AU2009287446A1 - Transgenic plants with enhanced growth characteristics - Google Patents
Transgenic plants with enhanced growth characteristics Download PDFInfo
- Publication number
- AU2009287446A1 AU2009287446A1 AU2009287446A AU2009287446A AU2009287446A1 AU 2009287446 A1 AU2009287446 A1 AU 2009287446A1 AU 2009287446 A AU2009287446 A AU 2009287446A AU 2009287446 A AU2009287446 A AU 2009287446A AU 2009287446 A1 AU2009287446 A1 AU 2009287446A1
- Authority
- AU
- Australia
- Prior art keywords
- plant
- transgene
- gpt
- seq
- transgenic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1096—Transferases (2.) transferring nitrogenous groups (2.6)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
- Enzymes And Modification Thereof (AREA)
Description
WO 2010/025466 PCT/US2009/055557 TRANSGENIC PLANTS WITH ENHANCED GROWTH CHARACTERISTICS STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER 5 FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under Contract No. W-7405 ENG-36 awarded by the United States Department of Energy to The Regents of The University of California, and Contract No. DE-AC52-06NA25396, awarded by the United States Department of Energy to Los Alamos National Security, LLC. 10 The government has certain rights in this invention. RELATED APPLICATIONS This application claims priority to United States Provisional Application No. 61/190,520 filed August 29, 2008. 15 BACKGROUND OF THE INVENTION As the human population increases worldwide, and available farmland continues to be destroyed or otherwise compromised, the need for more effective and sustainable agriculture systems is of paramount interest to the human race. 20 Improving crop yields, protein content, and plant growth rates represent major objectives in the development of agriculture systems that can more effectively respond to the challenges presented. In recent years, the importance of improved crop production technologies has only 25 increased as yields for many well-developed crops have tended to plateau. Many agricultural activities are time sensitive, with costs and returns being dependent upon rapid turnover of crops or upon time to market. Therefore, rapid plant growth is an economically important goal for many agricultural businesses that involve high-value crops such as grains, vegetables, berries and other fruits, 30 Genetic engineering has and continues to play an increasingly important yet controversial role in the development of sustainable agriculture technologies. A large number of genetically modified plants and related technologies have been developed in recent years, many of which are in widespread use today WO 2010/025466 PCT/US2009/055557 (Factsheet: Genetically Modified Crops in the United States, Pew Initiative on Food and Biotechnology, August 2004, (pewagbiotechorg/resources/factsheets). The adoption of transgenic plant varieties is now very substantial and is on the rise, with approximately 250 million acres planted with transgenic plants in 2006. 5 While acceptance of transgenic plant technologies may be gradually increasing, particularly in the United States, Canada and Australia, many regions of the World remain slow to adopt genetically modified plants in agriculture, notably Europe, Therefore, consonant with pursuing the objectives of responsible and sustainable 10 agriculture, there is a strong interest in the development of genetically engineered plants that do not introduce toxins or other potentially problematic substances into plants and/or the environment. There is also a strong interest in minimizing the cost of achieving objectives such as improving herbicide tolerance, pest and disease resistance, and overall crop yields. Accordingly, there remains a need for 15 transgenic plants that can meet these objectives. The goal of rapid plant growth has been pursued through numerous studies of various plant regulatory systems, many of which remain incompletely understood. In particular, the plant regulatory mechanisms that coordinate carbon and nitrogen 20 metabolism are not fully elucidated. These regulatory mechanisms are presumed to have a fundamental impact on plant growth and development. The metabolism of carbon and nitrogen in photosynthetic organisms must be regulated in a coordinated manner to assure efficient use of plant resources and 25 energy. Current understanding of carbon and nitrogen metabolism includes details of certain steps and metabolic pathways which are subsystems of larger systems. In photosynthetic organisms, carbon metabolism begins with C02 fixation, which proceeds via two major processes, termed C-3 and C~ metabolism. In plants with C~3 metabolism, the enzyme ribulose bisphosphate carboxylase (RuBisCo) 30 catalyzes the combination of C02 with ribulose bisphosphate to produce 3 phosphoglycerate, a three carbon compound (C-3) that the plant uses to synthesize carbon-containing compounds, In plants with C-4 metabolism, C02 is combined with phosphoenol pyruvate to form acids containing four carbons (C-4), 2 WO 2010/025466 PCT/US2009/055557 in a reaction catalyzed by the enzyme phosphoenol pyruvate carboxylase, The acids are transferred to bundle sheath cells, where they are decarboxylated to release C02, which is then combined with ribulose bisphosphate in the same reaction employed by C-3 plants. Numerous studies have found that various metabolites are important in plant regulation of nitrogen metabolism. These compounds include the organic acid palate and the amino acids glutamate and glutamine, Nitrogen is assimilated by photosynthetic organisms via the action of the enzyme glutamine synthetase (GS) 10 which catalyzes the combination of ammonia with glutamate to form glutamine. GS plays a key role in the assimilation of nitrogen in plants by catalyzing the addition of ammonium to glutamate to form glutamine in an ATP-dependent reaction (Miflin and Habash, 2002 Journal of Experimental Botany, Vol. 53, No, 370, pp. 979-987), GS also reassimilates ammonia released as a result of 15 photorespiration and the breakdown of proteins and nitrogen transport compounds. GS enzymes may be divided into two genera classes, one representing the cytoplasmic form (GS1) and the other representing the plastidic (i.e. chloroplastic) form (GS2). 20 Previous work has demonstrated that increased expression levels of GS1 result in increased levels of GS activity and plant growth, although reports are inconsistent. For example, Fuentes et al, reported that CaMV $35 promoter driven overexpression of Alfalfa GS1 (cytoplasmic form) in tobacco resulted in increased levels of GS expression and GS activity in leaf tissue, increased growth under 25 nitrogen starvation, but no effect on growth under optimal nitrogen fertilization conditions (Fuentes et aL, 2001, J. Exp. 'Botany 52: 1071-81), Temple et al. reported that transgenic tobacco plants overexpressing the full length Alfalfa GS1 coding sequence contained greatly &evated levels of GS transcript, and GS polypeptide which assembled into active enzyme, but did not report phenotypic 30 effects on growth (Temple et al., 1993, Molecular and General Genetics 236: 315 325). Corruzi et al. have reported that transgenic tobacco overexpressing a pea cytosolic GS1 transgene under the control of the CaMV S35 promoter show increased GS activity, increased cytosolic GS protein, and improved growth 3 WO 2010/025466 PCT/US2009/055557 characteristics (U.S. Patent No. 6,107,547). Unkefer et at. have more recently reported that transgenic tobacco plants overexpressing the Alfalfa GS1 in foliar tissues, which had been screened for increased leaf-to-root GS activity following genetic segregation by selling to achieve increased GS1 transgene copy number, 5 were found to produce increased 2-hydroxy-5-oxoproline levels in their foliar portions, which was found to lead to markedly increased growth rates over wildtype tobacco plants (see, U.S, Patent Nos. 6,555,500; 6,593,275; and 6,831,040). 10 Unkefer et a. have further described the use of 2-hydroxy-5-oxoproline (also known as 2-oxoglutaramate) to improve plant growth (U,S. Patent Nos, 6,555,500; 6,593,275; 6,831,040), In particular, Unkefer et al, disclose that increased concentrations of 2-hydroxy-5-oxoproline in foliar tissues (relative to root tissues) triggers a cascade of events that result in increased plant growth characteristics, 15 Unkefer et alt describe methods by which the foliar concentration of 2-hydroxy-5 oxoproline may be increased in order to trigger increased plant growth characteristics, specifically, by applying a solution of 2-hydroxy-5-oxoproline directly to the foliar portions of the plant and over-expressing glutamine synthetase preferentially in leaf tissues. 20 A number of transaminase and hydrolyase enzymes known to be involved in the synthesis of 2-hydroxy-5-oxoproline in animals have been identified in animal liver and kidney tissues (Cooper and Meister, 1977, CRC Critical Reviews in Biochemistry, pages 281-303; Meister, 1952, J, Biochem. 197: 304), In plants, 25 the biochemical synthesis of 2-hydroxy-5-oxoproline has been known but has been poorly characterized. Moreover, the function of 2-hydroxy-5-oxoproline in plants and the significance of its pool size (tissue concentration) are unknown. Finally, the art provides no specific guidance as to precisely what transaminase(s) or hydrolase(s) may exist and/or be active in catalyzing the synthesis of 2 30 hydroxy-5-oxoproline in plants, and no such plant transaminases have been reported, isolated or characterized. 4 WO 2010/025466 PCT/US2009/055557 SUMMARY OF THE INVENTION The invention relates to transgenic plants exhibiting dramatically enhanced growth rates, greater seed and fruit/pod yields, earlier and more productive flowering, 5 more efficient nitrogen utilization, increased tolerance to high salt conditions, and increased biomass yields. In one embodiment, transgenic plants engineered to over-express both glutamine phenylpyruvate transaminase (GPT) and glutamine synthetase (GS) are provided. The GPT+GS double-transgenic plants of the invention consistently exhibit enhanced growth characteristics, with TO generation 10 lines showing an increase in biomass over wild type counterparts of between 50% and 300%, Generations that result from sexual crosses and/or selfing typically perform even better, with some of the double-transgenic plants achieving an astounding four-fold biomass increase over wild type plants. Similarly, flower and fruit or pod yields are also tremendously improved, with TO generation lines 15 typically showing 50% to 70% increases over their wild type counterparts, and in some cases showing a 100% increase. Transgenic plants exhibiting such enhanced growth phenotypic characteristics have been successfully generated across a spectrum of individual plant species, using various transformation methodologies, different expression vectors and promoters, and heterologous and 20 homologous transgene sequences from a variety of species, as exemplified by the numerous working examples provided herein. This invention, therefore, provides a fundamental break-though technology that has the potential to transform virtually all areas of agriculture. 25 Applicants have identified the enzyme glutamine phenylpyruvate transaminase (GPT) as a catalyst of 2-hydroxy-5-oxoproline (2-oxoglutaramate) synthesis in plants. 2-oxoglutaramate is a powerful signal metabolite which regulates the function of a large number of genes involved in the photosynthesis apparatus, carbon fixation and nitrogen metabolism, The invention provides isolated nucleic 30 acid molecules encoding GPT, and discloses the novel finding that the encoded enzyme is directly involved in the synthesis of 2-hydroxy-5-oxoproline. This aspect of the invention is exemplified herein by the disclosure of GPT polynucleotides encoding GPTs from several species, including Arabidopsis, 5 WO 2010/025466 PCT/US2009/055557 Grape, Rice, Soybean, Barley, Bamboo and a non-plant homolog from Zebra fish, most of which have been expressed as recombinant GPTs and confirmed as having GPT activity, 5 The invention further provides transgenic plants which express both a GPT transgene and a GS transgene. The expression of these two transgenes in such "double-transgene" plants results in a substantially increased rate of carbon dioxide fixation and an extremely potent growth enhancing effect, as these plants exhibit very significantly and sometimes tremendously enhanced growth rates and 10 flower/fruit/pod/seed yields. Methods for the generation of such growth-enhanced transgenic plants are provided. By preferentially increasing the concentration of the signal metabolite 2 oxoglutaramate (i.e., in foliar tissues), the transgenic plants of the invention are 15 capable of producing higher overall yields over shorter periods of time, and therefore may provide agricultural industries with enhanced productivity across a wide range of crops Importantly, unlike many transgenic plants described to date, the invention utilizes natural plant genes encoding a natural plant enzyme. The enhanced growth characteristics of the transgenic plants of the invention is 20 achieved essentially by introducing additional GPT and GS capacity into the plant. Thus, the transgenic plants of the invention do not express any toxic substances, growth hormones, viral or bacterial gene products, and are therefore free of many of the concerns that have heretofore impeded the adoption of transgenic plants in certain parts of the World. 25 In one embodiment, the invention provides a transgenic plant comprising a GPT transgene and a GS transgene, wherein said GPT transgene and said GS transgene are operably linked to a plant promoter. In a specific embodiment, the GS transgene is a GS1 transgene. In another specific embodiment, the GPT 30 transgene encodes a polypeptide having an amino acid sequence selected from the group consisting of (a) SEQ ID NO: 2; SEQ ID NO: 9; SEQ ID NO: 15, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO 24, SEQ ID NO: 30, SEQ ID NO:31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ D NO: 35 and SEQ ID NO: 36, 6 WO 2010/025466 PCT/US2009/055557 and (b) an amino acid sequence that is at least 75% identical to any one of SEQ ID NO: 2; SEQ ID NO: 9; SEQ ID NO: 15, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO 24, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36 and has GPT activity. In yet 5 another specific embodiment, the GS transgene encodes a polypeptide having an amino acid sequence selected form the group consisting of (a) SEQ ID NO: 4 and SEQ ID NO. 7 from residue 11, and (b) an amino acid sequence that is at least 75% identical to SEQ ID NO: 4 or SEQ ID NO: 7. In some embodiments, the GPT and GS transgenes are incorporated into the genome of the plant. The transgenic 10 plant of the invention may be a monocotyledonous or a dicotyledonous plant. The invention also provides progeny of any generation of the transgenic plants of the invention, wherein said progeny comprises a GPT transgene and a GS transgene, as well as a seed of any generation of the transgenic plants of the 15 invention, wherein said seed comprises said GPT transgene and said GS transgene. The transgenic plants of the invention may display one or more enhanced growth characteristics rate when compared to an analogous wild-type or untransformed plant, including without limitation increased growth rate, biomass yield, seed yield, flower or flower bud yield, fruit or pod yield, larger 20 leaves, and may also display increased levels of GPT and/or GS activity, and/or increased levels of 2-oxoglutaramate, In some embodiments, the transgenic plants of the invention display increased nitrogen use efficiency or increased tolerance to salt or saline conditions. 25 Methods for producing the transgenic plants of the invention and seeds thereof are also provided, including methods for producing a plant having enhanced growth properties, increased nitrogen use efficiency and increased tolerance to germination or growth in salt or saline conditions, relative to an analogous wild type or untransformed plant. 30 7 WO 2010/025466 PCT/US2009/055557 BRIEF DESCRIPTION OF THE DRAWINGS The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. FIG. 1, Nitrogen assimilation and 2-oxoglutaramate biosynthesis: schematic of metabolic pathway. FIG. 2. Photograph showing comparison of transgenic tobacco plants over 10 expressing either GS1 or GPT, compared to wild type tobacco plant. From left to right: wild type plant, Alfalfa GS1 transgene, Arabidopsis GPT transgene. See Examples 3 and 5, infra, FIG. 3. Photograph showing comparison of transgenic Micro-Tom tomato plants 15 over-expressing either GSi or GPT, compared to wild type tomato plant. From left to right wild type plant, Alfalfa GS1 transgene, Arabidopsis GPT transgene. See Examples 4 and 6, infra. FIG. 4. Photographs showing comparisons of leaf sizes between wild type and 20 GS1 or GPT transgenic tobacco plants. A: Comparison between leaves from GS1 transgenic tobacco (bottom leaf) and wild type (top leaf), B: Comparison between leaves from GPT transgenic tobacco (bottom leaf) and wild type (top leaf). 25 FIG. 5. Photographs showing comparisons of transgenic tobacco plants generated from various crosses between GS1 and GPT transgenic tobacco lines with wild type and single transgene plants. A-C: Cross 2, 3 and 7, respectively. See Example 7, infra; 30 FIG. 6. Photographs showing comparisons of leaf sizes between wild type and crosses between GS1 and GPT transgenic tobacco plants. A: Comparison between leaves from GSXGPT Cross 3 (bottom leaf) and wild type (top leaf). B: Comparison between leaves from GSXGPT Cross 7 (bottom leaf) and wild type (top leaf). See Example 7, infra. 8 WO 2010/025466 PCT/US2009/055557 FIG, 7. Photograph of transgenic pepper plant (right) and wild type control pepper plant (left), showing larger pepper fruit yield in the transgenic plant relative to the wild type control plant. See Example 8, infra. FIG. 8. Transgenic bean plants compared to wild type control bean plants (several transgenic lines expressing Arabidopsis GPT and GS transgenes), Upper Left: plant heights on various days; Upper right: flower bud numbers; Lower left flower numbers; Lower right: bean pod numbers, Wildtype is the control, and 10 lines 2A, 4A and 5B are all transgenic plant lines. See Example 9, infra. FIG. 9. Photograph of transgenic bean plant (right) and wild type control bean plant (left), showing increased growth in the transgenic plant relative to the wild type control plant. Transgenic line expressing Arabidopsis GPT and GS 15 transgenes. See Example 9, infra, FIG. 10. Transgenic bean plants pods, flowers and flower buds compared to wild type control bean plants (transgenic line expressing grape GPT and Arabidopsis GS transgenes). See Example 10, infra, 20 FIG. 11, Photograph of transgenic bean plant (right) and wild type control bean plant (left), showing increased growth in the transgenic plant relative to the wild type control plant. Transgenic line expressing Grape GPT and Arabidopsis GS transgenes. See Example 10, infra, 25 FIG. 12. Transgenic Cowpea Line A plants compared to wild type control Cowpea plants (transgenic line expressing Arabidopsis GPT and GS transgenes), showing that the transgenic plants grow faster and flower and set pods sooner than wild type control plants. (A) Relative height and longest leaf measurements as of May 30 21, (B) Relative trifolate leafs and flower buds as of June 18, (C) Relative numbers of flowers, flower buds and pea pods as of June 22. See Example 11, infra, 9 WO 2010/025466 PCT/US2009/055557 FIG, 13. Photograph of transgenic Cowpea Line A plant (right) and wild type control Cowpea plant (left), showing increased growth in the transgenic plant relative to the wild type control plant. Transgenic line expressing Arabidopsis GPT and GS transgenes. See Example 11, infra. FIG. 14, Transgenic Cowpea Line G plants compared to wild type control Cowpea plants (transgenic line expressing Grape GPT and Arabidopsis GS transgenes), showing that the transgenic plants grow faster and flower and set pods sooner than wild type control plants, (A) plant heights, (B) flowers and pea pod numbers, 10 (C) leaf bud and trifolate numbers. See Example 12. infra. FIG. 15, Photograph of transgenic Cowpea Line G plant (right) and wild type control Cowpea plant (left), showing increased growth in the transgenic plant relative to the wild type control plant. Transgenic line expressing Grape GPT and 15 Arabidopsis GS transgenes. See Example 12, infra. FIG. 16. Photograph of transgenic Cantaloupe plant (right) and wild type control Cantaloupe plant (left), showing increased growth in the transgenic plant relative to the wild type control plant. Transgenic line expressing Arabidopsis GPT and 20 GS transgenes. See Example 14, infra. FIG. 17. Photograph of transgenic Pumpkin plants (right) and wild type control Pumpkin plants (left), showing increased growth in the transgenic plants relative to the wild type control plants. Transgenic lines expressing Arabidopsis GPT and 25 GS transgenes. See Example 15, infra. FIG. 18. Photograph of transgenic Arabidopsis plants (right) and wild type control Arabidopsis plants (left), showing increased growth in the transgenic plants relative to the wild type control plants. Transgenic lines expressing Arabidopsis 30 GPT and GS transgenes. See Example 16, infra. FIG. 19, Transgenic tomato plants expressing Arabidopsis GPT and GS transgenes compared to control tomato plants. (A) Photograph of transgenic 10 WO 2010/025466 PCT/US2009/055557 tomato plant leaves (right) vs. wild type control leaves (left) showing larger leaves in the transgenic plant, (B) Photograph of transgenic tomato plants (right) and wild type control plants (left), showing increased growth in the transgenic plants relative to the wild type control plants. See Example 17, infra. FIG. 20. Photograph of transgenic Camelina plant (right) and wild type control Camelina plant (left), showing increased growth in the transgenic plant relative to the wild type control plant. Transgenic line expressing Arabidopsis GPT and GS transgenes. See Example 18, infra, 10 DETAILED DESCRIPTION OF THE INVENTION DEFINITIONS 15 Unless otherwise defined, all terms of art, notations and other scientific terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not 20 necessarily be construed to represent a substantial difference over what is generally understood in the art. The techniques and procedures described or referenced herein are generally well understood and commonly employed using conventional methodology by those skilled in the art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al, 25 Molecular Cloning: A Laboratory Manual 3rd. edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Current Protocols in Molecular Biology (Ausbel et al., eds., John Wiley & Sons, Inc 2001; Transgenic Plants: Methods and Protocols (Leandro Pena, ed, Humana Press, 1s edition, 2004); and, Agrobacterium Protocols (Wan, ed., Humana Press, 2 edition, 2006). As 30 appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer defined protocols and/or parameters unless otherwise noted, The term "nucleic acid" refers to deoxyribonucleotides or ribonucleotides and 35 polymers thereof ("polynucleotides") in either single- or double-stranded form. 11 WO 2010/025466 PCT/US2009/055557 Unless specifically limited, the term "polynucleotide" encompasses nucleic acids containing known analogues of natural nucleotides which have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic 5 acid sequence also implicitly encompasses conservatively modified variants thereof (e.g. degenerate codon substitutions) and complementary sequences and as well as the sequence explicitly indicated, Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or 10 deoxyinosine residues (Batzer et al, 1991, Nucleic Acid Res. 19: 5081; Ohtsuka et al., 1985 J, Biol. Chem. 260: 2605-2608; and Cassol et al., 1992; Rossolini et al., 1994, Molt Cell. Probes 8: 91-98), The term nucleic acid is used interchangeably with gene, cDNA, and mRNA encoded by a gene. 15 The term "promoter" refers to an array of nucleic acid control sequences that direct transcription of an operably linked nucleic acid. As used herein, a "plant promoter" is a promoter that functions in plants. Promoters include necessary nucleic acid sequences near the start site of transcription, such as, in the case of a polymerase i type promoter, a TATA element. A promoter also optionally 20 includes distal enhancer or repressor elements, which can be located as much as several thousand base pairs from the start site of transcription. A "constitutive" promoter is a promoter that is active under most environmental and developmental conditions. An induciblee" promoter is a promoter that is active under environmental or developmental regulation. The term "operably linked" 25 refers to a functional linkage between a nucleic acid expression control sequence (such as a promoter, or array of transcription factor binding sites) and a second nucleic acid sequence, wherein the expression control sequence directs transcription of the nucleic acid corresponding to the second sequence. 30 The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic 12 WO 2010/025466 PCT/US2009/055557 of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers, The term "amino acid" refers to naturally occurring and synthetic amino acids, as 5 well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, eg., hydroxyproline, y-carboxyglutamate, and 0-phosphoserine. Amino acid analogs refers to compounds that have the same basic chemical structure as 10 a naturally occurring amino acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid 15 mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid. Amino acids may be referred to herein by either their commonly known three letter 20 symbols or by the one-letter symbols recommended by the IUPAC-lUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes. The term "plant" includes whole plants, plant organs (e.g., leaves, stems, flowers, 25 roots, etc.), seeds and plant cells and progeny thereof, The class of plants which can be used in the method of the invention is generally as broad as the class of higher plants amenable to transformation techniques, including angiosperms (monocotyledonous and dicotyledonous plants), as well as gymnosperms. It includes plants of a variety of ploidy levels, including polyploid, diploid, haploid 30 and hemizygous. The terms "GPT polynucleotide" and "GPT nucleic acid" are used interchangeably herein, and refer to a full length or partial length polynucleotide sequence of a 13 WO 2010/025466 PCT/US2009/055557 gene which encodes a polypeptide involved in catalyzing the synthesis of 2 oxoglutaramate, and includes polynucleotides containing both translated (coding) and un-translated sequences, as well as the complements thereof. The term "GPT coding sequence" refers to the part of the gene which is transcribed and 5 encodes a GPT protein, The term "targeting sequence" refers to the amino terminal part of a protein which directs the protein into a subcellular compartment of a cell, such as a chloroplast in a plant cell. GPT polynucleotides are further defined by their ability to hybridize under defined conditions to the GPT polynucleotides specifically disclosed herein, or to PCR products derived 10 therefrom, A "GPT transgene" is a nucleic acid molecule comprising a GPT polynucleotide which is exogenous to transgenic plant, or plant embryo, organ or seed, harboring the nucleic acid molecule, or which is exogenous to an ancestor plant, or plant 15 embryo, organ or seed thereof, of a transgenic plant harboring the GPT polynucleotide. The terms "GS polynucleotide" and "GS nucleic acid" are used interchangeably herein, and refer to a full length or partial length polynucleotide sequence of a 20 gene which encodes a glutamine synthetase protein, and includes polynucleotides containing both translated (coding) and un-translated sequences, as well as the complements thereof, The term "GS coding sequence" refers to the part of the gene which is transcribed and encodes a GS protein. The terms "GS1 polynucleotide" and "GS1 nucleic acid" are used interchangeably herein, and refer 25 to a full length or partial length polynucleotide sequence of a gene which encodes a glutamine synthetase isoform 1 protein, and includes polynucleotides containing both translated (coding) and un-translated sequences, as well as the complements thereof. The term "GS1 coding sequence" refers to the part of the gene which is transcribed and encodes a GS1 protein. 