EA040754B1 - CORN MON 87419 TRANSGENIC EVENT AND METHODS OF ITS APPLICATION - Google Patents

Description

Cross-reference to related applications

This application claims priority over U.S. Provisional Application No. 61/968,342, filed March 20, 2014.

Enabling a sequence listing

The sequence listing is contained in a file named MONS362WO.txt, 29.4 kb in size (measured on MS-Windows), created on March 9, 2015, is provided in this document in electronic form and is incorporated herein by reference.

Technical field

The present invention relates to recombinant DNA molecules unique to the MON 87419 maize transgenic event. The present invention also relates to transgenic maize plants, parts, seeds, cells, and agricultural products containing the MON 87419 maize event, as well as methods of using them. Transgenic corn plants containing corn event MON 87419 show resistance to dicamba and glufosinate herbicides.

State of the art

Corn (Zea mays) is an important crop in many parts of the world. For this crop, biotechnological methods have been applied to produce corn with desirable traits. One such desirable trait is herbicide resistance. Expression of a heterologous gene, also known as a transgene, for herbicide resistance in a plant can confer herbicide resistance throughout the plant. However, the expression of a transgene and, therefore, its efficiency can be influenced by many different factors, including such as the orientation and composition of the cassette, which is responsible for the expression of a single transgene placed in the plant chromosome, and chromosomal localization, as well as the genomic outcome of the transgene insertion. This is further complicated in transgenic plants with multiple molecularly linked transgenes that define individual traits. In such a situation, the correct expression of each of the molecularly related transgenes in the plant must be the result of the same transgene insertion (also referred to as a multigene event). In such cases, it is necessary to develop and test multiple expression cassettes, each with a different configuration of transgenes and expression elements, and then produce and analyze a large number of individual plant transformation events over multiple generations of plants in order to select transgenic events with superior properties in relation to each of the desired traits and the optimal phenotypic and agricultural characteristics necessary to make it suitable for commercial purposes. Such selection requires extensive molecular studies, as well as many years of greenhouse and field trials in various locations and under various conditions, so that a significant amount of agronomic, phenotypic and molecular data can be collected in this way. The resulting data and observations should then be analyzed by teams of scientists and agronomists in order to select an event suitable for commercial use in agriculture in a wide range of genetic traits and under various field conditions. Once selected, a commercial event that defines the desired trait can be introgressed into other background genotypes using plant breeding techniques, thereby producing a range of different crop varieties that contain the desired trait and are appropriately adapted to specific local growing conditions. In order to create a transgenic plant containing a single transformation target, a portion of the recombinant DNA construct is transferred into the corn cell genome using plant transformation techniques. This corn cell is subsequently used to produce a unique R ₀ plant, which can then be used to produce progeny of transgenic plants. The genome of progeny plants contains a unique event and these plants can be tested for the desired trait(s) as well as agronomic performance. Event efficiency may be influenced by cis and/or trans factors relative to the integration site in the transformation object. The phenotype determined by the event can also be influenced by the size and structure of the DNA construct, which can vary depending on the combination of genetic elements in the expression cassette, the number of transgenes, the number of expression cassettes, and the configuration of such elements and such cassettes. The productivity of this event can be further complicated by factors such as plant development-related, diurnal, seasonal, and spatial patterns of transgene expression; or external factors such as environmental conditions for plant growth, water availability, nitrogen availability, heat or stress. Thus, the ability to create an event that defines a desired set of phenotypic traits is not easily predictable.

Brief description of the essence of the invention

The present invention relates to a recombinant DNA molecule containing a sequence selected from the group consisting of SEQ ID NO: 1-10. The present invention also relates to recombinant DNA derived from a transgenic corn plant or seed containing the corn event MON 87419. A representative seed sample containing the corn event MON 87419 has been deposited with the ATCC under accession number PTA-120860. The present invention also relates to

- 1 040754 combinant DNA molecule, which is an amplicon for the purpose of diagnosing the presence of DNA obtained from the MON 87419 maize event. transgenic maize containing event maize MON 87419.

The present invention relates to a DNA molecule with a sufficient length of the contiguous DNA sequence of SEQ ID NO: 10 to function as a DNA probe that hybridizes under stringent hybridization conditions to a DNA molecule containing a DNA sequence selected from the group consisting of SEQ ID NO: 1-10 and does not hybridize under stringent hybridization conditions to a DNA molecule that does not contain a DNA sequence selected from the group consisting of SEQ ID NOs: 1-10. The invention also relates to a pair of DNA molecules containing a first DNA molecule and a second DNA molecule, while the first DNA molecule is a fragment of the sequence SEQ ID NO: 9, and the second DNA molecule is a fragment of genomic DNA of maize event maize MON 87419, while both the first and second DNA molecules contain a DNA sequence of contiguous nucleotides of sufficient length to function as DNA primers when used in an amplification reaction with DNA containing a MON 87419 maize event to generate an amplicon to diagnose a MON 87419 maize event in a sample .

The invention relates to a method for detecting the presence of a MON 87419 maize event in a DNA sample by bringing the sample into contact with a DNA probe, subjecting the sample and the DNA probe to stringent hybridization conditions, and detecting hybridization of the DNA probe to a DNA molecule in the sample, where hybridization of the DNA probe to the DNA molecule indicates for the presence of the MON 87419 maize event in the DNA sample.

The invention relates to a method for detecting the presence of a MON 87419 maize event in a DNA sample by contacting the sample with a pair of DNA primers, performing an amplification reaction sufficient to produce a DNA amplicon containing a sequence selected from the group consisting of SEQ ID NOS: 1-8 and SEQ ID NO: 10, and detecting the presence of the DNA amplicon in the reaction, wherein the presence of the DNA amplicon in the reaction indicates the presence of the MON 87419 maize event in the DNA sample.

The invention also relates to a DNA detection kit comprising at least one DNA molecule comprising a DNA sequence of contiguous nucleotides of SEQ ID NO: 10 long enough to function as a specific primer or probe for detecting the presence of a MON 87419 maize event in DNA sample.

The present invention relates to a recombinant corn plant, seed, cell, plant part or commercial product containing a DNA molecule with a DNA sequence selected from the group consisting of SEQ ID NO: 1-10. The present invention also relates to a transgenic corn plant, seed or cell resistant to the herbicides glufosinate or dicamba or glufosinate and dicamba. The present invention also relates to a transgenic corn plant, seed, cell, plant part or commercial product containing the MON 87419 corn event. corn event MON 87419.

The present invention relates to a method for controlling weeds in an area comprising planting a transgenic corn containing event corn MON 87419 in an area and applying an effective dose of dicamba or glufosinate or dicamba and glufosinate herbicides to control weeds in the area without damaging the transgenic corn. The invention also relates to a method of controlling weeds by applying an effective dose of glufosinate herbicide from about 0.1 pounds (0.045 kg) acid equivalent per acre (0.405 ha) to about 16 pounds (7.248 kg) acid equivalent per acre of glufosinate herbicide during the growing season . The invention also relates to a method of controlling weeds by applying an effective dose of glufosinate herbicide from about 0.4 pounds (0.181 kg) acid equivalent per acre (0.405 ha) to about 1.59 pounds (0.720 kg) acid equivalent per acre of glufosinate herbicide for vegetation period. The invention also relates to a method of controlling weeds by applying an effective dose of dicamba herbicide from about 0.1 pounds (0.045 kg) of acid equivalent per acre (0.405 ha) to about 16 pounds (7.248 kg) of acid equivalent per acre of dicamba herbicide during the growing season . The invention also relates to a method of controlling weeds by applying an effective dose of dicamba herbicide from about 0.5 pounds (0.227 kg) of acid equivalent per acre (0.405 ha) to about 2 pounds (0.906 kg) of acid equivalent per acre of dicamba herbicide during the growing season .

The present invention also relates to a method for producing a transgenic corn plant resistant to glufosinate and dicamba herbicides by sexually crossing a transgenic corn plant containing the MON 87419 maize event with a second corn plant, harvesting the resulting seeds, growing the seeds to produce plant progeny, treating the progeny plants herbicides glufosinate or dicamba or glufosinate and dicamba, as well as the selection of a descendant plant resistant to the herbicides glufosinate and dicamba. The present invention also relates to a method for producing a transgenic maize plant resistant to the herbicides glufosinate and dicamba by ca

- 2 040754 pollinating a transgenic corn plant containing event corn MON 87419, collecting the resulting seeds, growing the seeds to produce plant progeny, treating the progeny plants with glufosinate or dicamba or glufosinate and dicamba herbicides, and selecting a progeny plant resistant to glufosinate and dicamba herbicides .

Brief description of graphic materials

The drawing illustrates the organization of a transgenic insert into the genome of a maize plant containing the maize event MON 87419. The horizontal lines correspond to the relative positions of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 , SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10; the thick arrows labeled SQ26644 and SQ26645 indicate the approximate position of the primer pair used to identify the transgenic maize containing the MON 87419 maize event; thick, short lines numbered 14 to 22 represent the relative position of the unique sequences of the recombinant constructs within the DNA insert (SEQ ID NO: 9) and the number refers to the corresponding SEQ ID NO of each, respectively; thin horizontal arrows represent the relative organization of the two distinct insert DNA expression cassettes of the heterologous transgene, maize event MON 87419, and the boxes indicate the individual elements of the two expression cassettes; capital 'P' denotes promoter element, capital 'L' denotes leader element, capital 'I' denotes intron, capital 'TS' denotes chloroplast transit peptide, capital T denotes 3' transcription termination and polyadenylation element (3' UTR), pat denotes the protein coding region for phosphinothricin acetyltransferase (PAT); and dmo denotes the protein coding region for dicamba monooxygenase (DMO).

Brief description of the sequences

SEQ ID NO: 1 is a thirty nucleotide long DNA sequence showing the 5' junction of maize genomic DNA and the transgenic insert. SEQ ID NO: 1 corresponds to nucleotide positions 1232 to 1261 of SEQ ID NO: 10.

