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A new fluori-metric method for the determination of pyridine nucleotides in plant material and its use in following changes in the pyridine nucleotides during the respiration climacteric in apples


A new fluori-metric method for the determination of pyridine nucleotides in plant material and its use in following changes in the pyridine nucleotides during the respiration climacteric in apples



Phytochemistry 7(3): 337-353



ISSN/ISBN: 0031-9422

DOI: 10.1016/s0031-9422(00)90872-0

A method is described for the determination of the oxidized and reduced forms of pyridine nucleotides by measuring the increase in fluorescence due to the reduction of resazurin to its highly fluorescent reduced form, resorufin in a recycling system involving a specific dehydrogenase, its substrate, N-methyl phenazonium methosulphate and resazurin. The conditions of the assay were arranged so that the level of pyridine nucleotide was rate limiting and under these conditions it was possible to measure amounts of pyridine nucleotides as low as 10 /ijumole. Suitable investigations have shown that under the conditions of extraction and assay used, inhibitory effects due to the possible presence of enzyme inhibitors or quenchers of fluorescence in apple fruit extracts are negligible. A study of changes in pyridine nucleotides during the ripening of apple fruits detached from the tree was made. The most marked changes were in NADP and NADPH2 in the pulp, but small but significant changes also occurred in the NAD level both in the peel and pulp. The method of assay was applied to the peel and pulp of pears and to pea leaves in addition to apple tissue and the values obtained are compared with other published values for plant tissues.

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Accession: 024185413

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Related references

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