30 A "GS transgene" is a nucleic acid molecule comprising a GS polynucleotide which is exogenous to transgenic plant, or plant embryo, organ or seed, harboring the nucleic acid molecule, or which is exogenous to an ancestor plant, or plant 14 WO 2010/025466 PCT/US2009/055557 embryo, organ or seed thereof, of a transgenic plant harboring the GPT polynucleotide. A "GS1 transgene" is a nucleic acid molecule comprising a GS1 polynucleotide which is exogenous to transgenic plant, or plant embryo, organ or seed, harboring the nucleic acid molecule, or which is exogenous to an ancestor 5 plant, or plant embryo, organ or seed thereof, of a transgenic plant harboring the GPT polynucleotide. Exemplary GPT polynucleotides of the invention are presented herein, and include GPT coding sequences for Arabidopsis, Rice, Barley, Bamboo, Soybean, 10 Grape, and Zebra Fish GPTs. Partial length GPT polynucleotides include polynucleotide sequences encoding N or C-terminal truncations of GPT, mature GPT (without targeting sequence) as well as sequences encoding domains of GPT, Exemplary GPT polynucleotides 15 encoding N-terminal truncations of GPT include Arabidopsis -30, -45 and -56 constructs, in which coding sequences for the first 30, 45, and 56 respectively, amino acids of the full length GPT structure of SEQ ID NO: 2 are eliminated. In employing the GPT polynucleotides of the invention in the generation of 20 transformed cells and transgenic plants, one of skill will recognize that the inserted polynucleotide sequence need not be identical, but may be only "substantially identical" to a sequence of the gene from which it was derived, as further defined below, The term "GPT polynucleotide" specifically encompasses such substantially identical variants. Similarly, one of skill will recognize that 25 because of codon degeneracy, a number of polynucleotide sequences will encode the same polypeptide, and all such polynucleotide sequences are meant to be included in the term GPT polynucleotide. In addition, the term specifically includes those sequences substantially identical (determined as described below) with an GPT polynucleotide sequence disclosed herein and that encode 30 polypeptides that are either mutants of wild type GPT polypeptides or retain the function of the GPT polypeptide (e.g., resulting from conservative substitutions of amino acids in a GPT polypeptide). The term "GPT polynucleotide" therefore also includes such substantially identical variants, 15 WO 2010/025466 PCT/US2009/055557 The term "conservatively modified variants" applies to both amino acid and nucleic acid sequences. With respect to particular nucleic acid sequences, conservatively modified variants refers to those nucleic acids which encode identical or 5 essentially identical amino acid sequences, or where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. Because of the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine Thus, at every position where 10 an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are 'silent variations," which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic 15 acid, One of skill will recognize that each codon in a nucleic acid (except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan) can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence. 20 As to amino acid sequences, one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a 'conservatively modified 25 variantf where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles of the invention. 30 The following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 16 WO 2010/025466 PCT/US2009/055557 5) Isoleucine (1), Leucine (L), Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C), Methionine (M) (see, eg., Creighton, Proteins (1984)). 5 Macromolecular structures such as polypeptide structures can be described in terms of various levels of organization. For a general discussion of this organization, see, e.g.. Alberts et a.., Aolecular Biology of the Cell (3d ed., 1994) and Cantor and Schimmel, Biophysical Chemistry Part I: The Conformation of Biological Macromolecules (1980). "Primary structure" refers to the amino acid 10 sequence of a particular peptide. "Secondary structure" refers to locally ordered, three dimensional structures within a polypeptide. These structures are commonly known as domains. Domains are portions of a polypeptide that form a compact unit of the polypeptide and are typically 25 to approximately 500 amino acids long. Typical domains are made up of sections of lesser organization such 15 as stretches of p-sheet and cehelices. "Tertiary structure" refers to the complete three dimensional structure of a polypeptide monomer, "Quatemary structure" refers to the three dimensional structure formed by the noncovalent association of independent tertiary units. Anisotropic terms are also known as energy terms. 20 The term "isolated" refers to material which is substantially or essentially free from components which normally accompany the material as it is found in its native or natural state. However, the term "isolated" is not intended refer to the components present in an electrophoretic gel or other separation medium. An isolated component is free from such separation media and in a form ready for 25 use in another application or already in use in the new application/milieu. An "isolated" antibody is one that has been identified and separated and/or recovered from a component of its natural environment, Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and 30 other proteinaceous or non-proteinaceous solutes. In preferred embodiments, the antibody will be purified (1) to greater than 95% by weight of antibody as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal 17 WO 2010/025466 PCT/US2009/055557 amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain. Isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody's natural 5 environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step. The term "heterologous" when used with reference to portions of a nucleic acid indicates that the nucleic acid comprises two or more subsequences that are not 10 found in the same relationship to each other in nature. For instance, a nucleic acid is typically recombinantly produced, having two or more sequences from unrelated genes arranged to make a new functional nucleic acid, e.g., a nucleic acid encoding a protein from one source and a nucleic acid encoding a peptide sequence from another source. Similarly, a heterologous protein indicates that 15 the protein comprises two or more subsequences that are not found in the same relationship to each other in nature (e.g, a fusion protein). The terms "identical' or percent "identity, in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences 20 that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (iLe., about 70% identity, preferably 75%, 80%, 85%, 90%, or 95% identity over a specified region, when compared and aligned for maximum correspondence over a comparison window, or designated region as measured using a sequence comparison algorithms, or by manual alignment and 25 visual inspection. This definition also refers to the complement of a test sequence, which has substantial sequence or subsequence complementarity when the test sequence has substantial identity to a reference sequence. This definition also refers to the complement of a test sequence, which has substantial sequence or subsequence complementarity when the test sequence has substantial identity to 30 a reference sequence. When percentage of sequence identity is used in reference to polypeptides, it is recognized that residue positions that are not identical often differ by conservative 18 WO 2010/025466 PCT/US2009/055557 amino acid substitutions, where amino acids residues are substituted for other amino acid residues with similar chemical properties (e.g., charge or hydrophobicity) and therefore do not change the functional properties of the polypeptide. Where sequences differ in conservative substitutions, the percent 5 sequence identity may be adjusted upwards to correct for the conservative nature of the substitution, For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared, When using a sequence comparison 10 algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences 15 relative to the reference sequence, based on the program parameters. A "comparison window, as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of from 20 to 600, usually about 50 to about 200, more usually about 100 to about 20 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are weIl-known in the art, Optimal alignment of sequences for comparison can be conducted, e.g, by the local homology algorithm of Smith & Waterman, 1981, Adv. AppL. Math. 2:482, by 25 the homology alignment algorithm of Needleman & Wunsch, 1970, J. Mol. Biol. 48:443, by the search for similarity method of Pearson & Lipman, 1988, Proc. Nat'l Acad. Sci. USA 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or 30 by manual alignment and visual inspection (see, e,g., Current Protocols in Molecular Biology (Ausubel et al., eds. 1995 supplement)). 19 WO 2010/025466 PCT/US2009/055557 A preferred example of algorithm that is suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., 1977, Nuc. Acids Res. 25:3389 3402 and Altschul et al, 1990, J. Molt Biol. 215:403-410, respectively. BLAST 5 and BLAST 2.0 are used, typically with the default parameters described herein, to determine percent sequence identity for the nucleic acids and proteins of the invention. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information, This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of 10 length W in the query sequence, which either match or satisfy some positive valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al, supra), These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are 15 extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative 20 score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity 25 and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, an expectation (E) of 10, M=5, N=-4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a word length of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. NatL. Acad. Sc. USA 30 89:10915 (1989)) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands. 20 WO 2010/025466 PCT/US2009/055557 The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, 1993, Proc, Nat'l. Acad. Sci, USA 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by 5 which a match between two nucleotide or amino acid sequences would occur by chance, For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0,2, more preferably less than about 0.01, and most preferably less than about 0.001, 10 The phrase "stringent hybridization conditions" refers to conditions under which a probe will hybridize to its target subsequence, typically in a complex mixture of nucleic acid, but to no other sequences. Stringent conditions are sequence dependent and will be different in different circumstances, Longer sequences hybridize specifically at higher temperatures. An extensive guide to the 15 hybridization of nucleic acids is found in Tijssen, Techniques in Biochemistry and Molecular Biology-Hybridization with Nucleic Probes, "Overview of principles of hybridization and the strategy of nucleic acid assays" (1993). Generally, highly stringent conditions are selected to be about 5-10"C. lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength pH, Low 20 stringency conditions are generally selected to be about 15-30C, below the Tm. Tm is the temperature (under defined ionic strength, pH, and nucleic concentration) at which 50% of the probes complementary to the target hybridize to the target sequence at equilibrium (as the target sequences are present in excess, at Tm, 50% of the probes are occupied at equilibrium). Stringent 25 conditions will be those in which the salt concentration is less than about I .OM sodium ion, typically about 0,01 to 1.OM sodium ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30*C for short probes (e.g., 10 to 50 nucleotides) and at least about 60C for long probes (e.g., greater than 50 nucleotides), Stringent conditions may also be achieved with the addition of 30 destabilizing agents such as formamide. For selective or specific hybridization, a positive signal is at least two times background, preferably 10 times background hybridization, 21 WO 2010/025466 PCT/US2009/055557 Nucleic acids that do not hybridize to each other under stringent conditions are still substantially identical if the polypeptides which they encode are substantially identical. This occurs, for example, when a copy of a nucleic acid is created using the maximum codon degeneracy permitted by the genetic code. In such cased, 5 the nucleic acids typically hybridize under moderately stringent hybridization conditions. Genomic DNA or cDNA comprising GPT polynucleotides may be identified in standard Southern blots under stringent conditions using the GPT polynucleotide 10 sequences disclosed here. For this purpose, suitable stringent conditions for such hybridizations are those which include a hybridization in a buffer of 40% formamide, IM NaCl, 1% SDS at 37"C, and at least one wash in 0.2 X SSC at a temperature of at least about 50'C, usually about 55*C to about 60*C, for 20 minutes, or equivalent conditions. A positive hybridization is at least twice 15 background, Those of ordinary skill will readily recognize that alternative hybridization and wash conditions may be utilized to provide conditions of similar stringency. A further indication that two polynucleotides are substantially identical is if the 20 reference sequence, amplified by a pair of oligonucleotide primers, can then be used as a probe under stringent hybridization conditions to isolate the test sequence from a cDNA or genomic library, or to identify the test sequence in, e.g, a northern or Southern blot. 25 TRANSGENIC PLANTS; The invention provides novel transgenic plants exhibiting substantially enhanced agronomic characteristics, including faster growth, greater mature plant fresh weight and total biomass, earlier and more abundant flowering, and greater fruit, 30 pod and seed yields. The transgenic plants of the invention are generated by introducing into a plant one or more expressible genetic constructs capable of driving the expression of one or more polynucleotides encoding glutamine synthetase (GS) and glutamine phenylpyruvate transaminase (GPT). In an exemplary embodiment, single-transgene parental lines carrying either a GPT or 22 WO 2010/025466 PCT/US2009/055557 GS1 transgene coding sequence are generated, preferably selfed until homozygous for the transgene, then crossed to generate progeny plants containing both transgenes, 5 In stable transformation embodiments of the invention, one or more copies of the expressible genetic construct become integrated into the host plant genome, thereby providing increased GS and GPT enzyme capacity into the plant, which serves to mediate increased synthesis of 2-oxoglutaramate, which in turn signals metabolic gene expression, resulting in increased plant growth and the 10 enhancement other agronomic characteristics. 2-oxoglutara mate is a metabolite which is an extremely potent effector of gene expression, metabolism and plant growth (U.S, Patent No, 6,555,500), and which may play a pivotal role in the coordination of the carbon and nitrogen metabolism systems (Lancien et al, 2000, Enzyme Redundancy and the Impotdance of 2-Oxoglutamte in Higher Plants 15 Ammonium Assimilation, Plant PhysioL 123: 817-824). See, also, the schematic of the 2-oxoglutaramate pathway shown in FIG. 1. In one aspect of the invention, applicants have isolated a nucleic acid molecule encoding the Arabidopsis glutamine phenylpyruvate transaminase (GPT) enzyme 20 (see Example 1, infra), and have demonstrated for the first time that the expressed recombinant enzyme is active and capable of catalyzing the synthesis of the signal metabolite, 2-oxoglutaramate (Example 2, infra). Further, applicants have demonstrated for the first time that over-expression of the Arabidopsis glutamine transaminase gene in a transformed heterologous plant results in 25 enhanced C02 fixation rates and increased growth characteristics (Example 3, infra). Applicants' previous work demonstrated that over-expression of Alfalfa GS1 gene under the control of a strong constitutive promoter results in transgenic tobacco 30 plants with higher levels of GS activity in the leaves, These plants outgrow their wild-type counterparts, fix CO2 faster, contain increased concentrations of total protein, as well as increased concentrations of glutamine and 2-oxoglutaramate, and show increased rates of uptake of nitrate through their roots. 23 WO 2010/025466 PCT/US2009/055557 As disclosed herein (see Example 3, infra), over-expression of a transgene comprising the full-length Arabidopsis GPT coding sequence in transgenic tobacco plants also results in faster CO2 fixation, and increased levels of total 5 protein, glutamine and 2-oxoglutara mate. These transgenic plants also grow faster than wild-type plants (FIG, 2). Similarly, in preliminary studies conducted with tomato plants (see Example 4, infra), tomato plants transformed with the Arabidopsis GPT transgene showed significant enhancement of growth rate, flowering, and seed yield in relation to wild type control plants (FIG, 3 and 10 Example 4, infra). In one particular embodiment, exemplified herein by way of Examples 3, 5 and 7, infra, a first set of parental single-transgene tobacco plant lines carrying the Alfalfa GS1 gene, including 5' and 3' untranslated regions, were generated using 15 Agrobacterium mediated gene transformation, under selective pressure, together with screening for the fastest growing phenotype, and selfing to transgene/phenotype homozygosity (see Example 5, infra). A second set of parental single-transgene tobacco plant lines carrying the full length coding sequence of Arabidopsis GPT were generated in the same manner (Example 3, 20 infra). High growth rate performing plants from each of the parental lines were then sexually crossed to yield progeny lines (Example 7, infra). The resulting progeny from multiple crosses of Arabidopsis GS1 and GPT transgenic tobacco plants produce far better and quite surprising increases in 25 growth rates over the single-transgene parental lines as well as wildtype plants. FIG. 5 shows photographs of double-transgene progeny from single-transgene GS1 X GPT plant crosses, relative to wild type and single-transgene parental plants. FIG. 6 shows photographs comparing leaf sizes of double-transgene progeny and wild type plants. Experimentally observed growth rates in these 30 double transgenic plants ranged between 200% and 300% over wild-type plants (Exarnple 7, infra), Moreover, total biomass levels increased substantially in the double-transgene plants, with whole plant fresh weights typically being about two to three times the wild-type plant weights. Similarly, seed yields showed similar 24 WO 2010/025466 PCT/US2009/055557 increases in the double-transgene plants, with seed pod production typically two to three times the wild type average, and overall seed yields exceeding wild-type plant yields by 300-400% 5 In addition to the transgenic tobacco plants referenced above, various other species of transgenic plants comprising GPT and GS transgenes are specifically exemplified herein. As exemplified herein, transgenic plants showing enhanced growth characteristics have been generated in two species of Tomato (see Examples 4 and 17), Pepper (Example 8), Beans (Examples 9 and 10), Cowpea 10 (Examples 11 and 12), Alfalfa (Example 13), Cantaloupe (Example 14), Pumpkin (Example 15), Arabidopsis (Example 16) and Camilena (Example 18), These transgenic plants of the invention were generated using a variety of transformation methodologies, including Agrobacterium-mediated callus, floral dip, seed inoculation, pod inoculation, and direct flower inoculation, as well as combinations 15 thereof, and via sexual crosses of single transgene plants, as exemplified herein. Different GPT and GS transgenes were successfully employed in generating the transgenic plants of the invention, as exemplified herein. The invention also provides methods of generating a transgenic plant having 20 enhanced growth and other agronomic characteristics. In one embodiment, a method of generating a transgenic plant having enhanced growth and other agronomic characteristics comprises introducing into a plant cell an expression cassette comprising a nucleic acid molecule encoding a GPT transgene, under the control of a suitable promoter capable of driving the expression of the 25 transgene, so as to yield a transformed plant cell, and obtaining a transgenic plant which expresses the encoded GPT, In another embodiment, a method of generating a transgenic plant having enhanced growth and other agronomic characteristics comprises introducing into a plant cell one or more nucleic acid constructs or expression cassettes comprising nucleic acid molecules encoding a 30 GPT transgene and an GS transgene, under the control of one or more suitable promoters (and, optionally, other regulatory elements) capable of driving the 25 WO 2010/025466 PCT/US2009/055557 expression of the transgenes, so as to yield a plant cell transformed thereby, and obtaining a transgenic plant which expresses the GPT and GS transgenes. Based on the results disclosed herein, it is clear that any number of GPT and GS 5 polynucleotides may be used to generate the transgenic plants of the invention. Both GS1 and GPT proteins are highly conserved among various plant species, and it is evident from the experimental data disclosed herein that closely-related non-plant GPTs may be used as well (e.g., Danio rerio GPT), With respect to GPT, numerous GPT polynucleotides derived from different species have been 10 shown to be active and useful as GPT transgenes. Similarly, different GS polynucleotides may be used, including without limitation any plant GS1 encoding polynucleotide that generates GS activity in a host cell transformed with an expressible GS1 construct. 15 In a specific embodiment, the GPT transgene is a GPT polynucleotide encoding an Arabidopsis derived GPT, such as the GPT of SEQ ID NO: 2, SEQ ID NO: 21 and SEQ ID NO: 30, and the GS transgene is a GS polynucleotide encoding an Alfalfa derived GS1 (i e., SEQ ID NO: 4) or an Arabidopsis derived GS1 (SEQ ID NO: 7). The GPT transgene may be encoded by the nucleotide sequence of SEQ 20 ID NO: 1; a nucleotide sequence having at least 75% and more preferably at least 80% identity to SEQ ID NO: 1, and encoding a polypeptide having GPT activity; a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2, or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GPT activity; and a nucleotide sequence encoding the polypeptide of 25 SEQ ID NO: 2 truncated at its amino terminus by between 30 to 56 amino acid residues, or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GPT activity. The GS1 transgene may be encoded by the polynucleotide of SEQ ID NO: 3 or SEQ ID NO: 6 or a nucleotide sequence having at least 75% and more preferably at least 80% identity to SEQ 30 ID NO: 3 or SEQ ID NO: 6, and encoding a polypeptide having GPT activity; and a nucleotide sequence encoding the polypeptide of SEQ ID NO: 4 or 7, or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GS activity, 26 WO 2010/025466 PCT/US2009/055557 In another specific embodiment, the GPT transgene is a GPT polynuceotide encoding a Grape derived GPT, such as the Grape GPTs of SEQ ID NO; 9 and SEQ ID NO; 31, and the GS transgene is a GS1 polynucleotide. The GPT transgene may be encoded by the nucleotide sequence of SEQ ID NO: 8; a 5 nucleotide sequence having at least 75% and more preferably at least 80% identity to SEQ ID NO: 8, and encoding a polypeptide having GPT activity; a nucleotide sequence encoding the polypeptide of SEQ ID NO: 9 or SEQ ID NO; 31, or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GPT activity, 10 In yet another specific embodiment, the GPT transgene is a GPT polynucleotide encoding a Rice derived GPT, such as the Rice GPTs of SEQ ID NO; 11 and SEQ ID NO; 32, and the GS transgene is a GS1 polynucleotide. The GPT transgene may be encoded by the nucleotide sequence of SEQ ID NO: 10; a 15 nucleotide sequence having at least 75% and more preferably at least 80% identity to SEQ ID NO; 10, and encoding a polypeptide having GPT activity; a nucleotide sequence encoding the polypeptide of SEQ ID NO: 11 or SEQ ID NO: 32, or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GPT activity. 20 In yet another specific embodiment, the GPT transgene is a GPT polynucleotide encoding a Soybean derived GPT, such as the Soybean GPTs of SEQ ID NO; 13, SEQ IS NO; 33 or SEQ ID NO; 33 with a further Isoleucine at the N-terminus of the sequence, and the GS transgene is a GS1 polynucleotide. The GPT 25 transgene may be encoded by the nucleotide sequence of SEQ ID NO; 12; a nucleotide sequence having at least 75% and more preferably at least 80% identity to SEQ ID NO: 12, and encoding a polypeptide having GPT activity; a nucleotide sequence encoding the polypeptide of SEQ ID NO 13 or SEQ ID NO: 33 or SEQ ID NO: 33 with a further Isoleucine at the N-terminus of the sequence, 30 or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GPT activity. 27 WO 2010/025466 PCT/US2009/055557 In yet another specific embodiment, the GPT transgene is a GPT polynucleotide encoding a Barley derived GPT, such as the Barley GPTs of SEQ ID NO: 15 and SEQ ID NO; 34, and the GS transgene is a GS1 polynucleotide. The GPT transgene may be encoded by the nucleotide sequence of SEQ ID NO: 14; a 5 nucleotide sequence having at least 75% and more preferably at least 80% identity to SEQ ID NO: 10, and encoding a polypeptide having GPT activity; a nucleotide sequence encoding the polypeptide of SEQ ID NO: 15 or SEQ ID NO: 34, or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GPT activity, 10 In yet another specific embodiment, the GPT transgene is a GPT polynucleotide encoding a Zebra fish derived GPT, such as the Zebra fish GPTs of SEQ ID NO: 17 and SEQ ID NO: 35, and the GS transgene is a GS1 polynucleotide, The GPT transgene may be encoded by the nucleotide sequence of SEQ ID NO: 16; a 15 nucleotide sequence having at least 75% and more preferably at least 80% identity to SEQ ID NO; 16, and encoding a polypeptide having GPT activity; a nucleotide sequence encoding the polypeptide of SEQ ID NO: 17 or SEQ ID NO: 35, or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GPT activity. 