SEQ ID NO: 2 is a thirty nucleotide DNA sequence showing the 3' junction of maize genomic DNA and the transgenic insert. SEQ ID NO: 2 corresponds to nucleotide positions 7994 to 8023 of SEQ ID NO: 10.

SEQ ID NO: 3 is a sixty nucleotide long DNA sequence showing the 5' junction of maize genomic DNA and the transgenic insert. SEQ ID NO: 3 corresponds to nucleotide positions 1217 to 1276 of SEQ ID NO: 10.

SEQ ID NO: 4 is a sixty nucleotide long DNA sequence showing the 3' junction of the maize genomic DNA and the transgenic insert. SEQ ID NO: 4 corresponds to nucleotide positions 7979 to 8038 of SEQ ID NO: 10.

SEQ ID NO: 5 is a hundred nucleotide long DNA sequence showing the 5' junction of maize genomic DNA and the transgenic insert. SEQ ID NO: 5 corresponds to nucleotide positions 1197 to 1296 of SEQ ID NO: 10.

SEQ ID NO: 6 is a hundred nucleotide long DNA sequence showing the 3' junction of maize genomic DNA and the transgenic insert. SEQ ID NO: 6 corresponds to nucleotide positions 7959 to 8058 of SEQ ID NO: 10.

SEQ ID NO: 7 is a 1771 bp DNA sequence representing 1246 bp of maize 5' flanking genomic DNA and 525 bp of the 5' end of the transgene insert.

SEQ ID NO: 8 is a 1767 bp DNA sequence representing 516 bp of the 3' end of the transgene insert and 1251 bp of the 3' flanking maize genomic DNA.

SEQ ID NO: 9 is the 6762 nucleotide long DNA sequence corresponding to the MON 87419 maize event transgene insert.

SEQ ID NO: 10 is a DNA sequence of 9259 nucleotides in length representing the events of maize MON 87419; the sequence contains the 5' flanking genomic DNA sequence positions 1 to 1246, the transgenic insert DNA sequence positions 1247 to 8008, and the 3' flanking genomic DNA sequence positions 8009 to 9259.

SEQ ID NO: 11 is a 33 nucleotide long DNA sequence corresponding to the primer designated SQ26644, which is used to identify the MON 87419 maize event DNA in a sample; it corresponds to positions 7966 to 7998 of SEQ ID NO: 10.

SEQ ID NO: 12 is a 24 nucleotide long DNA sequence corresponding to the primer designated SQ26645, which is used to identify the MON 87419 maize event DNA in a sample; it corresponds to positions 8022 to 8045 of SEQ ID NO: 10.

SEQ ID NO: 13 is a 19 nucleotide long DNA sequence corresponding to the probe designated PB11207, which is used to identify the MON 87419 maize event DNA in a sample; it corresponds to positions 8002 to 8020 of SEQ ID NO: 10.

SEQ ID NO: 14-22 are DNA sequences corresponding to unique

- 3 040754 sequences within the transgenic insertion of event maize MON 87419.

Detailed description of the invention

The following definitions and methods are provided in order to better define the present invention and guide those of ordinary skill in the art in the practice of the present invention. Unless otherwise indicated, the terms are to be understood in accordance with normal usage by those of ordinary skill in the art.

Modern methods of plant transformation are used to create genetically modified plants. The term transgenic can also be used to refer to genetically engineered plants. In the process of creating a transgenic plant, foreign DNA is randomly inserted into the genome of a plant cell. During transformation, many individual cells are transformed. Due to random integration, a separate and unique DNA recombination event will occur within the genome of each individual transformed plant cell. The whole transgenic plant is then grown from one single transgenic cell, which necessarily results in each cell of the transgenic plant having a uniquely inserted DNA as a stable part of its genome. The herbicide-resistant transgenic corn containing the MON 87419 maize event accommodates one transgenic DNA insert in the maize germplasm chromosome/genome. Maize Event MON 87419 was generated by (i) transforming thousands of cells of a maize plant with a nucleic acid construct that includes the transgenes of interest;

(ii) regenerating a population of maize plants, each containing a unique transgenic event; and (iii) multi-year testing, screening and selection to select an event with the required agronomic properties, corn event MON 87419.

The MON 87419 maize event is characterized by a unique DNA sequence for inserting a transgene at a specific location in the maize plant genome.

The act of inserting transgenic DNA into the corn plant genome is accomplished by the act of plant transformation and results in the creation of a new transgenic genomic molecular sequence known as an event. This sequence is unique and event specific and can be easily identified by comparison with the original maize genomic sequence or other maize transgenics. Molecular analysis of the maize event MON 87419 identified a genomic insertion site for the inserted DNA (SEQ ID NO: 9) and a flanking maize genomic DNA sequence immediately adjacent to both sides of the inserted DNA (SEQ ID NO: 7 and SEQ ID NO: 8). This arrangement of the inserted DNA with respect to the surrounding DNA of the maize plant genome is thus specific and unique for herbicide-tolerant transgenic maize comprising the maize event MON 87419. This novel genomic molecular sequence (SEQ ID NO: 10) is also an integral part of the chromosome transgenic herbicide-resistant maize plants containing the maize event MON 87419, and thus is static in the plant and can be passed on to the progeny of the plant. The present invention also relates to the progeny of the original transformant containing the MON 87419 maize event. Such progeny can be obtained by self-pollination of a corn plant containing the MON 87419 maize event, or by sexual crossing between a corn plant containing the MON 87419 maize event and another plant that contains or does not contain the MON 87419 maize event. Such other plant may be a transgenic plant containing the same or different event(s), or a non-transgenic plant such as a plant of a different variety. Even after multiple backcrosses with the recurrent parent, the MON 87419 maize event derived from the transformed parent is present in the crossed progeny at the same genomic site.

As used herein, the term corn means Zea mays (also known as corn) and includes all plant species that can be crossed with corn.

The present invention relates to a herbicide-resistant transgenic corn plant containing corn event MON 87419, resistant to dicamba (3,6-dichloro-2-methoxybenzoic acid) and glufosinate (2-amino-4-(hydroxymethylphosphinyl)-butanoic acid) herbicides. Dicamba is a synthetic auxin herbicide used to control a wide range of broadleaf weeds. Glufosinate is an organophosphorous herbicide used to control a wide range of annual and perennial grasses and broadleaf weeds. Maize event MON 87419 contains the demethylase (dmo) gene from Stenotrophomonas maltophilia that expresses the dicamba mono-oxygenase (DMO) protein to confer resistance to the dicamba herbicide and the bialaphos resistance (pat) gene from Streptomyces viridochromogenes that expresses the phosphinothricin N-acetyltransferase protein ( PAT) to confer resistance to the herbicide glufosinate.

As used herein, the term recombinant refers to a non-natural DNA, protein, or organism that does not normally occur in nature and has been created by human

- 4 040754 interventions. As used herein, a recombinant DNA molecule is a DNA molecule containing a combination of DNA molecules that do not naturally occur together and is the result of human intervention, such as a DNA molecule that contains a combination of at least two DNA molecules that are heterologous to each other. to each other, such as a DNA molecule that contains the transgene and plant genomic DNA adjacent to the transgene. An example of a recombinant DNA molecule is a DNA molecule containing at least one sequence selected from SEQ ID NOs: 1-10. As used herein, a recombinant plant is one that does not normally exist in nature, is the result of human intervention, and contains a transgenic DNA molecule. As a result of this change in the genome, the recombinant plant is something new and markedly different from the corresponding wild-type plant. An example of a recombinant plant is a corn plant containing corn event MON 87419.

As used herein, the term transgene refers to a DNA molecule artificially inserted into the genome of an organism as a result of human intervention, such as through plant transformation techniques. The transgene may be heterologous with respect to the organism. The term transgenic insert as used herein refers to a transgene inserted by plant transformation techniques into the maize genome to produce maize event MON 87419. The sequence for this transgenic insert is provided as SEQ ID NO: 9. The term transgenic refers to something that contains the transgene, for example a transgenic plant refers to a plant containing the transgene.

As used herein, the term heterologous refers to a first molecule that is not normally associated with a second molecule or organism in nature. For example, a DNA molecule can be obtained from the first species and inserted into the genome of the second species. The DNA molecule will thus be heterologous with respect to the genome and the organism.

As used herein, the term chimeric refers to a single DNA molecule obtained by fusing a first DNA molecule with a second DNA molecule, where neither the first nor the second DNA molecule would normally be found in such a fused configuration with respect to the other. . A chimeric DNA molecule is thus a new DNA molecule that is not normally found in nature. An example of a chimeric DNA molecule is a DNA molecule containing at least one sequence selected from SEQ ID NOs: 1-10.

The present invention relates to DNA molecules and their corresponding DNA sequences. As used herein, the terms DNA and DNA molecule refer to a deoxyribonucleic acid (DNA) molecule. The DNA molecule can be of genomic or synthetic origin and is generally presented from the 5' (left) end to the 3' (right) end. As used herein, the term DNA sequence refers to the nucleotide sequence of a DNA molecule. The nomenclature used is that required by section 37 of the United States Code of Federal Regulations § 1.822 and is tabulated in WIPO Standard ST.25 (1998), Appendix 2, Table. 1 and 3. As is customary, the DNA sequences of the present invention and fragments thereof are described with reference to only one strand of the two complementary strands of the DNA sequence. Implicitly and intentionally, complementary sequences of the sequences proposed herein (complementary strand sequences), also referred to in the art as reverse complementary sequences, are within the scope of the present invention and are expressly intended within the claimed subject matter. Thus, as used herein, references to SEQ ID NOs: 1-10 and SEQ ID NOs: 14-22 and fragments thereof include and refer to the complementary chain sequence and fragments thereof.

