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Increased stress tolerance and enhanced yield in plantsRelated Patent Categories: Multicellular Living Organisms And Unmodified Parts Thereof And Related Processes, Method Of Introducing A Polynucleotide Molecule Into Or Rearrangement Of Genetic Material Within A Plant Or Plant Part, The Polynucleotide Confers Resistance To Heat Or Cold (e.g., Chilling, Etc.)Increased stress tolerance and enhanced yield in plants description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20070180585, Increased stress tolerance and enhanced yield in plants. Brief Patent Description - Full Patent Description - Patent Application Claims
BACKGROUND [0002] The non-protein amino acid .beta.-alanine is a precursor of pantothenate (vitamin B5) in all plants; and the osmoprotectant .beta.-alanine betaine in most members of Plumbaginaceae (Hanson et. al., 1991). .beta.-Alanine betaine is a product of three sequential methylations of .beta.-alanine (Rathinasabapathi et. al., 2001). While beta-alanine itself can be an osmoprotectant, in certain plants it is methylated to a more effective osmoprotectant called beta-alanine betaine. [0003] Bacteria make beta-alanine by decarboxylating aspartic acid. In Escherichia coli, this reaction is catalyzed by the product of panD gene encoding L-aspartate-.alpha.-decarboxylase. Aspartate decarboxylation reaction is not known in plants. Plants appear to use a variety of other ways to synthesize beta-alanine. Thus, engineering strategies to increase .beta.-alanine pool in plants has potential applications for improving nutritional quality, yield and abiotic stress tolerance of crops. SUMMARY [0004] It is an object of the present invention to provide methods and compositions for increasing stress tolerance in plants. [0005] It is another object of the present invention to provide plants and plant cells which have increased stress resistance. [0006] It is a further object of the present invention is to increase biomass yield in plants. Preferably, enhanced biomass yield is accomplished by increasing leaf carbon dioxide concentration. Preferably still, enhanced growth and biomass is achieved by transforming plants with a polynucleotide molecule that decarboxylates aspartic acid. [0007] Another object of the present invention is to enhance production of pantothenate in select plants. [0008] The objects of the present invention, and others, may be accomplished with a method of increasing stress resistance in a plant, comprising expressing an aspartate decarboxylase in the plant. More preferred, an embodiment of the invention pertains to transforming plants with the panD gene of E. coli. (SEQ. ID NO: 1) [0009] The objects of the present invention may also be accomplished with a method of increasing stress resistance in a plant cell, comprising expressing an aspartate decarboxylase in the plant cell. [0010] The objects of the present invention may also be accomplished with a plant or a plant cell transformed with a nucleic acid, which encodes an aspartate decarboxylase. [0011] Thus, the present invention also provides a method of producing such a plant or plant cell, by transforming a plant or plant cell with the nucleic acid which encodes the aspartate decarboxylase. [0012] The present invention also provides an isolated and purified aspartate decarboxylase having the amino acid sequence of SEQ ID NO: 2. [0013] The present invention also provides a method of producing the aspartate decarboxylase described above, comprising culturing host cells which have been transformed with a nucleic acid encoding the aspartate decarboxylase under conditions in which the aspartate decarboxylase is expressed, and isolating the aspartate decarboxylase. [0014] In another embodiment, the present invention provides an isolated and purified enzyme having aspartate decarboxylase activity, wherein the amino acid sequence of the enzyme has a homology of from 70% to less than 100% to SEQ ID NO: 2. [0015] The present invention also provides a method of producing the enzyme described above, comprising culturing host cells, which have been transformed with a nucleic acid encoding the enzyme under conditions in which the enzyme is expressed, and isolating the enzyme. BRIEF DESCRIPTION OF THE DRAWINGS [0016] FIG. 1. Diagram showing the generation of .beta.-alanine through the .alpha.-decarboxylation of L-aspartic acid by the bacterial L-aspartate-.alpha.-decarboxylase (ADC). [0017] FIG. 2. ADC self-processing and assembly. Auto-cleavage between Gly24 and Ser25 amino acid residue generates pyruvoyl active group at the .alpha.-subunit. Three .alpha.-subunits and three .beta.-subunits assemble together with one unprocessed protein to form the active ADC. [0018] FIG. 3. PCR screening for eight pMON-panD putative transgenic plants and two pMON-R5 putative transgenic plants. Two positive control TOPO-panD vector and one negative control no DNA. [0019] FIG. 4. Southern blot analysis for transgenic tobacco lines. 10 .mu.g genomic DNA was digested with Hind III and hybridized with 32P-panD probe. Seven transgenic plants had a single gene insertion arrows indicate lines with single gene insertion pattern. [0020] FIG. 5. Tobacco transgenic lines with different levels of AAC(3)-III and panD transcription. The lower panel shows ethidium bromide stained gel with equal load (20 .mu.g total RNA each lane) [0021] FIG. 6. ADC induction. E. coli strain BL21-DE3 harboring pETB-panD vector was induced for 3 h with different IPTG concentrations showing over-expression of recombinant ADC at 16.8 KDa. [0022] FIG. 7. ADC purification. Left, 10-20% Tris-tricine gel for the DEAE-Sepharose collected fractions. Right, Western blot for the DEAE-sepharose fractions using anti-His-CooH antibody. The antibody recognized the unprocessed .pi.-peptide (16.8 KDa) and the .alpha.-subunit (14 KDa) of the ADC. Continue reading about Increased stress tolerance and enhanced yield in plants... 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