20 In yet another specific embodiment, the GPT transgene is a GPT polynucleotide encoding a Bamboo derived GPT, such as the Bamboo GPT of SEQ ID NO: 36, and the GS transgene is a GS1 polynucleotide. The GPT transgene may be encoded by a nucleotide sequence encoding the polypeptide of SEQ ID NO; 36, 25 or a polypeptide having at least 75% and more preferably at least 80% sequence identity thereto which has GPT activity. Other GPT polynucleotides suitable for use as GPT transgenes in the practice of the invention may be obtained by various means, as will be appreciated by one 30 skilled in the art, tested for the ability to direct the expression of a GPT with GPT activity in a recombinant expression system (i.e., E coil (see Examples 20-23), in a transient in planta expression system (see Example 19), or in a transgenic plant (see Examples 1-18), 28 WO 2010/025466 PCT/US2009/055557 TRANSGENE CONSTRUCTS/EXPRESSION VECTORS In order to generate the transgenic plants of the invention, the gene coding sequence for the desired transgene(s) must be incorporated into a nucleic acid 5 construct (also interchangeably referred to herein as a (transgene) expression vector, expression cassette, expression construct or expressible genetic construct) which can direct the expression of the transgene sequence in transformed plant cells. Such nucleic acid constructs carrying the transgene(s) of interest may be introduced into a plant cell or cells using a number of methods 10 known in the art, including but not limited to electroporation, DNA bombardment or biolistic approaches, microinjection, and via the use of various DNA-based vectors such as Agrobacterium tumefaciens and Agrobacterium rhizogenes vectors. Once introduced into the transformed plant cell, the nucleic acid construct may direct the expression of the incorporated transgene(s) (i.e., GPT), either in a 15 transient or stable fashion. Stable expression is preferred, and is achieved by utilizing plant transformation vectors which are able to direct the chromosomal integration of the transgene construct. Once a plant cell has been successfully transformed, it may be cultivated to regenerate a transgenic plant. 20 A large number of expression vectors suitable for driving the constitutive or induced expression of inserted genes in transformed plants are known. In addition, various transient expression vectors and systems are known. To a large extent, appropriate expression vectors are selected for use in a particular method of gene transformation (see, infra) Broadly speaking, a typical plant expression 25 vector for generating transgenic plants will comprise the transgene of interest under the expression regulatory control of a promoter, a selectable marker for assisting in the selection of transformants, and a transcriptional terminator sequence. 30 More specifically, the basic elements of a nucleic acid construct for use in generating the transgenic plants of the invention are: a suitable promoter capable of directing the functional expression of the transgene(s) in a transformed plant cell, the transgene (s) (i.e., GPT coding sequence) operably linked to the 29 WO 2010/025466 PCT/US2009/055557 promoter, preferably a suitable transcription termination sequence (i.e., nopaline synthetic enzyme gene terminator) operably linked to the transgene, and typically other elements useful for controlling the expression of the transgene, as well as one or more selectable marker genes suitable for selecting the desired 5 transgenic product (i.e., antibiotic resistance genes). As Agrobacterium tumefaciens is the primary transformation system used to generate transgenic plants, there are numerous vectors designed for Agrobacterium transformation. For stable transformation, Agrobacterium systems 10 utilize "binary" vectors that permit plasmid manipulation in both E coli and Agrobacterium, and typically contain one or more selectable markers to recover transformed plants (Hellens et aL, 2000, Technical focus: A guide to Agrobacterium binary Ti vectors, Trends Plant Sci 5:446-451). Binary vectors for use in Agrobacterium transformation systems typically comprise the borders of T 15 DNA, multiple cloning sites, replication functions for Escherichia coil and A. tumefaciens, and selectable marker and reporter genes. So-called "super-binary" vectors provide higher transformation efficiencies, and generally comprise additional virulence genes from a Ti (Komari et al., 2006, 20 Methods Mol. Biol. 343; 15-41). Super binary vectors are typically used in plants which exhibit lower transformation efficiencies, such as cereals. Such additional virulence genes include without limitation virB, vrE, and virG (Vain et al., 2004, The effect of additional virulence genes on transformation efficiency, transgene integration and expression in rice plants using the pGreen/pSoup dual binary 25 vector system. Transgenic Res. 13: 593-603; Srivatanakul et al- 2000, Additional virulence genes influence transgene expression: transgene copy number, integration pattern and expression. J. Plant Physiol. 157, 685-690; Park et al, 2000, Shorter T-DNA or additional virulence genes improve Agrobacterium mediated transformation. Theory. Apple. Genet. 101, 1015-1020; Jin et aL, 1987, 30 Genes responsible for the supervirulence phenotype of Agrobacterium tumefaciens A281. J. Bacteriol. 169: 4417-4425). 30 WO 2010/025466 PCT/US2009/055557 In the embodiments exemplified herein (see Examples, infra), expression vectors which place the inserted transgene(s) under the control of the constitutive CaMV 35S promoter and the RuBisCo promoter are employed. A number of expression vectors which utilize the CaMV 358 and RuBsCo promoter are known and/or 5 commercially available and/or derivable using ordinary skill in the art PLANT PROMOTERS The term 'promoter' is used to designate a region in the genome sequence 10 upstream of a gene transcription start site (TSS), although sequences downstream of TSS may also affect transcription initiation as well, Promoter elements select the transcription initiation point, transcription specificity and rate. Depending on the distance from the TSS, the terms of 'proximal promoter' (several hundreds nucleotides around the TSS) and 'distal promoter' (thousands 15 and more nucleotides upstream of the TSS) are also used. Both proximal and distal promoters include sets of various elements participating in the complex process of cell-, issue-, organ-, developmental stage and environmental factors specific regulation of transcription. Most promoter elements regulating TSS selection are localized in the proximal promoter. 20 A large number of promoters which are functional in plants are known in the art. In constructing GPT and GS transgene constructs, the selected promoter(s) may be constitutive, non-specific promoters such as the Cauliflower Mosaic Virus 35S ribosomal promoter (CaMV 35$ promoter), which is widely employed for the 25 expression of transgenes in plants, Examples of other strong constitutive promoters include without limitation the rice actin 1 promoter, the CaMV 198 promoter, the Ti plasmid nopaline synthase promoter, the alcohol dehydrogenase promoter and the sucrose synthase promoter. 30 Alternatively, in some embodiments, it may be desirable to select a promoter based upon the desired plant cells to be transformed by the transgene construct, the desired expression level of the transgene, the desired tissue or subcellular compartment for transgene expression, the developmental stage targeted, and 31 WO 2010/025466 PCT/US2009/055557 the like, For example, when expression in photosynthetic tissues and compartments is desired, a promoter of the ribulose bisphosphate carboxylase (RuBisCo) gene 5 may be employed. In the Examples which follow, expressible nucleic acid constructs comprising GPT and G1 transgenes under the control of a tomato RuBisCo promoter were prepared and used in the generation of transgenic plants or to assay for GPT activity in plant or in E coli, 10 When the expression in seeds is desired, promoters of various seed storage protein genes may be employed. For expression in fruits, a fruit-specific promoter such as tomato 2A1 1 may be used. Examples of other tissue specific promoters include the promoters encoding lectin (Vodkin et at, 1983, Cell 34:1023-31; Lindstrom et at, 1990, Developmental Genetics 11:160-167), com alcohol 15 dehydrogenase 1 (Vogel et al, 1989, J. Cell. Biochem, (Suppl. 0) 13:Part D, Dennis et al, 1984, Nuc. Acids Res., 12(9): 3983-4000), corn light harvesting complex (Simpson, 1986, Science, 233: 34-38; Bansal et at, 1992, Proc. Natl. Acad. Sci. USA, 89: 3654-3658), corn heat shock protein (Odell et at., 1985, Nature, 313: 810-812; Rochester et at, 1986, EMBO J,, 5: 451-458), pea small 20 subunit RuBP carboxylase (Poulsen et al., 1986, Mol. Gen Genet., 205(2): 193 200; Cashmore et at., 1983, Gen. Eng, Plants, Plenum Press, New York, pp 29 38), Ti plasmid mannopine synthase and Ti plasmid nopaline synthase (Langridge et at, 1989, Proc. NatI. Acad, Sci. USA, 86: 3219-3223), petunia chalcone isomerase (Van Tunen et at., 1988, EMBO J. 7(5): 1257-1263), bean glycine rich 25 protein 1 (Keller et at., 1989, EMBO J, 8(5): 1309-1314), truncated CaMV 35s (Odell et at., 1985, supra), potato patatin (Wenzier et at, 1989, Plant Mol. Biol. 12: 41-50), root cell (Conkding et al, 1990, Plant Physiol. 93: 1203-1:211), maize zein (Reina et at., 1990, Nuc. Acids Res. 18(21): 6426; Kriz et al., 1987, Mol. Gen, Genet. 207(1): 90-98; Wandelt and Feix, 1989, Nuc, Acids Res. 17(6): 2354; 30 Langridge and Feix, 1983, Cell 34: 1015-1022; Reina et at., 1990, NucL. Acids Res. 18(21): 6426), globulin-I (Belanger and Kriz, 1991, Genetics 129: 863-872), c-tubulin (Carpenter et at., 1992, Plant Cell 4(5): 657-571; Uribe et at., 1998, Plant Molt Biol. 37(6): 1069-1078), cab (Sullivan, et at., 1989, Molt Gen. Genet, 215(3): 32 WO 2010/025466 PCT/US2009/055557 431-440), PEPCase (Hudspeth and Grula, 1989, Plant Mol Biol. 12: 579-589), R gene complex (Chandler et at, 1989, The Plant Cell 1: 1175-1183), chalcone synthase (Franken et al,, 1991, EMBO J, 10(9): 2605-2612) and glutamine synthetase promoters (U.S. Pat. No. 5,391,725; Edwards et at, 1990, Proc, Nati. 5 Acad. Sci, USA 87: 3459-3463; Brears et al, 1991, Plant J. 1(2): 235-244). In addition to constitutive promoters, various inducible promoter sequences may be employed in cases where it is desirable to regulate transgene expression as the transgenic plant regenerates, matures, flowers, etc. Examples of such 10 inducible promoters include promoters of heat shock genes, protection responding genes (ie, phenylalanine ammonia lyase; see, for example Bevan et at, 1989, EMBO J. 8(7): 899-906), wound responding genes (Le., cell wall protein genes), chemically inducible genes (Le., nitrate reductase, chitinase) and dark inducible genes (i.e., asparagine synthetase; see, for example U.S. Patent No. 5,256,558), 15 Also, a number of plant nuclear genes are activated by light, including gene families encoding the major chlorophyll a/b binding proteins (cab) as well as the small subunit of ribulose-1, 5-bisphosphate carboxylase (rbcS) (see, for example, Tobin and Silverthorne, 1985, Annu. Rev. Plant Physiol. 36: 569-593; Dean et at, 1989, Annu, Rev. Plant Physiol. 40: 415-439.), 20 Other inducible promoters include ABA- and turgor-inducible promoters, the auxin-binding protein gene promoter (Schwob et at, 1993, Plant J, 4(3): 423-432), the UDP glucose flavonoid glycosyl-transferase gene promoter (Ralston et al, 1988, Genetics 119(1): 185-197); the MPI proteinase inhibitor promoter (Cordero 25 et at., 1994, Plant J. 6(2): 141-150), the glyceraldehyde-3-phosphate dehydrogenase gene promoter (Kohler et al., 1995, Plant Molt Biol. 29(6): 1293 1298; Quigley at alt, 1989, J. Mol. Evol. 29(5): 412~421; Martinez et at.,1989, J. Mol. Biol. 208(4); 551-565) and light inducible plastid glutamine synthetase gene from pea (U.S. Pat No. 5,391,725; Edwards et al., 1990, supra). 30 For a review of plant promoters used in plant transgenic plant technology, see Potenza et al., 2004, In Vitro Cell. Devel. Biol - Plant, 40(1): 1-22. For a review of 33 WO 2010/025466 PCT/US2009/055557 synthetic plant promoter engineering, see, for example, Venter, M., 2007, Trends Plant Sci, 12(3): 118-124. GLUTAMINE PHENYLPYRUVATE TRANSAMINASE (GPT) TRANSGENE The present invention discloses for the first time that plants contain a glutamine phenylpyruvate transaminase (GPT) enzyme which is directly functional in the synthesis of the signal metabolite 2-hydroxy-5-oxoproline, Until now, no plant transaminase with a defined function has been described. Applicants have 10 isolated and tested GPT polynucleotide coding sequences derived from several plant and animal species, and have successfully incorporated the gene into heterologous transgenic host plants which exhibit markedly improved growth characteristics, including faster growth, higher foliar protein content, increased glutamine synthetase activity in foliar tissue, and faster CO 2 fixation rates. 15 In the practice of the invention, the GPT gene functions as one of at least two transgenes incorporated into the transgenic plants of the invention, the other being the glutamine sythetase gene (see infra). 20 It is expected that all plant species contain a GPT which functions in the same metabolic pathway, involving the biosynthesis of the signal metabolite 2-hydroxy S-oxoproline. Thus, in the practice of the invention, any plant gene encoding a GPT homolog or functional variants thereof may be useful in the generation of transgenic plants of this invention. Moreover, given the structural similarity 25 between various plant GPT protein structures and the putative ( and biologically active) GPT homolog from Danio rerio (Zebra fish) (see Example 22), other non plant GPT homologs may be used in preparing GPT transgenes for use in generating the transgenic plants of the invention. 30 When individually compared (by BLAST alignment) to the Arabidopsis mature protein sequence provided in SEQ ID NO: 30, the following sequence identities and homologies (BLAST "positives", including similar amino acids) were obtained for the following mature GPT protein sequences: 34 WO 2010/025466 PCT/US2009/055557 [SEQ ID] or FIG, NO, ORIGIN % IDENTITY %POSITIVE [31] Grape 84 93 [32] Rice 83 91 5 [33] Soybean 83 93 [34] Barley 82 91 [35] Zebra fish 83 92 [36] Bamboo 81 90 FIG, 2 Corn 79 90 10 FIG.2 Castor 84 93 FIG.2 Poplar 85 93 Underscoring the conserved nature of the structure of the GPT protein across most plant species, the conservation seen within the above plant species extends 15 to the non-human putative GPTs from Zebra fish and Chlamydomonas. In the case of Zebra fish, the extent of identity is very high (83% amino acid sequence idently with the mature Arabidopsis GPT of SEQ ID NO: 30, and 92% homologous taking similar amino acid residues into account). The Zebra fish mature GPT was confirmed by expressing it in E coi and demonstrating biological activity 20 (synthesis of 2-oxoglutara mate). In order to determine whether putative GPT homologs would be suitable for generating the growth-enhanced transgenic plants of the invention, one need initially express the coding sequence thereof in E coi or another suitable host 25 and determine whether the 2-oxoglutaramate signal metabolite is synthesized at increased levels (see Examples 19-23). Where such an increase is demonstrated, the coding sequence may then be introduced into both homologous plant hosts and heterologous plant hosts, and growth characteristics evaluated. Any assay that is capable of detecting 2-oxoglutaramate with 30 specificity may be used for this purpose, including without limitation the NMR and HPLC assays described in Example 2, infra. In addition, assays which measure GPT activity directly may be employed, such as the GPT activity assay described in Example 7, 35 WO 2010/025466 PCT/US2009/055557 Any plant GPT with 2-oxoglutaramate synthesis activity may be used to transform plant cells in order to generate transgenic plants of the invention. There appears to be a high level of structural homology among plant species, which appears to 5 extend beyond plants, as evidenced by the close homology between various plant GPT proteins and the putative Zebra fish GPT homolog. Therefore, various plant GPT genes may be used to generate growth-enhanced transgenic plants in a variety of heterologous plant species. In addition, GPT transgenes expressed in a homologous plant would be expected to result in the desired enhanced-growth 10 characteristics as well (i., rice glutamine transaminase over-expressed in transgenic rice plants), although it is possible that regulation within a homologous cell may attenuate the expression of the transgene in some fashion that may not be operable in a heterologous cell, 15 GLUTAMINE SYNTHETASE (GS) TRANSGENE: In the practice of the invention, the glutamine synthetase (GS) gene functions as one of at least two transgenes incorporated into the transgenic plants of the 20 invention (GPT being the other of the two). Glutamine synthetase plays a key role in nitrogen metabolism in plants, as well as in animals and bacteria. The GS enzyme catalyzes the addition of ammonium to glutamate to synthesize glutamine in an ATP-dependent reaction. GS enzymes 25 from assorted species show highly conserved amino acid residues considered to be important for active site function, indicating that GS enzymes function similarly (for review, see Eisenberg et al., Biochimica et Biophysica Acta, 1477:122 145, 2000), 30 GS is distributed in different subcellular locations (chloroplast and cytoplasm) and is found in various plant tissues, including leaf, root, shoot, seeds and fruits, There are two major isoforms of plant GS: the cystolic isoform (GS1) and the plastidic (chloroplastic) isoform (GS2). GS2 is principally found in leaf tissue and functions in the assimilation of ammonia produced by photorespiration or by 36 WO 2010/025466 PCT/US2009/055557 nitrate reduction, GS1 is mainly found in leaf and root tissue, typically exists in a number of different isoforms in higher plants, and functions to assimilate ammonia produced by all other physiological processes (Coruzzi, 1991, Plant Science 74: 145-155; McGrath and Coruzzi, 1991, Plant J. 1(3): 275-280; Lam et al., 1996, 5 Ann. Rev. Plant Physiol. Plant Mol. Biolt 47: 569-593; Stitt, 1999, Curr. Op. Plant BioL 2: 178-186; Oliveira et al., 2001, Brazilian J. Med. Biol. Res. 34: 567-575). Multiple GS genes are associated with a complex promoter repertoire which enable the expression of GS in an organ and tissue specific manner, as well as in an environmental factor-dependent manner 10 Plant glutamine synthetase consists of eight subunits, and the native enzyme in plants has a molecular mass ranging from 320 to 380 kD, each subunit having a molecular mass of between 38 and 45 kD. The GS1 genes of several plants, especially legumes, have been cloned and sequenced (Tischer et al,, 1986, Mol 15 Gen Genet. 203: 221-229; Gebhardt et al, 1986, EMBO J. 5:1429-1435; Tingey et al, 1987, EMBO J, 6: 1-9; Tingey et al., 1988, J Biol Chem. 263: 9651-9657; Bennett et al, 1989, Plant Mol Biol. 12: 553-565; Boron and Legocki, 1993, Gene 136: 95-102; Roche et al., 1993, Plant Mol Biol. 22: 971-983; Marsolier et al., 1995, Plant Mol Biol. 27: 1-15; Temple et aL, 1995., Mol Plant-Microbe Interact. 8: 20 218-227), All have been found to be encoded by nuclear genes (for review, see, Morey et alt, 2002, Plant Physiol. 128(1): 182-193), Chloroplastic GS2 appears to be encoded by a single gene, while various cystoloic GS1 isoforms are encoded within multigene families (Tingey et al., 1987, 25 supra; Sakamoto et al., 1989, Plant Mol. Biol. 13: 611-614; Brears et al, 1991, supra; Li et al, 1993, Plant Mol. Biolt, 23:401-407; Dubois et al, 1996, Plant Mol. Biot. 31:803-817; Lam et al,, 1996, supra), GS1 multigene families appear to encode different subunits which may combine to form homo- or hetero-octamers, and the different members show a unique expression pattern suggesting that the 30 gene members are differentially regulated, which may relate to the various functional roles of glutamine synthetase plays in overall nitrogen metabolism (Gebhardt et al., 1986, supra; Tingey et al., 1987, supra; Bennett et al., 1989, supra; Walker and Coruzzi, 1989, supra; Peterman and Goodman, 1991, Mol Gen 37 WO 2010/025466 PCT/US2009/055557 Genet. 1991;330:145-154.; Marsolier et al, 1995, supra; Temple et al, 1995, supra; Dubois et al, 1996, supra). In one embodiment, a GS1 gene coding sequence is employed to generate GS 5 transgene constructs, In particular embodiments, further described in the Examples, infra, the Alfalfa or Arabidopsis GS1 gene coding sequence is used to generate a transgene construct that may be used to generate a transgenic plant expressing the GS1 transgene. As an example, such a construct may be used to transform Agrobacteria. The transformed Agrobacteria are then used to generate 10 To transgenic plants. Example 5 demonstrates the generation of To GS1 transgenic tobacco plants using this approach. Similarly, Examples 6 and 17 demonstrates the generation of To GS1 transgenic tomato plants, Example 8 demonstrates the generation of T 0 GS1 transgenic pepper plants, Examples 9 and 10 demonstrate the generation of To GS1 transgenic bean plants, Examples 11 15 and 12 demonstrate the generation of To GS1 transgenic cowpea plants, Example 13 demonstrates the generation of T 0 GS1 transgenic alfalfa plants, Example 14 demonstrates the generation of To GS1 transgenic cantaloupe plants, Example 15 demonstrates the generation of T 0 GS1 transgenic pumpkin plants, Example 16 demonstrates the generation of Tf) GS1 transgenic Arabidopsis plants, and 20 Example 18 demonstrates the generation of To GS1 transgenic Cantaloupe plants. TRANSCRIPTION TERMINATORS: 25 In preferred embodiments, a 3' transcription termination sequence is incorporated downstream of the transgene in order to direct the termination of transcription and permit correct polyadenylation of the mRNA transcript. Suitable transcription terminators are those which are known to function in plants, including without limitation, the nopaline synthase (NOS) and octopine synthase (OCS) genes of 30 Agrobacterium tumefaciens, the T7 transcript from the octopine synthase gene, the 3' end of the protease inhibitor I or i1 genes from potato or tomato, the CaMV 35$ terminator, the tml terminator and the pea rbcS E9 terminator. In addition, a gene's native transcription terminator may be used. In specific embodiments, described by way of the Examples, infra, the nopaline synthase transcription 38 WO 2010/025466 PCT/US2009/055557 terminator is employed, SELECTABLE MARKERS: 5 Selectable markers are typically included in transgene expression vectors in order to provide a means for selecting transformants. While various types of markers are available, various negative selection markers are typically utilized, including those which confer resistance to a selection agent that inhibits or kills untransformed cells, such as genes which impart resistance to an antibiotic (such 10 as kanamycin, gentamycin, anamycin, hygromycin and hygromycinB) or resistance to a herbicide (such as sulfonylurea, gulfosinate, phosphinothricin and glyphosate). Screenable markers include, for example, genes encoding P glucuronidase (Jefferson, 1987, Plant Mol. Biol. Rep 5: 387-405), genes encoding luciferase (Ow et aL, 1986, Science 234: 856-859) and various genes encoding 15 proteins involved in the production or control of anthocyanin pigments (See, for example, US. Patent 6,573,432). The E coli glucuronidase gene (gus, gusA or uidA) has become a widely used selection marker in plant transgenics, largely because of the glucuronidase enzyme's stability, high sensitivity and ease of detection (e.g., fluorometric, spectrophotometric, various histochemical methods). 20 Moreover, there is essentially no detectable glucuronidase in most higher plant species, TRANSFORMATION METHODOLOGIES AND SYSTEMS: 25 Various methods for introducing the transgene expression vector constructs of the invention into a plant or plant cell are well known to those skilled in the art, and any capable of transforming the target plant or plant cell may be utilized. Agrobacterium-mediated transformation is perhaps the most common method 30 utilized in plant transgenics, and protocols for Agrobacterium-mediated transformation of a large number of plants are extensively described in the literature (see, for example, Agrobacterium Protocols, Wan, ed., Humana Press, 2 nd edition, 2006). Agrobacterium tumefaciens is a Gram negative soil bacteria that causes tumors (Crown Gall disease) in a great many dicot species, via the 39 WO 2010/025466 PCT/US2009/055557 insertion of a small segment of tumor-inducing DNA ( "T-DNA", 'transfer DNA') into the plant cell, which is incorporated at a semi-random location into the plant genome. and which eventually may become stably incorporated there. Directly repeated DNA sequences, called T-DNA borders, define the left and the right 5 ends of the T-DNA. The T-DNA can be physically separated from the remainder of the Ti-plasmid, creating a'binary vector' system. Agrobacterium transformation may be used for stably transforming dicots, monocots, and cells thereof (Rogers et al, 1986, Methods Enzymol, 118: 627 10 641; Hernalsteen et al., 1984, EMBO J, 3: 3039-3041; Hoykass-Van Slogteren et al., 1984, Nature, 311: 763-764; Grimsley et al, 1987, Nature 325: 167-1679; Boulton et al., 1989, Plant Mol. Biolt 12: 31-40; Gould et al., 1991, Plant Physiol. 95: 426-434). Various methods for introducing DNA into Agrobacteria are known, including electroporation, freeze/thaw methods, and triparental mating, The most 15 efficient method of placing foreign DNA into Agrobacterium is via electroporation (Wise et al, 2006, Three A4ethods for the Introduction of Foreign DNA into Agrobacterium, Methods in Molecular Biology, vol. 343: Agrobacterium Protocols, 2/e, volume 1; Ed., Wang, Humana Press Inc., Totowa, NJ, pp. 43-53), In addition, given that a large percentage of T-DNAs do not integrate, 20 Agrobacterum-mediated transformation may be used to obtain transient expression of a transgene via the transcriptional competency of unincorporated transgene construct molecules (Helens et al., 2005, Plant Methods 1:13), A large number of Agrobacterium transformation vectors and methods have been 25 described (Karimi et al., 2002, Trends Plant Sci. 7(5): 193-5), and many such vectors may be obtained commercially (for example, Invitrogen). In addition, a growing number of 'open-source" Agrobacterium transformation vectors are available (for example, pCambia vectors; Cambia, Canberra, Australia). See, also, subsection herein on TRANSGENE CONSTRUCTS, supra, In a specific 30 embodiment described further in the Examples, a pMON316-based vector was used in the leaf disc transformation system of Horsch et. al. (Horsch et al.,1995, Science 227:1229-1231) to generate growth enhanced transgenic tobacco and tomato plants. 