Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1993-06-17
1995-09-12
Fox, David T.
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
536 236, 536 253, 536 245, 4351721, 4351723, 800200, 800205, 800DIG65, A01H 500, C12N 1511, C12N 1529, C12N 1552
Patent
active
054497647
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
This application relates to novel DNA constructs, plant cells containing the constructs and plants derived therefrom. In particular it involves the use of antisense or sense RNA technology to control gene expression in plants.
SUMMARY OF THE INVENTION
Many physiological and developmental processes are controlled by ethylene in higher plants, including peach (Prunus persica). These processes include fruit ripening where ethylene may be involved in both the initiation and rate of continuation of many of the changes involved in fruit ripening. However the exact role of ethylene has hitherto not been fully understood. We have now isolated novel DNA involved in the generation of ethylene in peaches. In this invention, we provide such novel DNA, and methods of using it. One such use is a method for controlling the rate of production of ethylene in ripening peaches. In this way the rate of many of the ethylene-related changes associated with fruit ripening on a plant can be modified according to the characteristics that are required.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
As is well known, a cell manufactures protein by transcribing the DNA of the gene for that protein to produce messenger RNA (mRNA), which is then processed (eg by the removal of introns) and finally translated by ribosomes into protein. This process may be inhibited by the presence in the cell of "antisense RNA". By this term is meant an RNA sequence which is complementary to a sequence of bases in the mRNA in question: complementary in the sense that each base (or the majority of bases) in the antisense sequence (read in the 3' to 5' sense) is capable of pairing with the corresponding base (G with C, A with U) in the mRNA sequence read in the 5' to 3' sense. It is believed that this inhibition takes place by formation of a complex between the two complementary strands of RNA, preventing the formation of protein. How this works is uncertain: the complex may interfere with further transcription, processing, transport or translation, or degrade the mRNA, or have more than one of these effects. Such antisense RNA may be produced in the cell by transformation with an appropriate DNA construct arranged to transcribe backwards part of the coding strand (as opposed to the template strand) of the relevant gene (or of a DNA sequence showing substantial homology therewith).
The use of this technology to downregulate the expression of specific plant genes has been described, in for example European Patent publication no 271988 to ICI (corresponding to U.S. Ser. No. 119614). Reduction of gene expression has led to a change in the phenotype of the plant: either at the level of gross visible phenotypic difference e.g. lack of anthocyanin production in flower petals of petunia leading to colourless instead of coloured petals (van der Krol et al, Nature, 333, 866-869, 1988); or at a more subtle biochemical level e.g. change in the amount of polygalacturonase and reduction in depolymerisation of pectins during tomato fruit ripening (Smith et al, Nature, 334, 724-726, 1988; Smith et al., Plant Molecular Biology, 13, 303-311, 1990) Thus antisense RNA has been proven to be useful in achieving downregulation of gene expression in plants.
The present invention relates to clones of a gene which expresses ethylene-forming enzyme (EFE). EFE is involved in ethylene production, and hence in the ripening of peaches (Tonutte et al., J. Am. Soc. Hort. Sci., 116, p274, 1991). Fragments of this gone have been cloned and characterised. We postulate that they will be of use in modifying the ripening characteristics of peaches. The gene in question is partially encoded in the-clone P13-B, which has homology (83% encoded amino acid homology) with the clone pTOM13 from tomato disclosed by Holdsworth et al (Nucleic Acids Research 15, 731, 1987). It has been shown that the gene encoded by pTOM13 is involved in ethylene synthesis in tomatoes (Hamilton et al, Nature, 346, pp284, 1990), and its translation product is now believed to be ethylene-formi
REFERENCES:
patent: PP6409 (1988-11-01), Zaiger et al.
Potrykus, 1991, Annu. Rev. Plant Physiol. Plant & Mol. Biol. 1991, 42:205-225.
Von der Krol et al. 1988, Nature, 333:865-869.
Holdsworth et al. 1987a Nucleic Acids Research, 15(2):731-739.
Holdsworth et al. 1987b, Nucleic Acids Research 15(24):10600.
Holdsworth et al. 1988, Plant Molecular Biology, 11:81-88.
Hamilton et al. 1990, Nature, 346:284-287.
Callahan et al. 1990a, Journal of Cellular Biochemistry Suppl. 14E:346 Abstract #R505.
Callahan et al. 1990b, Hortscience 25(9):130-131 Abstract #478.
Morgens et al. 1990, In Current Topics in Plant Physiology vol. 5 Flores et al., eds. pp. 319-320.
Giovannori et al. 1989, The Plant Cell 1:53-63.
Bird Colin R.
Ray John A.
Schuch Wolfgang W.
Fox David T.
Veitenheimer Erich E.
Zeneca Limited
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