Summary
Tomato plants were transformed with a chimaeric polygalacturonase (PG) gene, designed to produce a truncated PG transcript constitutively. In these plants expression of the endogenous PG gene was inhibited during ripening, resulting in a substantial reduction in PG mRNA and enzyme accumulation. This inhibition was comparable to that achieved previously using antisense genes. The expression of the truncated gene in ripe fruit was substantially lower than its expression in green fruit. Thus expression of both the endogenous and truncated genes is reduced in ripe fruit in which both are active. The implication of this observation is discussed in relation to the possible mechanism whereby sense constructs inhibit gene expression.
This is a preview of subscription content, log in via an institution to check access.
References
Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721
Biggs MS, Harriman RW, Handa AK (1986) Changes in gene expression during tomato fruit ripening. Plant Physiol 81:395–403
Bird CR, Smith CJS, Ray J, Moureau P, Bevan MJ, 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: 651–662
Chomczynski P, Qasba PK (1984) Alkaline transfer of DNA to plastic membranes. Biochem Biophys Res Commun 122:340–344
DellaPenna D, Bennett AB (1988) In vitro synthesis and processing of tomato fruit polygalacturonase. Plant Physiol 86:1057–1063
DellaPenna D, Alexander DC, Bennett AB (1986) Molecular cloning of tomato fruit polygalacturonase: analysis of polygalacturonase levels during ripening. Proc Nat] Acad Sci USA 83:6420–6424
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21
Depicker A, Stachel S, Dhaese P, Zambryski P, Goodman HM (1982) Nopaline synthase: transcript mapping and DNA sequence. J Mol Appl Genet 1: 561–573
Grierson D, Slater A, Speirs J, Tucker GA (1985) The appearance of polygalacturonase mRNA in tomatoes: one of a series of changes in gene expression during development and ripening. Planta 163:263–271
Grierson D, Maunders MJ, Slater A, Ray J, Bird CR, Schuch W, Holdsworth MJ, Tucker GA, Knapp JE (1986a) Gene expression during tomato ripening. Phil Trans R See [B] 314:399–410
Grierson D, Tucker GA, Keen J, Ray J, Bird CR, Schuch W (1986b) Sequencing and identification of a cDNA clone for tomato polygalacturonase. Nucleic Acids Res 14:8595–8603
Guilley H, Dudley RK, Jonard G, Balazs E, Richards KE (1982) Transcription of cauliflower mosasic virus DNA: detection of promoter sequences, and characterization of transcripts. Cell 30:763–773
Hamilton AJ, Lycett GW, Grierson D (1990) Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346:284–287
Hattori J, Gottlob-McHugh SG, Johnson DA (1987) The isolation of high-molecular-weight DNA from plants. Anal Biochem 165:70–74
Lagrimini LM, Bradford S, Rothstein S (1990) Peroxidase-induced wilting in transgenic tobacco plants. Plant Cell 2:7–18
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325
Napoli C, Lemieux C, Jorgensen R (1990) Introduction of a chimeric chalcone synthase gene into petunia results in reverible co-suppression of homologous genes in trans. Plant Cell 2:279–289
Raeder U, Broda P (1985) Rapid preparation of DNA from filamentous fungi. Lett Appl Microbiol 7:17–20
Smith CJS, Slater A, Grierson D (1986) Rapid appearance of a mRNA correlated with ethylene synthesis encoding a protein of MW 35,000. Planta 168:94–100
Smith CJS, Watson CF, Ray J, Bird CR, Morris PC, Schuch W, Grierson D (1988) Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature 334:724726
Smith CJS, Watson CF, Morris PC, Bird CR, Seymour GB, Grey JE, Arnold C, Tucker GA, Schuch W, Harding S, Grierson D (1990) Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes. Plant Mol Biol 14:369–379
Tucker GA, Grierson D (1982) Synthesis of tomato polygalacturonase during tomato fruit ripening. Planta 155:64–67
Tucker GA, Robertson NG, Grierson D (1980) Changes in polygalacturonase isoenzymes during the ripening of normal and mutant tomato fruit. Eur J Biochem 112:119–124
Tucker GA, Robertson NG, Grierson D (1981) The conversion of tomato fruit polygalacturonase isoenzyme 2 into isoenzyme 1 in vitro. Eur J Biochem 115: 87–90
van der Krol AR, Mur LA, Beld M, Mol JMN, Stuitje AR (1990) Flavonoid genes in petunia: addition of a limited number of gene copies may lead to a suppression of gene expression. Plant Cell 2:291–299
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Smith, C.J.S., Watson, C.F., Bird, C.R. et al. Expression of a truncated tomato polygalacturonase gene inhibits expression of the endogenous gene in transgenic plants. Molec. Gen. Genet. 224, 477–481 (1990). https://doi.org/10.1007/BF00262443
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00262443