40 WO 2010/025466 PCT/US2009/055557 Other commonly used transformation methods that may be employed in generating the transgenic plants of the invention include without limitation microprojectile bombardment, or biollistic transformation methods, protoplast 5 transformation of naked DNA by calcium, polyethylene glycol (PEG) or electroporation (Paszkowski et a., 1984, EMBO J. 3: 2727-2722; Potrykus et al, 1985, Mol. Gen, Genet 199: 169-177; Fromm et at, 1985, Proc. Nat, Acad. Sci USA 82: 5824-5828; Shimamoto et al., 1989, Nature, 338: 274-276, 10 Biolistic transformation involves injecting millions of DNA-coated metal particles into target cells or tissues using a biolistic device (or "gene gun"), several kinds of which are available commercially; once inside the cell, the DNA elutes off the particles and a portion may be stably incorporated into one or more of the cell's chromosomes (for review, see Kikkert et at, 2005, Stable Transformation of Plant 15 Cells by Particle Bombardment/Biolistics, in: Methods in Molecular Biology, vol. 286: Transgenic Plants: Methods and Protocols, Ed. L. Peia, Humana Press Inc, Totowa, NJ), Electroporation is a technique that utilizes short, high-intensity electric fields to 20 permeabilize reversibly the lipid bilayers of cell membranes (see, for example, Fisk and Dandekar, 2005, Introduction and Expression of Transgenes in Plant Protoplasts, in: Methods in Molecular Biology, vol. 286: Transgenic Plants: Methods and Protocols, Ed. L. Pena, Humana Press Inc., Totowa, NJ, pp, 79-90; Fromm et al,1987, Electroporation of DNA and RNA into plant protoplasts, in 25 Methods in Enzymology, Vol. 153, Wu and Grossman, eds., Academic Press, London, UK, pp. 351-366; Joersbo and Brunstedt, 1991, Electroporation: mechanism and transient expression stable transformation and biological effects in plant protoplasts Physiol. Plant, 81, 256-264; Bates, 1994, Genetic transformation of plants by protoplast electroporation. Mol. Biotech. 2: 135-145; 30 Dillen et alk, 1998, Electroporation-mediated DNA transfer to plant protoplasts and intact plant tissues for transient gene expression assays, in Cell Biology, Vol. 4, ed., Celis, Academic Press, London, UK, pp. 92-99), The technique operates by creating aqueous pores in the bacterial membrane, which are of sufficiently large 41 WO 2010/025466 PCT/US2009/055557 size to allow DNA molecules (and other macromolecules) to enter the cell, where the transgene expression construct (as T-DNA) may be stably incorporated into plant genomic DNA, leading to the generation of transformed cells that can subsequently be regenerated into transgenic plants, Newer transformation methods include so-called "floral dip" methods, which offer the promise of simplicity, without requiring plant tissue culture, as is the case with all other commonly used transformation methodologies (Bent et al., 2006, Arabidopsis thaliana Floral Dip Transformation Method, Methods Mol Biol, vol. 10 343: Agrobacterium Protocols, 2/e, volume 1; Ed., Wang, Humana Press Inc, Totowa, NJ, pp. 87-103; Clough and Bent, 1998, Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana, Plant J. 16: 735-743). However, with the exception of Arabidopsis, these methods have not been widely used across a broad spectrum of different plant species, Briefly, 15 floral dip transformation is accomplished by dipping or spraying flowering plants in with an appropriate strain of Agrobacterium tumefaciens. Seeds collected from these To plants are then germinated under selection to identify transgenic T, individuals. Example 16 demonstrated floral dip inoculation of Arabidopsis to generate transgenic Arabidopsis plants. 20 Other transformation methods include those in which the developing seeds or seedlings of plants are transformed using vectors such as Agrobacterium vectors, For example, as exemplified in Example 8, such vectors may be used to transform developing seeds by injecting a suspension or mixture of the vector (ie, 25 Agrobactea) directly into the seed cavity of developing pods (i.e., pepper pods, bean pods, pea pods and the like). Seedlings may be transformed as described for Alfalfa in Example 13. Germinating seeds may be transformed as described for Camelina in Example 18. Intra-fruit methods, in which the vector is injected into fruit or developing fruit, may be used as described for Cantaloupe melons in 30 Example 14 and pumpkins in Example 15. Still other transformation methods include those in which the flower structure is targeted for vector inoculation, such as the flower inoculation methods described 42 WO 2010/025466 PCT/US2009/055557 for beans in Examples 9 and 10, peas in Examples 11 and 12 and tomatoes in Example 17. The foregoing plant transformation methodologies may be used to introduce 5 transgenes into a number of different plant cells and tissues, including without limitation, whole plants, tissue and organ explants including chloroplasts, flowering tissues and cells, protoplasts, meristem cells, callus, immature embryos and gametic cells such as microspores, pollen, sperm and egg cells, tissue cultured cells of any of the foregoing, any other cells from which a fertile 10 regenerated transgenic plant may be generated. Callus is initiated from tissue sources including, but not limited to, immature embryos, seedling apical meristems, microspores and the like. Cells capable of proliferating as callus are also recipient cells for genetic transformation: 15 Methods of regenerating individual plants from transformed plant cells, tissues or organs are known and are described for numerous plant species. As an illustration, transformed plantlets (derived from transformed cells or tissues) are cultured in a root-permissive growth medium supplemented with the selective 20 agent used in the transformation strategy (i.e., and antibiotic such as kanamycin). Once rooted, transformed plantlets are then transferred to soil and allowed to grow to maturity, Upon flowering, the mature plants are preferably selfed (self fertilized), and the resultant seeds harvested and used to grow subsequent generations. Examples 3 - 6 describe the regeneration of transgenic tobacco and 25 tomato plants.
T
0 transgenic plants may be used to generate subsequent generations (e g, TI,
T
2 , etc.) by selfing of primary or secondary transformants, or by sexual crossing of primary or secondary transformants with other plants (transformed or 30 untransformed). For example, as described in Example 7, infra, individual plants over expressing the Alfalfa GS1 gene and outperforming wildtype plants were crossed with individual plants over-expressing the Arabidopsis GPT gene and outperforming wildtype plants, by simple sexual crossing using manual pollen 43 WO 2010/025466 PCT/US2009/055557 transfer. Reciprocal crosses were made such that each plant served as the male in a set of crosses and each plant served as the female in a second set of crosses. During the mature plant growth stage, the plants are typically examined for growth phenotype, CO 2 fixation rate, etc. (see following subsection). SELECTION OF GROWTH-ENHANCED TRANSGENIC PLANTS: Transgenic plants may be selected, screened and characterized using standard methodologies, The preferred transgenic plants of the invention will exhibit one or 10 more phenotypic characteristics indicative of enhanced growth and/or other desirable agronomic properties. Transgenic plants are typically regenerated under selective pressure in order to select transformants prior to creating subsequent transgenic plant generations. In addition, the selective pressure used may be employed beyond To generations in order to ensure the presence of the 15 desired transgene expression construct or cassette, To transformed plant cells, calli, tissues or plants may be identified and isolated by selecting or screening for the genetic composition of and/or the phenotypic characteristics encoded by marker genes contained in the transgene expression 20 construct used for the transformation. For example, selection may be conducted by growing potentially-transformed plants, tissues or cells in a growth medium containing a repressive amount of antibiotic or herbicide to which the transforming genetic construct can impart resistance. Further, the transformed plant cells, tissues and plants can be identified by screening for the activity of marker genes 25 (such as P-glucuronidase) which may be present in the transgene expression construct. Various physical and biochemical methods may be employed for identifying plants containing the desired transgene expression construct, as is well known. 30 Examples of such methods include Southern blot analysis or various nucleic acid amplification methods (i., PCR) for identifying the transgene, transgene expression construct or elements thereof; Northern blotting, S1 RNase protection, reverse transcriptase PCR (RT-PCR) amplification for detecting and determining the RNA transcription products; and protein gel electrophoresis, Western blotting, 44 WO 2010/025466 PCT/US2009/055557 immunoprecipitation, enzyme immunoassay, and the like for identifying the protein encoded and expressed by the transgene. In another approach, expression levels of genes, proteins and/or metabolic 5 compounds that are know to be modulated by transgene expression in the target plant may be used to identify transformants. In one embodiment of the present invention, increased levels of the signal metabolite 2-oxoglutaramate may be used to screen for desirable transformants, as exemplified in the Examples. Similarly, increased levels of GPT and/or GS activity may be assayed, as exemplified in the 10 Examples. Ultimately, the transformed plants of the invention may be screened for enhanced growth and/or other desirable agronomic characteristics, Indeed, some degree of phenotypic screening is generally desirable in order to identify transformed lines 15 with the fastest growth rates, the highest seed yields, etc., particularly when identifying plants for subsequent selfing, cross-breeding and back-crossing. Various parameters may be used for this purpose, including without limitation, growth rates, total fresh weights, dry weights, seed and fruit yields (number, weight), seed and/or seed pod sizes, seed pod yields (e.g., number, weight), leaf 20 sizes, plant sizes, increased flowering, time to flowering, overall protein content (in seeds, fruits, plant tissues), specific protein content (i.e., GS), nitrogen content, free amino acid, and specific metabolic compound levels (i,e., 2-oxoglutaramate), Generally, these phenotypic measurements are compared with those obtained from a parental identical or analogous plant line, an untransformed identical or 25 analogous plant, or an identical or analogous wild-type plant (i.e., a normal or parental plant), Preferably, and at least initially, the measurement of the chosen phenotypic characteristic(s) in the target transgenic plant is done in parallel with measurement of the same characteristic(s) in a normal or parental plant. Typically, multiple plants are used to establish the phenotypic desirability and/or 30 superiority of the transgenic plant in respect of any particular phenotypic characteristic. 45 WO 2010/025466 PCT/US2009/055557 Preferably, initial transformants are selected and then used to generate T 1 and subsequent generations by selfing (self-fertilization), until the transgene genotype breeds true (i.e., the plant is homozygous for the transgene). In practice, this is accomplished by selfing for 3 or 4 generations, screening at each generation for 5 the desired traits and selling those individuals. As exemplified herein, transgenic plant lines propagated through at least one sexual generation (Tobacco, Arabidopsis, Tomato) demonstrated higher transgene product activities compared to lines that did not have the benefit of sexual reproduction and the concomitant increase in transgene copy number. 10 Stable transgenic lines may be crossed and back-crossed to create varieties with any number of desired traits, including those with stacked transgenes, multiple copies of a transgene, etc. Additionally, stable transgenic plants may be further modified genetically, by transforming such plants with further transgenes or 15 additional copies of the parental transgene. Also contemplated are transgenic plants created by single transformation events which introduce multiple copies of a given transgene or multiple transgenes. Various common breeding methods are well know to those skilled in the art (see, e.g., Breeding Methods for Cultivar Development. Wilcox J, ed., American Society of Agronomy, Madison Wis. 20 (1987)). In a another aspect, the invention provides transgenic plants characterized by increased nitrogen use efficiency. Nitrogen use efficiency may be expressed as plant yield per given amount of nitrogen, In the Examples provided herein, the 25 transgene and control plants all received the same nutrient solutions in the same amounts. The transgenic plants were consistently characterized by higher yields, and thus have higher nitrogen use efficiencies. In yet another aspect, the invention provides transgenic plants and seeds thereof 30 with increased tolerance to high salt growth conditions. This aspect of the invention is exemplified by Example 24, which describes the germination of transgenic tobacco plant seeds in very high salt conditions (200 mM NaCI), While counterpart wild type tobacco seeds germinated at a rate of only about 10%, on 46 WO 2010/025466 PCT/US2009/055557 average, the transgenic tobacco seeds achieved nearly the same rate of germination obtained under no salt conditions for both transgenic and wild type seeds, or about 92%. 5 EXAMPLES Various aspects of the invention are further described and illustrated by way of the several examples which follow, none of which are intended to limit the scope of the invention. 10 EXAMPLE 1: ISLOATION OF ARABIDOPSIS GLUAMINE PHENYLPYRUVATE TRANSAMINASE (GPT) GENE: In an attempt to locate a plant enzyme that is directly involved in the synthesis of the signal metabolite 2-oxoglutaramate, applicants hypothesized that the putative 15 plant enzyme might bear some degree of structural relationship to a human protein that had been characterized as being involved in the synthesis of 2 oxoglutaramate. The human protein, glutamine transaminase K (E.C. 2.6.1,64) (also referred in the literature as cysteine conjugate R -lyase, kyneurenine aminotransferase, glutamine phenylpyruvate transaminase, and other names), 20 had been shown to be involved in processing of cysteine conjugates of halogenated xenobiotics (Perry et al, 1996, FEBS Letters 360:277-280). Rather than having an activity involved in nitrogen metabolism, however, human cysteine conjugate 1-lyase has a detoxifying activity in humans, and in animals. Nevertheless, the potential involvement of this protein in the synthesis of 2 25 oxoglutaramate was of interest. Using the protein sequence of human cysteine conjugate C-lyase, a search against the TIGR Arabidopsis plant database of protein sequences identified one potentially related sequence, a polypeptide encoded by a partial sequence at the 30 Arabidopsis gene locus at Atiq77670, sharing approximately 36% sequence homology/identity across aligned regions. 47 WO 2010/025466 PCT/US2009/055557 The full coding region of the gene was then amplified from an Arabidopsis cDNA library (Stratagene) with the following primer pair: 5'-CCCATCGATGTACC TGGACATAAATGGTGTGATG-3' 5 5'- GATGGTACCTCAGACTTTTCTCTTAAGCTTCTGCTTC3' These primers were designed to incorporate Cla I (ATCGAT) and Kpn I (GGTACC) restriction sites to facilitate subsequent subcloning into expression vectors for generating transgenic plants. Takara ExTaq DNA polymerase enzyme 10 was used for high fidelity PCR using the following conditions: initial denaturing 94C for 4 minutes, 30 cycles of 94C 30 second, annealing at 55C for 30 seconds, extension at 72C for 90 seconds, with a final extension of 72C for 7 minutes. The amplification product was digested with Cla I and Kpn I restriction enzymes, isolated from an agarose gel electrophoresis and ligated into vector pMon316 15 (Rogers, et. a[. 1987 Methods in Enzymology 153:253-277) which contains the cauliflower mosaic virus (CaMV, also CMV) 35S constitutive promoter and the nopaline synthase (NOS) 3' terminator. The ligation was transformed into DH5a cells and transformants sequenced to verify the insert. 20 A 1.3 kb cDNA was isolated and sequenced, and found to encode a full length protein of 440 amino acids in length, including a putative chloroplast signal sequence. EXAMPLE 2: PRODUCTION OF BIOLOGICALLY ACTIVE RECOMBINANT 25 ARABIDOPSIS GLUTAMINE PHENYLPYRUVATE TRANSAMINASE (GPT): To test whether the protein encoded by the cDNA isolated as described in Example 1, supra. is capable of catalyzing the synthesis of 2- oxoglutaramate, the cDNA was expressed in E coli, purified, and assayed for its ability to synthesize 30 2-oxoglutaramate using a standard method, NMR Assay for 2-oxoglutaramate: Briefly, the resulting purified protein was added to a reaction mixture containing 150 mM Tris-HCl, pH 8.5, 1 mM beta mercaptoethanol, 200 mM glutamine, 100 48 WO 2010/025466 PCT/US2009/055557 mM glyoxylate and 200 rnicroM pyridoxal 5'-phosphate. The reaction mixture without added test protein was used as a control. Test and control reaction mixtures were incubated at 374C for 20 hours, and then clarified by centrifugation to remove precipitated material. Supernatants were tested for the presence and 5 amount of 2-oxoglutaramate using 1C NMR with authentic chemically synthesized 2-oxoglutaramate as a reference, The products of the reaction are 2 oxoglutaramate and glycine, while the substrates (glutamine and glyoxylate) diminish in abundance, The cyclic 2-oxoglutaramate gives rise to a distinctive signal allowing it to be readily distinguished from the open chain glutamine 10 precursor. HPLC Assay for 2-oxoglutaramate: An alternative assay for GPT activity uses HPLC to determine 2-oxoglutaramate production, following a modification of Calderon et al., 1985, J Bacteriol 161(2): 15 807-809. Briefly, a modified extraction buffer consisting of 25 mM Tris-HCI pH 8,5, 1 mM EDTA, 20 pM FAD! 10 mM Cysteine, and -1.5% (v/v) Mercaptoethanol. Tissue samples from the test material (i.e., plant tissue) are added to the extraction buffer at approximately a 1/3 ratio (w/v), incubated for 30 minutes at 37"C, and stopped with 2 0 0 pl of 20% TCA. After about 5 minutes, the assay 20 mixture is centrifuged and the supernatant used to quantify 2-oxoglutaramate by HPLC, using an ION-300 7.8mm ID X 30 cm L column, with a mobile phase in 0,01N h2SO4, a flow rate of approximately 0.2 ml/min, at 4&C. Injection volume is approximately 20 d, and retention time between about 38 and 39 minutes. Detection is achieved with 210nm UV light. 25 Results Using NMR Assay: This experiment revealed that the test protein was able to catalyze the synthesis of 2- oxoglutaramate. Therefore, these data indicate that the isolated cDNA encodes a glutamine phenylpyruvate transaminase that is directly involved in the 30 synthesis of 2-oxoglutaramate in plants, Accordingly, the test protein was designated Arabidopsis glutamine phenylpyruvate transaminase, or "GPT". 49 WO 2010/025466 PCT/US2009/055557 The nucleotide sequence of the Arabidopsis GPT coding sequence is shown in the Table of Sequences, SEQ ID NO. 1. The translated amino acid sequence of the GPT protein is shown in SEQ ID NO. 2, 5 EXAMPLE 3: CREATION OF TRANSGENIC TOBACCO PLANTS OVER EXPRESSING ARABIDOPSIS GPT: Generation of Plant Expression Vector pMON-PJU: 10 Briefly, the plant expression vector pMon316-PJU was constructed as follows. The isolated cDNA encoding Arabidopsis GPT (Example 1) was cloned into the Clal-Kpni polylinker site of the pMON316 vector, which places the GPT gene under the control of the constitutive cauliflower mosaic virus (CaMV) 35S promoter and the nopaline synthase (NOS) transcriptional terminator. A 15 kanamycin resistance gene was included to provide a selectable marker. Agrobacteriurn-Mediated Plant Transformations: pMON-PJU and a control vector pMon316 (without inserted DNA) were transferred to Agrobacterium tumefaciens strain pTiTT37ASE using a standard 20 electroporation method (McCormac et al, 1998, Molecular Biotechnology 9:155 159), followed by plating on LB plates containing the antibiotics spectinomycin (100 micro gm I ml) and kanamycin (50 micro gm I ml). Antibiotic resistant colonies of Agrobacterium were examined by PCR to assure that they contained plasmid, 25 Nicotiana tabacum cv. Xanthi plants were transformed with pMON-PJU transformed Agrobacteria using the leaf disc transformation system of Horsch et. al. (Horsch et al.,1995, Science 227:1229-1231). Briefly, sterile :leaf disks were inoculated and cultured for 2 days, then transferred to selective MS media 30 containing 100 pg/mI kanamycin and 500 pg/ml clafaran, Transformants were confirmed by their ability to form roots in the selective media. Generation of GPT Transgenic Tobacco Plants: Sterile leaf segments were allowed to develop callus on Murashige & Skoog 35 (M&S) media from which the transformant plantlets emerged. These plantlets 50 WO 2010/025466 PCT/US2009/055557 were then transferred to the rooting-permissive selection medium (M&S medium with kanamycin as the selection agent). The healthy, and now rooted, transformed tobacco plantlets were then transferred to soil and allowed to grow to maturity and upon flowering the plants were selfed and the resultant seeds were 5 harvested. During the growth stage the plants had been examined for growth phenotype and the C02 fixation rate was measured for many of the young transgenic plants. Production of TI and T2 Generation GPT Transgenic Plants: 10 Seeds harvested form the T 3 generation of the transgenic tobacco plants were germinated on M&S media containing kanamycin (100 mg / L) to enrich for the transgene, At least one fourth of the seeds did not germinate on this media (kanamycin is expected to inhibit germination of the seeds without resistance that would have been produced as a result of normal genetic segregation of the gene) 15 and more than half of the remaining seeds were removed because of demonstrated sensitivity (even mild) to the kanamycin. The surviving plants (T 1 generation) were thriving and these plants were then selfed to produce seeds for the T 2 generation. Seeds from the T generation were 20 germinated on MS media supplemented for the transformant lines with kanamycin (10mg/liter). After 14 days they were transferred to sand and provided quarter strength Hoagland's nutrient solution supplemented with 25 mM potassium nitrate, They were allowed to grow at 244C with a photoperiod of 16 h light and 8 hr dark with a light intensity of 900 micromoles per meter squared per second. They were 25 harvested 14 days after being transferred to the sand culture. Characterization of GPT Transgenic Plants: Harvested transgenic plants (both GPT transgenes and vector control transgenes) were analyzed for glutamine sythetase activity in root and leaf, whole plant fresh 30 weight, total protein in root and leaf, and C02 fixation rate (Knight et aL, 1988, Plant Physiol. 88: 333). Non-transformed, wild-type A. tumefaciens plants were also analyzed across the same parameters in order to establish a baseline control. 51 WO 2010/025466 PCT/US2009/055557 Growth characteristic results are tabulated below in Table . Additionally, a photograph of the GPT transgenic plant compared to a wild type control plant is shown in FIG. 2 (together with GS1 transgenic tobacco plant, see Example 5). Across all parameters evaluated, the GPT transgenic tobacco plants showed 5 enhanced growth characteristics. In particular, the GPT transgenic plants exhibited a greater than 50% increase in the rate of C02 fixation, and a greater than two-fold increase in glutamine synthetase activity in leaf tissue, relative to wild type control plants. In addition, the leaf-to-root GS ratio increased by almost three-fold in the transaminase transgenic plants relative to wild type control. 10 Fresh weight and total protein quantity also increased in the transgenic plants, by about 50% and 80% (leaf), respectively, relative to the wild type control. These data demonstrate that tobacco plants overexpressing the Arabidopsis GPT transgene achieve significantly enhanced growth and CO2 fixation rates. 15 Table I Protein mg/gram fresh weight Leaf Root Wild type - control 8.3 2.3 Une PN1-8a second control 8.9 2.98 Line PN9-9 13Y 3.2 Glutamine Synthetase activity, micromoles/min/mg protein Wild type (Ratio of leaf I root = 411) 4.3 1.1 PN1-8 (Ratio of leaf: root= 42: 1) 5.2 1.3 PN9-9 (Ratio of leaf : root= 10.9: 1) 10.5 0.97 Whole Plant Fresh Weig t g .................. PN 1-8 26.1 PN9-9 33.1 C02 Fixation Rate, umole/m21sec Wild type 8.4 PN1-8 8.9 PN9-9 12,9 Data= average of three plants Wild type - control plants; not regenerated or transformed. PNI lines were produced by regeneration after transformation using a construct without inserted gene. 20 A control against the processes of regeneration and transformation. PN 9 lines were produced by regeneration after transformation using a construct with the Arabidopsis GPT gene. 52 WO 2010/025466 PCT/US2009/055557 EXAMPLE 4: GENERATION OF TRANSGENIC TOMATO PLANTS CARRYING ARABIDOPSIS GPT TRANSGENE: Transgenic Lycopersicon esculenturn (Micro-Tom Tomato) plants carrying the 5 Arabidopsis GPT transgene were generated using the vectors and methods described in Example 3. To transgenic tomato plants were generated and grown to maturity, Initial growth characteristic data of the GPT transgenic tomato plants is presented in Table H. The transgenic plants showed significant enhancement of growth rate, flowering, and seed yield in relation to wild type control plants. In 10 addition, the transgenic plants developed multiple main stems, whereas wild type plants developed with a single main stem, A photograph of a GPT transgenic tomato plant compared to a wild type plant is presented in FIG. 3 (together with GS1 transgenic tomato plants, see Example 6). Is TABLE 11 Growth Wildtype GPT Transgenic Characteristics Tomato Tomato Stem height, cm 6 5 18, 12,11 major stems Stems 1 3 major, 0 other Buds 2 16 Flowers 8 12 Fruit 0 3 EXAMPLE 5: GENERATION OF TRANSGENIC TOBACCO PLANTS 20 OVEREXPRESSING ALFALFA GS1: Generation of Plant Expression Vector pGSI 11: Transgenic tobacco plants overexpressing the Alfalfa GS1 gene were generated as previously described (Temple et al., 1993, Mot Gen. Genetics 236: 315-325). 