate were increased by 16% to 17%, 36% to 52%, 38% to 49%, and 36% to 52%, respectively. In contrast to non-limed packages, limed packages had nodule attributes like the number of nodules per plant and nodular weight, which were 28% to 41% and 33% to 37% greater, respectively. Inoculation of native rhizobia strains with liming to acid soil increased 46% to 72% of leaf nitrogen content over non-limed rhizobia inoculated packages. In a field experiment, the adoption of soil test-based fertilizer application had an advantage of 25% in pod yield over farmers' practice. Acid soil amelioration with lime improved pod yield from 14% to 39% over non-limed packages. Farmers' field demonstration recorded the highest pod yield in the package where seeds were inoculated with S2 (RBHR-21) strain added with soil test-based fertilizers (STD) followed by STD + S1 (RBHR-15) with 98% and 84% increase over farmers' practice. However, experimental evidence favored using both strains for bio-inoculation of the French bean crop.

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Inducible UDP-glucose dehydrogenase from French bean (Phaseolus vulgaris L.) locates to vascular tissue and has alcohol dehydrogenase activity.

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