Reduced and oxidised glutathione and glutathione-reductase activity in tissues of Pisum sativum

In three-week-old pea plants (Pisum sativum L., cv. Little Marvel) grown in the light, total glutathione levels were highest in apex and expanding leaves (1.5 μmol·(g FW)-1), and lower (0.4–0.6 μmol·(g FW)-1) in older leaves and roots. In the light period, levels in expanded leaves increased by about 40%, compared with dark values, with lesser increases in roots and apex. In illuminated plants the proportion in the reduced form was 86–88% in leaves and 80% in roots, and these values fell during the dark period (to 82% and 69%, respectively). Reduced glutathione makes up 65–70% of the low-molecular-weight thiol in leaves, but over 95% in roots. Chloroplasts contained about 10% of the leaf glutathione, at a concentration of 1–2 mM; total glutathione in the chloroplasts, and the proportion of oxidised form (GSSG) increased in light, while NADP+/NADPH ratios decreased, indicating both synthesis and active oxidation of glutathione in light. Chloroplasts contained 52% (young leaf) to 75% (mature leaf) of the GSSG-reductase (EC 1.6.4.2) activity in the leaves. In roots, over 30% of the GSSG reductase was in the plastid fraction. Very little GSSG-reductase activity was associated with mitochondria in leaf or root.

[1]  L. J. Kok,et al.  EFFECTS OF FROST-HARDENING AND SALINITY ON GLUTATHIONE AND SULFHYDRYL LEVELS AND ON GLUTATHIONE-REDUCTASE ACTIVITY IN SPINACH LEAVES , 1983 .

[2]  S. Miyachi,et al.  [43] Cycling assay for nicotinamide adenine dinucleotides , 1980 .

[3]  B. Halliwell,et al.  The presence of glutathione and glutathione reductase in chloroplasts: A proposed role in ascorbic acid metabolism , 2004, Planta.

[4]  A. Keys,et al.  Increased levels of glutathione in a catalase-deficient mutant of barley (Hordeum vulgare L.) , 1984 .

[5]  R. Hampp,et al.  Determination of compartmented metabolite pools by a combination of rapid fractionation of oat mesophyll protoplasts and enzymic cycling. , 1984, Plant physiology.

[6]  H. Rennenberg Glutathione metabolism and possible biological roles in higher plants , 1980 .

[7]  B. Miflin,et al.  Intracellular localization of aspartate kinase and the enzymes of threonine and methionine biosynthesis in green leaves. , 1983, Plant physiology.

[8]  I. K. Smith,et al.  Sulfate transport in cultured tobacco cells. , 1975, Plant physiology.

[9]  L. Bergmann Aspects of S- and N-Metabolism in Tissue Cultures , 1981 .

[10]  H. Rennenberg,et al.  Phloem transport of sulfur in Ricinus , 1982, Planta.

[11]  G. Ellman,et al.  Tissue sulfhydryl groups. , 1959, Archives of biochemistry and biophysics.

[12]  K. Joy,et al.  A rapid method for isolation of purified, physiologically active chloroplasts, used to study the intracellular distribution of amino acids in pea leaves , 1980, Planta.

[13]  R. C. Fahey,et al.  Role of hydration state and thiol-disulfide status in the control of thermal stability and protein synthesis in wheat embryo. , 1980, Plant physiology.

[14]  K. Joy,et al.  Amino Acid Metabolism of Pea Leaves , 2022 .

[15]  E. Latzko,et al.  Partial Purification and Properties of Spinach Leaf Glutathione Reductase , 1978 .

[16]  J. Mclaren,et al.  Evidence for carrier-mediated transport of L-leucine into isolated pea (Pisum sativum L.) chloroplasts , 2004, Planta.

[17]  K. Joy,et al.  Properties of glutathione reductase from chloroplasts and roots of pea , 1986 .

[18]  B. Halliwell,et al.  Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. The effect of hydrogen peroxide and of Paraquat. , 1983, The Biochemical journal.

[19]  J. Burke,et al.  Chloroplast glutathione reductase: Purification and properties , 1984 .

[20]  D. Arnon COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. , 1949, Plant physiology.

[21]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[22]  N. Tolbert,et al.  Intracellular location of nitrate reductase and nitrite reductase. I. Spinach and tobacco leaves. , 1972, Biochimica et biophysica acta.

[23]  A. Keys,et al.  The regulation of the biosynthesis of glutathione in leaves of barley (Hordeum vulgare L.) , 1985 .

[24]  A. Lazarow,et al.  A microspectrophotometric method for the determination of cytochrome oxidase. , 1951, The Journal of biological chemistry.

[25]  H. Esterbauer,et al.  Seasonal Variation of Glutathione and Glutathione Reductase in Needles of Picea abies. , 1978, Plant physiology.

[26]  K. Joy,et al.  Amino Acid Metabolism of Pea Leaves: Diurnal Changes and Amino Acid Synthesis from 15N-Nitrate 1 , 1977 .

[27]  D. Chapman,et al.  Changes in Pool Sizes of Free Amino Acids and Amides in Leaves and Plastids of Zea mays during Leaf Development. , 1979, Plant physiology.

[28]  I. K. Smith,et al.  Stimulation of glutathione synthesis in photorespiring plants by catalase inhibitors. , 1985, Plant physiology.