Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or...
Reissue Patent
1998-02-17
2001-07-17
McElwain, Elizabeth F. (Department: 1649)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
C800S320100, C800S300000, C435S320100, C435S252300, C435S468000, C435S418000, C435S419000, C536S024100, C536S023400, C536S023200, C536S023700
Reissue Patent
active
RE037287
ABSTRACT:
BACKGROUND OF THE INVENTION
The present invention relates to novel transit peptide DNA sequences, to novel chimeric genes and to their use in plants for conferring to them an increased tolerance to herbicides in general especially to those of the phosphonomethylglycine family. It also relates to the plant cells transformed by means of these genes, to the transformed plants regenerated from these cells as well as to the plants derived from crossbreedings using these transformed plants.
Glyphosate, sulfosate or fosametine are broad-spectrum systemic herbicides of the phosphonomethyl-glycine family. They act essentially as competitive inhibitors of 5-(enolpyruvyl)shikimate-3-phosphate synthase (EC 2.5.1.19) or EPSPS in relation to PEP (phosphoenolpyruvate). After their application to the plant, they are translocated inside the plant where they accumulate in the rapidly growing parts, in particular the caulinary and root apexes, causing the deterioration and even the destruction of sensitive plants.
Plastidial EPSPS, the main target of these products, is an enzyme of the aromatic amino acid biosynthesis pathway which is encoded by one or more nuclear genes and synthesised in the form of a cytoplasmic precursor and then imported into the plastids where it accumulates in its natural form.
The tolerance of plants to glyphosate and to products of the family is obtained by the stable introduction inside their genome of an EPSPS gene of plant or bacterial origin mutant or nonmutant with respect to the characteristics of the inhibition of the product of this gene by glyphosate. Given the mode of action of glyphosate and the degree of tolerance to glyphosate of the product of the genes used, it is useful to be able to express the product of translation of this gene so as to permit its substantial accumulation in plastids.
It is known, for example from American Patent U.S. Pat. No. 4,535,060, to confer to a plant a tolerance to a herbicide of the abovementioned type, in particular N-(phosphonomethyl)glycine or glyphosate, by introducing into the plant genome a gene encoding an EPSPS carrying at least one mutation making this enzyme more resistant to its competitive inhibitor (glyphosate), after localisation of the enzyme in the plastidial compartment. However, these techniques need to be improved in order to achieve greater reliability in the use of these plants under agronomic conditions.
SUMMARY OF THE INVENTION
In the present description, “plant” is understood as meaning any differentiated multicellular organism capable of photosynthesis and “plant cell” any cell derived from a plant and capable of forming undifferentiated tissues such as calluses or differentiated tissues such as embryos or plant sections, plants or seeds.
The subject of the present invention is the production of transformed plants having an increased tolerance to herbicides in general and especially to those of the phosphonomethylglycine family by regenerating cells transformed by means of novel chimeric genes comprising a gene for tolerance to these herbicides. The invention also relates to these novel chimeric genes, to the novel transit peptides which they contain as well as to the plants containing them which are made more tolerant by an accumulation of the mutant enzyme, in its mature form, in the plants.
More particularly, the subject of the invention is a chimeric gene for conferring to plants an increased tolerance to a herbicide whose target is EPSPS, comprising, in the direction of transcription, a promoter region, a transit peptide region, a sequence of a gene encoding a glyphosate tolerance enzyme and an untranslated polyadenylation signal region in 3′, wherein the transit peptide region comprises, in the direction of transcription, a transit peptide of a plant gene encoding a plastid-localised enzyme, a partial sequence of the N-terminal mature pan of a plant gene encoding a plastid-localised enzyme and then a second transit peptide of a plant gene encoding a plastid-localised enzyme.
The invention also relates to any DNA sequence of the transit peptide region defined above.
The transit peptides which can be used in the transit peptide region may be known per se and may be of plant origin, for example, derived from maize, sunflower, peas, tobacco or the like. The first and the second transit peptides may be identical, analogous or different. They may in addition each comprise one or more transit peptide units. A sequence derived from the SSU of the ribulose 1,5-diphosphate carboxylase oxygenase (RuBisCO) gene is preferably used.
