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
1993-01-25
1995-12-19
Fox, David T.
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...
800200, 800255, 435 691, 435 698, 4351723, 435200, 4352404, 4352523, 530370, 530378, 536 231, 536 232, 536 236, 935 60, 935 67, 935 72, A01H 500, C12N 924, C12N 1529
Patent
active
054770017
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
The invention relates to a recombinant DNA coding for a novel protein having .beta.-1,3-glucanase activity, or for a precursor of this protein, to a bacterium containing this recombinant DNA, to a plant cell, plant or plant part, especially a plant seed, transformed by this recombinant DNA, as well as to this novel protein and to a method for preparing it.
Crop plants are known to be subjected to attack by-parasites such as phytopathogenic fungi, which are responsible for substantial harvest losses. The .main means at present for controlling these fungi lies in the use of chemical substances having fungicidal activity. It is now known that plants react naturally to such attack by various defence mechanisms, which are unfortunately, in general, triggered too late and at too low an intensity to be sufficiently effective.
One of these mechanisms comprises the induction of an enzyme known as .beta.-1,3-glucanase E.C.3.2.1.39 (Kombrink et al., 1988, Pr. Ntl. Acad. Sci. USA, 85,982-986 and Pegg et al., 1981, Physiol. Plant. Pathol. 19,371-382). This induction can be triggered artificially through the effect of elicitors, that is to say compounds of biological origin capable of inducing in a healthy plant the defence reactions which it deploys naturally during an infection by pathogenic agents, or during a hormonal imbalance caused by auxin, cytokinins or ethylene (Abeles et al., 1971, Plant Physiol. 47,129-134 and de Loose et al., 1988, Gene, 70,13-23).
.beta.-1,3-glucans are linear polysaccharide polymers consisting of glucose units linked via .beta.-(1.fwdarw.3) linkages, sometimes possessing .beta.-(1.fwdarw.4) or .beta.-(1.fwdarw.6) type branching (Farka 1982, in "Fungal protoplasts", Peberdy and Ferencry, published by Dekker Inc.). These polysaccharides constitute a typical component of the skeleton of the wall of most fungi, and in particular phytopathogenic fungi. .beta.-1,3-glucanases are capable of degrading them by fragmentation of the .beta.-1,3-glucan chains. Most known plant .beta.-1,3-glucanases are of the endo type.
It is known, moreover, that recent progress in so-called recombinant DNA technology and in the transformation of plant cells, as well as in the regeneration of whole plants from the latter, enable a gene of interest to be introduced into plant cells, a plant or a plant part so as to obtain an advantageous phenotype.
SUMMARY OF THE INVENTION
DNA sequences coding for several plant .beta.-1,3-glucanases, in particular a Nicotiana plumbaginifolia .beta.-1,3-glucanase (De Loose et al., 1988, Gene, 70,13-23) and a soybean .beta.-1,3-glucanase (Takeuchi et al., 1990, Plant Physiol., 93,673-682), have been isolated, cloned and determined. Patent Application EP-A-0,353,191 describes the isolation and cloning of different fragments of complementary DNA, the assembling of which enables the complementary DNA sequence coding for a tobacco .beta.-1,3-glucanase to be deduced, as well as of the genomic DNA sequence coding for this enzyme. The document EP-0,392,225 discloses, in particular, the construction of a chimeric gene coding for this tobacco .beta.-1,3-glucanase, the transformation of tobacco with the latter and the verification, by Western blot and visualisation using polyclonal antibodies, of the over-expression of this endogenous protein in the transformed plants. This patent application does not show that the recombinant tobacco .beta.-1,3-glucanase is biologically active, nor a fortiori that it confers a resistance of the said transformed plants to pathogenic agents.
It is known, moreover, that .beta.-1,3-glucanases such as, for example, Bacillus subtilis .beta.-1,3-glucanase are useful enzymes in the conversion of biomass, in particular in some sectors of the papermaking industry and in the agri-foodstuffs industry, especially brewing (Meaden, 1986, Brewers Guardian, 115, 7 and MacQueen, October 1987, New Scientist, 66).
It is known, lastly, that many recombinant proteins can be produced by eukaryotic cells and bacteria, following introduction into these of
REFERENCES:
Edington et al, "cDNA Cloning And Characterization of a Putative 1,3-.beta.-D-glucanase Transcript Induced By Fungal Elicitor in Bean Cell Suspension Cultures," Plant Molecular Biology, 16:81-94 (1991).
Takeuchi et al, "Molecular Cloning and Ethylene Induction of mRNA Encoding a Phytoalexin Elicotor-Releasing Factor, .beta.-1,3-Endoglucanase, in Soybean," Plant Physiol., 93:673-682, (1990).
Keen et al, ".beta.-1,3-Endoglucanase From Soybean Releases Elicotor-Active Carbohydrates From Fungus Cell Walls," Plant Physiol., 71:460-465, (1983).
Cline et al, "Host-Pathogen Interactions," Plant Physiol., 68:221-228, (1981).
De Loose et al, "Primary Structure of a Hormonally Regulated .beta.-Glucanase of Nicotiana Plumbaginifolia," Gene, 70:13-23, (1988).
Cline et al, "Host-Pathogen Interactions", Plant Physiol., 68:207-220, (1981).
M J Chrispeels (1991) Ann Rev Plant Physiol Plant Mol Biol 42:21-53.
I K Vasil (1990) Bio/technology 8:296-300.
J. Danecke et al (1990) Plant Cell 2:51-59.
Grison Rene
Leguay Jean-Jacques
Sass Catherine
Toppan Alain
Campell Bruce
Elf Sanofi
Fox David T.
Societe Nationale Elf Aquitaine
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