25 Briefly, the plant expression vector pGSI 11 was constructed by inserting the entire coding sequence together with extensive regions of both the 5' and 3' untranslated regions of the Alfalfa GS1 gene [SEQ ID NO: 3] (DasSarma at al, 1986, Science, Vol 232, Issue 4755, 1242-1244) into pMON316 (Rogers et at, 1987, supra), placing the transgene under the control of the constitutive 30 cauliflower mosaic virus (CaMV) 353 promoter and the nopaline synthase (NOS) 53 WO 2010/025466 PCT/US2009/055557 transcriptional terminator. A kanamycin resistance gene was included to provide a selectable marker. Generation of GS1 Transformants: 5 pGS111 was transferred to Agrobacterium tumefaciens strain pTiTT37ASE using triparental mating as described (Rogers et at, 1987, supra; Unkefer et al, U.S. Patent No. 6,555,500). Nicotiana tabacum cv, Xanthi plants were transformed with pGS111 transformed Agrobacteria using the leaf disc transformation system of Horsch et. at (Horsch et at,1995, Science 227:1229-1231). Transformants 10 were selected and regenerated on MS medium containing 100pg/ml kanamycin. Shoots were rooted on the same medium (with kanamycin, absent hormones) and transferred to potting soilperlite:vermiculite (3:1:1), grown to maturity, and allowed to self. Seeds were harvested from this To generation, and subsequence generations produced by selfing and continuing selection with kanamycin. The 15 best growth performers were used to yield a T3 for crossing with the best performing GPT over-expressing lines identified as described in Example 3, A photograph of the GS1 transgenic plant compared to a wild type control plant is shown in FIG, 2 (together with GPT transgenic tobacco plant, see Example 3) 20 EXAMPLE 6: GENERATION OF TRANSGENIC TOMATO PLANTS CARRYING ALFALFA GS1 TRANSGENE: Transgenic Lycopersicon esculentum (Micro-Tom Tomato) plants carrying the 25 Alfalfa GS1 transgene were generated using the vector described in Example 5 and a transformation protocol essentially as described (Sun et at, 2006. Plant Cell Physiol. 46(3) 426-31). To transgenic tomato plants were generated and grown to maturity. Initial growth characteristic data of the GPT transgenic tomato plants is presented in Table ll. The transgenic plants showed significant enhancement of 30 growth rate, flowering, and seed yield in relation to wild type control plants. In addition, the transgenic plants developed multiple main stems, whereas wild type plants developed with a single main stem. A photograph of a GS1 transgenic tomato plant compared to a wild type plant is presented in FIG. 3 (together with GPT transgenic tomato plant, see Example 4). 35 54 WO 2010/025466 PCT/US2009/055557 TABLE Ill Growth Wildtype GS1 Transgenic Characteristics Tomato Tomato Stem height, cm 6,5 16, 7, 5 major stems Stems 1 3 major, 3 med, I sm Buds 2 2 Flowers 8 13 Fruit 0 4 5 EXAMPLE 7: GENERATION OF DOUBLE TRANSGENIC TOBACCO PLANTS CARRYING GS1 AND GPT TRANSGENES: In an effort to determine whether the combination of GS1 and GPT transgenes in a single transgenic plant might improve the extent to which growth and other 10 agronomic characteristics may be enhanced, a number of sexual crosses between high producing lines of the single transgene (GS1 or GPT) transgenic plants were carried out. The results obtained are dramatic, as these crosses repeatedly generated progeny plants having surprising and heretofore unknown increases in growth rates, biomass yield, and seed production. 15 Materials and Methods: Single-transgene, transgenic tobacco plants overexpressing GPT or GS1 were generated as described in Examples 3 and 4, respectively. Several of fastest growing T 2 generation GPT transgenic plant lines were crossed with the fastest 20 growing T3 generation GS1 transgenic plant lines using reciprocal crosses. The progeny were then selected on kanamycin containing M&S media as described in Example 3, and their growth, flowering and seed yields examined. Tissue extractions for GPT and GS activities; GPT activity was extracted from 25 fresh plant tissue after grinding in cold 100 mM Tris-HCI, pH 7.6, containing 1 mm ethylenediaminetetraacetic, 200 mM pyridoxal phosphate and 6 mM mercaptoethanol in a ratio of 3 ml per gram of tissue. The extract was clarified by centrifugation and used in the assay. GS activity was extracted from fresh plant tissue after grinding in cold 50 mM Imidazole, pH 7.5 containing 10 mM MgC12, 30 and 12.5 mM mercaptoethano in a ratio of 3 ml per gram of tissue. The extract 55 WO 2010/025466 PCT/US2009/055557 was clarified by centrifugation and used in the assay, GPT activity was assayed as described in Calderon and Mora, 1985, Journal Bacteriology 161:807-809. GS activity was measured as described in Shapiro and Stadtmann, 1970, Methods in Enzymology 17A: 910-922. Both assays involve an incubation with substrates 5 and cofactor at the proper pH. Detection was by HPLC. Results: The results are presented in two ways, First, specific growth characteristics are 10 tabulated in Tables IVA and IV.B (biomass, seed yields, growth rate, GS activity, GPT activity, 2-oxoglutaramate activity, etc). Second, photographs of progeny plants and their leaves are shown in comparison to single-transgene and wild type plants and leaves are presented in FIG. 5 and FIG. 6, which show much larger whole plants, larger leaves, and earlier and/or more abundant flowering in 15 comparison to the parental single-transgene plants and wild type control plants. Referring to Table IV.A, double-transgene progeny plants form these crosses showed tremendous increases total biomass (fresh weight), with fresh weights ranging from 45-89 grams per individual progeny plant, compared to a range of 20 only 19-24 grams per individual wild type plant, representing on average, about a two- to three-fold increase over wild type plants, and representing at the high end, an astounding four-fold increase in biomass over wild type plants. Taking the 24 individual double-transgene progeny plants evaluated, the average individual plant biomass was about 2.75 times that of the average wild type control plant. 25 Four of the progeny lines showed approximately 2.5 fold greater average per plant fresh weights, while two lines showed over three-fold greater fresh weights in comparison to wild type plants. In comparison to the single-transgene parental lines, the double-transgene 30 progeny plants also showed far more than an additive growth enhancement. Whereas GPT single-transgene lines show as much as about a 50% increase over wild type biomass, and GS1 single-transgene lines as much as a 66% increase, progeny plants averaged almost a 200% increase over wild type plants. 56 WO 2010/025466 PCT/US2009/055557 Similarly, the double transgene progeny plants flowered earlier and more prolifically than either the wild type or single transgene parental lines, and produced a far greater number of seed pods as well as total number of seeds per plant, Referring again to Table IV.A, on average, the double-transgene progeny 5 produced over twice the number of seed pods produced by wild type plants, with two of the high producer plants generating over three times the number of seed pods compared to wild type. Total seed yield in progeny plants, measured on a per plant weight basis, ranged from about double to nearly quadruple the number produced in wild type plants, 57 WO 2010/025466 PCT/US2009/055557 S-112,983 TABLE IV.A PLANT LINE FRESH WEIGHT SEED PODS SEED YIELD GS ACTIVITY glwhole plant #pods/plant glplant LEAF ROOT LIR RATIO Wild Type Tobacco______ Wild type 1 18.73 26 0.967 Wild type 2 24.33 24 1.07 Wild type 3 23.6 32 0.9 Wild type 4 18.95 32 1.125 W7 Average 21.4025 28.5 1.0155 7.75 145 5.34 Cross I X1L1a x PA9-9ff 1 59.21 62 2.7811 - 2 65.71 56 3 55.36 72 4 46.8 56 Cross I Average 56.77 61.5 14.98 1.05 14.27 Compared to WT +265% +216% +274% +193% -28% +267% Cross 2 PA9-2 x L9 1 70.83 61 1.76 2 49.17 58 3.12 3 50.23 90 NA 4 45.77 Cross 2 Average 54 58.3 2A4 16.32 1.81 9.02 Compared to WT +252% +205% +240% +211% +125% +169% Cross 3 PA9-9ff xLla 1 89.1 77 3.687 2 78.18 3 58.34 4 61.79 Cross 3 Average 71.85 77 (one plant) 3.678 (one plant) 15.92 1.38 11.54 Compared to WT +336% +270% +362% +205% -5% +216% 58 WO 2010/025466 PCT/US2009/055557 PLANT LINE FRESH WEIGHT SEED PODS SEED YIELD GS ACTIVITY g/whole plant #pods/plant g/plant LEAF ROOT LIR RATIO Cross 5 PA9-1Oaa x LIa 1 65.34 45 2.947 2 53.28 64 3,3314 3 49.85 42 1.5667 4 44.63 42 2.5013 Cross 5 Average 53.275 48.25 2.86928 13.03 1.8 7.24 Compared to WT +244% +169% +283% +168% Cross 6 PA9-17b x Lia 1 56.7 64 2.492 2 55.05 66 2.162 3 51.51 59 1.8572 4 45.38 72 4.742 Cross 6 Average 52.16 65.25 2.8133 14.114,7 1.1.1124 13.29 Compared to WT +244% +229% +277% 52 Cross 7 PA9-20aa x L1b 1 76.26 67 2.0535 2 66.27 42 1.505 3 72.26 72 2.3914 4 63.91 91 2.87 Cross 7 Average 69.675 68 2.204975 14.12 1.24 11.39 Compared to WT +326% +239% +217% Control PA9-9ff 1 32.18 N/A 2 32.64 N/A 3 34.67 N/A 4 25.18 N/A Average 31.17 N/A 11.57 1.14 10.15 Compared to WT +148% 59 WO 2010/025466 PCT/US2009/055557 PLANT LINE FRESH WEIGHT SEED PODS SEED YIELD GS ACTIVITY g/whole plant #pods/plant g/plant LEAF ROOT L/R RATIO Control GS Lia 1 41.74 N/A 2 36.24 N/A 3 33.8 N/A 4 30.48 N/A Average 35.57 N/A 13.15 1.23 10.69 Compared to WT +166% 1 1 1 1 _ 1 60 WO 2010/025466 PCT/US2009/055557 Table IV.B shows growth rate, biomass and yield, and biochemical characteristics of Line XX (Line 3 further selfed) compared to the single transgene line expressing GS1 and wild type control tobacco. All parameters are greatly increased in the double transgenic plant (Line XX). Notably, 2-oxoglutaramate activity was almost 17-fold 5 higher, and seed yield and foliar biomass was three-fold higher, in Line XX plants versus control plants. TABLE IV.B Specific GS GPT 2 Plant Growth oliar Fruit / Seed Activity Activ oxoglu- Trans Type Rath Biomass Flowers umol/ taramate Gene Type mg/g/d FWt, g /Buds Yield g min/gFW /gFWt nmol/gF Assay m// Wt wt Wildtype, 228 21.40 28.5 1.02 7.75 16.9 68.9 No Line 1 GS 269 35.57 NM NM 11.6 NM 414 Yes Line XX 339 59.71 62.9 2.94 16.3 243.9 1,153.6 Yes NM Not Measured 10 EXAMPLE 8: GENERATION OF DOUBLE TRANSGENIC PEPPER PLANTS CARRYING GS1 AND GPT TRANSGENES: 15 In this example, Big Jim chili pepper plants (New Mexico varietal) were transformed with the Arabidopsis GPT full length coding sequence of SEQ ID NO: 1 under the control of the CMV 35S promoter, and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter, using Agrobacterium-mediated transfer to seed pods. After 3 days, seeds were harvested and used to generate TO 20 plants and screened for transformants. The resulting double-transgenic plants showed higher pod yields, faster growth rates, and greater biomass yields in comparison to the control plants. 61 WO 2010/025466 PCT/US2009/055557 Materials and Methods: Solanaceae Capisicurn Pepper plants ("Big Jim" varietal) were transformed with the Arabidopsis GPT full length coding sequence of SEQ ID NO: 1 under the control of 5 the CMV 35S promoter within the expression vector pMON (see Example 3), and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCambia 1201 (Tomato rubisco rbcS3C promoter: Kyozulka et al,, 1993, Plant Physiol. 103: 991-1000; SEQ ID NO: 22; vector construct of SEQ ID NO: 6), using Agrobacterium-mediated transfer to seed 10 pods. For this and all subsequent examples, the Cambia 1201 or 1305.1 vectors were constructed according to standard cloning methods (Sambrook et aL, 1989, supra, Saiki et al., 1988, Science 239: 487-491). The vector is supplied with a 35S CaMV 15 promoter; that promoter was replaced with RcbS-3C promoter from tomato to control the expression of the target gene. The Cambia 1201 vectors contain bacterial chlorophenicol and plant hygromycin resistance selectable marker genes. The Cambia 1305.1 vectors contain bacterial chlorophenicol and hygromycin resistance selectable marker genes. 20 The transgene expression vectors pMON (GPT transgene) and pCambia 1201 (GS transgene) were transferred to separate Agrobacterium tumefaciens strain LBA4404 cultures using a standard electroporation method (McCormac et al, 1998, Molecular Biotechnology 9:155-159). Transformed Agrobacteriurn were selected on media 25 containing 50 pg/ml of either streptamycin for pMON constructs or chloroamphenicol for the Cambia constructs, Transformed Agrobacterium cells were grown in LB culture media containing 25 pg/mI of antibiotic for 36 hours, At the end of the 36 hr growth period cells were collected by centrifugation and cells from each transformation were resuspended in 100 ml LB broth without antibiotic. 30 62 WO 2010/025466 PCT/US2009/055557 Pepper plants were then transformed with a mixture of the resulting Agrobacterium cell suspensions using a transformation protocol in which the Agrobacterium is injected directly into the seed cavity of developing pods. Briefly, developing pods were injected with the 200 ml mixture in order to inoculate immature seeds with the 5 Agrobacteria essentially as described (Wang and Waterhouse, 1997, Plant Mol. Biol. Reporter 15: 209-215). In order to induce Agrobacteria virulence and improve transformation efficiencies, 10 pg/ml acetosyringonone was added to the Agrobacteria cultures prior to pod inoculations (see, Sheikholeslam and Weeks, 1986, Plant Mot Bio. 8: 291-298). 10 Using a syringe, pods were injected with a liberal quantity of the Agrobacterium vector mixture, and left to incubate for about 3 days. Seeds were then harvested and germinated, and developing plants observed for phenotypic characteristics including growth and antibiotic resistance. Plants carrying the transgenes were green, whereas 15 untransformed plants showed signs of chlorosis in leaf tips, Vigorous growing transformants were grown and compared to wild type pepper plants grown under identical conditions. 20 Results: The results are presented in FIG. 7 and Table V. FIG. 7 shows a photograph of a GPT+GS double transgenic pepper plant compared to a control plant grown for the same time under identical conditions, This photograph shows tremendous pepper 25 yield in the transgenic line compared to the control plant, Table V presents biomass yield and GS activity, as well as transgene genotyping, in the transgenic lines compared to the wild type control. Referring to Table V, double transgene progeny plants showed tremendous increases total biomass (fresh 30 weight), with fresh weights, ranging from 393 - 662 grams per individual transgenic plant, compared to an average of 328 grams per wild type plant. Transgenic line A5 63 WO 2010/025466 PCT/US2009/055557 produced more than twice the total biomass of the controls. Moreover, pepper yields in the transgenic lines were greatly improved over wild type plants, and were 50% greater than control plants (on average). Notably, one of the transgene lines produced twice as many peppers as the control plant average. 5 TABLE V: TRANSGENIC PEPPER GROWTH/BIOMASS AND REPRODUCTION Biomass, Yield GS activity Transgene Plant type Foliar Fresh Peppers, g Umoles/min Presence Wt, g DWt /gFWt Assay Wildtype, avg 328,2 837 1.09 Negative Line A2 457,3 184,2 1.57 GPT- Yes Line A5 661.7 148.1 1,8 GPT - Yes Line B1 493,4 141,0 1.3 GPT - Yes Line B4 393,1 136,0 1.6 GPT - Yes Line C1 509A 152.9 1.55 GPT - Yes FWt Fresh Weight; DWt Dry Weight 10 EXAMPLE 9: GENERATION OF DOUBLE TRANSGENIC BEAN PLANTS CARRYING ARABIDOPSIS GS1 AND GPT TRANSGENES: 15 In this example, yellow wax bean plants (Phaseolus vulgaris) were transformed with the Arabidopsis GPT full length coding sequence of SEQ ID NO: 1 under the control of the CMV 35S promoter within the expression vector pCambia 1201, and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCambia 1201, using Agrobacterium-mediated 20 transfer into flowers. Materials and Methods: The transgene expression vectors pCambia 1201-GPT (vector construct of SEQ ID 25 NO: 27) and pCambia 1201-GS (vector construct of SEQ ID NO: 6) were transferred to separate Agrobacterium tumefaciens strain LBA4404 cultures using a standard 64 WO 2010/025466 PCT/US2009/055557 electroporation method (McCormac et al., 1998, Molecular Biotechnology 9:155-159). Transformed Agrobacterium were selected on media containing 50 pg/mI of chloroamphenicol. Transformed Agrobacterium cells were grown in LB culture media containing 25 pg/ml of antibiotic for 36 hours. At the end of the 36 hr growth period 5 cells were collected by centrifugation and cells from each transformation were resuspended in 100 ml LB broth without antibiotic. Bean plants were then transformed with a mixture of the resulting Agrobacterium cell suspensions using a transformation protocol in which the Agrobacteria is injected 10 directly into the flower structure (Yasseem, 2009, Plant Mol. Biol. Reporter 27: 20 28), In order to induce Agrobacteria virulence and improve transformation efficiencies, 10 pg/mI acetosyringonone was added to the Agrobacteria cultures prior to flower inoculation. :Briefly, once flowers bloomed, the outer structure encapsulating the reproductive organs was gently opened with forceps in order to 15 permit the introduction of the Agrobacteria mixture, which was added to the flower structure sufficient to flood the anthers, Plants were grown until bean pods developed, and seeds were harvested and used to generate transgenic plants. Transgenic plants were then grown together with 20 control bean plants under identical conditions, photographed and phenotypically characterized. Growth rates were measured for both transgenic and control plants. In this and all examples, Glutamine synthetase (GS) activity was assayed according to the methods in Shapiro and Stadtmann, 1970, Methods in Enzymology 17A: 910 922; and, Glutamine phenylpyruvate transaminase (GPT) activity was assayed 25 according to the methods in Calderon et aL, 1985, J. Bacteriol. 161: 807-809. See details in Example 7, Methods, supra, Results: 30 The results are presented in FIG. 8, FIG. 9 and Table VI, 65 WO 2010/025466 PCT/US2009/055557 FIG. 8 shows GPT+GS transgenic bean line A growth rate data relative to control plants, including plant heights on various days into cultivation, as well as numbers of flower buds, flowers, and bean pods. These data show that the GPT+GS double 5 transgenic bean plants outgrew their counterpart control plants. The transgenic plants grew taller, flowered earlier and produced more flower buds and flowers, and developed bean pods and produced more bean pods that the wild type control plants. TABLE Vi: TRANSGENIC BEANS LINE A 10 GPT Activity S Activity Plant Type Bean Pod nmoles/h/gF umoles/min Antibiotic Yield FWt, g Wt /gFWt Resistance Wildtype, avg 126.6 101.9 25.2 Negative 2A 211.5 NM NM + 4A 207,7 NM NM + 58 205.7 984.7 101.3 + WT Wildtype; FWt Fresh Weight; NM Not Measured Table VI presents bean pod yield, GPT and GS activity, as well as antibiotic 15 resistance status, in the transgenic lines compared to the wild type control (average of several robust control plants; control plants that did not grow well were excluded from the analyses), Referring to Table VI, double-transgene progeny plants showed substantial bean pod biomass increases (fresh pod weight) in comparison to the control plants, with bean pod biomass yields consistently above 200 grams per 20 individual transgenic plant, compared to an average of 127 grams per wild type plant, representing an over 60% increase in pod yield in the double transgene lines relative to control plant(s). 66 WO 2010/025466 PCT/US2009/055557 Lastly, FIG, 9 shows a photograph of a GPT+GS double transgenic bean plant compared to a control plant grown for the same time under identical conditions, showing increased growth in the transgenic plant. 5 EXAMPLE 10: GENERATION OF DOUBLE TRANSGENIC BEAN PLANTS CARRYING ARABIDOPSIS GS1 AND GRAPE GPT TRANSGENES: In this example, yellow wax bean plants (Phaseolus vulgaris) were transformed with 10 the Grape GPT full length coding sequence of SEQ ID NO: 8 under the control of the RuBisCo promoter within the expression vector pCambia 1305.1, and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCambia 1201, using Agrobacterium-mediated transfer into developing pods. 15 Materials and Methods: The transgene expression vectors pCambia 1201-GPT(grape) (vector construct of SEQ ID NO: 8) and pCambia 1201-GS (vector construct of SEQ ID NO: 6) were 20 transferred to separate Agrobacterium tumefaciens strain LBA4404 cultures using a standard electroporation method (McCormac et al., 1998, Molecular Biotechnology 9:155-159), Transformed Agrobacterium were selected on media containing 50 pg/mI of chloroamphenicol. Transformed Agrobacterium cells were grown in LB culture media containing 25 pg/mI of antibiotic for 36 hours. At the end of the 36 hr 25 growth period cells were collected by centrifugation and cells from each transformation were resuspended in 100 ml LB broth without antibiotic, Bean plants were then transformed with a mixture of the resulting Agrobacterium cell suspensions using a transformation protocol in which the Agrobacteria is injected 30 directly into the flower structure. In order to induce Agrobacteria virulence and improve transformation efficiencies, 10 pg/mI acetosyringonone was added to the 67 WO 2010/025466 PCT/US2009/055557 Agrobacteria cultures prior to flower inoculation. Briefly, once flowers bloomed, the outer structure encapsulating the reproductive organs was gently opened with forceps in order to permit the introduction of the Agrobacteria mixture, which was added to the flower structure sufficient to flood the anthers. 5 Plants were grown until bean pods developed, and seeds were harvested and used to generate transgenic plants, Transgenic plants were then grown together with control bean plants under identical conditions, photographed and phenotypically characterized, Growth rates were measured for both transgenic and control plants. 10 Results: The results are presented in FIG, 10, FIG. 11 and Table VII. 15 FIG. 10 shows GPT+GS transgenic bean line G growth rate data relative to control plants, specifically including numbers of flower buds, flowers, and bean pods. These data show that the GPT+GS double transgenic bean plants outgrew their counterpart control plants. Notably, the transgenic plants produced substantially more bean pods that the wild type control plants 20 TABLE VII: TRANSGENIC BEANS LINE G: POD YIELDS Plant Type Bean Pod Yield FWt, g Antibiotic Resistance Wild type, avg 157.9 Negative GI 200,5 + G2 178.3 + WT Wildtype; FWt Fresh Weight; NM Not Measured 25 Table VII presents bean pod yield and antibiotic resistance status, in the transgenic lines compared to the wild type control (average of several robust control plants; control plants that did not grow well were excluded from the analyses) Referring to Table VI, double-transgene progeny plants showed substantial bean pod biomass 68 WO 2010/025466 PCT/US2009/055557 increases (fresh pod weight) in comparison to the control plants, with bean pod biomass yields of 200.,5 (line GI) and 178 grams (line G2) per individual transgenic plant, compared to an average of 158 grams per individual wild type plant, representing approximately a 27% increase in pod yield in the double transgene lines 5 relative to control plants. Lastly, FIG. 11 shows a photograph of a GPT+GS double transgenic bean plant compared to a control plant grown for the same time under identical conditions. The transgenic plant shows substantially increased size and biomass, larger leaves and a 10 more mature flowering compared to the control plant. EXAMPLE 11: GENERATION OF DOUBLE TRANSGENIC COWPEA PLANTS CARRYING ARABIDOPSIS GS1 AND GPT TRANSGENES: 15 In this example, common Cowpea plants were transformed with the Arabidopsis GPT full length coding sequence of SEQ ID NO: 1 under the control of the CMV 35S promoter within the expression vector pMON, and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCambia 1201, using Agrobacterium-mediated transfer into 20 flowers. Materials and methods were as in Example 9, supra. Results: The results are presented in FIGS. 12 and 13, and Table VIL FIG. 12 shows relative 25 growth rates for the GPT+GS transgenic Cowpea line A and wild type control Cowpea at several intervals during cultivation, including (FIG. 12A) height and longest leaf measurements, (FIG. 12B) trfolate leafs and flower buds, and (FIG. 12C) flowers, flower buds and pea pods. These data show that the GPT+GS double transgenic Cowpea plants outgrew their counterpart control plants, The transgenic 69 WO 2010/025466 PCT/US2009/055557 plants grew faster and taller, had longer leaves, and set flowers and pods sooner than wild type control plants. TABLE Vill: TRANSGENIC COWPEA LINE A 5 Pea Pod GPT Activity GS Activity Plant Type Yield, ramoles/h/gF umol/min/gF Antibiotic FWt, g Wt Wt Resistance Wildtype, avg 74.7 44,4 28.3 Negative 4A 112.8 NM 41.3 + 8B 113,8 736,2 54.9 + WT Wildtype; FWt Fresh Weight; NM Not Measured Table Vill presents pea pod yield, GPT and GS activity, as well as antibiotic resistance status, in the transgenic lines compared to the wild type control (average of several robust control plants; control plants that did not grow well were excluded 10 from the analyses). Referring to Table Vill, double-transgene progeny plants showed substantial pea pod biomass increases (fresh pod weight) in comparison to the control plants, with average transgenic plant pea pod biomass yields nearly 52% greater than the yields measured in control plant(s). 15 Lastly, FIG. 13 shows a photograph of a GPT+GS double transgenic bean plant compared to a control plant grown for the same time under identical conditions, showing increased biomass and pod yield in the transgenic plant relative to the wild type control plant. 20 EXAMPLE 12: GENERATION OF DOUBLE TRANSGENIC COWPEA PLANTS CARRYING ARABIDOPSIS GS1 AND GRAPE GPT TRANSGENES: In this example, common Cowpea plants were transformed with the Grape GPT full 25 length coding sequence of SEQ ID NO: 8 under the control of the RuBisCo promoter within the expression vector pCambia 1305.1 (vector construct of SEQ ID NO: 8), 70 WO 2010/025466 PCT/US2009/055557 and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCambia 1201 (vector construct of SEQ ID NO: 6), using Agrobacterium-mediated transfer into flowers. Materials and methods were as in Example 11, supra. 5 Results: The results are presented in FIGS. 14 and 15, and Table IX. 10 FIG, 14 shows relative growth rates for the GPT+GS transgenic Cowpea line G and wild type control Cowpea, These data show that the transgenic plants are consistently higher (FIG. 14A), produce substantially more flowers, flower buds and pea pods (FIG, 148), and develop trifolates and leaf buds faster (FIG. 14C). 15 TABLE IX: TRANSGENIC COWPEA LINE G Pod Yield, GPT Activity GS Activity Antibiotic Plant Type FWt, g nmoles/h/gF umol/min/gF Resistance WT Wt Wildtype, avg 597 44.4 26.7 Negative G9 102,0 555,6 34.5 + WT Wildtype; FWt Fresh Weight;NM Not Measured Table IX presents pea pod yield, GPT and GS activity, as well as antibiotic resistance 20 status, in the transgenic lines compared to the wild type control (average of several robust control plants; control plants that did not grow well were excluded from the analyses). Referring to Table IX, double-transgene progeny plants showed substantial pea pod biomass increases (fresh pod weight) in comparison to the control plants, with average pea pod biomass yields 70% greater in the transgenic 25 plants compared to control plant(s). 71 WO 2010/025466 PCT/US2009/055557 Lastly, FIG. 15 shows a photograph of a GPT+GS double transgenic pea plant compared to a control plant grown for the same time under identical conditions, showing increased height, biomass and leaf size in the transgenic plant relative to the wild type control plant, 5 EXAMPLE 13: GENERATION OF DOUBLE TRANSGENIC ALFALFA PLANTS CARRYING ARABIDOPSIS GSI AND GPT TRANSGENES: 10 In this example, Alfalfa plants (Medicago sativa, var Ladak) were transformed with the Arabidopsis GPT full length coding sequence of SEQ ID NO: 1 under the control of the CMV 35S promoter within the expression vector pMON316 (see Example 3, supra), and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCambia 1201 (vector 15 construct of SEQ ID NO: 6), using Agrobacterium-mediated transfer into seedling plants. Agrobacterium vectors and mixtures were prepared for seedling inoculations as described in Example 11, supra. Seedlings noculations: 20 When Alfalfa seedlings were still less than about 1/2 inch tall, they were soaked in paper toweling that had been flooded with the Agrobacteria mixture containing both transgene constructs, The seedlings were left in the paper toweling for two to three days, removed and then planted in potting soil. Resulting TO and control plants were then grown for the first 30 days in a growth chamber, thereafter cultivated in a 25 greenhouse, and then harvested 42 days after sprouting. At this point, only the transgenic Alfalfa line displayed flowers, as the wild type plants only displayed immature flower buds. The plants were characterized as to flowering status and total biomass. 