As used herein, the term fragment refers to a small part of a whole. For example, fragments of SEQ ID NO: 10 will include sequences that are at least about 20 contiguous nucleotides, at least about 25 contiguous nucleotides, at least about 30 contiguous nucleotides, at least about 35 contiguous nucleotides, at least about 40 consecutive nucleotides, at least about 45 consecutive nucleotides, at least about 50 consecutive nucleotides, at least about 60 consecutive nucleotides, at least about 70 consecutive nucleotides, at least about 80 consecutive nucleotides, at least about 90 consecutive nucleotides or at least about 100 consecutive nucleotides of the complete sequence of SEQ ID NO: 10.

The DNA sequence corresponding to the complete DNA sequence of the transgene insert and the significant DNA segments of the maize genome flanking either end of the transgene insert is shown as SEQ ID NO: 10. the transgenic insert is shown as SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7. The DNA sequence of maize genomic DNA is physically linked by a phosphodiester linkage and thus the 3' flanking end of the transgenic insert is shown as SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8.

- 5 040754

The transgenic maize containing the MON 87419 maize event contains two regions, referred to as junctions. A junction is the site at which one end of the transgenic insert is joined to the genomic DNA. The junction spans or stretches at the junction of the transgenic insert and its adjacent flanking genomic DNA, and as such is the junction point of the two as one contiguous molecule. One connection at the 5' end of the transgenic insert and one at the 3' end of the transgenic insert, referred to herein as the 5' and 3' connection, respectively. A junction sequence refers to a DNA sequence of any length that spans the 5' or 3' junction of an event. The MON 87419 corn event junction sequences are apparent to those skilled in the art using SEQ ID NO: 10. Examples of MON 87419 corn event junction sequences are provided as SEQ ID NO: 1-8. The drawing illustrates the physical arrangement of SEQ ID NO: 1-10, located from 5' to 3'. Thus, the present invention relates to a DNA molecule that contains at least one of the DNA sequences shown in SEQ ID NO: 1-8.

Maize event junction sequences MON 87419 may be present as part of the genome of the transgenic corn plant, seed or cell containing the corn event MON 87419. Identification of any one or more of SEQ ID NOs: 1-8 in a sample taken from the transgenic corn plant, plant part, seed or cell indicates that the DNA was derived from transgenic maize containing the MON 87419 maize event and is diagnostic of the presence of the MON 87419 maize event.

Maize event MON 87419 contains sequences that are unique to the transgenic insert, in particular SEQ ID NOS: 14-22. These sequences are unique to a particular chimeric configuration of various promoters, introns, chloroplast-targeted peptides (CTPs), 3' termination signal, pat and dmo genes within a transgene insert event. Figure 1 illustrates the relative position of each of these unique transgene insertion sequences with respect to SEQ ID NO: 9. Provided are examples of DNA molecules that can be used as primers or probes to diagnose the presence of DNA derived from MON 87419 maize event in a sample. Such primers or probes are specific for the target nucleic acid sequence and as such can be used to identify the MON 87419 maize event using the methods described herein.

A primer is a DNA molecule that is intended for use in annealing or hybridization methods that involve an amplification reaction. The amplification reaction is an in vitro reaction that amplifies template DNA to produce an amplicon. As used herein, an amplicon is a DNA molecule that has been synthesized using amplification techniques. Amplicons according to the present invention have a DNA sequence containing one or more of SEQ ID NO: 1-10 or fragments thereof. The primer pair can be used with a template DNA, such as a sample of maize genomic DNA, in an amplification reaction, such as polymerase chain reaction (PCR), to generate an amplicon, whereby the resulting amplicon will have a DNA sequence corresponding to that of the template DNA located between the two areas where the primers hybridize with the template. The primer is typically designed to hybridize to a complementary target DNA strand to form a hybrid between the primer and the target DNA strand. The presence of the primer is the recognition point for the polymerase to begin primer extension using the target DNA strand as a template. Primer pairs refer to the use of two primers linking opposite strands of a double-stranded nucleotide segment to amplify the nucleotide segment in between. Example primer sequences are shown as SEQ ID NO: 11 and SEQ ID NO: 12. The primer pair shown as SEQ ID NO: 11 and SEQ ID NO: 12 can be used as the first DNA molecule and the second DNA molecule, provided that the first DNA molecule is a fragment of SEQ ID NO: 9, and the second DNA molecule is a fragment of the corn genomic DNA sequence of SEQ ID NO: 10, and each is of sufficient length to function as DNA primers when used together in an amplification reaction with DNA, containing the MON 87419 maize event to generate an amplicon for diagnosing the MON 87419 maize event in the sample. The genomic DNA sequence of maize event maize MON 87419 is shown as positions 1-1246 and 8009-9259 of SEQ ID NO: 10.

A probe is a nucleic acid molecule that is complementary to the target nucleic acid strand and is used in hybridization detection methods. The probes of the present invention include not only deoxyribonucleic or ribonucleic acids, but also polyamides and other probe materials that specifically bind to a target DNA sequence, and detection of such binding can be used to detect the presence or absence of a target DNA sequence. The probe may be attached to a conventional detectable label or reporter, such as a radioactive isotope, ligand, chemiluminescent agent, or enzyme. As an example, a DNA sequence used as a probe for detecting a

- 6 040754 maize MON 87419, presented as SEQ ID NO: 13.

Methods for constructing and using primers and probes are well known in the art, and DNA molecules containing fragments of SEQ ID NO: 1-10 and used as primers and probes for detecting the MON 87419 maize event can be easily designed by a person skilled in the art. technology. The DNA molecules and their corresponding DNA sequences provided herein are thus used to identify the MON 87419 maize event in transgenic maize plants, cells, seeds, or plant parts; selecting corn varieties or hybrids containing corn events MON 87419; and detecting the presence or absence of the MON 87419 maize event in the sample. As used herein, the term isolated refers to the separation of a molecule from other molecules normally associated with it in its native or natural state. The term isolated, therefore, may refer to a DNA molecule that has been separated from other DNA molecule(s) that would normally be associated with it in its native or native state. Such a DNA molecule may be present in a recombinant state, such as a recombinant DNA molecule. Thus, DNA molecules fused to regulatory or coding sequences to which they are not normally associated, for example as a result of recombinant technologies, are considered isolated, even when inserted as a transgene into a cell chromosome or present with other DNA molecules.

The present invention relates to corn plants, progeny, seeds, plant cells and plant parts containing corn event MON 87419, and commercial products obtained using them. A representative sample of transgenic herbicide-tolerant corn seed containing the MON 87419 event is deposited under the Budapest Treaty with the American Type Culture Collection (ATCC®). The ATCC repository for the seed of a transgenic herbicide resistant corn plant containing the corn event MON 87419 has assigned patent deposit designation PTA-120860. Plants, progeny, seeds, plant cells, plant parts and commercial products of the present invention contain a detectable amount of DNA having at least one of the sequences shown as SEQ ID NO: 1-10 and SEQ ID NO: 14-22. The plants, progeny, seeds, plant cells, and plant parts of the present invention may also contain one or more additional transgenic traits, in particular those introduced by crossing a maize plant containing the MON 87419 maize event with another plant containing the additional transgenic trait( And). Such traits include, but are not limited to, improved insect resistance, improved water use efficiency, improved crop productivity, improved drought tolerance, improved seed quality, improved nutritional performance, hybrid seed production, and/or improved herbicide tolerance, wherein the trait measured relative to a corn plant that does not have that transgenic trait.

The plants of the present invention can be used to produce progeny that contain the MON 87419 maize event. As used herein, the term progeny includes any plant, seed, and plant cell containing the MON 87419 maize event inherited from an ancestor plant, designated a plant containing a DNA molecule having at least one sequence selected from SEQ ID NO: 1-10. Plants, progeny, and seeds may be homozygous or heterozygous for the MON 87419 maize event. Plant progeny may be grown from seeds produced by a corn plant containing the MON 87419 maize event, or from seeds produced by a corn plant fertilized with pollen containing the MON corn event. 87419.

As used herein, the term plant part of the present invention is any part derived from a transgenic maize plant containing the MON 87419 maize event. Plant parts include, but are not limited to, pollen, ovules, pod, flower, roots, stems, fibers, and leaves. Plant parts may or may not be viable. The invention relates to a commercial product that is derived from transgenic maize containing the MON 87419 maize event. Commercial products of the present invention contain a detectable amount of DNA containing a DNA sequence selected from the group consisting of SEQ ID NOs: 1-10. As used herein, the term commercial product refers to any composition or product that consists of material derived from a transgenic corn plant, corn seed, corn plant cell, or corn plant part containing the MON 87419 corn event. Commercial products include, but are not limited to , processed seeds, grains, plant parts and food. Transgenic corn containing corn event MON 87419 can be used to produce any commercial product, typically derived from corn. A commercial product of the present invention will contain a detectable amount of DNA corresponding to corn event MON 87419. Detection of one or more of these DNAs in a sample can be used to determine the content or source of the commercial product. You can use any standard method for detecting DNA molecules, including the methods of detection described in this document. The present invention relates to methods of weed control using

- 7 040754 herbicides glufosinate or dicamba or glufosinate and dicamba with transgenic corn containing corn event MON 87419. It is envisaged that a method of controlling weeds in an area, such as a field, consists of planting transgenic corn plants containing corn event MON 87419 in the area and applying a herbicide-effective dose of glufosinate or dicamba or glufosinate and dicamba herbicides to a site to control weeds in the site without damaging transgenic corn plants containing corn event MON 87419. Such application of glufosinate or dicamba or glufosinate and dicamba herbicides can be pre-emergence ( any time after the transgenic corn seed containing the MON 87419 corn event has been planted and before the transgenic corn plant containing the MON 87419 corn event has emerged) or post-emergence (any time after the transgenic corn plant containing the MON 87419 corn event has grown). The herbicidally effective dose of glufosinate for use in a weed control area should be in the range of about 0.1 lb (0.045 kg) acid equivalent per acre (a.e./acre) up to about 16 lb (7.248 kg) a.e. ./acre of glufosinate during the growing season. The herbicidally effective dose of dicamba for use in the weed control area should be in the range of about 0.1 lb (0.045 kg) ae/acre up to about 16 lb (7.248 kg) ae/acre dicamba for vegetation period. A variety of applications of glufosinate or dicamba or glufosinate and dicamba herbicides can be used during the growing season, for example two applications (such as application before planting and application after emergence or pre-emergence application and application after emergence) or three applications (such as application before planting, pre-emergence application and post-emergence application).

