Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters fat – fatty oil – ester-type wax – or...
Patent
1995-07-26
1998-10-13
Robinson, Douglas W.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters fat, fatty oil, ester-type wax, or...
435 9131, 4351721, 4351723, 536 231, 536 232, 536 241, 5361245, C12N 1500, C07H 2104, C12P 1934
Patent
active
058213988
DESCRIPTION:
BRIEF SUMMARY
This application is a 371 PCT/AU93/00654, filed on Dec. 15, 1993, published as WO94/13797 Jun. 23, 1994.
This invention relates to inducible plant promoter DNA sequences, DNA sequences encoding tomato alcohol dehydrogenase 2 (ADH2) enzyme, and hybrid DNA molecules incorporating such sequences. In a particular application of the invention, these hybrid DNA molecules are used to transform plants to enable the control of the condition and quality of fruit.
In the soft fruit industry (e.g. strawberries, peaches, plums and tomatoes), substantial losses are incurred during transport, storage and marketing because of the susceptibility of softer fruit to mechanical damage and invasion by microorganisms. To limit such losses, plant breeders have selected lines which are less soft, and for many fruits firm cultivars now dominate national and international trade. Unfortunately, while marketing losses may be lower when fruit is less soft, the firmer lines may have reduced market appeal and also tend to be deficient in flavour.
In some fruits, such as tomatoes, avocadoas and peaches, softening arises from the degradation of cell wall components by enzymes deposited in response to signals associated with ripening. Ripening signals typically include ethylene and the fruit response to ethylene and other signals can be strongly affected by temperature, and by oxygen and carbon dioxide concentrations. In firm fruit cultivars, the response to ripening signals has usually been damped so that less enzyme is produced; softening is correspondingly protracted and unless the signal response can be enhanced, by the time the fruits are acceptably soft the tissue is often senescent and lacking in flavour and aroma.
In higher organisms, structural genes, including those involved in softening and the development of flavour and aroma in fruit, are switched on by DNA sequences known as promoters, which recognise internal signals and "promote" transcription of the adjacent functional genes. Promoters which respond to oxygen stress or to heat shock have been characterised in plants, for example alcohol dehydrogenase (ADH) accumulates in plants after 5 to 10 hours of oxygen stress, and heat shock proteins can be detected after only brief periods of high temperature. In each case the new chemical entities are produced through the response of promoters to environmental signals. Thus, these kinds of inducible promoters permit the construction of hybrid DNA molecules in which a structural gene encoding, for example, a polypeptide involved in fruit softening, is brought under the control of an inducible promoter sequence, so that the structural gene is transcribed when the promoter is subjected to an activating signal.
Accordingly, in a first aspect the present invention provides an isolated DNA molecule comprising a nucleotide sequence encoding a soft fruit promoter or functional portion thereof, wherein said promoter or functional portion thereof is characterised in that it can be activated by environmental agents or conditions and/or is activated, or primarily activated, during a late stage of normal soft fruit ripening.
Preferably the soft fruit promoter or functional portion thereof is activated thermally, chemically or by light. In the case of chemical activation, preferred promoters and functional portions thereof will be responsive to particular levels of gases such as oxygen, carbon dioxide or carbon monoxide. Alternatively, chemical activation may be achieved through exposure to some organic acids.
The soft fruit promoter or functional portion thereof may be isolated from grapes, strawberries, peaches, plums or tomatoes. Preferably the soft fruit promoter or functional portion thereof is activated by environmental agents or conditions, and is further characterised by also being activated, or primarily activated, during a late stage of normal ripening. For example, for tomato, during the period of ripening following 5-6 days after ripening-onset. More preferably, the promoter or functional portion thereof is the tomato alcohol dehydrog
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Brady Colin John
Hinde Richard
Lee Elizabeth
Longhurst Terrence James
Speirs James
Commonwealth Scientific & Industrial Research Org.
Robinson Douglas W.
Wai Thanda
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