30 72 WO 2010/025466 PCT/US2009/055557 Results: The results are presented in Table X, The data shows that the transgenic Alfalfa plants grew faster, flowered sooner, and yielded on average about a 62% biomass 5 increase relative to the control plants. TABLE X: TRANSGENIC ALFALFA VS. CONTROL Plant Type Biomass at Sacrifice, g Flowering Stage Small defined buds Wildtype, avg 6.03 No buds swelling. No flowers Transgene #5 10.38 4 Open flowers Transgene # 11 9.03 Flower buds swelling Transgene #13 9,95 Flower buds swelling 10 EXAMPLE 14: GENERATION OF DOUBLE TRANSGENIC CANTALOUPE PLANTS CARRYING ARABIDOPSIS GS1 AND GPT TRANSGENES: 15 In this example, Cantaloupe plants (Cucumis melo var common) were transformed with the Arabidopsis GPT full length coding sequence of SEQ ID NO: 1 under the control of the CMV 35S promoter within the expression vector pMON316 (see Example 3, supra), and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 20 under the control of the RuBisCo promoter within the expression vector pCambia 1201 (vector construct of SEQ ID NO: 6), using Agrobacterium-mediated transfer via injection into developing melons. Agrobacterium vectors and mixtures were prepared for intra-melon inoculations as described in Example 8, supra. Inoculations into developing melons were carried out essentially as described in Example 8. The 25 plants were characterized as to flowering status and total biomass relative to control melon plants grown under identical conditions. 73 WO 2010/025466 PCT/US2009/055557 The results are presented in FIG, 16 and Table Xl. Referring to Table XI, the transgenic plants showed substantial foliar plant biomass increases in comparison to the control plants, with an average increase in biomass of 63%. Moreover, a 5 tremendous increase in flower and flower bud yields was observed in all five transgenic lines. Control plants displayed no flowers and only 5 buds at sacrifice, on average. In sharp contrast, the transgenic plants displayed between 2 and 5 flowers per plant, and between 21 and 30 flower buds, per plant, indicating a substantially higher growth rate and flower yield, Increased flower yield would be expected to 10 translate into correspondingly higher melon yields in the transgenic plants. Referring to FIG. 16 (a photograph comparing transgenic Cantaloupe plants to control Cantaloupe plants), the transgenic Cantaloupe plants show dramatically increased height, overall biomass and flowering status relative to the control plants, 15 TABLE XI: TRANGENIC CANTALOUPE VERSUS CONTROL Biomass Flowers / Flower Antibiotic Foliar FWt, g Buds at Sacrifice Resistance Wildtype, avg 22,8 0/5 Negative Line 1 37.0 3/21 + Line 2 35,0 2/30 + Line 3 37,1 3/27 + Line 4 40,6 5/26 + Line 5 35.7 4/30 + FWt Fresh Weight 20 EXAMPLE 15: GENERATION OF DOUBLE TRANSGENIC PUMPKIN PLANTS CARRYING ARABIDOPSIS GS1 AND GPT TRANSGENES: 25 In this example, common Pumpkin plants (Cucurbita maxima) were transformed with the Arabidopsis GPT full length coding sequence of SEQ ID NO: I under the control 74 WO 2010/025466 PCT/US2009/055557 of the CMV 35S promoter within the expression vector pMON316 (see Example 3, supra), and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCambia 1201 (vector construct of SEQ ID NO: 6), using Agrobacterium-mediated transfer via injection into 5 developing pumpkins, essentially as described in Example 14, supra. The transgenic and control pumpkin plants were grown under identical conditions until the emergence of flower buds in the control plants, then all plants were characterized as to flowering status and total biomass. 10 The results are presented in FIG. 17 and Table XI. Referring to Table XII, the transgenic plants showed substantial foliar plant biomass increases in comparison to the control plants, with an increase in average biomass yield of 67% over control plants. Moreover, an increase in flower bud yields was observed in four of the five transgenic lines in comparison to control. Control plants displayed only 4 buds at 15 sacrifice (average). In contrast, four transgenic plant lines displayed between 8 and 15 flowers buds per plant, representing a two- to nearly four-fold yield increase, TABLE XII: TRANGENIC PUMPKIN VERSUS CONTROL Biomass Flower Buds at Antibiotic Plant Type Foliar FWt, g Sacrifice Resistance Wildtype, avg 477 42 Negative Line 1 (Photo) 82.3 8 Line 2 74,3 8 + Line 3 80,3 9 Line 4 (Photo) 77.8 4 Line 5 84,5 15 + 20 FWt Fresh Weight; Referring to FIG. 17 (a photograph comparing transgenic pumpkin plants to control plants), the transgenic pumpkin plants show substantially increased plant size, overall biomass and leaf sizes and numbers relative to the control plants. 25 75 WO 2010/025466 PCT/US2009/055557 EXAMPLE 16: GENERATION OF DOUBLE TRANSGENIC ARABIDOPSIS PLANTS CARRYING ARABIDOPSIS GS1 AND GPT TRANSGENES: In this example, Arabidopsis thaliana plants were transformed with the truncated 5 Arabidopsis GPT coding sequence of SEQ ID NO: 18 under the control of the CMV 35S promoter within the expression vector pMON316 (see Example 3, supra), and transgenic plants thereafter transformed with the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCarnbia 1201 (vector construct of SEQ ID NO: 6), using Agrobacterium 10 mediated "floral dip" transfer as described (Harrison et al_ 2006, Plant Methods 2:19 23; Clough and Bent, 1998, Plant J. 16:735-743). Agrobacterium vectors pMON316 carrying GPT and pCambia 1201 carrying GS1 were prepared as described in Examples 3 and 11, respectively, 15 Transformation of two different cultures of Agrobacterium with either a pMon 316 + Arabidopsis GTP construct or with a Cambia 1201 + Arabidopsis GS construct was done by electroporation using the method of Weigel and Glazebrook 2002. The transformed Agrobacterium were then grown under antibiotic selection, collected by centrifugation resuspended in LB broth with antibiotic and used in the floral dip of 20 Arabidopsis inflorescence. Floral dipped Arabidopsis plants were taken to maturity and self-fertilized and seeds were collected. Seeds from twice dipped plants were first geminated on a media containing 20ug/ml of kanamycin and by following regular selection procedures surviving seedlings were transferred to media containing 20 ug of hygromycin. Plants (3) surviving the selection process on both antibiotics were 25 self-fertilized and seeds were collected. Seeds from the TI generation were germinated on MS media containing 20 ug/mI of hygromycin and surviving seedlings were taken to maturity, self-fertilized and seeds collected. This seed population the T2 generation was then used for subsequent growth studies. 30 The results are presented in FIG. 18 and Table X11. Referring to Table XII, which shows data from 6 wild type and 6 transgenic Arabidopsis plants (averaged), the 76 WO 2010/025466 PCT/US2009/055557 transgenic plants displayed increased levels of both GPT and GS activity. GPT activity was over twenty-fold higher than the control plants. Moreover, the transgenic plant fresh foliar weight average was well over four-fold that of the wild type control plant average, A photograph of young transgene Arabidopsis plants in comparison 5 to wild type control Arabidopsis plants grown under identical conditions is shown in FIG. 18, and reveals a consistent and very significant increase in transgenic plants relative to the control plants. TABLE XIIl: TRANSGENIC ARABIDOPSIS VERSUS CONTROL GS Activity Atboi Plant type Biomass, g GPT Activity uo Antibiotic Fresh foliar wt nmol/h/gFWt Wi Resistance wt Wildtype, avg 0246 18 4 7.0 Negative Transgene 1,106 395.6 18.2 Positive 10 EXAMPLE 17: GENERATION OF TRANSGENIC TOMATO PLANTS CARRYING ARABIDOPSIS GPT AND GS1 TRANSGENES: 15 In this example, tomato plants (Solanum Jycopersicon, "money Maker" variety) were transformed with the Arabidopsis GPT full length coding sequence of SEQ ID NO: 1 under the control of the CMV 35S promoter within the expression vector pMON316 (see Example 3, supra), and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 20 under the control of the RuBisCo promoter within the expression vector pCambia 1201 (vector construct of SEQ ID NO: 6). Single transgene (GPT) transgenic tomato plants were generated and grown to flowering essentially as described in Example 4. The Arabidopsis GS1 transgene was then introduced into the single-transgene TO plants using Agrobacterium-mediated transfer via injection directly into flowers (as 25 described in Example 8). The transgenic and control tomato plants were grown under identical conditions and characterized as to growth phenotype characteristics, Resulting TO double-transgene plants were then grown to maturity, photographed along with control tomato plants, and phenotypically characterized. 77 WO 2010/025466 PCT/US2009/055557 The results are presented in FIG, 19 and in Table IXX. Referring to Table IXX, double-transgene tomato plants showed substantial foliar plant biomass increases in comparison to the control plants, with an increase in average biomass yield of 45% over control. Moreover, as much as a 70% increase in tomato fruit yield was 5 observed in the transgenic lines compared to control plants (e.g,, 51 tomatoes harvested from Line 4C, versus and average of approximately 30 tomatoes from control plants). A much higher level of GPT activity was observed in the transgenic plants (e.g., line 4C displaying an approximately 32-fold higher GPT activity in comparison to the average GPT activity measured in control plants). GS activity was 10 also higher in the transgenic plants relative to control plants (almost double in Line 4C). With respect to growth phenotype, and referring to FIG, 19, the transgenic tomato plants displayed substantially larger leaves compared to control plants (FIG 19A). In 15 addition, it can be seen that the transgenic tomato plants were substantially larger, taller and of a greater overall biomass (see FIG. 198). 20 TABLE IXX: TRANSGENIC TOMATO GROWTH AND REPRODUCTION Total GPT GS Activity Biomass Tomatoes Activity umoles/mi Transgene Plant Type Foliar FWt, Harvested nmoles/h n Presence g until /gFWt /gFWt Assay Sacrifice Wildtype, 891 30.2 287 14.27 Negative avg Line 6C 1288 43 9181 18.3 + Line 4C 1146 51 1718 26.4 + 25 78 WO 2010/025466 PCT/US2009/055557 EXAMPLE 18: GENERATION OF TRANSGENIC CAMILENA PLANTS CARRYING ARABIDOPSIS GPT AND GS1 TRANSGENES: In this example, Camelina plants (Camelina sativa, Var MT 303) were transformed 5 with the Arabidopsis GPT full length coding sequence of SEQ ID NO: 1 under the control of the RuBisCo promoter within the expression vector pCambia 1201, and the Arabidopsis GS1 coding sequence of SEQ ID NO: 6 under the control of the RuBisCo promoter within the expression vector pCambia 1201, using Agrobacterium-mediated transfer into germinating seeds according to the method described in Chee et al, 10 1989, Plant Physiol. 91: 1212-1218. Agrobacterium vectors and mixtures were prepared for seed inoculations as described in Example 11, supra. Transgenic and control Camelina plants were grown under identical conditions (30 days in a growth chamber and then moved to greenhouse cultivation) for 39 days, 15 and characterized as to biomass, growth characteristics and flowering stage, The results are presented in Table XX and FIGs 20. Referring to Table XX, it can be seen that total biomass in the transgenic plants was, on average, almost double control plant biomass. Canopy diameter was also significantly improved in the 20 transgenic plants. FIG. 20 shows a photograph of transgenic Camelina compared to control The transgenic plant is noticeably larger and displays more advanced flowering status, TABLE XX: TRANSGENIC CAMELINA VERSUS CONTROL 25 Plant Type Height I Canopy Biomass Flowering Stage Diameter, inches g Wildtype, avg 14/4 8.35 Partial flowering Transgene C-1 155 1 5 16,54 Full flowering Transgene C-3 14/ 7 14.80 Initial flowering 79 WO 2010/025466 PCT/US2009/055557 EXAMPLE 19: ACTIVITY OF BARLEY GPT TRANSGENE IN PLANTA In this example, the putative coding sequence for Barley GPT was isolated and expressed from a transgene construct using an in planta transient expression assay. 5 Biologically active recombinant Barley GPT was produced, and catalyzed the increased synthesis of 2- oxoglutaramate, as confirmed by HPLC. The Barley (Hordeum vulgare) GPT coding sequence was determined and synthesized. The DNA sequence of the Barley GPT coding sequence used in this 10 example is provided in SEQ ID NO: 14, and the encoded GPT protein amino acid sequence is presented in SEQ ID NO: 15. The coding sequence for Barley GPT was inserted into the 1305,1 cambia vector, and transferred to Agrobacterium tumefaciens strain LBA404 using a standard 15 electroporation method (McCormac et al, 1998, Molecular Biotechnology 9:155-159), followed by plating on LB plates containing hygromycin (50 micro gm I ml). Antibiotic resistant colonies of Agrobacterium were selected for analysis. The transient tobacco leaf expression assay consisted of injecting a suspension of 20 transformed Agrobacterium (1,5-2.0 OD 650) into rapidly growing tobacco leaves, Intradermal injections were made in a grid across the leaf surface to assure that a significant amount of the leaf surface would be exposed to the Agrobacterium. The plant was then allowed to grow for 3-5 days when the tissue was extracted as described for all other tissue extractions and the GPT activity measured. 25 GPT activity in the inoculated leaf tissue (1217 nanomoles/gFWt/h) was three-fold the level measured in the control plant leaf tissue (407 nanomoles/gFWt/h), indicating that the Hordeum GPT construct can direct the expression of functional GPT in a transgenic plant, 80 WO 2010/025466 PCT/US2009/055557 EXAMPLE 20: ISOLATION AND EXPRESSION OF RECOMBINANT RICE GPT GENE CODING SEQUENCE AND ANALYSIS OF BIOLOGICAL ACTIVITY In this example, the putative coding sequence for rice GPT was isolated and 5 expressed in E. coli: Biologically active recombinant rice GPT was produced, and catalyzed the increased synthesis of 2- oxoglutaramate, as confirmed by HPLC. Materials and Methods: Rice GPT coding sequence and expression in E coli: 10 The rice (Oryza sativia) GPT coding sequence was determined and synthesized, inserted into a PET28 vector, and expressed in E. coli. Briefly, E. coli cells were transformed with the expression vector and transformants grown overnight in LB broth diluted and grown to OD 04, expression induced with isopropy-B-D thiogalactoside (0.4 micromolar), grown for 3 hr and harvested. A total of 25 X 106 15 cells were then assayed for biological activity using the NMR assay, below. Untransformed, wild type E coli cells were assayed as a control An additional control used E coli cells transformed with an empty vector. The DNA sequence of the rice GPT coding sequence used in this example is 20 provided in SEQ ID NO: 10, and the encoded GPT protein amino acid sequence is presented in SEQ ID NO: 11, HPLC Assay for 2-oxoglutaramate: HPLC was used to determine 2-oxoglutaramate production in GPT-overexpressing E 25 coli cells, following a modification of Calderon et al, 1985, J Bacteriol 161(2): 807 809, Briefly, a modified extraction buffer consisting of 25 mM Tris-HCI pH 8,5 1 mM EDTA, 20 pIM Pyridoxal phosphate, 10 mM Cysteine, and -1,5% (v/v) Mercaptoethanoi was used. Samples (lysate from E. coli cells, 25 X 106 cells) were added to the extraction buffer at approximately a 1/3 ratio (w/v), incubated for 30 30 minutes at 374C, and stopped with 200pd of 20% TCA. After about 5 minutes, the 81 WO 2010/025466 PCT/US2009/055557 assay mixture is centrifuged and the supernatant used to quantify 2-oxoglutaramate by HPLC, using an ION-300 7.8mm ID X 30 cm L column, with a mobile phase in 0.01N h2SO4, a flow rate of approximately 0.2 ml/min, at 40'C. Injection volume is approximately 20 d, and retention time between about 38 and 39 minutes. Detection 5 is achieved with 21Onm UV light, NMR analysis comparison with authentic 2-oxoglutaramate was used to establish that the Arabidopisis full length sequence expresses a GPT with 2-oxoglutaramate synthesis activity. Briefly, authentic 2-oxoglutarmate (structure confirmed with NMR) 10 made by chemical synthesis to validate the HPLC assay, above, by confirming that the product of the assay (molecule synthesized in response to the expressed GPT) and the authentic 2-oxoglutaramate elute at the same retention time. In addition, when mixed together the assay product and the authentic compound elute as a single peak. Furthermore, the validation of the HPLC assay also included monitoring 15 the disappearance of the substrate glutamine and showing that there was a 1:1 molar stoechiometry between glutamine consumed to 2-oxoglutaramte produced. The assay procedure always included two controls, one without the enzyme added and one without the glutamine added. The first shows that the production of the 2 oxoglutaramate was dependent upon having the enzyme present, and the second 20 shows that the production of the 2-oxoglutaramate was dependent upon the substrate glutamine. Results: 25 Expression of the rice GPT coding sequence of SEQ ID NO: 10 resulted in the over expression of recombinant GPT protein having 2-oxoglutaramate synthesis catalyzing bioactivity. Specifically, 1.72 nanomoles of 2-oxoglutaramate activity was observed in the E. coli cells overexpressing the recombinant rice GPT, compared to only 0.02 nanomoles of 2-oxoglutaramate activity in control E. coli cells, an 86-fold 30 activity level increase over control. 82 WO 2010/025466 PCT/US2009/055557 EXAMPLE 21: ISOLATION AND EXPRESSION OF RECOMBINANT SOYBEAN GPT GENE CODING SEQUENCE AND ANALYSIS OF BIOLOGICAL ACTIVITY In this example, the putative coding sequence for soybean GPT was isolated and 5 expressed in E coli. Biologically active recombinant soybean GPT was produced, and catalyzed the increased synthesis of 2~ oxoglutaramate, as confirmed by HPLC. Materials and Methods: 10 Soybean GPT coding sequence and expression in E. coli The soybean (Glycine max) GPT coding sequence was determined and synthesized, inserted into a PET28 vector, and expressed in E. coli. Briefly, E. coli cells were transformed with the expression vector and transformants grown overnight in LB broth diluted and grown to OD 0.4, expression induced with isopropyl-B-D 15 thiogalactoside (0.4 micromolar), grown for 3 hr and harvested. A total of 25 X 106 cells were then assayed for biological activity using the NMR assay, below, Untransformed, wild type E. coli cells were assayed as a control. An additional control used E coli cells transformed with an empty vector. 20 The DNA sequence of the soybean GPT coding sequence used in this example is provided in SEQ ID NO: 12, and the encoded GPT protein amino acid sequence is presented in SEQ ID NO: 13. H PLC Assay for 2-oxoglutaramate: 25 HPLC was used to determine 2-oxoglutaramate production in GPT-overexpressing E. coli cells, as described in Example 20, supra. Results: 30 Expression of the soybean GPT coding sequence of SEQ ID NO: 12 resulted in the over-expression of recombinant GPT protein having 2-oxoglutaramate synthesis 83 WO 2010/025466 PCT/US2009/055557 catalyzing bioactivity. Specifically, 31.9 nanomoles of 2-oxoglutaramate activity was observed in the E, coli cells overexpressing the recombinant soybean GPT, compared to only 0A02 nanomoles of 2-oxoglutaramate activity in control E. coli cells, a nearly 1,600-fold activity level increase over control. 5 EXAMPLE 22: ISOLATION AND EXPRESSION OF RECOMBINANT ZEBRA FISH GPT GENE CODING SEQUENCE AND ANALYSIS OF BIOLOGICAL ACTIVITY 10 In this example, the putative coding sequence for Zebra fish GPT was isolated and expressed in E coli. Biologically active recombinant Zebra fish GPT was produced, and catalyzed the increased synthesis of 2- oxoglutaramate, as confirmed by NMR. Materials and Methods: 15 Zebra fish GPT coding sequence and expression in E. coi: The Zebra fish (Danio rerio) GPT coding sequence was determined and synthesized, inserted into a PET28 vector, and expressed in E. co/. Briefly, E. coli cells were transformed with the expression vector and transformants grown overnight in LB 20 broth diluted and grown to OD 0.4, expression induced with isopropyl-B-D thiogalactoside (0.4 micromolar), grown for 3 hr and harvested, A total of 25 X 106 cells were then assayed for biological activity using the NMR assay, below. Untransformed, wild type E. coli cells were assayed as a control. An additional control used E coli cells transformed with an empty vector. 25 The DNA sequence of the Zebra fish GPT coding sequence used in this example is provided in SEQ ID NO: 16, and the encoded GPT protein amino acid sequence is presented in SEQ ID NO: 17, 30 84 WO 2010/025466 PCT/US2009/055557 H PLC Assay for 2-oxoglutaramate: HPLC was used to determine 2-oxoglutaramate production in GPT-overexpressing E coli cells, as described in Example 20, supra. 5 Results: Expression of the Zebra fish GPT coding sequence of SEQ ID NO: 16 resulted in the over-expression of recombinant GPT protein having 2-oxoglutaramate synthesis catalyzing bioactivity. Specifically, 28.6 nanomoles of 2-oxoglutaramate activity was 10 observed in the E coli cells overexpressing the recombinant Zebra fish GPT, compared to only 0.02 nanomoles of 2-oxoglutaramate activity in control E coli cells, a more than 1,400-fold activity level increase over control. EXAMPLE 23: GENERATION AND EXPRESSION OF RECOMBINANT 15 TRUNCATED ARABIDOPSIS GPT GENE CODING SEQUENCES AND ANALYSIS OF BIOLOGICAL ACTIVITY In this example, two different truncations of the Arabidopsis GPT coding sequence were designed and expressed in E coli, in order to evaluate the activity of GPT 20 proteins in which the putative chloroplast signal peptide is absent or truncated. Recombinant truncated GPT proteins corresponding to the full length Arabidopsis GPT amino acid sequence SEQ ID NO: 1, truncated to delete either the first 30 amino-terminal amino acid residues, or the first 45 amino-terminal amino acid residues, were successfully expressed and showed biological activity in catalyzing 25 the increased synthesis of 2- oxoglutaramate, as confirmed by NMR. Materials and Methods: Truncated Arabidoosis GPT coding sequences and expression in E. coli: 30 The DNA coding sequence of a truncation of the Arabidopsis thaliana GPT coding sequence of SEQ ID NO: I was designed, synthesized, inserted into a PET28 vector, 85 WO 2010/025466 PCT/US2009/055557 and expressed in E col The DNA sequence of the truncated Arabidopsis GPT coding sequence used in this example is provided in SEQ ID NO: 20 (-45 AA construct), and the corresponding truncated GPT protein amino acid sequence is provided in SEQ ID NO: 21. Briefly, E coli cells were transformed with the 5 expression vector and transformants grown overnight in LB broth diluted and grown to OD 0.4, expression induced with isopropyl-B-D-thiogalactoside (0.4 micromolar), grown for 3 hr and harvested. A total of 25 X 103 cells were then assayed for biological activity using HPLC as described in Example 20. Untransformed, wild type E. coli cells were assayed as a control, An additional control used E coli cells 10 transformed with an empty vector. Expression of the truncated -45 Arabidopsis GPT coding sequence of SEQ ID NO: 20 resulted in the over-expression of biologically active recombinant GPT protein (2 oxoglutaramate synthesis-catalyzing bioactivity). Specifically, 16.1 nanomoles of 2 15 oxoglutaramate activity was observed in the E. coli cells overexpressing the truncated -45 GPT, compared to only 0,02 nanomoles of 2-oxoglutaramate activity in control E coli cells, a more than 800-fold activity level increase over control. For comparison, the full length Arabidopsis gene coding sequence expressed in the same E. coli assay generated 2.8 nanomoles of 2-oxoglutaramate activity, or roughly 20 less than one-fifth the activity observed from the truncated recombinant GPT protein. EXAMPLE 24: GPT + GS TRANSGENIC TOBACCO SEED GERMINATION TOLERATES HIGH SALT CONCENTRATIONS 25 In this example, seeds form the double transgene tobacco line XX-3 (Cross 3 in Table 4, see Example 7) were tested in a seed germination assay designed to evaluate tolerance to high salt concentrations. 30 86 WO 2010/025466 PCT/US2009/055557 Materials and Methods: Tobacco seeds from the wild type and XX-3 populations were surfaced sterilized (5% bleach solution for 5 minutes followed by a 10% ethanol wash for 3 minutes) and 5 rinsed with sterile distilled water. The surface sterilized seeds were then spread on Murashige and Skoog media (10% agarose) without sucrose and containing either 0 or 200 mM NaCl. The seeds were allowed to germinate in darkness for 2 days followed by 6 days under a 16:8 photoperiod at 24C, On day eight the rate of germination was determined by measuring the percentage of seeds from the control 10 or transgene plants that had germinated. Results: The results are tabulated in Table XXI below, The rate of germination of the 15 transgenic plant line seeds under zero salt conditions was the same as observed with wild type control plant seeds. In stark contrast, the germination rate of the transgenic plant line seeds under very high salt conditions far exceeded the rate seen in wild type control seeds. Whereas over 81% of the transgenic plant seeds had germinated under the high salt conditions, only about 9% of the wild type control plant seeds had 20 germinated by the same time point. These data indicate that the transgenic seeds are capable of germinating very well under high salt concentrations, an important trait for plant growth in areas of increasingly high water and/or soil salinity. 25 TABLE XXI: TRANSGENIC TOBACCO PLANTS GERMINATE AND TOLERATE HIGH SALT Plant type Control 0 mM NaC-) Test (200 mM NaCi a % Germination % Germination Wild type 92,87,94 9, 11,8 Transgene line XX-3 92, 91, 94 84, 82, 78 87 WO 2010/025466 PCT/US2009/055557 All publications, patents, and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. 5 The present invention is not to be limited in scope by the embodiments disclosed herein, which are intended as single illustrations of individual aspects of the invention, and any which are functionally equivalent are within the scope of the invention. Various modifications to the models and methods of the invention, in addition to those described herein, will become apparent to those skilled in the art 10 from the foregoing description and teachings, and are similarly intended to fall within the scope of the invention. Such modifications or other embodiments can be practiced without departing from the true scope and spirit of the invention. 