As used herein, the term active ingredient or a.i. is the component of the herbicidal formulation responsible for herbicidal activity, often measured in pounds per gallon or applied in pounds per acre (0.405 ha). For herbicides that are acids (e.g., molecules that have a carboxyl group as part of their structure) in which the acid group is often converted to (may be replaced with desired ions to form) a salt or (reacted with an alcohol to form) ester in the process of composition. This can change not only the chemical characteristics of a particular herbicide molecule, but also the mass. However, the corresponding acid is the herbicidally active part of the formulation and the equivalence of herbicidal activity between different active ingredients can be calculated using the acid equivalent as the standard unit of measure. The term acid equivalent or a.e. means that part of the active ingredient in the formulation, which can theoretically be converted back to the corresponding acid. Herbicide application rates can be expressed as acid equivalent per acre (abbreviated as ae/acre) or as active ingredient per acre (abbreviated as a.i./acre).

Methods are provided for producing a transgenic herbicide-resistant corn plant containing corn event MON 87419. The progeny obtained by these methods may be varietal or hybrid plants; can be grown from seeds produced by a transgenic corn plant containing the MON 87419 corn event, or from seeds produced by a corn plant fertilized with pollen from a transgenic corn plant containing the MON 87419 corn event; and may be homozygous or heterozygous for the MON 87419 maize event. Plants may be self-pollinated (also known as self-pollination) or cross-pollinated (also known as cross-pollination). Transgenic maize plants containing the MON 87419 maize event can be self-pollinated to create a true line for breeding plants that are homozygous for the MON 87419 maize event. Selfing results in offspring known as inbred and is used to produce inbred lines that are genetically homogeneous. Alternatively, transgenic corn plants containing the MON 87419 corn event can be outcrossed (crossed with another plant that is transgenic or non-transgenic) to produce varietal or hybrid seed. Seeds and plant progeny produced by the methods of the present invention will contain corn event MON 87419 and may then be treated with glufosinate or dicamba or glufosinate and dicamba herbicides. Herbicide treatment with glufosinate or dicamba or glufosinate and dicamba can be used to select resistant offspring. Alternatively, these progeny plants can be analyzed using diagnostic methods to select plants or seeds containing the MON 87419 maize event. The plants, progeny, seeds, plant cells, and plant parts of the present invention may also contain one or more additional transgenic traits. corn, in particular those introduced by crossing a corn plant containing the MON 87419 corn event with another corn plant containing the additional transgenic trait(s). Such transgenic traits in corn include, but are not limited to, increased insect resistance, increased water use efficiency, increased crop productivity, increased drought tolerance, improved seed quality, improved nutritional performance, hybrid seed production, and herbicide tolerance, where the trait is measured in relation to the corn plant, not

- 8 040754 having such a transgenic trait. Such transgenic maize traits are known to the person skilled in the art; for example, a list of such traits is provided by the United States Department of Agriculture (USDA) Animal and Plant Health Inspection Services (APHIS) and can be found on their website: http://www.aphis.usda.gov. Two transgenic plants can thus be crossed to produce offspring that contain two or more individual transgenic traits that are independent of each other. Backcrossing with a parent plant and outcrossing with a non-transgenic plant are also considered, as is vegetative propagation.

Descriptions of breeding methods that are commonly used for various traits and crops can be found in one of several references, such as Fehr, Breeding Methods for Cultivar Development, Wilcox J. ed., American Society of Agronomy, Madison WI (1987).

The plants, seeds, cells, plant parts, and commercial products of the present invention can be used to detect DNA and protein molecules indicative of the presence of the MON 87419 maize event. Such detection can be performed using the DNA sequence provided herein and the corresponding protein sequences. DMO and PAT encoded by the transgene insert shown as SEQ ID NO: 9. Detection of the presence of the MON 87419 maize event can be performed using methods known in the art such as thermal nucleic acid amplification, nucleic acid hybridization methods (such as, Northern blot and Southern analysis), protein detection methods (such as Western blot, immunoprecipitation, and ELISA)) or using the detection methods and/or detection kits provided herein. One method involves contacting a DNA sample with a primer pair that is capable of producing an amplicon from transgenic maize DNA containing the MON 87419 maize event, performing amplification reactions and thereby producing a DNA amplicon containing at least one of the DNA sequences shown as SEQ ID NOs: 1-10 and SEQ ID NOs: 14-22, and then detecting the presence or absence of an amplicon molecule and possibly confirming within the amplicon sequence a sequence containing at least one of the sequences shown as SEQ ID NOs: 1-10 and SEQ ID NOs: 14-22. The presence of such an amplicon is diagnostic of the presence of specific DNA for the transgenic maize containing the MON 87419 maize event, and thus the biological material in the sample, which is derived from the transgenic maize containing the MON 87419 maize event. Another method involves contacting the DNA sample with the DNA probe, subjecting the probe and the DNA sample to stringent hybridization conditions, and then detecting hybridization between the probe and the target DNA sample. The detection of hybridization is diagnostic of the presence of specific DNA for transgenic maize containing the MON 87419 maize event in the DNA sample.

DNA detection kits for the corn event MON 87419 are available.

Variations of such kits can also be developed using the compositions and methods described herein and DNA detection methods well known in the art. DNA detection kits can be applied to methods for breeding transgenic corn plants containing the MON 87419 corn event. Such kits contain DNA primers or probes containing fragments of SEQ ID NOs: 1-10. One example of such a kit contains at least one DNA molecule from adjacent nucleotides of SEQ ID NO: 10 of sufficient length to function as a DNA probe used to detect the presence and/or absence of DNA derived from transgenic herbicide-resistant maize plants in a sample. containing the MON 87419 maize event. Provides a DNA molecule sufficient to be used as a DNA probe that is used to determine, detect, or diagnose the presence and/or absence of a transgenic herbicide-resistant corn DNA containing the MON 87419 maize event shown as SEQ ID NO: 13. Other probes can be easily designed by one of skill in the art and should contain at least about fifteen nucleotides of SEQ ID NO: 10 and be sufficiently unique for herbicide-tolerant transgenic corn containing MON 87419 maize event DNA to target detection of DNA derived from maize event MON 87419. Another type of n the kit contains a pair of primers used to generate an amplicon that is used to detect the presence or absence of a MON 87419 maize event in a sample. Such a kit would use a method comprising contacting a target DNA sample with a pair of primers and then performing a nucleic acid amplification reaction, sufficient to obtain an amplicon containing a DNA molecule having at least one sequence selected from SEQ ID NO: 1-10, and then detecting the presence or absence of the amplicon. Such a method may also include sequencing the amplicon or a fragment thereof. Other primer pairs can be easily designed by one of skill in the art and should contain at least twenty nucleotides of SEQ ID NO: 10 and be sufficiently unique for herbicide-tolerant transgenic maize containing the MON 87419 maize event to detect the MON 87419 maize event. of the present invention may also optionally include reagents to perform

- 9 040754 detection or diagnostic reactions described in this document or instructions for use of the kit and its contents.

As used herein, the term containing means including, but not limited to.

Deposit information.

A representative seed sample of transgenic corn containing corn event MON 87419 was deposited under the Budapest Treaty with the American Type Culture Collection (ATCC), at 10801 University Boulevard, Manassas, Virginia, USA, zip code 20110. ATCC with patent deposit designation ( access number) for seed containing corn event MON 87419 is PTA-120860 and the deposit date was January 17, 2014. The deposit will be held at the depository for 30 or 5 years after the last request or for the duration of the patent, whichever which period is longer.

Examples

The following examples are included to more fully describe the present invention. Those skilled in the art will appreciate that various modifications can be made to the specific examples that are described, but still produce a similar result. Certain agents that are both chemically and physiologically similar may be used in place of the agents described herein, achieving the same or similar results. All such substitutions and modifications obvious to those skilled in the art are considered to be within the scope of the present invention.

Example 1: Receive and select MON event 87419.

This example describes the production, analysis and selection of transgenic corn containing event MON 87419, an event that can provide resistance to both dicamba and glufosinate herbicides. In summary, definitive selection of the MON 87419 maize event requires the acquisition and analysis of tens of thousands of individual plants over many years through rigorous molecular, phenotypic, and field testing.

Transformation vectors containing various expression cassettes have been developed and tested to confirm their usefulness for the expression of the dmo and pat genes. Using these data, combinations of expression elements were selected and eight different transformation vectors were constructed and transformed into maize. These vectors were used to test promoters and terminators in various combinations with two coding regions of the pat and dmo genes (Table 1). The resulting plants were analyzed for protein expression and two vectors (designated A and B in Table 1) were selected for commercial transformation of maize.