The partial sequence of the N-terminal mature part is derived from a plant gene encoding a plastid-localised enzyme, such as for example a maize, sunflower or pea gene or the like, it being possible for the original plant species to be identical, analogous or different from that from which the first and second transit peptides are derived respectively. Furthermore, the partial sequence of the mature part may comprise a varying number of amino acids, generally from 10 to 40, preferably from 18 to 33. A sequence derived from the SSU of the ribulose 1,5-diphosphate carboxylase oxygenase (RuBisCO) gene is preferably used.
Construction of the entire transit region may be carded out in a manner known per se, in particular by fusion or any other suitable means. The role of this characteristic region is to enable the release of a mature, native protein with a maximum efficiency.
The coding sequence for herbicide tolerance which may be used in the chimeric gene according to the invention encodes a mutant EPSPS having a degree of glyphosate tolerance. This sequence, obtained in particular by mutation of the EPSPS gene, may be of bacterial origin, for example derived from
Salmonella typhymurium
(and called in the text which follows “AroA gene”), or of plant origin, for example from petunia or from tomatoes. This sequence may comprise one or more mutations, for example the Pro 101 to Ser mutation or alternatively the Gly 96 to Ala mutations.
The promoter region of the chimeric gene according to the invention may consist advantageously of at least one promoter on a fragment thereof of a gene which is expressed naturally in plants, that is to say promoters of vital origin such as that of 35S RNA of the cauliflower mosaic virus (CaMV35S) or of plant origin such as the small subunit of the ribulose 1,5-diphosphate carboxylase (RuBisCO) gene of a crop such as maize or sunflower.
The untranslated polyadenylation signal region in 3′ of the chimeric gene according to the invention may be of any origin, for example bacterial, such as the nopaline synthase gene, or of plant origin, such as the small subunit of the maize or sunflower RuBisCO.
The chimeric gene according to the invention may comprise, in addition to the above essential pans, an untranslated intermediate region (linker) between the promoter region and the coding sequence which may be of any origin, bacterial, vital or plant.
REFERENCES:
patent: 4535060 (1985-08-01), Comai
patent: 4801540 (1989-01-01), Hiatt et al.
patent: 4940835 (1990-07-01), Shah et al.
patent: 4962028 (1990-10-01), Bedbrook et al.
patent: 5073677 (1991-12-01), Heimer et al.
patent: 5188642 (1993-02-01), Shah et al.
patent: 5254799 (1993-10-01), De Greve et al.
patent: 5258300 (1993-11-01), Glassman et al.
patent: 5312910 (1994-05-01), Kishore et al.
patent: 5349123 (1994-09-01), Shewmaker
patent: 5463175 (1995-10-01), Barry et al.
patent: 5489520 (1996-02-01), Adams et al.
patent: 5498830 (1996-03-01), Barry et al.
patent: 5530188 (1996-06-01), Ausich et al.
patent: 5538878 (1996-07-01), Thomas et al.
patent: 5554798 (1996-09-01), Lundquist et al.
patent: 5559024 (1996-09-01), Leroux et al.
patent: 5597717 (1997-01-01), Guerineau et al.
patent: 5608149 (1997-03-01), Barry et al.
patent: 5633444 (1997-05-01), Guerineau et al.
patent: 0 189 707 B1 (1986-08-01), None
patent: 0 218 571 B1 (1987-04-01), None
patent: 0 337 899 (1989-10-01), None
patent: WO 88/02402 (1988-04-01), None
B. Reiss et al (1989), “Effect of Mutations on the Binding and Translocatio
Lebrun Michel
Leroux Bernard
Sailland Alain
Aventis Cropscience S.A.
Connolly Bove & Lodge & Hutz LLP
McElwain Elizabeth F.
LandOfFree
Chimeric gene for the transformation of plants does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Chimeric gene for the transformation of plants, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Chimeric gene for the transformation of plants will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2449290