Planta

, Volume 200, Issue 2, pp 265–272

The impact of reduced vacuolar invertase activity on the photosynthetic and carbohydrate metabolism of tomato

  • Julie Scholes
  • Nicholas Bundock
  • Robin Wilde
  • Stephen Rolfe
Article

Abstract

The impact of reduced vacuolar invertase activity on photosynthetic and carbohydrate metabolism was examined in tomato (Solanum lycopersicon L.). The introduction of a co-suppression construct (derived from tomato vacuolar invertase cDNA) produced plants containing a range of vacuolar invertase activities. In the leaves of most transgenic plants from line INV-B, vacuolar invertase activity was below the level of detection, whereas leaves from line INV-A and untransformed wild-type plants showed considerable variation. Apoplasmic invertase activity was not affected by the co-suppression construct. It has been suggested that, in leaves, vacuolar invertase activity regulates sucrose content and its availability for export, such that in plants with high vacuolar invertase activity a futile cycle of sucrose synthesis and degradation takes place. In INV-B plants with no detectable leaf vacuolar invertase activity, sucrose accumulated to much higher levels than in wild-type plants, and hexoses were barely detectable. There was a clear threshold relationship between invertase activity and sucrose content, and a linear relationship with hexose content. From these data the following conclusions can be drawn. (i) In INV-B plants sucrose enters the vacuole where it accumulates as hydrolysis cannot take place. (ii) There was not an excess of vacuolar invertase activity in the vacuole; the rate of sucrose hydrolysis depended upon the concentration of the enzyme. (iii) The rate of import of sucrose into the vacuole is also important in determining the rate of sucrose hydrolysis. The starch content of leaves was not significantly different in any of the plants examined. In tomato plants grown at high irradiance there was no impact of vacuolar invertase activity on the rate of photosynthesis or growth. The impact of the cosuppression construct on root vacuolar invertase activity and carbohydrate metabolism was less marked.

Key words

Carbohydrate metabolism Invertase (co-suppression construct) Photosynthesis Lycopersicon Vacuole 