Table 1

Trance cassette configuration judging vectors Vector cassette 1 (PAT) cassette 2 (DMO) promoter Interest gene Terminator promoter Interest gene Terminator A AND.ge.Ubql RAT Os.Ara5 PCSV/1-Actl CTP4/DMO Hspl7.5 IN R-1 RAT T-1 PCSV/1-Actl CTP4/DMO Hspl7.5 WITH AND.ge.Ubql RAT Os Aga5 R-1 CTP4/DMO Hspl7.5 D R-1 RAT T-1 AND.ge.Ubql CTP4/DMO Hspl7.5 E AND.ge.Ubql RAT Os.Ara5 R-2 CTP4/DMO Hspl7.5 F AND.ge.Ubql RAT Os.Ara5 R-3 CTP4/DMO Hspl7.5 G R-1 RAT T-1 R-2 CTP4/DMO Hspl7.5 H R-1 RAT T-1 R-3 CTP4/DMO Hspl7.5

Over thirteen thousand unique transformed maize plants were generated using two vectors (A and B) that were selected. Plants often show significant differences in expression levels of the inserted gene among individual events, and gene expression can be directly positively or negatively correlated with the phenotype of the plant containing the event. The expression of foreign genes in plants is known to be influenced, among other things, by their position on the chromosome. For this reason, it was necessary to sample a large number of individual plants containing random insertions of events over many years and localizations in order to determine the optimal event. The maize transgenic event MON 87419 was generated by Agrobacterium-mediated LH244 transformation of immature maize germs. Methods for transforming corn are known in the art. Corn cells were transformed and regenerated into whole corn plants. Rooted plants with normal phenotypic characteristics were selected. Thousands of individual independent events

- 10040754 was then transferred to soil for growth and further evaluation.

table 2

Event selection process

Stage Vector A Vector B Total Advanced Events Advanced Events Unique advanced events R0 estimate Received transgenic events 5236 8413 13649 Passing the first analysis with one copy 1300 1698 2998 Spraying R0 642 798 1440 Primary detailed molecular analysis 85 99 184 Southern R0 54 58 112 Early tackles (R1 and R2) R1 tests and molecular analysis 22 22 44 R2 tests and molecular analysis 20 22 42 Advanced field testing and molecular analysis 1 year UAM 7 10 17 1 year USA 5 6 AND 2 year UAM 2 3 5 Detailed assessment 2 0 2 Final event selection 1 0 1

Throughout the event selection process, molecular analysis was performed, often simultaneously, as well as field trials to assess phenotype, agronomy, and event efficacy (Table 2). 13,000 unique R0-transformed maize plants were analyzed initially by PCR to select events with one copy of the transgenic insert (passing the first one copy analysis). This resulted in a promotion of 2998 events. The next selection for resistance R0 to herbicide spraying was carried out in the greenhouse. Plants were tested simultaneously for resistance to glufosinate (Ignite® 280 herbicide) and dicamba (Clarity® herbicide) using a tank mix of glufosinate (0.9 lb.a.i./acre) and dicamba (2.0 lb.a.e./acre). ) sputtered at the V1/V2 growth stage. Plants showing >15% damage were discarded and 1440 events were selected for further analysis. An initial detailed molecular analysis was performed which included sequence identification and confirmation and a second check for copy number and lack of backbone. This analysis resulted in 184 events containing only one copy of the transgenic insert selected for promotion. Southern analysis on DNA extracted from R0 events was performed to further confirm the copy number of the transgenic insert and to confirm the absence of backbone transformation. Based on this, 112 events were selected to progress to R1 for additional analysis. R0 plants self-pollinated and seeds were harvested for testing with R1.

Simultaneously with all field trials, an additional molecule was under development.

- 11 040754 lar analysis. Northern analysis was performed to detect and measure mRNA transcripts of the pat and dmo genes. N-terminal protein sequencing of PAT and DMO proteins purified from transgenic plants containing selected events was performed in order to confirm the sequence of the recombinant protein. Western analysis for the detection of PAT and DMO proteins was performed with transgenic plant samples. The entire transgene and both 5' and 3' ends of the insert were sequenced and subsequently used to develop methods for detecting individual events. Detailed Southern analysis was performed on R1 plants to confirm copy number and lack of backbone.

For the field trial of R1 early selections, among 112 events selected from the R0 screening, 82 events were selected based on seed germination and nursery size. R1 plants were separated and thus were null, hemizygous or homozygous for the event. Eighty-two events were evaluated in R1 plants when selected for effectiveness in a field with high levels of herbicide application. Plant damage was assessed after treatment with various combinations of glufosinate (Ignite 280) and dicamba (Clarity) (up to 20x glufosinate and up to 16x dicamba of the indicated application rates) and considering the time of application at various growth stages from V1/V2 to V8/V10 . The degree of damage was assessed 10-14 days after herbicide application. Damage standards include scoring for chlorosis, malformations, and interbreeding. Overall averages across multiple plants containing the same event were used to select an event for promotion. Herbicide applications at 2-fold typically result in less than 10% damage, while herbicide applications at 16-fold and 20-fold result in more than 10% damage. In addition to efficacy testing, agronomic scoring was performed for each plant and correlated with the event it contains. The agronomic scoring criteria that were evaluated included plant height, spike height, moisture percentage, weight test, days to 50% pollination, and days to 50% silk. Based on the analysis of data collected during the R1 field trials and molecular analysis, 44 events were selected to progress towards R2 for additional analysis. R1 plants self-pollinated and seeds were harvested for testing with R2.

In the R2 field study of early selections, R2 and F1 events (from R1 outcrosses) were evaluated in field performance selections at three locations (two states and Puerto Rico). Plant damage was assessed after treatment with various combinations of glufosinate and dicamba application rates and taking into account the time of application. R2 plants are homozygous for the event and the rate of failed herbicide resistance after herbicide treatment was low, confirming the results of R1. Agronomic data were collected and scored as in field trials R1. Additional molecular analysis, including gene expression analysis, was also used to select plants containing the best events. Based on the analysis of data collected during the R2 and F1 field trials and molecular analysis, 42 events were selected to progress towards R3 for additional analysis. R2 plants self-pollinated and seeds were harvested for testing with R3.

For advanced field trials, both hybrid and inbred performance field trials and hybrid and inbred agronomic field trials were conducted. Agronomic field trials were conducted during the same season as the efficacy field trials. All field trials used full group randomization and were conducted at multiple locations. For field trials of efficacy and agronomic field trials, agronomic scoring was conducted throughout the trial season and yield (crop efficiency or agronomic yield) was determined at the end of the season. Field efficacy trials were conducted to assess crop damage 10 to 14 days after herbicide application, crop damage and crop damage. Target crop damage score was less than 10% for event promotion. For agronomic field trials, the plots were kept weed free and no glufosinate or dicamba herbicides were used during the growing season. Hybrid agronomic field trials included controls of a comparison hybrid (hybrid control) produced using the same maize parent lines used to produce the transgenic hybrid cross, but not containing the transgenic event. Inbred controls were comparable in inbredity with transgenic inbred lines.

Meta-analysis was performed using a set of multi-seasonal multi-location data from field trials. In table. 3 shows the number of repetitions for which the observation was repeated for a particular type of field study for plants containing one of the events. For each of these two events, there were 135 values recorded for agronomic implementations of the hybrid, 933 values for hybrid efficiency, 179 values for inbred agronomy, 30 values for inbred efficiency, and 16 values for testing the event by testing herbicide application rates for corn containing the event.

- 12 040754

Table 3

Field test replays for the two final events

Description Repeats for each object hybrid agronomy 135 reps Hybrid efficiency 933 repeats Inbred agronomy 179 repeats Inbred efficiency 30 repetitions Testing an event with loads 16 repetitions Total number of retries for each event 1293

Hybrid plants each containing one of 23 selected events (a subset of 42 R2 field trial events) were evaluated in South America (SAAM) in year 1 of the efficacy field trial contra-season and agronomic field trials. The trials were conducted at six sites randomized to full groups with 4 treatments and 2 repetitions of each treatment. In field trials of efficacy, the formulation of dicamba was the herbicide Banvel® 4SL and the formulation of glufosinate was Liberty® 1.67SL. Herbicide treatments consisted of the following:

(1) raw control;

(2) 2 lb (0.906 kg) ae dicamba/acre (acre) PRE (with PRE defined as at planting or pre-harvest) followed by 1 lb (0.45 kg) ae dicamba ./acre applied to each of VE-V2 followed by V4 followed by V8;

(3) glufosinate applied at 0.8 lb (0.363 kg) ai/ac to VE-V2 followed by V4 followed by V8; and (4) glufosinate applied at 0.8 lb (0.363 kg) ai/acre plus dicamba applied at 1 lb (0.454 kg) ae/acre to VE-V2 followed by V4 followed by V8 .

The degree of damage was assessed 10-14 days after herbicide application. Overall averages across multiple plants containing the same event were used to select events for promotion. The target crop damage score was less than 10% and the observed damage score was less than 1%. Based on hybrid damage scoring, agronomic scoring of yield efficiency, agronomic yield and additional molecular analysis, 17 events were selected for promotion.

Inbreds and hybrids from 17 events advanced from the 1 year UAM contra-season field trial were then further evaluated in the 1 year efficacy field trial and agronomic field trial in the United States (USA). These tests were conducted in 2012, which was a severe drought season in the United States. Field trials of hybrid efficacy were conducted in 12 locations, 2 states randomized to full groups with 6 treatments and 3 replicates of each treatment. Hybrid plants containing the glyphosate resistance transgenic event were crossed from a male parent. In field efficacy testing, the glyphosate formulation was Roundup PowerMAX® 4.5SL, the dicamba formulation was Clarity 4SL, and the glufosinate formulation was Ignite 280 2.34SL. Herbicide treatments consisted of the following:

(1) raw control;

(2) 3 lb (1.361 kg) ae/acre glyphosate applied to V4 followed by V8;

(3) glufosinate at 0.8 lb (0.363 kg) ai/acre applied to V2 followed by V4 followed by V8;

(4) 2 lb (0.906 kg) ai/acre dicamba applied to PRE and then applied again to V4, followed by V8;

(5) glyphosate at 3 lb (1.361 kg) ae/acre plus dicamba at 1.5 lb (0.68 kg) ae/acre applied to V2 followed by V4 followed by V8;

(6) 2 lb (0.906 kg) ae/acre dicamba applied to V2 followed by 0.8 lb (0.363 kg) ai/acre glufosinate plus 1 lb (0.454 kg) dicamba to .ae/acre applied in growth stage V4 followed by 3 lb (1.361 kg) ae/acre glyphosate plus 1.5 lb (0.68 kg) ae/acre dicamba applied on growth stages V8.

