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. 1989 Jan;1(1):53–63. doi: 10.1105/tpc.1.1.53

Expression of a chimeric polygalacturonase gene in transgenic rin (ripening inhibitor) tomato fruit results in polyuronide degradation but not fruit softening.

J J Giovannoni 1, D DellaPenna 1, A B Bennett 1, R L Fischer 1
PMCID: PMC159736  PMID: 2535467

Abstract

Tomato fruit ripening is accompanied by extensive degradation of pectic cell wall components. This is thought to be due to the action of a single enzyme, polygalacturonase, whose activity is controlled, at least in part, at the level of gene expression. At the onset of tomato fruit ripening, polygalacturonase enzyme activity, mRNA levels, and relative rate of gene transcription all increase dramatically. To elucidate the role of polygalacturonase during tomato fruit ripening, we utilized a pleiotropic genetic mutation, rin, that blocks many aspects of ripening, including the activation of polygalacturonase gene transcription. The polygalacturonase structural gene was ligated to a promoter that is inducible in mature rin fruit and inserted into the fruit genome, and plants were regenerated. This allowed expression of the polygalacturonase gene in transgenic rin fruit at a time corresponding to ripening in wild-type fruit. Expression of this gene resulted in the accumulation of active polygalacturonase enzyme and the degradation of cell wall polyuronides in transgenic rin fruit. However, no significant effect on fruit softening, ethylene evolution, or color development was detected. These results indicate that polygalacturonase is the primary determinant of cell wall polyuronide degradation, but suggest that this degradation is not sufficient for the induction of softening, elevated rates of ethylene biosynthesis, or lycopene accumulation in rin fruit.

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Selected References

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  1. Biggs M. S., Harriman R. W., Handa A. K. Changes in Gene Expression during Tomato Fruit Ripening. Plant Physiol. 1986 Jun;81(2):395–403. doi: 10.1104/pp.81.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blumenkrantz N., Asboe-Hansen G. New method for quantitative determination of uronic acids. Anal Biochem. 1973 Aug;54(2):484–489. doi: 10.1016/0003-2697(73)90377-1. [DOI] [PubMed] [Google Scholar]
  3. Burg S. P., Burg E. A. Molecular requirements for the biological activity of ethylene. Plant Physiol. 1967 Jan;42(1):144–152. doi: 10.1104/pp.42.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crookes P. R., Grierson D. Ultrastructure of tomato fruit ripening and the role of polygalacturonase isoenzymes in cell wall degradation. Plant Physiol. 1983 Aug;72(4):1088–1093. doi: 10.1104/pp.72.4.1088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davis K. R., Darvill A. G., Albersheim P., Dell A. Host-Pathogen Interactions : XXIX. Oligogalacturonides Released from Sodium Polypectate by Endopolygalacturonic Acid Lyase Are Elicitors of Phytoalexins in Soybean. Plant Physiol. 1986 Feb;80(2):568–577. doi: 10.1104/pp.80.2.568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deikman J., Fischer R. L. Interaction of a DNA binding factor with the 5'-flanking region of an ethylene-responsive fruit ripening gene from tomato. EMBO J. 1988 Nov;7(11):3315–3320. doi: 10.1002/j.1460-2075.1988.tb03202.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dellapenna D., Alexander D. C., Bennett A. B. Molecular cloning of tomato fruit polygalacturonase: Analysis of polygalacturonase mRNA levels during ripening. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6420–6424. doi: 10.1073/pnas.83.17.6420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dellapenna D., Kates D. S., Bennett A. B. Polygalacturonase Gene Expression in Rutgers, rin, nor, and Nr Tomato Fruits. Plant Physiol. 1987 Oct;85(2):502–507. doi: 10.1104/pp.85.2.502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fuchs Y., Anderson J. D. Purification and characterization of ethylene inducing proteins from cellulysin. Plant Physiol. 1987 Jul;84(3):732–736. doi: 10.1104/pp.84.3.732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grierson D., Tucker G. A., Keen J., Ray J., Bird C. R., Schuch W. Sequencing and identification of a cDNA clone for tomato polygalacturonase. Nucleic Acids Res. 1986 Nov 11;14(21):8595–8603. doi: 10.1093/nar/14.21.8595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hobson G. E. Polygalacturonase in normal and abnormal tomato fruit. Biochem J. 1964 Aug;92(2):324–332. doi: 10.1042/bj0920324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lincoln J. E., Cordes S., Read E., Fischer R. L. Regulation of gene expression by ethylene during Lycopersicon esculentum (tomato) fruit development. Proc Natl Acad Sci U S A. 1987 May;84(9):2793–2797. doi: 10.1073/pnas.84.9.2793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lincoln J. E., Fischer R. L. Diverse mechanisms for the regulation of ethylene-inducible gene expression. Mol Gen Genet. 1988 Apr;212(1):71–75. doi: 10.1007/BF00322446. [DOI] [PubMed] [Google Scholar]
  14. Lincoln J. E., Fischer R. L. Regulation of Gene Expression by Ethylene in Wild-Type and rin Tomato (Lycopersicon esculentum) Fruit. Plant Physiol. 1988 Oct;88(2):370–374. doi: 10.1104/pp.88.2.370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McMurchie E. J., McGlasson W. B., Eaks I. L. Treatment of fruit with propylene gives information about the biogenesis of ethylene. Nature. 1972 May 26;237(5352):235–236. doi: 10.1038/237235a0. [DOI] [PubMed] [Google Scholar]
  16. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Themmen A. P., Tucker G. A., Grierson D. Degradation of isolated tomato cell walls by purified polygalacturonase in vitro. Plant Physiol. 1982 Jan;69(1):122–124. doi: 10.1104/pp.69.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tong C. B., Labavitch J. M., Yang S. F. The induction of ethylene production from pear cell culture by cell wall fragments. Plant Physiol. 1986 Jul;81(3):929–930. doi: 10.1104/pp.81.3.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tucker G. A., Robertson N. G., Grierson D. Changes in polygalacturonase isoenzymes during the 'ripening' of normal and mutant tomato fruit. Eur J Biochem. 1980 Nov;112(1):119–124. doi: 10.1111/j.1432-1033.1980.tb04993.x. [DOI] [PubMed] [Google Scholar]
  20. Van Haute E., Joos H., Maes M., Warren G., Van Montagu M., Schell J. Intergeneric transfer and exchange recombination of restriction fragments cloned in pBR322: a novel strategy for the reversed genetics of the Ti plasmids of Agrobacterium tumefaciens. EMBO J. 1983;2(3):411–417. doi: 10.1002/j.1460-2075.1983.tb01438.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Walker-Simmons M., Ryan C. A. Proteinase inhibitor I accumulation in tomato suspension cultures : induction by plant and fungal cell wall fragments and an extracellular polysaccharide secreted into the medium. Plant Physiol. 1986 Jan;80(1):68–71. doi: 10.1104/pp.80.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wallner S. J., Bloom H. L. Characteristics of tomato cell wall degradation in vitro: implications for the study of fruit-softening enzymes. Plant Physiol. 1977 Aug;60(2):207–210. doi: 10.1104/pp.60.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wallner S. J., Walker J. E. Glycosidases in Cell Wall-degrading Extracts of Ripening Tomato Fruits. Plant Physiol. 1975 Jan;55(1):94–98. doi: 10.1104/pp.55.1.94. [DOI] [PMC free article] [PubMed] [Google Scholar]

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