88 WO 2010/025466 PCT/US2009/055557 TABLE OF SEQUENCES: SEQ ID NO: 1 Arabidopsis glutamine phenylpyruvate transaminase DNA coding sequence: ATGTACCTGGACATAAATGGTGTGATGATCAAACAGTTTAGCTTCAAAGCCTCTC TTCTCCCATTCTCTTCTAATTTCCGACAAAGCTCCGCCAAAATCCATCGTCCTAT CGGAGCCACCATGACCACAGTTTCGACTCAGAACGAGTCTACTCAAAAACCCGT CCAGGTGGCGAAGAGATTAGAGAAGTTCAAGACTACTATTTTCACTCAAATGAG 10 CATATTGGCAGTTAAACATGGAGCGATCAATTTAGGCCAAGGCTTTCCCAATTTC GACGGTCCTGATTTTGTTAAAGAAGCTGCGATCCAAGCTATTAAAGATGGTAAAA ACCAGTATGCTCGTGGATACGGCATTCCTCAGCTCAACTCTGCTATAGCTGCGC GGTTTCGTGAAGATACGGGTCTTGTTGTTGATCCTGAGAAAGAAGTTACTGTTAC ATCTGGTTGCACAGAAGCCATAGCTGCAGCTATGTTGGGTTTAATAAACCCTGG 15 TGATGAAGTCATTCTCTTTGCACCGTTTTATGATTCCTATGAAGCAACACTCTCTA TGGCTGGTGCTAAAGTAAAAGGAATCACTTTACGTCCACCGGACTTCTCCATCC CTTTGGAAGAGCTTAAAGCTGCGGTAACTAACAAGACTCGAGCCATCCTTATGA ACACTCCGCACAACCCGACCGGGAAGATGTTCACTAGGGAGGAGCTTGAAACC ATTGCATCTCTCTGCATTGAAAACGATGTGCTTGTGTTCTCGGATGAAGTATACG 20 ATAAGCTTGCGTTTGAAATGGATCACATTTCTATAGCTTCTCTTCCCGGTATGTA TGAAAGAACTGTGACCATGAATTCCCTGGGAAAGACTTTCTCTTTAACCGGATG GAAGATCGGCTGGGCGATTGCGCCGCCTCATCTGACTTGGGGAGTTCGACAAG CACACTCTTACCTCACATTCGCCACATCAACACCAGCACAATGGGCAGCCGTTG CAGCTCTCAAGGCACCAGAGTCTTACTTCAAAGAGCTGAAAAGAGATTACAATG 25 TGAAAAAGGAGACTCTGGTTAAGGGTTTGAAGGAAGTCGGATTTACAGTGTTCC CATCGAGCGGGACTTACTTTGTGGTTGCTGATCACACTCCATTTGGAATGGAGA ACGATGTTGCTTTCTGTGAGTATCTTATTGAAGAAGTTGGGGTCGTTGCGATCC CAACGAGCGTCTTTTATCTGAATCCAGAAGAAGGGAAGAATTTGGTTAGGTTTG CGTTCTGTAAAGACGAAGAGACGTTGCGTGGTGCAATTGAGAGGATGAAGCAG 30 AAGCTTAAGAGAAAAGTCTGA SEQ ID NO: 2 Arabidopsis GPT amino acid sequence 35 MYLDINGVMIKQFSFKASLLPFSSNFRQSSAKIHRPIGATMTTVSTQNESTQKPVQV AKRLEKFKTTIFTQMSILAVKHGAINLGQGFPNFDGPDFVKEAAIQAIKDGKNQYARG YGIPQLNSAIAARFREDTGLVVDPE KEVTVTSGCTEAIAAAMLGLINPGDEVILFAPFY DSYEATLSMAGAKVKGITLRPPDFSIPLEELKAAVTNKTRAILMNTPHNPTGKMFTRE ELETIASLCIENDVLVFSDEVYDKLAFEMDHISIASLPGMYERTVTMNSLGKTFSLTG 40 WKIGWAIAPPHLTWGVRQAHSYLTFATSTPAQWAAVAALKAPESYFKELKRDYNVK KETLVKGLKEVGFTVFPSSGTYFVVADHTPFGMENDVAFCEYLIEEVGVVAIPTSVF YLNPEEGKNLVRFAFCKDEETLRGAIERMKQKLKRKV 89 WO 2010/025466 PCT/US2009/055557 SEQ ID NO: 3 Alfalfa GS1 DNA coding sequence (upper case) with 5' and 3' untranslated sequences (indicated in lower case), atttccgttttcgttttcatttg attca ttg aatcaa a tcg a atcg aatctttag gattca ata cag 5 attccttagattttactaagtttgaaaccaaaaccaaaacATGTCTCTCCTTTCAGAT CTTATCAACCTTGACCTCTCCGAAACCACCGAGAAAATCATCG CCG AATACATATGGATTGGTGGATCTGGTTTGGACTTGAGGAGCAAAGC AAGGACTCTACCAGGACCAGTTACTGACCCTTCACAGCTTCCCAAG TGGAACTATGATGGTTCCAGCACAGGTCAAGCTCCTGGAGAAGAT 10 AGTGAAGTTATTATCTACCCACAAGCCATTTTCAAGGACCCATTTA GAAG GGGTAACAATATCTTGGTTATGTGTGATGCATACACTCCAGC TGGAGAGCCCATTCCCACCAACAAGAGACATGCAGCTGCCAAGAT TTTCAGCCATCCTGATGTTGTTGCTGAAGTACCATGGTATGGTATT GAGCAAGAATACACCTTGTTGCAGAAAGACATCAATTGGCCTCTTG 15 GTTGGCCAGTTGGTGGTTTTCCTGGACCTCAGGGACCATACTATTG TGGAGCTGGTGCTGACAAGGCATTTGGCCGTGACATTGTTGACTC ACATTACAAAGCCTGTCTTTATGCCGGCATCAACATCAGTGGAATC AATG GTG AAG TGATGC CTG GTCAATGG GAATTCCAAGTTGGTCC CT CAGTTGGTATCTCTGCTGGTGATGAGATATGGGTTGCTCGTTACAT 20 TTTGGAGAGGATCACTGAGGTTGCTGGTGTGGTGCTTTCCTTTGAC C CAAAAC CAATTAAG G GTG ATTG GAATG GTG CTG GTG CTCACAGAA ATTACAGCACCAAGTCTATGAGAGAAGATGGTGGCTATGAAGTCAT CTTGAAAGCAATTGAGAAGCTTGGGAAGAAGCACAAGGAGCACAT TGCTGCTTATGGAGAAGGCAACGAGCGTAGATTGACAGGGCGACA 25 TGAGACAGCTGACATTAACACCTTCTTATGGGGTGTTGCAAACCGT GGTGCGTCGATTAGAGTTGGAAGGGACACAGAGAAAGCAGGGAAA GGTTATTTCGAGGATAGGAGGCCATCATCTAACATGGATCCATATG TTGTTACTTCCATGATTGCAGACACCACCATTCTCTGGAAACCATA Ag cca cca ca ca ca catg ca ttg a agtatttg a a a gtca ttg ttg attccg ca ttag a atttg g 30 tcattgttttttctaggatttggatttgtgttattgttatggttcacactttgtttgtttgaatttgaggc cttgttatagg tttcatatttctttctcttgttcta ag ta aatgtcag aa taataatgta at SEQ ID NO: 4 Alfalfa GS1 amino acid sequence 35 MSLLSDLINLDLSETTEKIIAEYIWIGGSGLDLRSKARTLPGPVTDPSQLPKWNYDGS STGQAPGEDSEVIIYPQAIFKDPFRRGNNILVMCDAYTPAGEPIPTNKRHAAAKIFSH PDWAEVPWYGIEQEYTLLQKDINWPLGWPVGGFPGPQGPYYCGAGADKAFGRDI VDSHYKACLYAGINISGINGEVMPGQWEFQVGPSVGISAGDEIWVARYILERITEVA 40 GVVLSFDPKPIKGDWNGAGAHTNYSTKSMREDGGYEVILKAIEKLGKKHKEHIAAYG EGNERRLTGRHETADINTFLWGVANRGASI RVGRDTEKAGKGYFEDRRPSSN MDP YVVTSMIADTTILWKP 90 WO 2010/025466 PCT/US2009/055557 SEQ ID NO: 5 Alfalfa GS1 DNA coding sequence (upper case) with 5' and 3' untranslated sequences (indicated in lower case) and vector sequences from Clal to Smal/Sspl and Sspl/Smal to Sal/Xhol (lower case, underlined). 5 atcqatqaattcpaqctcqqtacccatttccgttttcgttttcatttgattcattgaatcaaatcga atcgaatctttag gattcaatacagattccttagattttactaagtttgaaaccaaaa ccaaaa cATGTCTCTCCTTTCAGATCTTATCAACCTTGACCTCTCCGAAACCA CCGAGAAAATCATCGCCGAATACATATGGATTGGTGGATCTGGTTT GGACTTGAGGAGCAAAGCAAGGACTCTACCAGGACCAGTTACTGA 10 CCCTTCACAGCTTCCCAAGTGGAACTATGATGGTTCCAGCACAGGT CAAGCTCCTGGAGAAGATAGTGAAGTTATTATCTACCCACAAGCCA TTTTCAAGGACCCATTTAGAAGGGGTAACAATATCTTGGTTATGTG TGATGCATACACTCCAGCTGGAGAGCCCATTCCCACCAACAAGAG ACATGCAGCTGCCAAGATTTTCAGCCATCCTGATGTTGTTGCTGAA 15 GTACCATGGTATGGTATTGAGCAAGAATACACCTTGTTGCAGAAAG ACATCAATTGGCCTCTTGGTTGGCCAGTTGGTGGTTTTCCTGGACC T CAG G G AC CATA CTATT GTG GAG CTG GTGCT GAC AAGG CATTTGG CCG TGA CATT GT TGAC TCACATTA C AAAG C CTG TT TTATG C CG GC ATCAACATCAGTGGAATCAATGGTGAAGTGATGCCTGGTCAATGGG 20 AATTCCAAGTTGGTCCCTCAGTTGGTATCTCTGCTGGTGATGAGAT ATGGGTTGCTCGTTACATTTTGGAGAGGATCACTGAGGTTGCTGGT GTGGTGCTTTCCTTTGACCCAAAACCAATTAAGGGTGATTGGAATG GTGCTGGTGCTCACACAAATTACAGCACCAAGTCTATGAGAGAAGA TGGTGGCTATGAAGTCATCTTGAAAGCAATTGAGAAGCTTGGGAAG 25 AAGCACAAGGAGCACATTGCTGCTTATGGAGAAGGCAACGAGCGT AGATTGACAGGGCGACATGAGACAGCTGACATTAACACCTTCTTAT GGGGTGTTGCAAACCGTGGTGCGTCGATTAGAGTTGGAAGGGACA CAGAGAAAGCAGGGAAAGGTTATTTCGAGGATAGGAGGCCATCAT CTAACATGGATCCATATGTTGTTACTTCCATGATTGCAGACACCAC 30 CATTCTCTGGAAACCATAAgccaccacacacacatgcattgaagtatttgaaagtc attgttg attccgcattag aa tttggtcattgttttttctagg atttgg atttgtgttattgttatggttc acactttgtttgtttgaatttgaggccttgttataggtttcatatttctttctcttgttctaagtaaatg tc ag aat aat aat g t a at p at c ct c tap at c pa 35 SEQ ID NO: 6 Arabidopsis GS1 coding sequence Cambia 1201 vector + rbcS3C+arabidopsis GS1 Bold ATG is the start site, AAAAAAGAAAAAAAAAACATATCTTGTTTGTCAGTATGGGAAGTTTGAGATAAGG 40 ACGAGTGAGGGGTTAAAATTCAGTGGCCATTGATTTTGTAATGCCAAGAACCAC AAAATCCAATGGTTACCATTCCTGTAAGATGAGGTTTGCTAACTCTTTTTGTCCG TTAGATAGGAAGCCTTATCACTATATATACAAGGCGTCCTAATAACCTCTTAGTA ACCAATTATTTCAGCACCA TGTCTCTGCTCTCAGATCTCGTTAACCTCAACCTCA CCGATGCCACCGGGAAAATCATCGCCGAATACATATGGATCGGTGGATCTGGA 91 WO 2010/025466 PCT/US2009/055557 ATGGATATCAGAAGCAAAGCCAGGACACTACCAGGACCAGTGACTGATCCATCA AAGCTTCCCAAGTGGAACTACGACGGATCCAGCACCGGTCAGGCTGCTGGAGA AGACAGTGAAGTCATTCTATACCCTCAGGCAATATTCAAGGATCCCTTCAGGAAA GGCAACAACATCCTGGTGATGTGTGATGGTTACACACCAGCTGGTGATCCTATT 5 CCAACCAACAAGAGGCACAACGCTGCTAAGATCTTCAGCCACCCCGACGTTGC CAAGGAGGAGCCTTGGTATGGGATTGAGCAAGAATACACTTTGATGCAAAAGGA TGTGAACTGGCCAATTGGTTGGCCTGTTGGTGGCTACCCTGGCCCTCAGGGAC CTTACTACTGTGGTGTGGGAGCTGACAAAGCCATTGGTCGTGACATTGTGGATG CTCACTACAAGGCCTGTCTTTACGCCGGTATTGGTATTTCTGGTATCAATGGAGA 10 AGTCATGCCAGGCCAGTGGGAGTTCCAAGTCGGCCCTGTTGAGGGTATTAGTT CTGGTGATCAAGTCTGGGTTGCTCGATACCTTCTCGAGAGGATCACTGAGATCT CTGGTGTAATTGTCAGCTTCGACCCGAAACCAGTCCCGGGTGACTGGAATGGA GCTGGAGCTCACTGCAACTACAGCACTAAGACAATGAGAAACGATGGAGGATTA GAAGTGATCAAGAAAGCGATAGGGAAGCTTCAGCTGAAACACAAAGAACACATT 15 GCTGCTTACGGTGAAGGAAACGAGCGTCGTCTCACTGGAAAGCACGAAACCGC AGACATCAACACATTCTCTTGGGGAGTCGGCGAACCGTGGAGCGTCAGTGAGAG TGGGACGTGACACAGAGAAGGAAGGTAAAGGGTACTTCGAAGACAGAAGGCCA GCTTCTAACATGGATCCTTACGTTGTCACCTCCATGATCGCTGAGACGACCATA CTCGGTTGA 20 SEQ ID NO: 7 Arabidopsis GS1 amino acid sequence Vector sequences at N-terminus in italics 25 MVDLRNRRTSMSLLSDLVNLNLTDATGKIIAEYIWIGGSGMDI RSKARTLPGPVTDPS KLPKWNYDGSSTGQAAGEDSEVILYPQAIFKDPFRKGNNILVMCDAYTPAGDPIPTN KRHNAAKIFSHPDVAKEEPWYGIEQEYTLMQKDVNWPIGWPVGGYPGPQGPYYC GVGADKAIGRDIVDAHYKACLYAGIGISGINGEVMPGQWEFQVGPVEGISSGDQVW VARYLLERITEISGVIVSFDPKPVPGDWNGAGAHCNYSTKTMRNDGGLEVIKKAIGK 30 LQLKHKEHIAAYGEGNERRLTGKHETADINTFSWGVANRGASVRVGRDTEKEGKG YFEDRRPASNMDPYVVTSMIAETTILG SEQ ID NO: 8 Grape GPT DNA sequence 35 Showing Cambia 1305.1 with (3' end of) rbcS3C+Vitis (Grape). Bold ATG is the start site, parentheses are the cati intron and the underlined actagt is the spel cloning site used to splice in the hordeum gene. 40 AAAAAAGAAAAAAAAAACATATCTTGTTTGTCAGTATGGGAAGTTTGAGATAAGG ACGAGTGAGGGGTTAAAATTCAGTGGCCATTGATTTTGTAATGCCAAGAACCAC AAAATCCAATGGTTACCATTCCTGTAAGATGAGGTTTGCTAACTCTTTTTGTCCG TTAGATAGGAAGCCTTATCACTATATATACAAGGCGTCCTAATAACCTCTTAGTA ACCAATTATTTCAGCACCATGGTAGATCTGAGG(GTAAATTTCTAGTTTTTCTCCT 92 WO 2010/025466 PCT/US2009/055557 TCATTTTCTTGGTTAGGACCCTTTTCTCTTTTTATTTTTTTGAGCTTTGATCTTTCT TTAAACTGATCTATTTTTTAATTGATTGGTTATGGTGTAAATATTACATAGCTTTAA CTGATAATCTGATTACTTTATTTCGTGTGTCTATGATGATGATGATAGTTACAG)A ACCGACGAACTAGTATGCAGCTCTCTCAATGTACCTGGACATTCCCAGAGTTGC 5 TTAAAAGACCAGCCTTTTTAAGGAGGAGTATTGATAGTATTTCGAGTAGAAGTAG GTCCAGCTCCAAGTATCCATCTTTCATGGCGTCCGCATCAACGGTCTCCGCTCC AAATACGGAGGCTGAGCAGACCCATAACCCCCCTCAACCTCTACAGGTTGCAAA GCGCTTGGAGAAATTCAAAACAACAATCTTTACTCAAATGAGCATGCTTGCCATC AAACATGGAGCAATAAACCTTGGCCAAGGGTTTCCCAACTTTGATGGTCCTGAG 10 TTTGTCAAAGAAGCAGCAATTCAAGCCATTAAGGATGGGAAAAACCAATATGCTC GTGGATATGGAGTTCCTGATCTCAACTCTGCTGTTGCTGATAGATTCAAGAAGG ATACAGGACTCGTGGTGGACCCCGAGAAGGAAGTTACTGTTACTTCTGGATGTA CAGAAGCAATTGCTGCTACTATGCTAGGCTTGATAAATCCTGGTGATGAGGTGA TCCTCTTTGCTCCATTTTATGATTCCTATGAAGCCACTCTATCCATGGCTGGTGC 15 CCAAATAAAATCCATCACTTTACGTCCTCCGGATTTTGCTGTGCCCATGGATGAG CTCAAGTCTGCAATCTCAAAGAATACCCGTGCAATCCTTATAAACACTCCCCATA ACCCCACAGGAAAGATGTTCACAAGGGAGGAACTGAATGTGATTGCATCCCTCT GCATTGAGAATGATGTGTTGGTGTTTACTGATGAAGTTTACGACAAGTTGGCTTT CGAAATGGATCACATTTCCATGGCTTCTCTTCCTGGGATGTACGAGAGGACCGT 20 GACTATGAATTCCTTAGGGAAAACTTTCTCCCTGACTGGATGGAAGATTGGTTG GACAGTAGCTCCCCCACACCTGACATGGGGAGTGAGGCAAGCCCACTCATTCC TCACGTTTGCTACCTGCACCCCAATGCAATGGGCAGCTGCAACAGCCCTCCGG GCCCCAGACTCTTACTATGAAGAGCTAAAGAGAGATTACAGTGCAAAGAAGGCA ATCCTGGTGGAGGGATTGAAGGCTGTCGGTTTCAGGGTATACCCATCAAGTGG 25 GACCTATTTTGTGGTGGTGGATCACACCCCATTTGGGTTGAAAGACGATATTGC GTTTTGTGAGTATCTGATCAAGGAAGTTGGGGTGGTAGCAATTCCGACAAGCGT TTTCTACTTACACCCAGAAGATGGAAAGAACCTTGTGAGGTTTACCTTCTGTAAA GACGAGGGAACTCTGAGAGCTGCAGTTGAAAGGATGAAGGAGAAACTGAAGCC TAAACAATAGGGGCACGTGA 30 SEQ ID NO: 9 Grape GPT amino acid sequence MVDLRNRRTSMQLSQCTWTFPELLKRPAFLRRSIDSISSRSRSSSKYPSFMASAST 35 VSAPNTEAEQTHNPPQPLQVAKRLEiKFKTTIFTQMSMLAIKHGAINLGQGFPNFDGP EFVKEAAIQAIKDGKNQYARGYGVPDLNSAVADRFKKDTGLVVDPEKEVTVTSGCT EAIAATMLGLINPG DEVILFAPFYDSYEATLSMAGAQIKSITLRPPDFAVPMDELKSA SKNTRAILINTPHNPTGKMFTREELNVIASLCIENDVLVFTDEVYDKLAFEMDHISMAS LPGMYERTVTMNSLGKTFSLTGWKIGWTVAPPHLTWGVRQAHSFLTFATCTPMQW 40 AAATALRAPDSYYEELKRDYSAKKAILVEGLKAVGFRVYPSSGTYFVVVDHTPFGLK DDIAFCEYLIKEVGVVAIPTSVFYLHPEDGKNLVRFTFCKDEGTLRAAVERMKEKLKP KQ 93 WO 2010/025466 PCT/US2009/055557 SEQ ID NO: 10 Rice GPT DNA sequence Rice GPT codon optimized for E. coli expression; untranslated sequences shown in lower case 5 atgtggATGAACCTGGCAGGCTTTCTGGCAACCCCGGCAACCGCAACCGCAACCC GTCATGAAATGCCG CTGAACCCGAGCAG GAG CGC GAGCTTTCTGCTGAGGAGC CTGCGTCGTAGCCTGGTGGCGAGCCTGCGTAAAGCGAGCCCGGCAGCAGCAG CAGCACTGAGCCCGATGGCAAGCGCAAGCACCGTGGCAGCAGAAAACGGTGC AGCAAAAGCAGCAGCAGAAAAACAGCAGCAGCAGCCGGTGCAGGTGGCGAAA 10 CGTCTGGAAAAATTTAAAACCACCATTTTTACCCAGATGAGCATGCTGGCGATTA AACATGGCGCGATTAACCTGGGCCAGGGCTTTCC GAACTTTGATGGCCCGGATTTTGTGAAAGAAGCGGCGATTCAGGCGATTAACGC GGGCAAAAACCAGTATGCGCGTGGCTATGGCGTGCCGGAACTGAACAGCGCGA TTGCGGAACGTTTTCTGAAAGATAGCGGCCTGCAGGTGGATCCGGAAAAAGAA 15 GTGACCGTGACCAGCGGCTGCACCGAAGCGATTGCGGCGACCATTCTGGGCCT GATTAACCCGGGCGATGAAGTGATTCTGTTTGCGCCGTTTTATGATAGCTATGA AGCGACCCTGAGCATGGCGGGCGCGAACGTGAAAGCGATTACCCTGCGTCCG CCGGATTTTAGCGTGCCGCTGGAAGAACTGAAAGCGGCCGTGAGCAAAAACAC CCGTGCGATTATGATTAACACCCCGCATAACCCGACCGGCAAAATGTTTACCCG 20 TGAAGAACTGGAATTTATTGCGACCCTGTGCAAAGAAAACGATGTGCTGCTGTT TGCGGATGAAGTGTATGATAAACTGGCGTTTGAAGCGGATCATATTAGCATGGC GAGCATTCCGGGCATGTATGAACGTACCGTGACCATGAACAGCCTGGGCAAAA CCTTTAGCCTGACCGGCTGGAAAATTGGCTGGGCGATTGCGCCGCCGCATCTG ACCTGGGGCGTGCGTCAGGCACATAGCTTTCTGACCTTTGCAACCTGCACCCC 25 GATGCAGGCAGCCGCCGCAGCAGCACTGCGTGCACCGGATAGCTATTATGAAG AACTGCGTCGTGATTATGGCGCGAAAAAAGCGCTGCTGGTGAACGGCCTGAAA GATGCGGGCTTTATTGTGTATCCGAGCAGCGGCACCTATTTTGTGATGGTGGAT CATACCCCGTTTGGCTTTGATAACGATATTGAATTTTGCGAATATCTGATTCGTG AAGTGGGCGTGGTGGCGATTCCGCCGAGCGTGTTTTATCTGAACCCGGAAGAT 30 GGCAAAAACCTGGTGCGTTTTACCTTTTGCAAAGATGATGAAACCCTGCGTGCG GCGGTGGAACGTATGAAAACCAAACTGCGTAAAAAAAAGCTTgcggccgcactogagc accaccaccaccaccactga 35 SEQ ID NO: 11 Rice GPT amino acid sequence Includes amino terminal amino acids MWfor cloning and His tag sequences from pet28 vector in italics. MWMNLAGFLATPATATATRHEMPLNPSSSASFLLSSLRRSLVASLRKASPAAAAAL 40 SPMASASTVAAENGAAKAAAEKQQQQPVQVAKRLEKFKTTIFTQMSMLAIKHGAINL GQGFPNFDGPDFVKEAAlQAI NAGKNQYARGYGVPELNSAIAERFLKDSGLQVDPE KEVTVTSGCTEAIAATILGLINPGDEVILFAPFYDSYEATLSMAGANVKAITLRPPDFS VPLEELKAAVSKNTRAIMINTPHNPTGKMFTREELEFIATLCKENDVLLFADEVYDKL AFEADHISMASIPGMYERTVTMNSLGKTFSLTGWKIGWAIAPPHLTWGVRQAHSFL 94 WO 2010/025466 PCT/US2009/055557 TFATCTPMQAAAAAALRAPDSYYEELRRDYGAKKALLVNGLKDAGFIVYPSSGTYF VMVDHTPFGFDNDIEFCEYLIREVGVVAIPPSVFYLNPEDGKNLVRFTFCKDDETLR AAVERMKTKLR KKKLAAALEHHHHHH 5 SEQ ID NO: 12 Soybean GPT DNA sequence TOPO 151 D WITH SOYBEAN for E coli expression From starting codon. Vector sequences are italicized 10 ATGCATCA TCACCA TCACCA TGG TAAGCCTATCCCTAACCCTCTCCTCGGTCTC GA TTCTACGGAAAACCTGTA TTTTCAGGGAATTGATCCCTTCACCGCGAAACGT CTGGAAAAATTTCAGACCACCATTTTTACCCAGATGAGCCTGCTGGCGATTAAAC ATGGCGCGATTAACCTGGGCCAGGGCTTTCCGAACTTTGATGGCCCGGAATTT GTGAAAGAAGCGGCGATTCAGGCGATTCGTGATGGCAAAAACCAGTATGCGCG 15 TGGCTATGGCGTGCCGGATCTGAACATTGCGATTGCGGAACGTTTTAAAAAAGA TACCGGCCTGGTGGTGGATCCGGAAAAAGAAATTACCGTGACCAGCGGCTGCA CCGAAGCGATTGCGGCGACCATGATTGGCCTGATTAACCCGGGCGATGAAGTG ATTATGTTTGCGCCGTTTTATGATAGCTATGAAGCGACCCTGAGCATGGCGGGC GCGAAAGTGAAAGGCATTACCCTGCGTCCGCCGGATTTTGCGGTGCCGCTGGA 20 AGAACTGAAAAGCACCATTAGCAAAAACACCCGTGCGATTCTGATTAACACCCC GCATAACCCGACCGGCAAAATGTTTACCCGTGAAGAACTGAACTGCATTGCGAG CCTGTGCATTGAAAACGATGTGCTGGTGTTTACCGATGAAGTGTATGATAAACT GGCGTTTGATATGGAACATATTAGCATGGCGAGCCTGCCGGGCATGTTTGAACG TACCGTGACCCTGAACAGCCTGGGCAAAACCTTTAGCCTGACCGGCTGGAAAAT 25 TGGCTGGGCGATTGCGCCGCCGCATCTGAGCTGGGGCGTGCGTCAGGCGCAT GCGTTTCTGACCTTTGCAACCGCACATCCGTTTCAGTGCGCAGCAGCAGCAGCA CTGCGTGCACCGGATAGCTATTATGTGGAACTGAAACGTGATTATATGGCGAAA CGTGCGATTCTGATTGAAGGCCTGAAAGCGGTGGGCTTTAAAGTGTTTCCGAGC AGCGGCACCTATTTTGTGGTGGTGGATCATACCCCGTTTGGCCTGGAAAACGAT 30 GTGGCGTTTTGCGAATATCTGGTGAAAGAAGTGGGCGTGGTGGCGATTCCGAC CAGCGTGTTTTATCTGAACCCGGAAGAAGGCAAAAACCTGGTGCGTTTTACCTT TTGCAAAGATGAAGAAACCATTCGTAGCGCGGTGGAACGTATGAAAGCGAAACT GCGTAAAGTCGACTAA 35 SEQ ID NO: 13 Soybean GPT amino acid sequence Translated protein product, vector sequences italicized MHHHHHHGKPIPNPLLGLDSTENLYFQGIDPFTAKRLEKFQTT IFTQMSLLAIKHGAI 40 NLGQGFPNFDGPEFVKEAAQAIRDGKNQYARGYGVPDLNIAIAERFKKDTGLVVDP EKEITVTSGCTEAIAATMIGLINPGDEVIMFAPFYDSYEATLSMAGAKVKGITLRPPDF AVPLEELKSTISKNTRAILINTPHNPTGKMFTREELNCIASLCIENDVLVFTDEVYDKL AFDMEH:ISMASLPGMFERTVTLNSLGKTFSLTGWKIGWAIAPPHLSWGVRQAHAFL TFATAHPFQCAAAAALRAPDSYYVELKRDYMAKRAILIEGLKAVGFKVFPSSGTYFV 95 WO 2010/025466 PCT/US2009/055557 VVDHTPFGLENDVAFCEYLVKEVGVVAIPTSVFYLNPEEGKNLVRFTFCKDEETIRS AVERMKAKLRKVD 5 SEQ ID NO: 14 Barley GPT DNA sequence Coding sequence from start with intron removed A TGGTAGATCTGAGGAACCGACGAA CTAGTATGGCATCCGCCCCCGCCTCCGC CTCCGCGGCCCTCTCCACCGCCGCCCCCGCCGACAACGGGGCCGCCAAGCCC 10 ACGGAGCAGCGGCCGGTACAGGTGGCTAAGCGATTGGAGAAGTTCAAAACAAC AATTTTCACACAGATGAGCATGCTCGCAGTGAAGCATGGAGCAATAAACCTTGG ACAGGGGTTTCCCAATTTTGATGGCCCTGACTTTGTCAAAGATGCTGCTATTGA GGCTATCAAAGCTGGAAAGAATCAGTATGCAAGAGGATATGGTGTGCCTGAATT GAACTCAGCTGTTGCTGAGAGATTTCTCAAGGACAGTGGATTGCACATCGATCC 15 TGATAAGGAAGTTACTGTTACATCTGGGTGCACAGAAGCAATAGCTGCAACGAT ATTGGGTCTGATCAACCCTGGGGATGAAGTCATACTGTTTGCTCCATTCTATGAT TCTTATGAGGCTACACTGTCCATGGCTGGTGCGAATGTCAAAGCCATTACACTC CGCCCTCCGGACTTTGCAGTCCCTCTTGAAGAGCTAAAGGCTGCAGTCTCGAA GAATACCAGAGCAATAATGATTAATACACCTCACAACCCTACCGGGAAAATGTTC 20 ACAAGGGAGGAACTTGAGTTCATTGCTGATCTCTGCAAGGAAAATGACGTGTTG CTCTTTGCCGATGAGGTCTACGACAAGCTGGCGTTTGAGGCGGATCACATATCA ATGGCTTCTATTCCTGGCATGTATGAGAGGACCGTCACTATGAACTCCCTGGGG AAGACGTTCTCCTTGACCGGATGGAAGATCGGCTGGGCGATAGCACCACCGCA CCTGACATGGGGCGTAAGGCAGGCACACTCCTTCCTCACATTCGCCACCTCCA 25 CGCCGATGCAATCAGCAGCGGCGGCGGCCCTGAGAGCACCGGACAGCTACTT TGAGGAGCTGAAGAGGGACTACGGCGCAAAGAAAGCGCTGCTGGTGGACGGG CTCAAGGCGGCGGGCTTCATCGTCTACCCTTCGAGCGGAACCTACTTCATCATG GTCGACCACACCCCGTTCGGGTTCGACAACGACGTCGAGTTCTGCGAGTACTT GATCCGCGAGGTCGGCGTCGTGGCCATCCCGCCAAGCGTGTTCTACCTGAACC 30 CGGAGGACGGGAAGAACCTGGTGAGGTTCACCTTCTGCAAGGACGACGACACG CTAAGGGCGGCGGTGGACAGGATGAAGGCCAAGCTCAGGAAGAAATGA SEQ ID NO: 15 Barley GPT amino acid sequence 35 Translated sequence from start site (intron removed) MVDLRN:RRTSMASAPASASAALSTAAPADNGAAKPTEQRPVQVAKRLEKFKTTIFT QMSMLAVKHGAINLGQGFPNFDGPDFVKDAAIEA KAGKNQYARGYGVPELNSAVA ERFLKDSGLHIDPDKEVTVTSGCTEAIAATILGLINPGDEVILFAPFYDSYEATLSMAG 40 ANVKAITLRPPDFAVPLEELKAAVSKNTRA MINTPHNPTGKMFTREELEFIADLCKE N DVLLFADEVYDKLAFEADHISMASIPGMYERTVTMNSLGKTFSLTGWKIGWAIAPP H LTWGVRQAHSFLTFATSTPMQSAAAAALRAPDSYFEELKRDYGAKKALLVDGLKA AGFIVYPSSGTYFIMVDHTPFGFDNDVEFCEYLIREVGVVAIPPSVFYLNPEDGKNLV RFTFCKDDDTLRAAVDRMKAKLRKK 96 WO 2010/025466 PCT/US2009/055557 SEQ ID NO: 16 Zebra fish GPT DNA sequence Danio rerio sequence designed for expression in E colt Bold, italicized nucleotides added for cloning or from pET28b vector. ATGTCCGTGGCGAAACGTCTGGAAAAATTTAAAACCACCATTTTTACCCAGATGA GCATGCTGGCGATTAAACATGGCGCGATTAACCTGGGCCAGGGCTTTCCGAAC TTTGATGGCCCGGATTTTGTGAAAGAAGCGGCGATTCAGGCGATTCGTGATGGC AACAACCAGTATGCGCGTGGCTATGGCGTGCCGGATCTGAACATTGCGATTAG 10 CGAACGTTATAAAAAAGATACCGGCCTGGCGGTGGATCCGGAAAAAGAAATTAC CGTGACCAGCGGCTGCACCGAAGCGATTGCGGCGACCGTGCTGGGCCTGATT AACCCGGGCGATGAAGTGATTGTGTTTGCGCCGTTTTATGATAGCTATGAAGCG ACCCTGAGCATGGCGGGCGCGAAAGTGAAAGGCATTACCCTGCGTCCGCCGG ATTTTGCGCTGCCGATTGAAGAACTGAAAAGCACCATTAGCAAAAACACCCGTG 15 CGATTCTGCTGAACACCCCGCATAACCCGACCGGCAAAATGTTTACCCCGGAAG AACTGAACACCATTGCGAGCCTGTGCATTGAAAACGATGTGCTGGTGTTTAGCG ATGAAGTGTATGATAAACTGGCGTTTGATATGGAACATATTAGCATTGCGAGCCT GCCGGGCATGTTTGAACGTACCGTGACCATGAACAGCCTGGGCAAAACCTTTA GCCTGACCGGCTGGAAAATTGGCTGGGCGATTGCGCCGCCGCATCTGACCTGG 20 GGCGTGCGTCAGGCGCATGCGTTTCTGACCTTTGCAACCAGCAACCCGATGCA GTGGGCAGCAGCAGTGGCACTGCGTGCACCGGATAGCTATTATACCGAACTGA AACGTGATTATATGGCGAAACGTAGCATTCTGGTGGAAGGCCTGAAAGCGGTG GGCTTTAAAGTGTTTCCGAGCAGCGGCACCTATTTTGTGGTGGTGGATCATACC CCGTTTGGCCATGAAAACGATATTGCGTTTTGCGAATATCTGGTGAAAGAAGTG 25 GGCGTGGTGGCGATTCCGACCAGCGTGTTTTATCTGAACCCGGAAGAAGGCAA AAACCTGGTGCGTTTTACCTTTTGCAAAGATGAAGGCACCCTGCGTGCGGCGGT GGATCGTATGAAAGAAAAACTGCGTAAAGTCGACAAGCTTGCGGCCGCACTCG AGCACCACCACCACCACCACTGA 30 SEQ ID NO: 17 Zebra fish GPR amino acid sequence Amino acid sequence of Danio rerio cloned and expressed in E. coli (bold, italicized amino acids are added from vector/ cloning and His tag on C-terminus) 35 MSVAKRLEKFKTTIFTQMSMLAIKHGAINLGQGFPNFDGPDFVKEAAQAIRDGNNQ YARGYGVPDLNIAISERYKKDTGLAVDPEKEITVTSGCTEAIAATVLGLINPGDEVIVF APFYDSYEATLSMAGAKVKGITLRPPDFALPIEELKSTISKNTRAILLNTPHNPTGKMF TPEELNTIASLCI EN:DVLVFSDEVYDKLAFDMEHISIASLPGMFERTVTMNSLGKTFSL TGWKIGWAIAPPHLTWGVRQAHAFLTFATSNPMQWAAAVALRAPDSYYTELKRDY 40 MAKRSILVEGLKAVGFKVFPSSGTYFVVVDHTPFGHENDIAFCEYLVKEVGVVAIPT
SVFYLNPEEGKNLVRFTFCKDEGTLRAAVDRMKEKLRKVDKLAAALEHHHHHH
97 WO 2010/025466 PCT/US2009/055557 SEQ ID NO: 18 Arabidopsis truncated GPT -30 construct DNA sequence Arabidopsis GPT with 30 amino acids removed from the targeting sequence. ATGGCCAAAATCCATCGTCCTATCGGAGCCACCATGACCACAGTTTCGACTCAG 5 AACGAGTCTACTCAAAAACCCGTCCAGGTGGCGAAGAGATTAGAGAAGTTCAAG ACTACTATTTTCACTCAAATGAGCATATTGGCAGTTAAACATGGAGCGATCAATT TAGGCCAAGGCTTTCCCAATTTCGACGGTCCTGATTTTGTTAAAGAAGCTGCGA TCCAAGCTATTAAAGATGGTAAAAACCAGTATGCTCGTGGATACGGCATTCCTCA GCTCAACTCTGCTATAGCTGCGCGGTTTCGTGAAGATACGGGTCTTGTTGTTGA 10 TCCTGAGAAAGAAGTTACTGTTACATCTGGTTGCACAGAAGCCATAGCTGCAGC TATGTTGGGTTTAATAAACCCTGGTGATGAAGTCATTCTCTTTGCACCGTTTTAT GATTCCTATGAAGCAACACTCTCTATGGCTGGTGCTAAAGTAAAAGGAATCACTT TACGTCCACCGGACTTCTCCATCCCTTTGGAAGAGCTTAAAGCTGCGGTAACTA ACAAGACTCGAGCCATCCTTATGAACACTCCGCACAACCCGACCGGGAAGATGT 15 TCACTAGGGAGGAGCTTGAAACCATTGCATCTCTCTGCATTGAAAACGATGTGC TTGTGTTCTCGGATGAAGTATACGATAAGCTTGCGTTTGAAATGGATCACATTTC TATAGCTTCTCTTCCCGGTATGTATGAAAGAACTGTGACCATGAATTCCCTGGGA AAGACTTTCTCTTTAACCGGATGGAAGATCGGCTGGGCGATTGCGCCGCCTCAT CTGACTTGGGGAGTTCGACAAGCACACTCTTACCTCACATTCGCCACATCAACA 20 CCAGCACAATGGGCAGCCGTTGCAGCTCTCAAGGCACCAGAGTCTTACTTCAAA GAGCTGAAAAGAGATTACAATGTGAAAAAGGAGACTCTGGTTAAGGGTTTGAAG GAAGTCGGATTTACAGTGTTCCCATCGAGCGGGACTTACTTTGTGGTTGCTGAT CACACTCCATTTGGAATGGAGAACGATGTTGCTTTCTGTGAGTATCTTATTGAAG AAGTTGGGGTCGTTGCGATCCCAACGAGCGTCTTTTATCTGAATCCAGAAGAAG 25 GGAAGAATTTGGTTAGGTTTGCGTTCTGTAAAGACGAAGAGACGTTGC GTGGTGCAATTGAGAGGATGAAGCAGAAGCTTAAGAGAAAAGTCTGA SEQ ID NO: 19 Arabidopsis truncated GPT ~30 construct amino acid sequence 30 MAKIHRPIGATMTTVSTQNESTQKPVQVAKRLEKFKTTIFTQMSILAVKHGAINLGQG FPNFDGPDFVKEAAIQAIKDGKNQYARGYGIPQLNSAIAARFREDTGLVVDPEKEVT VTSGCTEAIAAAMLGLINPGDEVILFAPFYDSYEATLSMAGAKVKGITLRPPDFSI PLE ELKAAVTNKTRAILMNTPHNPTGKMFTREELETIASLCI ENDVLVFSDEVYDKLAFEM 35 DHISIASLPGMYERTVTMNSLGKTFSLTGWKIGWAIAPPHLTWGVRQAHSYLTFATS TPAQWAAVAALKAPESYFKELKRDYNVKKETLVKGLKEVGFTVFPSSGTYFVVADH TPFGMENDVAFCEYLIEEVGVVAIPTSVFYLNPEEGKNLVRFAFCKDEETLRGAIER MKQKLKRKV 40 98 WO 2010/025466 PCT/US2009/055557 SEQ ID NO: 20: Arabidopsis truncated GPT -45 construct DNA sequence Arabidopsis GPT with 45 residues in the targeting sequence removed ATGGCGACTCAGAACGAGTCTACTCAAAAACCCGTCCAGGTGGCGAAGAGATTA 5 GAGAAGTTCAAGACTACTATTTTCACTCAAATGAGCATATTGGCAGTTAAACATG GAGCGATCAATTTAGGCCAAGGCTTTCCCAATTTCGACGGTCCTGATTTTGTTAA AGAAGCTGCGATCCAAGCTATTAAAGATGGTAAAAACCAGTATGCTCGTGGATA CGGCATTCCTCAGCTCAACTCTGCTATAGCTGCGCGGTTTCGTGAAGATACGGG TCTTGTTGTTGATCCTGAGAAAGAAGTTACTGTTACATCTGGTTGCACAGAAGCC 10 ATAGCTGCAGCTATGTTGGGTTTAATAAACCCTGGTGATGAAGTCATTCTCTTTG CACCGTTTTATGATTCCTATGAAGCAACACTCTCTATGGCTGGTGCTAAAGTAAA AGGAATCACTTTACGTCCACCGGACTTCTCCATCCCTTTGGAAGAGCTTAAAGC TGCGGTAACTAACAAGACTCGAGCCATCCTTATGAACACTCCGCACAACCCGAC CGGGAAGATGTTCACTAGGGAGGAGCTTGAAACCATTGCATCTCTCTGCATTGA 15 AAACGATGTGCTTGTGTTCTCGGATGAAGTATACGATAAGCTTGCGTTTGAAATG GATCACATTTCTATAGCTTCTCTTCCCGGTATGTATGAAAGAACTGTGACCATGA ATTCCCTGGGAAAGACTTTCTCTTTAACCGGATGGAAGATCGGCTGGGCGATTG CGCCGCCTCATCTGACTTGGGGAGTTCGACAAGCACACTCTTACCTCACATTCG CCACATCAACACCAGCACAATGGGCAGCCGTTGCAGCTCTCAAGGCACCAGAG 20 TCTTACTTCAAAGAGCTGAAAAGAGATTACAATGTGAAAAAGGAGACTCTGGTTA AGGGTTTGAAGGAAGTCGGATTTACAGTGTTCCCATCGAGCGGGACTTACTTTG TGGTTGCTGATCACACTCCATTTGGAATGGAGAACGATGTTGCTTTCTGTGAGTA TCTTATTGAAGAAGTTGGGGTCGTTGCGATCCCAACGAGCGTCTTTTATCTGAAT CCAGAAGAAGGGAAGAATTTGGTTAGGTTTGCGTTCTGTAAAGACGAAGAGACG 25 TTGCGTGGTGCAATTGAGAGGATGAAGCAGAAGCTTAAGAGAAAAGTCTGA SEQ ID NO: 21: Arabidopsis truncated GPT 45 construct amino acid sequence 30 MATQNESTQKPVQVAKRLEKFKTTIFTQMSILAVKHGAINLGQGFPNFDGPDFVKEA AIQAIKDGKNQYARGYGI PQLNSAIAARFREDTGLVVDPEKEVTVTSGCTEAIAAAML GLINPGDEVILFAPFYDSYEATLSMAGAKVKGITLRPPDFSIPLEELKAAVTNKTRAIL MNTPHNPTGKMFTREELETIASLCIENDVLVFSDEVYDKLAFEMDHISIASLPGMYER TVTMNSLGKTFSLTGWKIGWAIAPPHLTWGVRQAHSYLTFATSTPAQWAAVAALKA 35 PESYFKELKRDYNVKKETLVKGLKEVGFTVFPSSGTYFVVADHTPFGMENDVAFCE YLIEEVGVVAIPTSVFYLNPEEGKNLVRFAFCKDEETLRGAIERMKQKLKRKV SEQ ID NO: 22: Tomato Rubisco promoter 40 TOMATO RuBisCo rbcS3C promoter sequence from KpnI to Ncol GGTACCGTTTGAATCCTCCTTAAAGTTTTTCTCTGGAGAAACTGTAGTAATTTTAC TTTGTTGTGTTCCCTTCATCTTTTGAATTAATGGCATTTGTTTTAATACTAATCTGC TTCTGAAACTTGTAATGTATGTATATCAGTTTCTTATAATTTATCCAAGTAATATCT 99 WO 2010/025466 PCT/US2009/055557 TCCATTCTCTATGCAATTGCCTGCATAAGCTCGACAAAAGAGTACATCAACCCCT CCTCCTCTGGACTACTCTAGCTAAACTTGAATTTCCCCTTAAGATTATGAAATTG ATATATCCTTAACAAACGACTCCTTCTGTTGGAAAATGTAGTACTTGTCTTTCTTC TTTTGGGTATATATAGTTTATATACACCATACTATGTACAACATCCAAGTAGAGTG 5 AAATGGATACATGTACAAGACTTATTTGATTGATTGATGACTTGAGTTGCCTTAG GAGTAACAAATTCTTAGGTCAATAAATCGTTGATTTGAAATTAATCTCTCTGTCTT AGACAGATAGGAATTATGACTTCCAATGGTCCAGAAAGCAAAGTTCGCACTGAG GGTATACTTGGAATTGAGACTTGCACAGGTCCAGAAACCAAAGTTCCCATCGAG CTCTAAAATCACATCTTTGGAATGAAATTCAATTAGAGATAAGTTGCTTCATAGCA 10 TAGGTAAAATGGAAGATGTGAAGTAACCTGCAATAATCAGTGAAATGACATTAAT ACACTAAATACTTCATATGTAATTATCCTTTCCAGGTTAACAATACTCTATAAAGT AAGAATTATCAGAAATGGGCTCATCAAACTTTTGTACTATGTATTTCATATAAGGA AGTATAACTATACATAAGTGTATACACAACTTTATTCCTATTTTGTAAAGGTGGAG AGACTGTTTTCGATGGATCTAAAGCAATATGTCTATAAAATGCATTGATATAATAA 15 TTATCTGAGAAAATCCAGAATTGGCGTTGGATTATTTCAGCCAAATAGAAGTTTG TACCATACTTGTTGATTCCTTCTAAGTTAAGGTGAAGTATCATTCATAAACAGTTT TCCCCAAAGTACTACTCACCAAGTTTCCCTTTGTAGAATTAACAGTTCAAATATAT GGCGCAGAAATTACTCTATGCCCAAAACCAAACGAGAAAGAAACAAAATACAGG GGTTGCAGACTTTATTTTCGTGTTAGGGTGTGTTTTTTCATGTAATTAATCAAAAA 20 ATATTATGACAAAAACATTTATACATATTTTTACTCAACACTCTGGGTATCAGGGT GGGTTGTGTTCGACAATCAATATGGAAAGGAAGTATTTTCCTTATTTTTTTAGTTA ATATTTTCAGTTATACCAAACATACCTTGTGATATTATTTTTAAAAATGAAAAACTC GTCAGAAAGAAAAAGCAAAAGCAACAAAAAAATTGCAAGTATTTTTTAAAAAAGA AAAAAAAAACATATCTTGTTTGTCAGTATGGGAAGTTTGAGATAAGGACGAGTGA 25 GGGGTTAAAATTCAGTGGCCATTGATTTTGTAATGCCAAGAACCACAAAATCCAA TGGTTACCATTCCTGTAAGATGAGGTTTGCTAACTCTTTTTGTCCGTTAGATAGG AAGCCTTATCACTATATATACAAGGCGTCCTAATAACCTCTTAGTAACCAATTATT TCAGCACCA TGG 30 SEQ ID NO: 23: Bamboo GPT DNA sequence ATGGCCTCCGCGGCCGTCTCCACCGTCGCCACCGCCGCCGACGGCGTCGCGA AGCCGACGGAGAAGCAGCCGGTACAGGTCGCAAAGCGTTTGGAAAAGTTTAAG 35 ACAACAATTTTCACACAGATGAGCATGCTTGCCATCAAGCATGGAGCAATAAAC CTCGGCCAGGGCTTTCCGAATTTTGATGGCCCTGACTTTGTGAAAGAAGCTGCT ATTCAAGCTATCAATGCTGGGAAGAATCAGTATGCAAGAGGATATGGTGTGCCT GAACTGAACTCGGCTGTTGCTGAAAGGTTCCTGAAGGACAGTGGCTTGCAAGTC GATCCCGAGAAGGAAGTTACTGTCACATCTGGGTGCACGGAAGCGATAGCTGC 40 AACGATATTGGGTCTTATCAACCCTGGCGATGAAGTGATCTTGTTTGCTCCATTC TATGATTCATACGAGGCTACGCTGTCGATGGCTGGTGCCAATGTAAAAGCCATT ACTCTCCGTCCTCCAGATTTTGCAGTCCCTCTTGAGGAGCTAAAGGCCACAGTC TCTAAGAACACCAGAGCGATAATGATAAACACACCACACAATCCTACTGGGAAA ATGTTTTCTAGGGAAGAACTTGAATTCATTGCTACTCTCTGCAAGAAAAATGATG 100 WO 2010/025466 PCT/US2009/055557 TGTTGCTTTTTGCTGATGAGGTCTATGACAAGTTGGCATTTGAGGCAGATCATAT ATCAATGGCTTCTATTCCTGGCATGTATGAGAGGACTGTGACTATGAACTCTCTG GGGAAGACATTCTCTCTAACAGGATGGAAGATCGGTTGGGCAATAGCACCACCA CACCTGACATGGGGTGTAAGGCAGGCACACTCATTCCTCACATTTGCCACCTGC 5 ACACCAATGCAATCGGCGGCGGCGGCGGCTCTTAGAGCACCAGATAGCTACTA TGGGGAGCTGAAGAGGGATTACGGTGCAAAGAAAGCGATACTAGTCGACGGAC TCAAGGCTGCAGGTTTTATTGTTTACCCTTCAAGTGGAACATACTTTGTCATGGT CGATCACACCCCGTTTGGTTTCGACAATGATATTGAGTTCTGCGAGTATTTGATC CGCGAAGTCGGTGTTGTCGCCATACCACCAAGCGTATTTTATCTCAACCCTGAG 10 GATGGGAAGAACTTGGTGAGGTTCACCTTCTGCAAGGATGATGATACGCTGAGA GCCGCAGTTGAGAGGATGAAGACAAAGCTCAGGAAAAAATGA SEQ ID NO: 24: Bamboo GPT amino acid sequence 15 MASAAVSTVATAADGVAKPTEKQPVQVAKRLEKFKTTIFTQMSMLAIKHGAINLGQG FPNFDGPDFVKEAAIQAINAGKNQYARGYGVPELNSAVAERFLKDSGLQVDPEKEV TVTSGCTEAIAATILGLINPGDEVILFAPFYDSYEATLSMAGANVKAITLRPPDFAVPL EELKATVSKNTRAIMINTPHNPTGKMFSREELEFIATLCKKNDVLLFADEVYDKLAFE 20 ADHISMASIPGMYERTVTMNSLGKTFSLTGWKIGWAIAPPHLTWGVRQAHSFLTFA TCTPMQSAAAAALRAPDSYYGELKRDYGAKKAILVDGLKAAGFIVYPSSGTYFVMV DHTPFGFDNDIEFCEYLIREVGVVAIPPSVFYLNPEDGKNLVRFTFCKDDDTLRAAVE RMKTKLRKK 25 SEQ ID NO: 25: 1305.1 +rbcS3C promoter + catl intron with rice GPT gene. Cambial305.1 with (3' end of) rbcS3C+rice GPT. Underlined ATG is start site, parentheses are the catl intron and the underlined actagt is the spel cloning site used 30 to splice in the rice gene. AAAAAAGAAAAAAAAAACATATCTTGTTTGTCAGTATGGGAAGTTTGAGATAAGG ACGAGTGAGGGGTTAAAATTCAGTGGCCATTGATTTTGTAATGCCAAGAACCAC AAAATCCAATGGTTACCATTCCTGTAAGATGAGGTTTGCTAACTCTTTTTGTCCG 35 TTAGATAGGAAGCCTTATCACTATATATACAAGGCGTCCTAATAACCTCTTAGTA ACCAATTATTTCAGCACCATGGTAGATCTGAGG(GTAAATTTCTAGTTTTTCTCCT TCATTTTCTTGGTTAGGACCCTTTTCTCTTTTTATTTTTTTGAGCTTTGATCTTTCT TTAAACTGATCTATTTTTTAATTGATTGGTTATGGTGTAAATATTACATAGCTTTAA CTGATAATCTGATTACTTTATTTCGTGTGTCTATGATGATGATGATAGTTACAG)A 40 ACCGACGAACTAGTATGAATCTGGCCGGCTTTCTCGCCACGCCCGCGACCGCG ACCGCGACGCGGCATGAGATGCCGTTAAATCCCTCCTCCTCCGCCTCCTTCCTC CTCTCCTCGCTCCGCCGCTCGCTCGTCGCGTCGCTCCGGAAGGCCTCGCCGG CGGCGGCCGCGGCGCTCTCCCCCATGGCCTCCGCGTCCACCGTCGCCGCCGA GAACGGCGCCGCCAAGGCGGCGGCGGAGAAGCAGCAGCAGCAGCCTGTGCA 101 WO 2010/025466 PCT/US2009/055557 GGTTGCAAAGCGGTTGGAAAAGTTTAAGACGACCATTTTCACACAGATGAGTAT GCTTGCCATCAAGCATGGAGCAATAAACCTTGGCCAGGGTTTTCCGAATTTCGA TGGCCCTGACTTTGTAAAAGAGGCTGCTATTCAAGCTATCAATGCTGGGAAGAA TCAGTACGCAAGAGGATATGGTGTGCCTGAACTGAACTCAGCTATTGCTGAAAG 5 ATTCCTGAAGGACAGCGGACTGCAAGTCGATCCGGAGAAGGAAGTTACTGTCA CATCTGGATGCACAGAAGCTATAGCTGCAACAATTTTAGGTCTAATTAATCCAGG CGATGAAGTGATATTGTTTGCTCCATTCTATGATTCATATGAGGCTACCCTGTCA ATGGCTGGTGCCAACGTAAAAGCCATTACTCTCCGTCCTCCAGATTTTTCAGTC CCTCTTGAAGAGCTAAAGGCTGCAGTCTCGAAGAACACCAGAGCTATTATGATA 10 AACACCCCGCACAATCCTACTGGGAAAATGTTTACAAGGGAAGAACTTGAGTTT ATTGCCACTCTCTGCAAGGAAAATGATGTGCTGCTTTTTGCTGATGAGGTCTAC