The degree of damage was assessed 10-14 days after herbicide application. Overall averages across multiple plants containing the same event were used to select events for promotion. The target crop damage score was less than 10% and the observed damage score was less than 1%. Based on hybrid damage scoring, agronomic scoring of crop efficiency, agronomic yield and additional molecular analysis, 11 events were selected for promotion. Field trial year 2 contra-season in South America (SAAM) to evaluate hybrid and inbred agronomic performance

- 13 040754 STIs were then run on plants containing these 11 events. Hybrid plants containing the glyphosate resistance transgenic event were crossed from a male parent. Field trials of hybrid efficacy were carried out essentially as described for the South American Field Trial Year 1 contra-season (SAAM), but with the following herbicide treatments:

(1) untreated control;

(2) 3 lb (1.361 kg) ae/acre glyphosate applied to V4 followed by V8;

(3) 0.8 lb (0.363 kg) ai/acre glufosinate applied to V4 followed by V8;

(4) 2 lb (0.906 kg) ae/acre dicamba applied to PRE and then applied again to V4 followed by V8;

(5) glyphosate at 3 lb (1.361 kg) ae/acre plus dicamba at 1.5 lb (0.68 kg) ae/acre applied to V4 followed by V8;

(6) glufosinate at 0.8 lb (0.363 kg) a.i./acre plus dicamba at 1 lb. (0.454 kg) ae/acre applied to V4 followed by glyphosate at 3 lb. (1.361 kg) to .ae/acre plus dicamba at 1.5 lb (0.68 kg) ae/acre applied to a V8.

The degree of damage was assessed 10-14 days after herbicide application. Overall averages across multiple plants containing the same event were used to select events for promotion. Based on hybrid injury scoring, agronomic scoring of hybrid yield efficiency, inbred agronomic yield, and additional molecular analysis, 5 events were selected for promotion. An additional molecular analysis was then performed for these 5 events. Long-term field and molecular data for each of the 5 events, including hybrid performance trait field trials, hybrid and inbred yield measurements, agronomic scoring, and molecular information, were subsequently reviewed and two events were selected for additional analysis. Both of these events were generated using the same transformation vector and thus had the same transgene insert but not the same genomic location or flanking sequence.

Year 2 field trials in the United States (USA) of inbred efficacy, and hybrid and inbred agronomic field trials were conducted to evaluate these two events. Field trials of hybrid performance were conducted in a similar manner to Year 1 US field trials, but with different spray patterns. Efficiency was measured by the degree of damage and yield efficiency of the hybrid. Then additional molecular analysis was performed for these events. Hybrid plants containing the glyphosate resistance transgenic event were crossed from a male parent. Field trials 1 and 2 hybrid performance were conducted at 12 locations across two states and field trials 3 hybrid performance were conducted at thirty-three locations across four states. In these efficacy field trials, the glyphosate formulation was Roundup PowerMAX 4.5SL, the dicamba formulation was Clarity 4SL, and the glufosinate formulation was Ignite 280 2.34SL. The herbicides for potency 1 and potency 2 (crossing two separate inbred lines) were (1) untreated control;

(2) 0.4 lb (0.181 kg) ai/acre glufosinate applied to VE-V2 followed by V6;

(3) 0.8 lb (0.363 kg) ai/acre glufosinate applied to VE-V2 followed by V6;

(4) 0.5 lb (0.227 kg) ae/acre dicamba applied to V4 followed by V8;

(5) 1.0 lb ae/acre dicamba applied to V4 followed by V8; and (6) glyphosate at 2.25 lb ae/acre plus dicamba at 1.0 lb ae/acre applied to V4 followed by V8.

The herbicides for efficacy trials 3 (representing a hybrid obtained from crossing with a third inbred line) were (1) an untreated control;

(2) 0.5 lb (0.227 kg) ae/acre dicamba applied to VE-V2 followed by 0.4 lb (0.181 kg) ai/acre glufosinate applied to V6; and (3) 1.0 lb ae/acre dicamba applied to VE-V2 followed by 0.8 lb a.i./acre glufosinate applied to V6.

The extent of damage was assessed 10-14 days after herbicide application and a plurality of plants containing the same event were used to select events for promotions. Yield data and agronomic data were collected. To compare the degree of damage to the hybrid for the two events, a meta-analysis of numerous field trials of the effectiveness of the hybrid was carried out. (Table 4). The extent of damage was assessed at V8 (whereas the V8 analysis covers cumulative damage from V2, V4, V6 and V8 herbicide applications) with the statistical least significant difference (LSD at p<0.05). Tester 1, Tester 2 and Tester 3 are crosses with 3 independent inbred maize parent lines that were used to create a hybrid for the indicated field trial. For none of the trials, a statistical difference was found in the degree of damage between hybrids obtained using the parental transgenic

- 14 040754 corn containing one of two events.

Table 4

Meta-analysis of the extent of damage from hybrid efficacy field trials

Field test transgenic corn Damage V8 LSD (P<0.05) 1 year combined USA and USA MON 87419 0.43 0.3 1 year combined USA and USA event 2 0.49 0.3 Hybrid efficiency 2 years UAM MON 87419 2.58 3 Hybrid efficiency 2 years UAM event 2 1.98 3 Tester 1 hybrid 2 years UAM MON 87419 0 0 Tester 1 hybrid 2 years UAM event 2 0 0 Tester 2 hybrids 2 years USA MON 87419 0 0 Tester 2 hybrids 2 years USA event 2 0 0 Tester 3 hybrids 2 years USA MON 87419 0.31 0.42 Tester 3 hybrids 2 years USA event 2 0.12 0.42

A meta-analysis of hybrid yield efficiency (bushels/acre) from multiple efficacy field trials was performed compared to hybrid yields containing each of the two events (Table 5). For none of the trials, a statistical difference was found in hybrid yield efficiency between hybrids produced using parental transgenic corn containing one of the two events.

Table 5

Yield meta-analysis of hybrid efficacy field trials

Field test corn hybrid Yield (bushels/acre) LSD (P<0.05) 1 year combined USA and USA M0N 87419 171.07 4 1 year combined with UAM event 2 172.90 4 and USA 2 year UAM M0N 87419 233.66 14 2 year UAM event 2 231.59 14 Tester 1 2 years USA M0N 87419 217.43 10 Tester 1 2 years USA event 2 220.06 10 Tester 2 2 years USA M0N 87419 219.68 7 Tester 2 2 years USA event 2 221.21 7 Tester 3 2 years USA M0N 87419 208.76 5.67 Tester 3 2 years USA event 2 213.02 5.67

Field trials of stress testing were also conducted with hybrid transgenic corn containing one of the two events. In herbicide loading tests, either glufosinate (Ignite 280, 2.34SL) or dicamba (Clarity 4SL) was applied at non-commercially high levels. For typical field trials, 1-fold glufosinate was 0.4 lb (0.181 kg) ai/acre and 1-fold dicamba was 0.5 lb (0.22 kg) ae/acre. For glufosinate loading testing field testing, glufosinate was used on VE-V2 followed by V4 followed by V8 with the following values:

(1) 1 lb (0.454 kg) ai/acre (2.5x);

(2) 2 lb (0.906 kg) ai/acre (5x);

(3) 4 lb ai/acre (10x); and (4) 8 lb (3.629 kg) ai/acre (20x).

For field trials of dicamba load testing, dicamba was applied to

- 15 040754

VE-V2 followed by V4 followed by V8 with the following values:

(1) 2 lb (0.906 kg) ae/acre (4x);

(2) 4 lb ae/acre (8x);

(3) 8 lb (3.629 kg) ae/acre (16x); and (4) 16 lb (7.248 kg) ae/acre (32x).

At the end of the season, field trials of the hybrid load testing were collected and yield (bushels/acre or bushels/acre) was determined. Yield data analysis was compared with hybrids containing either of these two events. For none of the trials, a statistical difference was found in yield for any of the herbicide application values between hybrids produced using parental transgenic corn containing one of the two events.

Table 6

Yields from field tests of the effectiveness of testing the hybrid by stress testing __________________

Field test Transgenic hybrid corn Yield (bushels/acre) LSD (P<0.05) 2.520x Glufosinate Load Test Test MON 87419 207.31 40 2.520x Glufosinate Load Test Test events 2 201.65 40 4-32x Dicamba Load Test Test MON 87419 239.93 40 4-32x Dicamba Load Test Test events 2 234.60 40

Agronomic field trials of the hybrids were carried out in 3 complexes of 15 sites in each complex in 21 locations across 3 states and were conducted during the same season with field trials of hybrid performance. Agronomic measurements were collected during the trial field season and agronomic yields were determined at the end of the season. A multi-season meta-analysis of multi-location agronomic field trials of the hybrid was used to compare with the yields of the hybrid control and hybrids containing one of the two events. No statistical difference for the agronomic yield of the hybrid was found either between the transgenic hybrids or when compared to the control hybrids (Table 7).

Table 7

Meta-analysis of yields from agronomic field trials of the hybrid

Field test corn hybrid Yield (bushels/acre) LSD (P<0.05) 1 year combined USA and USA Control 180.28 6 1 year combined USA and USA MON 87419 179.01 6 1 year combined USA and USA event 2 184.88 6 2 year UAM Control 231.70 10 2 year UAM MON 87419 232.47 10 2 year UAM event 2 225.07 10 Tester 1 2 years USA Control 185.80 17.40 Tester 1 2 years USA MON 87419 187.09 17.40 Tester 1 2 years USA event 2 192.28 17.40

- 16040754

Tester 2 2 years USA Control 219.12 12.83 Tester 2 2 years USA MON 87419 219.56 12.83 Tester 2 2 years USA event 2 221.22 12.83 Tester 3 2 years USA Control 197.62 8.66 Tester 3 2 years USA MON 87419 191.49 8.66 Tester 3 2 years USA event 2 195.48 8.66

Field trials of inbred efficacy were conducted using full group randomization at 6 locations in 1 state. In these field trials, the dicamba formulation was Clarity 4SL and the glufosinate formulation was Ignite 280 2.34SL. Herbicide applications for inbred efficacy field trials were glufosinate at 0.8 lb ae/acre and dicamba at 2.0 lb ae/acre applied at VE-V2 followed by glufosinate at 0.8 lb a. i./acre and dicamba at 2.0 lb ae/acre applied on a V8. Yield was measured at the end of the season. For each of the trials, a statistical difference was found in inbred yield efficiency compared to the inbred yield obtained in those trials from transgenic corn containing one of the two events (Table 8). These data indicate higher productivity of transgenic corn containing the MON 87419 maize event.