Abbreviations

CaMV

Cauliflower Mosaic Virus

WT

wild type

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Benhamou N, Grenier J, Chrispeels MJ (1991) Accumulation of β-fructosidase in the cell walls of tomato roots following infection by a fungal wilt pathogen. Plant Physiol 97: 739–750Google Scholar
  2. Bevan MW (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acid Res 12: 8711–8721Google Scholar
  3. Bird CR, Smith CJS, Ray JA, Moureau P, Bevan MW, Bird AS, Hughes S, Morris PC, Grierson D, Schuch W (1988) The tomato polygalacturonase gene and ripening-specific expression in transgenic plants. Plant Mol Biol 11: 651–662Google Scholar
  4. Chen JQ, Black CC (1992) Biochemical and immunological properties of alkaline invertase isolated from sprouting soybean hypocotyls. Arch Biochem Biophys 295: 61–69Google Scholar
  5. Daie J (1984) Characterisation of sugar transport in storage tissue of carrot. J Am Soc Hort Sci 109: 718–722Google Scholar
  6. Dickinson CD, Altabella T, Chrispeels MJ (1991) Slow-growth phenotype of transgenic tomato expressing apoplastic invertase. Plant Physiol 95: 420–425Google Scholar
  7. Elliott KJ, Butler WO, Dickinson CD, Konno Y, Vedvick TS, Fitzmaurice L, Mirkov TE (1993) Isolation and characterisation of fruit vacuolar invertase genes from two tomato species and temporal differences in mRNA levels during fruit ripening. Plant Mol Biol 21: 515–524Google Scholar
  8. Eschrich W (1980) Free space invertase, its possible role in phloem unloading. Ber Dtsch Bot Ges 93: 363–378Google Scholar
  9. Foyer CH (1987) The basis for source-sink interaction in leaves. Plant Physiol Biochem 25: 649–657Google Scholar
  10. Goldschmidt EE, Huber SC (1992) Regulation of photosynthesis by end-product accumulation in leaves of plants storing starch, sucrose and hexose sugars. Plant Physiol 99: 1443–1448Google Scholar
  11. Hammond JBW, Burton KS, Shaw AF, Ho LC (1984) Sourcesink relationships and carbon metabolism in tomato leaves. 2. Carbohydrate pools and catabolic enzymes. Ann Bot 53: 307–314Google Scholar
  12. Heineke D, Sonnewald U, Büssis D, Günter G, Leidreiter K, Wilke I, Raschke K, Willmitzer L, Heldt HW (1992) Apoplastic expression of yeast-derived invertase in potato. Effects on photosynthesis, leaf solute composition, water relations and tuber composition. Plant Physiol 100: 301–308Google Scholar
  13. Heineke D, Wildenberger K, Sonnewald U, Willmitzer L, Heldt HW (1994) Accumulation of hexoses in leaf vacuoles: studies with transgenic tobacco plants expressing yeast-derived invertase in the cytosol, vacuole or apoplasm. Planta 194: 29–33Google Scholar
  14. Hole CC, Dearman J (1994) Sucrose uptake by the phloem parenchyma of carrot storage root. J Exp Bot 45: 7–15Google Scholar
  15. Huber SC (1989) Biochemical mechanism for regulation of sucrose accumulation in leaves during photosynthesis. Plant Physiol 91: 656–662Google Scholar
  16. Jang J-C, Sheen J (1994) Sugar sensing in higher plants. Plant Cell 6: 1665–1679Google Scholar
  17. Kuhlemeier C, Green PJ, Chua N-H (1987) Regulation of gene expression in plants. Annu Rev Plant Physiol 38: 221–257Google Scholar
  18. Lichtenthaler AK, Wellburn LR (1983) Determination of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans 11: 591–592Google Scholar
  19. Matsushita K, Uritani I (1974) Changes in invertase activity of sweet potato in response to wounding and purification and properties of its invertases. Plant Physiol 54: 60–66Google Scholar
  20. Morris DA, Arthur ED (1985) Effect of gibberellic acid on patterns of carbohydrate distribution and acid invertase activity in Phaseolus vulgaris. Physiol Plant 65: 257–262Google Scholar
  21. Myrback K (1960) Invertases. In: Boyer PD, Lardy H, Myrback K (eds) The enzymes (2nd edn). Academic Press, New York, pp 379–396Google Scholar
  22. Ohyama A, Ito H, Sato T, Nishimura S, Imai T, Hiria M (1995) Suppression of acid invertase activity by antisense RNA modifies the sugar composition of tomato fruit. Plant Cell Physiol 36: 369–376Google Scholar
  23. Ramloch-Lorenz K, Knudsen S, Sturm A (1993) Molecular characterisation of the gene for carrot cell wall β-fructosidase. Plant J 4: 545–554Google Scholar
  24. Ranwala AP, Suematsu C, Masuda H (1992) Soluble and wallbound invertases in strawberry fruit. Plant Sci 84: 59–64Google Scholar
  25. Ricardo CPP, ap Rees T (1970) Invertase activity during the development of carrot roots. Phytochemistry 9: 239–247Google Scholar
  26. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual (2nd edn). Cold Spring Habor Laboratory Press, New YorkGoogle Scholar
  27. Scholes JD, Lee PJ, Horton P, Lewis DH (1994) Invertase: understanding changes in the photosynthetic and carbohydrate metabolism of barley leaves infected with powdery mildew. New Phytol 126: 213–222Google Scholar
  28. Sheen J (1990) Metabolic repression of transcription in higher plants. Plant Cell 2: 1027–1038CrossRefPubMedGoogle Scholar
  29. Sturm A, Chrispeels MJ (1990) cDNA cloning of carrot extracellular β-fructosidase and its expression in response to wounding and bacterial infection. Plant Cell 2: 1107–1119Google Scholar
  30. Sturm A, Sebkova V, Lorenz L, Hardegger M, Lienhard S, Unger C (1995) Development- and organ-specific expression of genes for sucrose synthase and three isoenzymes of acid β-fructofuranosidase in carrot. Planta 195: 601–610Google Scholar
  31. Sonnewald U, Brauer M, von Schaewen A, Stitt M, Willmitzer L (1991) Transgenic tobacco plants expressing yeast-derived invertase in either the cytosol, vacuole or apoplast: a powerful tool for studying sucrose metabolism and sink/source interactions. Plant J 1: 95–106Google Scholar
  32. Tang X, Ruffner H-P, Scholes JD, Rolfe SA (1996) Purification and characterisation of soluble invertases from leaves of Arabidopsis thaliana. Planta 198: 17–23Google Scholar
  33. Tang X, Rolfe SA, Scholes JD (1996) The effect of Albugo Candida (white blister rust) on the photosynthetic and carbohydrate metabolism of leaves of Arabidopsis thaliana. Plant Cell Environ, in pressGoogle Scholar
  34. Tetlow IJ, Farrar JF (1992) Sucrose-metabolizing enzymes from leaves of barley infected with brown rust (Puccinia hordei Otth). New Phytol 120: 475–480Google Scholar
  35. Unger C, Hardegger M, Lienhard S, Sturm A (1994) cDNA cloning of carrot (Daucus carota) soluble acid β-fructofuranosidases and comparison of the cell wall isoenzyme. Plant Physiol 104: 1351–1357Google Scholar
  36. Weber H, Borisjuk L, Heim U, Buchner P, Wobus U (1995) Seed coat-associated invertases of fava bean control both unloading and storage functions: cloning of cDNAs and cell-type-specific expression. Plant Cell 7: 1835–1846Google Scholar
  37. Wright DP, Baldwin BC, Shephard MC, Scholes JD (1995) Source-sink relationships in wheat leaves infected with powdery mildew. I. Alterations in carbohydrate metabolism. Physiol Mol Plant Pathol 47: 237–253Google Scholar
  38. Wu LL, Song I, Karuppiah N, Kaufman PB (1993) Kinetic induction of oat shoot pulvinus invertase mRNA by gravistimulation and partial cDNA cloning by the polymerase chain reaction. Plant Mol Biol 21: 1175–1179Google Scholar
  39. von Schaewen A, Stitt M, Schmidt R, Sonnewald U, Willmitzer L (1990) Expression of a yeast-derived invertase in the cell wall of tobacco and Arabidopsis plants leads to accumulation of carbohydrate and inhibition of photosynthesis and strongly influences growth and phenotype of transgenic tobacco plants. EMBO J 9: 3033–3044Google Scholar
  40. Zrenner R, Schüler K, Sonnewald U (1995) Soluble acid invertase determines the hexose-to-sucrose ratio in cold-stored potato tubers. Planta 198: 246–252Google Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Julie Scholes
    • 1
  • Nicholas Bundock
    • 1
  • Robin Wilde
    • 2
  • Stephen Rolfe
    • 1
  1. 1.Department of Animal and Plant SciencesUniversity of SheffieldSheffieldUK
  2. 2.Zeneca Plant Science, Jealott's Hill Research StationBracknellUK

Personalised recommendations