GACAAGTTAGCTTTTGAGGCAGATCATATATCAATGGCTTCTATTCCTGGCATGT ATGAGAGGACCGTGACCATGAACTCTCTTGGGAAGACATTCTCTCTTACAG GAT GGAAGATCGGTTGGGCAATCGCACCGCCACACCTGACATGGGGTGTAAGGCAG 15 GCACACTCATTCCTCACGTTTGCGACCTGCACACCAATGCAAGCAGCTGCAGCT GCAGCTCTGAGAGCACCAGATAGCTACTATGAGGAACTGAGGAGGGATTATGG AGCTAAGAAGGCATTGCTAGTCAACGGACTCAAGGATGCAGGTTTCATTGTCTA TCCTTCAAGTGGAACATACTTCGTCATGGTCGACCACACCCCATTTGGTTTCGA CAATGATATTGAGTTCTGCGAGTATTTGATTCGCGAAGTCGGTGTTGTCGCCATA 20 CCACCTAGTGTATTTTATCTCAACCCTGAGGATGGGAAGAACTTGGTGAGGTTC ACCTTTTGCAAGGATGATGAGACGCTGAGAGCCGCGGTTGAGAGGATGAAGAC AAAGCTCAGGAAAAAATGA 25 SEQ ID NO: 26: HORDEUM GPT SEQUENCE IN VECTOR Cambia1305.1 with (3' end of) rbcS3C+hordeum ID14. Underlined ATG is start site, parentheses are the cati intron and the underlined actagt is the spel cloning site used to splice in the hordeurn gene. 30 AAAAAAGAAAAAAAAAACATATCTTGTTTGTCAGTATGGGAAGTTTGAGATAAGG ACGAGTGAGGGGTTAAAATTCAGTGGCCATTGATTTTGTAATGCCAAGAACCAC AAAATCCAATGGTTACCATTCCTGTAAGATGAGGTTTGCTAACTCTTTTTGTCCG TTAGATAGGAAGCCTTATCACTATATATACAAGGCGTCCTAATAACCTCTTAGTA ACCAATTATTTCAGCACCATGGTAGATCTGAGG(GTAAATTTCTAGTTTTTCTCCT 35 TCATTTTCTTGGTTAGGACCCTTTTCTCTTTTTATTTTTTTGAGCTTTGATCTTTCT TTAAACTGATCTATTTTTTAATTGATTGGTTATGGTGTAAATATTACATAGCTTTAA CTGATAATCTGATTACTTTATTTCGTGTGTCTATGATGATGATGATAGTTACAG)A ACCGACGAACTAGTATGGCATCCGCCCCCGCCTCCGCCTCCGCGGCCCTCTCC ACCGCCGCCCCCGCCGACAACGGGGCCGCCAAGCCCACGGAGCAGCGGCCG 40 GTACAGGTGGCTAAGCGATTGGAGAAGTTCAAAACAACAATTTTCACACAGATG AGCATGCTCGCAGTGAAGCATGGAGCAATAAACCTTGGACAGGGGTTTCCCAAT TTTGATGGCCCTGACTTTGTCAAAGATGCTGCTATTGAGGCTATCAAAGCTGGA AAGAATCAGTATGCAAGAGGATATGGTGTGCCTGAATTGAACTCAGCTGTTGCT GAGAGATTTCTCAAGGACAGTGGATTGCACATCGATCCTGATAAGGAAGTTACT 102 WO 2010/025466 PCT/US2009/055557 GTTACATCTGGGTGCACAGAAGCAATAGCTGCAACGATATTGGGTCTGATCAAC CCTGGGGATGAAGTCATACTGTTTGCTCCATTCTATGATTCTTATGAGGCTACAC TGTCCATGGCTGGTGCGAATGTCAAAGCCATTACACTCCGCCCTCCGGACTTTG CAGTCCCTCTTGAAGAGCTAAAGGCTGCAGTCTCGAAGAATACCAGAGCAATAA 5 TGATTAATACACCTCACAACCCTACCGGGAAAATGTTCACAAGGGAGGAACTTG AGTTCATTGCTGATCTCTGCAAGGAAAATGACGTGTTGCTCTTTGCCGATGAG:G TCTACGACAAGCTGGCGTTTGAGGCGGATCACATATCAATGGCTTCTATTCCTG GCATGTATGAGAGGACCGTCACTATGAACTCCCTGGGGAAGACGTTCTCCTTGA CCGGATGGAAGATCGGCTGGGCGATAGCACCACCGCACCTGACATGGGGCGT 10 AAGGCAGGCACACTCCTTCCTCACATTCGCCACCTCCACGCCGATGCAATCAGC AGCGGCGGCGGCCCTGAGAGCACCGGACAGCTACTTTGAGGAGCTGAAGAGG GACTACGGCGCAAAGAAAGCGCTGCTGGTGGACGGGCTCAAGGCGGCGGGCT TCATCGTCTACCCTTCGAGCGGAACCTACTTCATCATGGTCGACCACACCCCGT TCGGGTTCGACAACGACGTCGAGTTCTGCGAGTACTTGATCCGCGAGGTCGGC 15 GTCGTGGCCATCCCGCCAAGCGTGTTCTACCTGAACCCGGAGGACGGGAAGAA CCTGGTGAGGTTCACCTTCTGCAAGGACGACGACACGCTAAGGGCGGCGGTG GACAGGATGAAGGCCAAGCTCAGGAAGAAATGATTGAGGGGCGCACGTGTGA 20 SEQ ID NO: 27 Cambia 1201 + Arabidopsis GPT sequence (35S promoter from CaMV in italics) CATGGAGTCAAAGATTCAAATAGAGGACCTAACAGAACTCGCCGTAAAGACTGG CGAACAGTTCA TACAGAGTCTCTTACGACTCAA TGACAA GAAGAAAA TCTTCG TC 25 AACATGGTGGAGCACGACACACTTGTCTACTCCAAAAATATCAAAGATACAGTCT CAGAAGACCAAAGGGCAATTGAGACTTTTCAACAAAGGGTAATATCCGGAAACC TCCTCGGA TTCCA TTGCCCAGC TA TCTGTCACTTTA TTGTGAAGA TAGTGGAAAA GGAAGGTGGCTCCTACAAATGCCATCATTGCGATAAAGGAAAGGCCATCGTTGA AGA TGCCTCTGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGCA 30 TCGTGGAAAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTG ATATCTCCACTGACGTAAGGGATGACGCACAATCCCACTATCCTTCGCAAGACC C TTCCTC TA TA TAAGGAAGTTCA TTTCA TTTGGA GA GAACACGGGGGACTCTTGA CCATGTACCTGGACATAAATGGTGTGATGATCAAACAGTTTAGCTTCAAAGCCTC TCTTCTCCCATTCTCTTCTAATTTCCGACAAAGCTCCGCCAAAATCCATCGTCCT 35 ATCGGAGCCACCATGACCACAGTTTCGACTCAGAACGAGTCTACTCAAAAACCC GTCCAGGTGGCGAAGAGATTAGAGAAGTTCAAGACTACTATTTTCACTCAAATG AGCATATTGGCAGTTAAACATGGAGCGATCAATTTAGGCCAAGGCTTTCCCAATT TCGACGGTCCTGATTTTGTTAAAGAAGCTGCGATCCAAGCTATTAAAGATGGTAA AAACCAGTATGCTCGTGGATACGGCATTCCTCAGCTCAACTCTGCTATAGCTGC 40 GCGGTTTCGTGAAGATACGGGTCTTGTTGTTGATCCTGAGAAAGAAGTTACTGT TACATCTGGTTGCACAGAAGCCATAGCTGCAGCTATGTTGGGTTTAATAAACCCT GGTGATGAAGTCATTCTCTTTGCACCGTTTTATGATTCCTATGAAGCAACACTCT CTATGGCTGGTGCTAAAGTAAAAGGAATCACTTTACGTCCACCGGACTTCTCCA TCCCTTTGGAAGAGCTTAAAGCTGCGGTAACTAACAAGACTCGAGCCATCCTTA 103 WO 2010/025466 PCT/US2009/055557 TGAACACTCCGCACAACCCGACCGGGAAGATGTTCACTAGGGAGGAGCTTGAA ACCATTGCATCTCTCTGCATTGAAAACGATGTGCTTGTGTTCTCGG:ATGAAGTAT ACGATAAGCTTGCGTTTGAAATGGATCACATTTCTATAGCTTCTCTTCCCGGTAT G TATGAAAGAACTGTGACCATGAATTCCCTGGGAAAGACTTTCTCTTTAACCGGA 5 TGGAAGATCGGCTGGGCGATTGCGCCGCCTCATCTGACTTGGGGAGTTCGACA AGCACACTCTTACCTCACATTCGCCACATCAACACCAGCACAATGGGCAGCCGT TGCAGCTCTCAAGGCACCAGAGTCTTACTTCAAAGAGCTGAAAAGAGATTACAA TGTGAAAAAGGAGACTCTGGTTAAGGGTTTGAAGGAAGTCGGATTTACAGTGTT CCCATCGAGCGGGACTTACTTTGTGGTTGCTGATCACACTCCATTTGGAATGGA 10 GAACGATGTTGCTTTCTGTGAGTATCTTATTGAAGAAGTTGGGGTCGTTGCGAT CCCAACGAGCGTCTTTTATCTGAATCCAGAAGAAGGGAAGAATTTGGTTAGGTT TGCGTTCTGTAAAGACGAAGAGACGTTGCGTGGTGCAATTGAGAGGATGAAGC AGAAGCTTAAGAGAAAAGTCTGA 15 SEQ ID NO: 28 Cambia p1 305,1 with (3 end of) rbcS3C+Arabidopsis GPT. Underlined ATG is start site, parentheses are the catl intron and the underlined actagt is the spel cloning site used to splice in the Arabidopsis gene. 20 AAAAAAGAAAAAAAAAACATATCTTGTTTGTCAGTATGGGAAGTTTGAGATAAGG ACGAGTGAGGGGTTAAAATTCAGTGGCCATTGATTTTGTAATGCCAAGAACCAC AAAATCCAATGGTTACCATTCCTGTAAGATGAGGTTTGCTAACTCTTTTTGTCCG TTAGATAGGAAGCCTTATCACTATATATACAAGGCGTCCTAATAACCTCTTAGTA ACCAATTATTTCAGCACCATGGTAGATCTGAGG(GTAAATTTCTAGTTTTTCTCCT 25 TCATTTTCTTGGTTAGGACCCTTTTCTCTTTTTATTTTTTTGAGCTTTGATCTTTCT TTAAACTGATCTATTTTTTAATTGATTGGTTATGGTGTAAATATTACATAGCTTTAA CTGATAATCTGATTACTTTATTTCGTGTGTCTATGATGATGATGATAGTTACAG)A ACCGACGAA CTAGTATGTACCTGGACATAAATGGTGTGATGATCAAACAGTTTA GCTTCAAAGCCTCTCTTCTCCCATTCTCTTCTAATTTCCGACAAAGCTCCGCCAA 30 AATCCATCGTCCTATCGGAGCCACCATGACCACAGTTTCGACTCAGAACGAGTC TACTCAAAAACCCGTCCAGGTGGCGAAGAGATTAGAGAAGTTCAAGACTACTAT TTTCACTCAAATGAGCATATTGGCAGTTAAACATGGAGCGATCAATTTAGGCCAA GGCTTTCCCAATTTCGACGGTCCTGATTTTGTTAAAGAAGCTGCGATCCAAGCTA TTAAAGATGGTAAAAACCAGTATGCTCGTGGATACGGCATTCCTCAGCTCAACT 35 CTGCTATAGCTGCGCGGTTTCGTGAAGATACGGGTCTTGTTGTTGATCCTGAGA AAGAAGTTACTGTTACATCTGGTTGCACAGAAGCCATAGCTGCAGCTATGTTGG GTTTAATAAACCCTGGTGATGAAGTCATTCTCTTTGCACCGTTTTATGATTCCTAT GAAGCAACACTCTCTATGGCTGGTGCTAAAGTAAAAGGAATCACTTTACGTCCA CCGGACTTCTCCATCCCTTTGGAAGAGCTTAAAGCTGCGGTAACTAACAAGACT 40 CGAGCCATCCTTATGAACACTCCGCACAACCCGACCGGGAAGATGTTCACTAG GGAGGAGCTTGAAACCATTGCATCTCTCTGCATTGAAAACGATGTGCTTGTGTT CTCGGATGAAGTATACGATAAGCTTGCGTTTGAAATGGATCACATTTCTATAGCT TCTCTTCCCGGTATGTATGAAAGAACTGTGACCATGAATTCCCTGGGAAAGACTT TCTCTTTAACCGGATGGAAGATCGGCTGGGCGATTGCGCCGCCTCATCTGACTT 104 WO 2010/025466 PCT/US2009/055557 GGGGAGTTCGACAAGCACACTCTTACCTCACATTCGCCACATCAACACCAGCAC AATGGGCAGCCGTTGCAGCTCTCAAGGCACCAGAGTCTTACTTCAAAGAGCTGA AAAGAGATTACAATGTGAAAAAGGAGACTCTGGTTAAGGGTTTGAAGGAAGTCG GATTTACAGTGTTCCCATCGAGCGGGACTTACTTTGTGGTTGCTGATCACACTC 5 CATTTGGAATGGAGAACGATGTTGCTTTCTGTGAGTATCTTATTGAAGAAGTTGG GGTCGTTGCGATCCCAACGAGCGTCTTTTATCTGAATCCAGAAGAAGGGAAGAA TTTGGTTAGGTTTGCGTTCTGTAAAGACGAAGAGACGTTGCGTGGTG:CAATTGA GAGGATGAAGCAGAAGCTTAAGAGAAAAGTCTGA 10 SEQ ID NO: 29 Arabidpsis GPT coding sequence (mature protein, no targeting sequence) GTGGCGAAGAGATTAGAGAAGTTCAAGACTACTATTTTCACTCAAATGAGCATAT 15 TGGCAGTTAAACATGGAGCGATCAATTTAGGCCAAGGCTTTCCCAATTTCGACG GTCCTGATTTTGTTAAAGAAGCTGCGATCCAAGCTATTAAAGATGGTAAAAACCA GTATGCTCGTGGATACGGCATTCCTCAGCTCAACTCTGCTATAGCTGCGCGGTT TCGTGAAGATACGGGTCTTGTTGTTGATCCTGAGAAAGAAGTTACTGTTACATCT GGTTGCACAGAAGCCATAGCTGCAGCTATGTTGGGTTTAATAAACCCTGGTGAT 20 GAAGTCATTCTCTTTGCACCGTTTTATGATTCCTATGAAGCAACACTCTCTATGG CTGGTGCTAAAGTAAAAGGAATCACTTTACGTCCACCGGACTTCTCCATCCCTTT GGAAGAGCTTAAAGCTGCGGTAACTAACAAGACTCGAGCCATCCTTATGAACAC TCCGCACAACCCGACCGGGAAGATGTTCACTAGGGAGGAGCTTGAAACCATTG CATCTCTCTGCATTGAAAACGATGTGCTTGTGTTCTCGGATGAAGTATACGATAA 25 GCTTGCGTTTGAAATGGATCACATTTCTATAGCTTCTCTTCCCGGTATGTATGAA AGAACTGTGACCATGAATTCCCTGGGAAAGACTTTCTCTTTAACCGGATGGAAG ATCGGCTGGGCGATTGCGCCGCCTCATCTGACTTGGGGAGTTCGACAAGCACA CTCTTACCTCACATTCGCCACATCAACACCAGCACAATGGGCAGCCGTTGCAGC TCTCAAGGCACCAGAGTCTTACTTCAAAGAGCTGAAAAGAGATTACAATGTGAAA 30 AAGGAGACTCTGGTTAAGGGTTTGAAGGAAGTCGGATTTACAGTGTTCCCATCG AGCGGGACTTACTTTGTGGTTGCTGATCACACTCCATTTGGAATGGAGAACGAT GTTGCTTTCTGTGAGTATCTTATTGAAGAAGTTGGGGTCGTTGCGATCCCAACG AGCGTCTTTTATCTGAATCCAGAAGAAGGGAAGAATTTGGTTAGGTTTGCGTTCT GTAAAGACGAAGAGACGTTGCGTGGTGCAATTGAGAGGATGAAGCAGAAGCTT 35 AAGAGAAAAGTCTGA SEQ ID NO: 30 Arabidpsis GPT amino acid sequence (mature protein, no targeting sequence) 40 VAKRLEKFKTTIFTQMSILAVKHGAINLGQGFPNFDGPDFVKEAAQAIKDGKNQYAR GYGIPQLNSAIAARFREDTGLVVDPEKEVTVTSGCTEAIAAAMLGLINPGDEVILFAP FYDSYEATLSMAGAKVKGITLRPPDFSIPLEELKAAVTNKTRAILMNTPHNPTGKMFT REELETIASLCIENDVLVFSDEVYDKLAFEMDHISIASLPGMYERTVTMNSLGKTFSL 105 WO 2010/025466 PCT/US2009/055557 TGWKIGWAIAPPHLTWGVRQAHSYLTFATSTPAQWAAVAALKAPESYFKELKRDYN VKKETLVKGLKEVGFTVFPSSGTYFVVADHTPFGMENDVAFCEYLIEEVGVVAIPTS VFYLNPEEGKNLVRFAFCKDEETLRGAIERMKQKLKRKV SEQ ID NO: 31 Grape GPT amino acid sequence (mature protein, no targeting sequence) VAKRLEKFKTTIFTQMSMLAIKHGAINLGQGFPNFDGPEFVKEAAIQAIKDGKNQYAR 10 GYGVPDLNSAVADRFKKDTGLVVDPEKEVTVTSGCTEAIAATMLGLINPGDEVILFA PFYDSYEATLSMAGAQIKSITLRPPDFAVPMDELKSAISKNTRAILINTPHNPTGKMFT REELNVIASLCIENDVLVFTDEVYDKLAFEMDHISMASLPGMYERTVTMNSLGKTFS LTGWKIGWTVAPPHLTWGVRQAHSFLTFATCTPMQWAAATALRAPDSYYEELKRD YSAKKAILVEGLKAVGFRVYPSSGTYFVVVDHTPFGLKDDIAFCEYLIKEVGVVAIPT 15 SVFYLHPEDGKNLVRFTFCKDEGTLRAAVERMKEKLKPKQ SEQ ID NO: 32 Rice GPT amino acid sequence (mature protein, no targeting sequence) 20 VAKRLEKFKTTIFTQMSMLAIKHGAINLGQGFPNFDGPDFVKEAAQAINAGKNQYAR GYGVPELNSAIAERFLKDSGLQVDPEKEVTVTSGCTEAIAATILGLI NPGDEVI LFAPF YDSYEATLSMAGANVKAITLRPPDFSVPLEELKAAVSKNTRAIMINTPHNPTGKMFT REELEFIATLCKENDVLLFADEVYDKLAFEADHISMASIPGMYERTVTMNSLGKTFSL 25 TGWKIGWAIAPPHLTWGVRQAHSFLTFATCTPMQAAAAAALRAPDSYYEELRRDY GAKKALLVNGLKDAGFIVYPSSGTYFVMVDHTPFGFDNDIEFCEYLIREVGVVAIPPS VFYLNPEDGKNLVRFTFCKDDETLRAAVERMKTKLRKK 30 SEQ ID NO: 33 Soybean GPT amino acid sequence (-I mature protein, no targeting sequence) AKRLEKFQTTIFTQMSLLAI KHGAINLGQGFPNFDGPEFVKEAAIQAIRDGKNQYARG YGVPDLN IAIAERFKKDTGLVVD PEKEITVTSGCTEAIAATMIGLI NPGDEVI MFAPFY 35 DSYEATLSMAGAKVKGITLRPPDFAVPLEELKSTISKNTRAILINTPHNPTGKMFTRE ELNCIASLCIEN DVLVFTDEVYDKLAFDMEH ISMASLPGM FERTVTLN$LGKTFSLTG WKIGWAIAPPHLSWGVRQAHAFLTFATAHPFQCAAAAALRAPDSYYVELKRDYMAK RAILIEGLKAVGFKVFPSSGTYFVVVDHTPFGLENDVAFCEYLVKEVGVVAIPTSVFY LN PEEGKNLVRFTFCKDEETIRSAVERMKAKLRKVD 40 106 WO 2010/025466 PCT/US2009/055557 SEQ ID NO: 34 Barley GPT amino acid sequence (mature protein, no targeting sequence) VAKRLEKFKTTIFTQMSMLAVKHGAINLGQGFPNFDGPDFVKDAAIEAIKAGKNQYA 5 RGYGVPELNSAVAERFLKDSGLHIDPDKEVTVTSGCTEAIAATILGLINPGDEVI LFAP FYDSYEATLSMAGANVKAITLRPPDFAVPLEELKAAVSKNTRAIMINTPHNPTGKMFT REELEFIADLCKENDVLLFADEVYDKLAFEADHISMASIPGMYERTVTMNSLGKTFSL TGWKIGWAIAPPHLTWGVRQAHSFLTFATSTPMQSAAAAALRAPDSYFEELKRDYG AKKALLVDGLKAAGFIVYPSSGTYFIMVDHTPFGFDNDVEFCEYLIREVGVVAIPPSV 10 FYLNPEDGKNLVRFTFCKDDDTLRAAVDRMKAKLRKK SEQ ID NO: 35 Zebra fish GPT amino acid sequence (mature protein, no targeting sequence) 15 VAKRLEKFKTTIFTQMSMLAIKHGAINLGQGFPNFDGPDFVKEAAQAIRDGNNQYA RGYGVPDLNIAISERYKKDTGLAVDPEKEITVTSGCTEAIAATVLGLINPGDEVIVFAP FYDSYEATLSMAGAKVKGITLRPPDFALPIEELKSTISKNTRAILLNTPHNPTGKMFTP EELNTIASLCIEN:DVLVFSDEVYDKLAFDMEHISIASLPGMFERTVTMNSLGKTFSLT 20 GWKIGWAIAPPHLTWGVRQAHAFLTFATSNPMQWAAAVALRAPDSYYTELKRDYM AKRSILVEGLKAVGFKVFPSSGTYFVVVDHTPFGHENDIAFCEYLVKEVGVVAIPTSV FYLNPEEGKNLVRFTFCKDEGTLRAAVDRMKEKLRK 25 SEQ ID NO: 36 Bamboo GPT amino acid sequence (mature protein, no targeting sequence) VAKRLEKFKTTIFTQMSMLAIKHGAINLGQGFPNFDGPDFVKEAAQAINAGKNQYAR GYGVPELNSAVAERFLKDSGLQVDPEKEVTVTSGCTEAIAATILGLINPGDEVILFAP 30 FYDSYEATLSMAGANVKAITLR.PPDFAVPLEELKATVSKNTRAIMINTPHNPTGKMFS REELEFIATLCKKNDVLLFADEVYDKLAFEADHISMASIPGMYERTVTMNSLGKTFSL TGWKIGWAIAPPHLTWGVRQAHSFLTFATCTPMQSAAAAALRAPDSYYGELKRDY GAKKAILVDGLKAAGFIVYPSSGTYFVMVDHTPFGFDNDIE FCEYLIREVGVVAIPPS VFYLNPEDGKNLVRFTFCKDDDTLRAAVERMKTKLRKK 35 107
Claims (28)
1. A transgenic plant comprising a GPT transgene and a GS transgene, wherein said 5 GPT transgene and said GS transgene are operably linked to a plant promoter.
2. The transgenic plant of claim 1, wherein the GS transgene is a GSI transgene.
3. The transgenic plant of claim 1 or 2, wherein the GPT transgene encodes a 10 polypeptide having an amino acid sequence selected from the group consisting of (a) SEQ ID NO: 2; SEQ ID NO: 9; SEQ ID NO: 15, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO 24, SEQ ID NO: 30, SEQ ID NO:31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36, and (b) an amino acid sequence that is at least 75% identical to any one of SEQ ID NO: 2; SEQ ID NO: 9; 15 -SEQ ID NO: 15, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO 24, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36 and has GPT activity.
4. The transgenic plant of claim 3, wherein the GS transgene encodes a polypeptide 20 having an amino acid sequence selected form the group consisting of (a) SEQ ID NO: 4 and SEQ ID NO: 7 from residue 11, and (b) an amino acid sequence that is at least 75% identical to SEQ ID NO: 4 or SEQ ID NO: 7.
5. The transgenic plant according to claims 1-4, wherein the GPT and GS transgenes 25 are incorporated into the genome of the plant.
6. The transgenic plant of claim 5, further defined as a monocotyledonous plant.
7. The transgenic plant of claim 5, further defined as a dicotyledonous plant. 30 108 WO 2010/025466 PCT/US2009/055557
8. A progeny of any generation of the transgenic plant of claim 5, wherein said progeny comprises said GPT transgene and said GS transgene.
9. A seed of any generation of the transgenic plant of claim 5, wherein said seed 5 comprises said GPT transgene and said GS transgene.
10. The transgenic plant of claim 5 which displays an enhanced growth rate when compared to an analogous wild-type or untransformed plant. 10
11. The transgenic plant of claim 5 which displays increased biomass yield when compared to an analogous wild-type or untransformed plant.
12. The transgenic plant of claim 5 which displays increased seed yield when compared to an analogous wild-type or untransformed plant. 15
13. The transgenic plant of claim 5 which displays increased flower or flower bud yield when compared to an analogous wild-type or untransformed plant.
14. The transgenic plant of claim 5 which displays increased fruit or pod yield when 20 compared to an analogous wild-type or untransformed plant.
15. The transgenic plant of claim 5 which displays larger leaves when compared to an analogous wild-type or untransformed plant. 25
16. The transgenic plant of claim 5 which displays increased GPT activity when compared to an analogous wild-type or untransformed plant.
17. The transgenic plant of claim 5 which displays increased GS activity when compared to an analogous wild-type or untransformed plant. 109 WO 2010/025466 PCT/US2009/055557
18. The transgenic plant of claim 5 which displays increased 2-oxoglutaramate levels when compared to an analogous wild-type or untransformed plant.
19. The transgenic plant of claim 5 which displays increased nitrogen use efficiency 5 when compared to an analogous wild-type or untransformed plant.
20. The transgenic plant of claim 5 which displays increased tolerance to salt or saline conditions when compared to an analogous wild-type or untransformed plant. 10
21. A method for producing a plant having enhanced growth properties relative to an analogous wild type or untransformed plant, comprising: (a) introducing a GPT transgene into the plant; (b) introducing a GS transgene into the plant or a progeny of the plant; (c) expressing the GPT transgene and the GS transgene in the plant or the 15 progeny of the plant; and, (d) selecting a plant having an increased growth characteristic relative to a plant of the same species that does not comprise a GPT transgene or a GS transgene. 20
22. A method for producing a plant having enhanced growth properties relative to an analogous wild type or untransformed plant, comprising: (a) introducing a GS transgene into the plant; (b) introducing a GPT transgene into the plant or a progeny of the plant; (c) expressing the GS transgene and the GPT transgene in the plant or the 25 progeny of the plant; and, (d) selecting a plant having an increased growth characteristic relative to a plant of the same species that does not comprise a GS transgene or a GPT transgene. 110 WO 2010/025466 PCT/US2009/055557
23. The method according to claim 21 or 22, wherein the increased growth characteristic is selected from the group consisting of increased biomass, earlier flowering, earlier budding, increased plant height, increased flowering, increased budding, larger leaves, increased fruit or pod yield and increased seed yield. 5
24. A method of producing a plant having increased nitrogen use efficiency relative to an analogous wild type or untransformed plant, comprising: (a) introducing a GPT transgene into the plant; (b) introducing a GS transgene into the plant or a progeny of the plant; 10 (c) expressing the GPT transgene and the GS transgene in the plant or the progeny of the plant; and, (d) selecting a plant having an increased nitrogen use efficiency relative to a plant of the same species that does not comprise a GPT transgene or a GS transgene. 15
25. A method of producing a plant having increased nitrogen use efficiency relative to an analogous wild type or untransformed plant, comprising: (a) introducing a GS transgene into the plant; (b) introducing a GPT transgene into the plant or a progeny of the plant; 20 (c) expressing the GS transgene and the GPT transgene in the plant or the progeny of the plant; and, (d) selecting a plant having an increased nitrogen use efficiency relative to a plant of the same species that does not comprise a GS transgene or a GPT transgene. 25
26. A method of producing a plant seed having increased tolerance to germination or growth in salt or saline conditions relative to an analogous wild type or untransformed plant, comprising: (a) introducing a GPT transgene into the plant; 111 WO 2010/025466 PCT/US2009/055557 (b) introducing a GS transgene into the plant or a progeny of the plant; (c) expressing the GPT transgene and the GS transgene in the plant or the progeny of the plant; (d) selecting a plant having an increased growth characteristic relative to a 5 plant of the same species that does not comprise a GPT transgene or a GS transgene; and, (e) harvesting seeds from said plant and selecting for seeds that demonstrate increased germination in high salt conditions. 10
27. A method of producing a plant seed having increased tolerance to germination in salt or saline conditions relative to an analogous wild type or untransformed plant, comprising: (a) introducing a GS transgene into the plant; (b) introducing a GPT transgene into the plant or a progeny of the plant; 15 (c) expressing the GS transgene and the GPT transgene in the plant or the progeny of the plant; (d) selecting a plant having an increased growth characteristic relative to a plant of the same species that does not comprise a GS transgene or a GPT transgene; and, 20 (e) harvesting seeds from said plant and selecting a seed that demonstrates increased germination in high salt conditions.
28. The method according to claim 26 or 27, further comprising propagating a plant from the seed so selected and harvesting a seed therefrom. 25 112
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU2016202733A AU2016202733B2 (en) | 2008-08-29 | 2016-04-28 | Transgenic plants with enhanced growth characteristics |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US19052008P | 2008-08-29 | 2008-08-29 | |
US61/190,520 | 2008-08-29 | ||
PCT/US2009/055557 WO2010025466A2 (en) | 2008-08-29 | 2009-08-31 | Transgenic plants with enhanced growth characteristics |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2016202733A Division AU2016202733B2 (en) | 2008-08-29 | 2016-04-28 | Transgenic plants with enhanced growth characteristics |
Publications (3)
Publication Number | Publication Date |
---|---|
AU2009287446A1 true AU2009287446A1 (en) | 2010-03-04 |
AU2009287446B2 AU2009287446B2 (en) | 2016-01-28 |
AU2009287446C1 AU2009287446C1 (en) | 2016-08-11 |
Family
ID=41722345
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2009287446A Ceased AU2009287446C1 (en) | 2008-08-29 | 2009-08-31 | Transgenic plants with enhanced growth characteristics |
AU2016202733A Ceased AU2016202733B2 (en) | 2008-08-29 | 2016-04-28 | Transgenic plants with enhanced growth characteristics |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2016202733A Ceased AU2016202733B2 (en) | 2008-08-29 | 2016-04-28 | Transgenic plants with enhanced growth characteristics |
Country Status (14)
Country | Link |
---|---|
EP (1) | EP2334166A4 (en) |
JP (2) | JP5779095B2 (en) |
CN (1) | CN102405289A (en) |
AU (2) | AU2009287446C1 (en) |
BR (1) | BRPI0917919A2 (en) |
CA (1) | CA2735646A1 (en) |
CL (1) | CL2011000396A1 (en) |
CO (1) | CO6341510A2 (en) |
IL (1) | IL211421A (en) |
MX (3) | MX2011002110A (en) |
NZ (1) | NZ591185A (en) |
RU (2) | RU2582260C2 (en) |
WO (1) | WO2010025466A2 (en) |
ZA (1) | ZA201102266B (en) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110030104A1 (en) | 2008-08-29 | 2011-02-03 | Los Alamos National Security, Llc | Nucleic acids encoding plant glutamine phenylpyruvate transaminase (GPT) and uses thereof |
US20110030089A1 (en) | 2008-08-29 | 2011-02-03 | Los Alamos National Security, Llc | Transgenic plants with enhanced growth characteristics |
WO2010025466A2 (en) * | 2008-08-29 | 2010-03-04 | Los Alamos National Security, Llc | Transgenic plants with enhanced growth characteristics |
PE20121693A1 (en) | 2010-01-22 | 2012-12-01 | Bayer Ip Gmbh | COMBINATION OF SPIROMESIPHENE AND ABAMECTIN AS INSECTICIDES |
EP2539456B1 (en) * | 2010-02-28 | 2017-04-19 | Los Alamos National Security, LLC | Increasing plant growth by modulating omega-amidase expression in plants |
CN103717076B (en) | 2011-08-10 | 2016-04-13 | 拜耳知识产权股份有限公司 | Active compound combinations containing specific tetramic acid derivatives |
RU2696201C2 (en) * | 2014-03-10 | 2019-07-31 | Зе Стейт Оф Израил, Министри Оф Эгрикалче Энд Рурал Девелопмент, Эгрикалчерал Рисёч Организейшн (Аро) | Melon plants with increased yield of fruits |
CN113564184B (en) * | 2021-07-16 | 2023-04-18 | 昆明理工大学 | Gastrodia elata glutamine synthetase gene and application thereof |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE722494T1 (en) * | 1993-10-06 | 1997-04-03 | Univ New York | TRANSGENIC PLANTS THAT HAVE INCREASED NITROGEN CAPTURE |
US6107547A (en) * | 1993-10-06 | 2000-08-22 | New York University | Transgenic plants that exhibit enhanced nitrogen assimilation |
RU2109446C1 (en) * | 1993-10-26 | 1998-04-27 | Евгения Яковлевна Яковенко | Method for producing natural plant growth regulator |
US20070011783A1 (en) * | 1999-05-06 | 2007-01-11 | Jingdong Liu | Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement |
EP1228222A2 (en) * | 1999-11-04 | 2002-08-07 | Incyte Genomics, Inc. | Human transferase molecules |
US6831040B1 (en) * | 2000-01-27 | 2004-12-14 | The Regents Of The University Of California | Use of prolines for improving growth and other properties of plants and algae |
EP2489726A3 (en) * | 2005-01-12 | 2012-11-28 | Monsanto Technology LLC | Genes and uses for plant improvement |
US20080005810A1 (en) * | 2006-05-18 | 2008-01-03 | The Governors Of The University Of Alberta | Method of conferring multiple stress tolerance and early flowering in plants |
WO2008070179A2 (en) * | 2006-12-06 | 2008-06-12 | Monsanto Technology, Llc | Genes and uses for plant improvement |
US8362325B2 (en) * | 2007-10-03 | 2013-01-29 | Ceres, Inc. | Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics |
WO2010025466A2 (en) * | 2008-08-29 | 2010-03-04 | Los Alamos National Security, Llc | Transgenic plants with enhanced growth characteristics |
-
2009
- 2009-08-31 WO PCT/US2009/055557 patent/WO2010025466A2/en active Application Filing
- 2009-08-31 BR BRPI0917919-4A patent/BRPI0917919A2/en not_active Application Discontinuation
- 2009-08-31 RU RU2011111344/10A patent/RU2582260C2/en not_active IP Right Cessation
- 2009-08-31 RU RU2015151684A patent/RU2015151684A/en not_active Application Discontinuation
- 2009-08-31 AU AU2009287446A patent/AU2009287446C1/en not_active Ceased
- 2009-08-31 MX MX2011002110A patent/MX2011002110A/en active IP Right Grant
- 2009-08-31 CN CN2009801343364A patent/CN102405289A/en active Pending
- 2009-08-31 NZ NZ591185A patent/NZ591185A/en not_active IP Right Cessation
- 2009-08-31 EP EP09810728A patent/EP2334166A4/en not_active Withdrawn
- 2009-08-31 MX MX2012014837A patent/MX357276B/en unknown
- 2009-08-31 CA CA2735646A patent/CA2735646A1/en not_active Abandoned
- 2009-08-31 JP JP2011525278A patent/JP5779095B2/en not_active Expired - Fee Related
- 2009-08-31 MX MX2013014031A patent/MX357045B/en unknown
-
2011
- 2011-02-23 CO CO11022122A patent/CO6341510A2/en unknown
- 2011-02-24 IL IL211421A patent/IL211421A/en not_active IP Right Cessation
- 2011-02-24 CL CL2011000396A patent/CL2011000396A1/en unknown
- 2011-03-28 ZA ZA2011/02266A patent/ZA201102266B/en unknown
-
2015
- 2015-04-24 JP JP2015089867A patent/JP6163514B2/en active Active
-
2016
- 2016-04-28 AU AU2016202733A patent/AU2016202733B2/en not_active Ceased
Also Published As
Publication number | Publication date |
---|---|
MX357276B (en) | 2018-07-03 |
IL211421A (en) | 2015-03-31 |
CO6341510A2 (en) | 2011-11-21 |
JP2012501191A (en) | 2012-01-19 |
JP6163514B2 (en) | 2017-07-12 |
EP2334166A2 (en) | 2011-06-22 |
ZA201102266B (en) | 2012-10-31 |
MX357045B (en) | 2018-06-22 |
CL2011000396A1 (en) | 2012-03-30 |
JP5779095B2 (en) | 2015-09-16 |
MX2011002110A (en) | 2011-08-03 |
AU2009287446B2 (en) | 2016-01-28 |
RU2582260C2 (en) | 2016-04-20 |
RU2015151684A (en) | 2019-01-15 |
WO2010025466A2 (en) | 2010-03-04 |
CN102405289A (en) | 2012-04-04 |
AU2016202733A1 (en) | 2016-05-19 |
WO2010025466A3 (en) | 2010-04-29 |
NZ591185A (en) | 2013-05-31 |
CA2735646A1 (en) | 2010-03-04 |
JP2015130896A (en) | 2015-07-23 |
RU2011111344A (en) | 2012-10-10 |
BRPI0917919A2 (en) | 2015-08-18 |
EP2334166A4 (en) | 2012-01-25 |
AU2009287446C1 (en) | 2016-08-11 |
AU2016202733B2 (en) | 2017-04-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20100186121A1 (en) | Transgenic Plants with Enhanced Growth Characteristics | |
AU2016202733B2 (en) | Transgenic plants with enhanced growth characteristics | |
JP6117837B2 (en) | Plant glutamine phenylpyruvate transaminase gene and transgenic plant carrying it | |
WO2011106794A1 (en) | Increasing plant growth by modulating omega-amidase expression in plants | |
US9862964B2 (en) | Transgenic plants with enhanced growth characteristics | |
WO2011106734A1 (en) | Transgenic soybean and rice plants with enhanced growth characteristics | |
US20100170009A1 (en) | Nucleic acids encoding plant glutamine phenylpyruvate transaminase (GPT) and uses thereof | |
US20100263090A1 (en) | Plant Glutamine Phenylpyruvate Transaminase Gene and Transgenic Plants Carrying Same | |
US10119127B2 (en) | Nucleic acids encoding plant glutamine phenylpyruvate transaminase (GPT) and uses thereof | |
US20130232641A1 (en) | Plant glutamine phenylpyruvate transaminase gene and transgenic plants carrying same | |
US20170121728A1 (en) | Plant glutamine phenylpyruvate transaminase gene and transgenic plants carrying same | |
WO2010025463A2 (en) | Nucleic acids encoding plant glutamine phenylpyruvate transaminase (gpt) and uses thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
DA2 | Applications for amendment section 104 |
Free format text: THE NATURE OF THE AMENDMENT IS AS SHOWN IN THE STATEMENT(S) FILED 07 MAR 2016 . |
|
DA3 | Amendments made section 104 |
Free format text: THE NATURE OF THE AMENDMENT IS AS SHOWN IN THE STATEMENT(S) FILED 07 MAR AND 28 APR 2016 |
|
FGA | Letters patent sealed or granted (standard patent) | ||
MK14 | Patent ceased section 143(a) (annual fees not paid) or expired |