Table 8

Meta-analysis of yields from field trials of inbred efficacy

Field test Inbred ^to U ^to URUza Yield (bushels/acre) LSD (P<0.05) Inbred Year 2 US MON 87419 110.95 7 Inbred Year 2 USA event 2 98.89 7

Inbred agronomic field trials were conducted during the same season as the US Year 1 Efficacy Field Trial, SAAM Year 1 Efficiency Field Trial, and US Year 2 Efficacy Field Trial (11 locations, 1 state). Sites were established by full group randomization conducted at multiple sites, and trials included control of a comparable inbred line with transgenic inbred lines. A multi-season meta-analysis of multi-location inbred agronomic field trials was performed comparing the yields of a paired control and a transgenic inbred plant using one of two events. For inbred agronomic yield, no statistical difference was found between control and transgenic corn containing event MON 87419 (Table 9). In contrast, there was a statistically significant reduction in yield of transgenic corn containing event 2 compared to either control or transgenic corn containing event MON 87419. These data further indicate higher productivity of transgenic corn containing event MON 87419 corn.

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Table 9

Yield meta-analysis of inbred agronomic field trials

Field test inbred corn General (bushels/acre) LSD (P<0.05) Inbred 1 year USA Control 65.87 31 Inbred 1 year USA MON 87419 60.33 31 Inbred 1 year USA event 2 43.10 31 Inbred 2 years UAM Control 95.88 7 Inbred 2 years UAM MON 87419 98.77 7 Inbred 2 years UAM event 2 75.85 7 Inbred 2 years USA Control 108.90 6 Inbred 2 years USA MON 87419 109.22 6 Inbred 2 years USA event 2 96.88 6

Example 2 DNA sequence characterization with MON 87419 maize event.

This example describes the extensive molecular characterization that has been performed on maize event MON 87419. The transgene insert of maize event MON 87419 in the 5' to 3' direction contains (i) a promoter (P-ANDge.Ubq1), a leader and an intron (L-1 -ANDge.Ubq1) ubiquitin gene (Ubq) from Andropogon gerardii; operably linked to the pat gene from Streptomyces viridochromogenes (CR-STRvi.pat), which encodes phosphinothricin N-acetyltransferase (PAT), which confers resistance to the herbicide glufosinate; operably linked to a polyadenylation signal (also known as a terminator that directs mRNA polyadenylation) from the RA5B precursor gene from Oryza sativa (T-Os.Ara5); and (ii) a full-length peanut chlorotic streak virus transcript promoter with a doubled enhancer site (PClSV); a gene from Triticum aestivum (L-Ta.Lhcb1) functionally linked to the leader of the b1 light harvesting complex; functionally linked to the first intron of actin gene 1 from Oryza sativa (I-Os.Act1); operably linked to an N-terminal chloroplast transit peptide from a petunia 5-enolpyruvylshikimate-3-phosphate synthase gene fusion (TS-Ph.ShkG-CTP4); a dmo-operated Stenotrophomonas maltophilia gene optimized for expression in monocots (CR-STEma.DMO), which encodes dicamba monooxygenase (DMO), conferring resistance to dicamba herbicide; operably linked to the heat shock protein 17 polyadenylation signal from Triticum aestivum (T-Ta.Hsp17).

The 5' end of the transgene insert is flanked by the right border of Agrobacterium tumifaciens and the 3' end of the transgene insert is flanked by the left border of Agrobacterium tumifaciens. Southern blot analysis was performed to confirm that the transgenic maize containing the MON 87419 maize event contained a single intact copy of the entire transgenic insert without any vector backbone. Flanking sequences were separated from both the 5' and 3' ends of the insert, and the corresponding compounds were identified using reverse PCR and/or genome walking methods. The chromosomal location of the MON 87419 maize event insert was determined by reverse PCR to amplify the genomic DNA beyond the site of interest. The flanking sequences of the MON 87419 maize event were mapped to the known physical population of the maize genome, and it was confirmed that the MON 87419 maize event was not within any of the known genes. This sequence information was used to develop TAQMAN® event-specific endpoint assays to detect the presence of the MON 87419 maize event in a sample. Information about the sequence of the insertion site was also used for bioinformatic analysis of the chromosomal localization of the event. Wholeness

- 18 040754 Insertion site viability was determined by PCR through the wild-type allele using primers specific to the flanking regions of the MON 87419 maize event. To ensure that no changes or mutations were introduced into any region of the transgene during transformation, the entire transgenic insertion of the MON 87419 maize event was isolated from the plant and sequenced.

N-terminal protein sequencing of the expressed PAT and DMO proteins was performed using immunopurified protein extracts obtained from transgenic maize grain containing the MON 87419 maize event. This sequence was then used to authenticate the N-terminal amino acid sequence. Western analysis was performed on protein extracts obtained from transgenic corn kernels containing the MON 87419 maize event. This confirmed that a single protein of the expected size was produced for PAT and DMO, respectively. ELISA assays were performed to determine protein levels in various tissue types of transgenic maize (leaves, seeds, roots and pollen) for PAT or DMO protein expressed from maize event MON 87419. Northern analysis was performed on polyadenylated RNA isolated from transgenic maize grain. containing the maize event MON 87419. This confirmed the transcript size and the number of pat and dmo mRNA products. RNA expression levels were also measured by real-time PCR using samples obtained from transgenic maize containing the MON 87419 maize event.

Example 3 TAQMAN® Event Specific Endpoint Assays.

This example describes an event-specific TAQMAN® endpoint thermal amplification method designed to identify transgenic maize containing the MON 87419 maize event in a sample. The DNA primers used in the endpoint assay are primers SQ26644 (SEQ ID NO: 11), SQ26645 (SEQ ID NO: 12) and 6-FAM™ labeled probe PB11207 (SEQ ID NO: 13). 6-FAM™ is a fluorescent dye product from Applied Biosystems (Foster City, CA) attached to a DNA probe. For TAQMAN® MGB™ 5' probes, the exonuclease activity of Taq DNA polymerase cleaves the probe at the 5' end between the fluorophore and quencher. When hybridized to the target DNA strand, the quencher and fluorophore are separated enough to produce a fluorescent signal, thereby releasing fluorescence. SQ26644 and SQ26645, when used with these reaction methods and PB 11207, produce a DNA amplicon that is diagnostic for the MON 87419 maize event.

Controls for this assay should include a positive control containing the MON 87419 maize event, a negative control from non-transgenic maize, and a negative control that contains no template DNA. In addition, the control of the PCR reaction should optimally include internal control primers and an internal control probe specific to one copy of the gene in the maize genome. These assays are optimized for use with either the Applied Biosystems GeneAmp® PCR System 9700 (run at maximum speed) or the MJ Research DNA Engine PTC-225 thermal cycler, but other equipment can be used.

An example of the conditions that are used with the TAQMAN® endpoint assay method used to detect the MON 87419 maize event is as follows.

Stage 1: 18 megaohm water was adjusted to a final volume of 10 μl.

Stage 2: 5.0 μl of 2x Universal Master Mix (dNTP, enzyme, buffer) was adjusted to 1x final concentration.

Step 3: 0.5 µl event primer-1 (SQ26644) and event primer-2 (SQ26645). The mixture (resuspended in 18 MΩ water to a concentration of 20 μM of each primer) was adjusted to 1.0 μM final concentration (e.g. in a microcentrifuge tube, to achieve 500 μl of a final concentration of 20 μM, the following must be added: 100 μl of primer SQ26644 at a concentration of 100 µM; 100 µl primer SQ26645 at a concentration of 100 µM; 300 µl 18 megaohm water).

Step 4: 0.2 µl 6-FAM™ MGB PB11207 Event Probe (10 µM) (resuspended in 18 MΩ water to 10 to 0.2 µM final concentration.

Step 5: 0.5 µl of a mixture of internal control primer-1 and internal control primer-2 (resuspended in 18 megohm water to a concentration of 20 µM for each primer) was adjusted to 1.0 µM final concentration.

Step 6: 0.2 µl VIC™ internal control probe (10 µM) was adjusted to 0.2 µM final concentration (resuspended in 18 MΩ water to 10 µM concentration).

Step 7: 3.0 µl of isolated DNA (template) for each sample with one of the following containing

1) leaf samples for analysis;

2) negative control (non-transgenic DNA);

3) negative water control (without matrix);

4) positive control of transgenic maize containing maize event DNA

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MON 87419.

Step 8: The thermal cycler conditions are as follows:

one cycle at 50°C for 2 min;

one cycle at 95°C for 10 min;

ten cycles at 95°C for 15 s, then 64°C for 1 min at (-1°O/cycle);

thirty cycles at 95°C for 15 s, then 54°C for 1 min, optional additional 10 to 20 cycles (95°C for 15 s, then 64°C for 1 min (-1°C/cycle) may provide a clearer population separation in an EndPoint TaqMan® assay);

final cycle at 10°C.

Example 4 Zygosity Analysis.

