Marker free transgenic plants engineering the chloroplast genome without the use of antibiotic selection

a technology of transgenic plants and chloroplasts, applied in the direction of oxidoreductases, biochemistry apparatus and processes, fermentation, etc., can solve the problems of reducing the european market of canadian organic farmers, bt resistance genes serve no useful purpose, and several lawsuits, so as to increase the risk of pests developing bt resistance, reduce bt resistance development, and increase bt expression levels

US20020137214A1Inactive Publication Date: 2002-09-26UNIV OF CENT FLORIDA +1
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Patent Information

Authority / Receiving Office
US · United States
Current Assignee / Owner
Publication Date
2002-09-26
Estimated Expiration
Not applicable · inactive patent

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Abstract

The present invention provides for a method to circumvent the problem of using antibiotic resistant selectable markers. In particular, target plants are transformed using a plastid vector which contains heterologous DNA sequences coding for a phytotoxin detoxifying enzyme or protein. The selection process involves converting an antibiotic-free phytotoxic agent by the expressed phytotoxin detoxifying enzyme or protein to yield a nontoxic compound. The invention provides for various methods to use antibiotic-free selection in chloroplast transformation.
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Description

CROSS-REFERENCES TO RELATED APPLICATIONS

[0001] This patent application claims the benefit of U.S. Provisional Application Nos. 60 / 208,763, filed Jun. 06, 2000, No. 60 / 257,406, filed Dec. 22, 2000 and No. 60 / 259,154, filed Dec. 28, 2000, Ser. No. 60 / 186,308, filed Mar. 02, 2000. All applications are hereby incorporated by reference.

[0003] This application pertains to the field of genetic engineering of plant plastid genomes, particularly chloroplasts, and to methods of and engineered plants without the use of antibiotics.

[0004] This application relates in particular to a method of selecting genetically engineered or transformed plants without the use of antibiotics as a selectable marker. The application also relates to a method of transforming plants to confer drought tolerance and to the transformed plants which are drought tolerant.DESCRIPTION OF THE RELATED ART

[0005] Publications

[0006] Various methods of selection of plants that employ antibiotic-free selectable marker, or non-an...

Examples

example 2

[0095] Other antibiotic-free phytotoxic agents include phytotoxic aldehydes such as acetaldehyde, formaldehyde, proprionaldehyde, and butyraldehyde; herbicides such as triazines and cyanamide, including those listed in Molecular Biotechnology by Glick and Pasternak, page 459, Table 18.4. Also useful is light selection.

example 3

[0096] Other Genes of interest may be isolated from other organisms such as Sugar Beet and E. Coli.

example 4

[0097] Other Promoters can be used to drive expression of the genes, including the psbA promoter, the accD promoter, the 16SrRNA promoter, and those listed in U.S. Pat. No. 5,693,507 and International Publication No. WO99 / 10513, both to Daniell.