Method of increasing foreign protein expression

Details Method of increasing foreign protein expression Method of increasing foreign protein expression

1998-08-21

2002-06-18

McElwain, Elizabeth F. (Department: 1638)

Multicellular living organisms and unmodified parts thereof and

Plant, seedling, plant seed, or plant part, per se

Higher plant, seedling, plant seed, or plant part

C800S278000, C800S288000, C800S295000, C800S298000, C800S314000, C536S023710, C536S024100, C435S069100, C435S440000

Reexamination Certificate

active

06407316

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to a method of increasing foreign protein expression in plants, a method of controlling insects at a locus, a method of increasing the time during which an herbicide may be applied to a herbicide tolerant plant, a plant in which the expression of a foreign protein has been increased by the application of aldicarb and a method of growing crops.
BACKGROUND OF THE INVENTION
It is known in the field of plant biotechnology that heterologous genes encoding foreign proteins which impart herbicide tolerance, disease resistance or toxicity to insects (insect resistance), or improve the quality of the crops, can be incorporated into the genome of certain plants. These genetically transformed plants can express these proteins for the added protection of the plants, or higher quality of the crops. Genes coding for protein imparting herbicide tolerance are known in the art, including genes imparting tolerance to oxynil herbicides (U.S. Pat. Nos. 4,810,648 and 5,559,024), genes imparting tolerance to glyphosate and EPSPS inhibitor herbicides (U.S. Pat. Nos. 4,535,060, 4,769,061, 5,094,945, 4,940,835, 5,188,642, 4,971,908, 5,145,783, 5,312,910, 5,310,667, 5,633,435, 5,627,061, 5,554,798, 5,633,448, WO 97/04103), genes imparting tolerance to glufosinate (EP 242 236), as well as genes imparting tolerance to HPPD inhibitors (WO 96/38567 and WO 98/02562). Genes coding for proteins imparting disease resistance are known in the art, including lytic peptides, oxalate oxydase genes for tolerance to sclerotinia or chitinases. Genes imparting insect resistance are also known in the art, including genes which encode for
Bacillus thuringiensis
(Bt) endotoxins (Inter alia, U.S. Pat. Nos. 5,460,963, 5,683,691, 5,545,565, 5,530,197, 5,317,096) etc., or for insecticidal toxins isolated from Photorhabdus (WO 97/17432 or WO 98/08932). In general, these heterologous genes have been placed in a variety of plants including cotton.
It is also known that the levels of protein encoded by the heterologous genes in the transgenic plants can vary according to inter alia environmental conditions. For example, the grower of the said plants can be led to believe that the said plants are capable of tolerance to herbicide without being substantially damaged by the herbicide, or capable of resistance to diseases or withstanding attack by insect pests without further assistance from other pesticides. However, it is a severe problem for the grower if the said plants do not express the said protein at the correct time, and in an effective amount, particularly a pesticidally effective amount, more particularly an insecticidally effective amount in case of insect resistance. For example, in the case of Bt endotoxin, if there is not enough in the said plants, then the plants are susceptible to damage from pests, particularly lepidopteran pests, including bollworms. Cotton is such a plant that may be severely affected by bollworm attack. For another example, in the case of cotton seeds or plants comprising a gene encoding a Type II EPSPS, like Roundup Ready® Cotton, the window for application of EPSPS inhibitor herbicides is very strict and narrow (before the 4 leaves stage), and bolls may be damaged when the level of EPSPS expression is not sufficient to impart a proper level of tolerance to glyphosate, leading to an unacceptable loss of yield for the farmer.
Although it is known that aldicarb is a systemically acting insecticide, acaricide and nematicide and generally provide plants with a plant growth promoting effect, aldicarb is not generally effective against lepidopteran species when applied at the seed. Aldicarb is also known as 2-methyl-2-(methylthio)propionaldehyde O-(methylcarbamoyl)oxime.
SUMMARY OF THE INVENTION
An object of the present invention is to enhance the expression of heterologous proteins encoded by genes in plants. Another object of the present invention is to enhance the expression of
Bacillus thuringiensis
endotoxin in plants.
Another object of the present invention is to enhance the expression of proteins imparting herbicide tolerance in plants.
Another object of the present invention is to provide an improved method for the control of pests at a locus.
Another object of the present invention is to provide an improved method of application of an herbicide to a plant tolerant to the said herbicide.
Another object of the invention is to provide an improved plant protected from predacious insects.
Another object of the invention is to provide a plant with improved protection from herbivorous insects.
Another object of the invention is to provide an improved plant comprising a foreign gene which encodes for a foreign protein.
Another object of the invention is to provide an improved seed comprising a foreign gene which encodes for a foreign protein.
These objects are met in whole or in part by the present invention.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to a method of increasing foreign protein expression encoded by a foreign gene in plants comprising treating the plant or a seed from which it grows with aldicarb.
By the term “heterologous gene” or “foreign gene” is meant a gene that has been transformed or introduced into the plant, particularly integrated into the transformed plant genome, for example, transformed into the plant or an ancestor thereof, more particularly a gene that is not naturally present in the transformed plant, or even more particularly a gene which contains a sequence coding for a heterologous or foreign protein, including regulatory elements appropriate for controlling the expression of the said coding sequence in the plant cells, for example, promoters, terminators, pre-pro peptides, or transit peptides, the latter driving the expression of the said heterologous protein in a specific targeted region of the plant cell. Promoters controlling the expression of a gene in plant cells are well known in the field of plant biotechnology, including any promoter sequence of a gene naturally expressed in plants or plant cells, form plant, viral or bacterial origin. Suitable such promoters are disclosed in Weising & al (1988, Annual Rev. Genet., 22:241, the subject matter of which is incorporated herein by reference). The following is a partial representative list of promoters suitable for use herein: regulatory sequences from the T-DNA of
A. tumefaciens
, including mannopine synthase, nopaline synthase and octopine synthase; regulatory sequences from plant origin, including alcohol deshydrogenase promoter from corn, light inducible promoters such as ribulose-biscarboxylase/oxygenase small subunit promoters (SSU RuBisCO) from genes of a variety of species and the major chlorophyl a/b binding gene promoters, histone promoters (EP 507 698), actin promoters, maize ubiquitin 1 promoters (Christenses & al., 1996, Transgenic Res., 5:213); regulatory sequences from viral origins, such as 19S or 35S promoters of the cauliflower mosaic virus, (U.S. Pat. Nos. 5,352,605; 5,530,196); developmentally regulated promoters such as waxy, zein, or bronze promoters from maize ; as well as synthetic or other natural promoters which are either inducible or constitutive, including those promoters exhibiting organ specific expression or expression at specific development stage(s) of the plant, like the promoter of napin (EP 255 378) or the alpha-tubulin promoter (U.S. Pat. No. 5,635,618).
By the term “heterologous protein” or “foreign protein” is meant a protein of interest produced by the expression of the foreign gene in the plants. It may be aprotein naturally present in the transformed plant specie but expressed at levels or in specific plant tissues or targeted regions of the plant cells, not naturally occurring in the said transformed plant specie, or a protein not naturally present in the transformed plant specie.
The invention provides a method of increasing foreign gene expression in transgenic plants which method comprises treating the plant or a seed from which it grows with aldicarb.
The transgenic plants according t

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