Zygosity analysis can be used to determine whether a plant containing the MON 87419 maize event is heterozygous or homozygous for the event or wild-type allele. An amplification reaction assay can be designed using the sequence information provided herein. For example, such a PCR assay would include the construction of at least three primers: primer-1, primer-2, and primer-3, with primer-1 specific for maize genomic DNA at the 3' flanking end of the MON 87419 maize event; primer-2 is specific for the MON 87419 maize event transgene insert; and primer-3 is specific for the wild-type allele. When used as a primer pair in an amplification reaction of primer-1 with primer-2, a PCR amplicon specific for the MON 87419 maize event will be obtained. When used as a primer pair in an amplification reaction of primer-1 with primer-3, a PCR amplicon specific for for the wild-type allele. In a PCR reaction performed on maize genomic DNA, the corresponding PCR amplicons generated with primer-1+primer-2 and primer-1+primer-3 will differ in amplicon sequence and size. When the three primers are included in a PCR reaction with DNA isolated from a plant homozygous for the MON 87419 maize event, only the primer-1+primer-2 amplicon will be synthesized. When the three primers are included in a PCR reaction with DNA isolated from a plant heterozygous for the MON 87419 maize event, a primer-1+primer-2 amplicon and a primer-1+primer-3 amplicon will be synthesized. When the three primers are mixed together in a PCR reaction with DNA isolated from a plant null for the MON 87419 event maize (one that is wild type), only the primer-1+primer-3 amplicon will be synthesized. Another way to determine the zygosity of a corn plant for corn event MON 87419 is the TaqMan® endpoint thermal amplification test. For this type of assay, in addition to the primers as described above, the assay will include two fluorescently labeled probes. Probe-1 will be specific to the MON 87419 maize event and probe-2 will be specific to a corn plant that is null for the MON 87419 (wild type) maize event, with the two probes containing different fluorescent labels, such as the 6-FAM™ label or VIC™ tag. When using TAQMAN®, the thermal amplification reaction at the primer-1+primer-2+probe-1 endpoint will produce the first fluorescent signal specific to the MON 87419 maize event. When using TAQMAN®, the thermal amplification reaction at the endpoint of primer-1+primer- 3+ probe-2 will produce a second fluorescent signal specific to wild-type corn. When three primers and two probes are included in a TAQMAN® endpoint thermal amplification reaction with DNA isolated from a plant homozygous for the MON 87419 maize event, only the first fluorescent signal will be generated (primer-1+primer-2+probe-1 specific) . When the three primers are included in a TAQMAN® endpoint thermal amplification reaction with DNA isolated from a plant heterozygous for the MON 87419 maize event, a first fluorescent signal (primer-1+primer-2+probe-1 specific) and a second fluorescent signal will be generated. (specific to primer-1+primer-3+probe-2). When the three primers are mixed together in a TAQMAN® endpoint thermal amplification reaction with DNA isolated from a plant that is null for the MON 87419 maize event (wild type), only a second fluorescent signal (specific to primer-1+primer-3+) will be generated. probe-2).

Another way to determine the zygosity of a corn plant for the MON 87419 event would be the Southern analysis. One of ordinary skill in the art will understand how to develop a Southern hybridization probe(s) specific for the corn event MON 87419 and a second Southern hybridization probe(s) specific for the corn plant that is null for the corn event. MON 87419 (wild type). Using Southern analysis, a signal detected only from the first hybridization probe for Southern analysis would indicate that the plant is homozygous for the MON 87419 maize event; a signal detected from both the first Southern hybridization probe and the second Southern hybridization probe would indicate that the plant is heterozygous for the MON 87419 maize event; and a signal detected only from the second hybridization probe for Southern analysis would indicate that the DNA was isolated from the plant

- 20 040754 which is null for corn event MON 87419 (wild type).

Claims (28)

CLAIM 1. Recombinant DNA molecule containing a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 , SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 and SEQ ID NO: 10 or complementary to them, and the recombinant DNA molecule is a marker of a transgenic event that provides tolerance to the herbicides glufosinate or dicamba or glufosinate and dicamba, and a representative semen sample containing this event was deposited with the ATCC under accession number PTA-120860. 2. A recombinant DNA molecule according to claim 1, characterized in that the DNA molecule is obtained from a transgenic corn plant or seed containing a transgenic corn event. 3. Recombinant DNA molecule according to claim 1, characterized in that the DNA molecule is an amplicon for diagnosing the presence of maize transgenic event DNA. 4. DNA molecule containing SEQ ID NO: 1 or SEQ ID NO: 2, where the DNA molecule is a DNA probe. 5. A pair of DNA primers for detecting a recombinant DNA molecule according to claim 1, where the pair of DNA primers contains the first DNA and the second DNA primers, and the first DNA primer contains a sequence of sufficient length of consecutive nucleotides within 1247-8008 of the sequence of SEQ ID NO: 10 or its complement to function as a DNA primer and the second DNA primer contains a sequence of sufficient length from consecutive nucleotides within 1-1246 or 8009-9259 of the sequence of SEQ ID NO: 10 or its complement to function as a DNA primer, and DNA primers when they are joint used in an amplification reaction with a DNA sample containing the recombinant DNA molecule according to claim 1, an amplicon is produced that contains SEQ ID NO: 1 or SEQ ID NO: 2. 6. A method for detecting the presence of a recombinant DNA molecule according to claim 1 in a DNA sample, the method comprising bringing the sample into contact with the DNA molecule according to claim 4; application of stringent hybridization conditions to the sample and DNA molecule; and detecting hybridization of the DNA molecule with the target DNA molecule in the sample, wherein the hybridization of the DNA molecule with the target DNA molecule indicates the presence of said recombinant DNA molecule according to claim 1 in the DNA sample. 7. A method for detecting the presence of a recombinant DNA molecule according to claim 1 in a DNA sample, and the method includes bringing the sample into contact with a pair of DNA primers according to claim 5; performing an amplification reaction sufficient to obtain a DNA amplicon containing a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8; and detecting the amplicon DNA in the reaction, wherein the presence of the amplicon DNA in the reaction indicates the presence of said recombinant DNA molecule according to claim 1 in the DNA sample. 8. DNA detection kit containing a) a DNA molecule containing SEQ ID NO: 1 or SEQ ID NO: 2; or b) a pair of DNA primers according to claim 5; And c) instructions for use of said kit, wherein the kit detects the presence of said maize transgenic event, wherein a representative seed sample containing said event is deposited with ATCC under accession number PTA-120860. 9. A transgenic maize plant comprising a recombinant DNA molecule according to claim 1, wherein the presence of the recombinant DNA molecule indicates the presence of a maize transgenic event conferring tolerance to the herbicides glufosinate or dicamba or glufosinate and dicamba, wherein a representative seed sample containing said event is deposited in ATCC under the access number PTA-120860. 10. Transgenic seed of a corn plant, containing a recombinant DNA molecule according to claim 1, wherein the presence of a recombinant DNA molecule indicates the presence of a transgenic corn event that provides tolerance to the herbicides glufosinate or dicamba or glufosinate and dicamba, and a representative seed sample containing the specified event is deposited in ATCC under the access number PTA-120860. - 21 040754 11. A transgenic corn plant cell comprising a recombinant DNA molecule according to claim 1, wherein the presence of the recombinant DNA molecule indicates the presence of a transgenic corn event conferring tolerance to glufosinate or dicamba or glufosinate and dicamba herbicides, wherein a representative seed sample containing said event is deposited in ATCC under the access number PTA-120860. 12. A transgenic part of a corn plant containing a recombinant DNA molecule according to claim 1, wherein the presence of a recombinant DNA molecule indicates the presence of a transgenic corn event that provides tolerance to the herbicides glufosinate or dicamba or glufosinate and dicamba, and a representative seed sample containing the specified event is deposited in ATCC under the access number PTA-120860. 13. A commercial product containing a recombinant DNA molecule according to claim 1, wherein said commercial product contains a detectable amount of said DNA molecule, and wherein the presence of the recombinant DNA molecule indicates the presence of a transgenic event that provides tolerance to the herbicides glufosinate or dicamba or glufosinate and dicamba, and a representative seed sample containing said event is deposited with the ATCC under accession number PTA-120860, where said commercial product includes processed seeds, grains, plant parts, or cornmeal. 14. A transgenic corn plant according to claim 9, characterized in that the corn plant is a hybrid having at least one parent plant that contains a transgenic corn event, and a representative seed sample containing the specified event is deposited with the ATCC under the PTA access number -120860. 15. The transgenic seed according to claim 10, characterized in that the corn seed is a hybrid having at least one parent plant that contains the corn transgenic event, and a representative seed sample containing the specified event is deposited with the ATCC under the accession number PTA- 120860. 16. A method for controlling weeds at a site, comprising planting a transgenic corn containing a transgenic corn event that provides tolerance to glufosinate or dicamba or glufosinate and dicamba herbicides at the site and applying an effective dose of dicamba or glufosinate or dicamba and glufosinate herbicides to control weeds at plot without damaging the transgenic corn, wherein a representative seed sample containing said event is deposited with the ATCC under accession number PTA-120860, said transgenic corn event containing a sequence selected from the group consisting of SEQ ID NOs: 1-10. 17. The method of claim 16, wherein the effective dose of glufosinate herbicide is generally from about 0.1 pounds (0.045 kg) of acid equivalent per acre (0.405 ha) to about 16 pounds (7.248 kg) of acid equivalent per acre of herbicide glufosinate during the growing season. 18. The method of claim 16, wherein the effective dose of glufosinate herbicide is generally from about 0.4 pounds (0.181 kg) acid equivalent per acre (0.405 ha) to about 1.59 pounds (0.720 kg) acid equivalent per an acre of glufosinate herbicide during the growing season. 19. The method of claim 16, wherein the effective dose of dicamba herbicide is generally from about 0.1 pounds (0.045 kg) of acid equivalent per acre (0.405 ha) to about 16 pounds (7.248 kg) of acid equivalent per acre of herbicide dicamba during the growing season. 20. The method of claim 16 wherein the effective dose of dicamba herbicide is generally from about 0.5 pounds (0.22 kg) acid equivalent per acre (0.405 ha) to about 2 pounds (0.906 kg) acid equivalent per an acre of dicamba herbicide during the growing season. 21. A method for producing a transgenic corn plant resistant to glufosinate and dicamba herbicides, the method comprising sexually crossing a transgenic corn plant containing a corn transgenic event that provides tolerance to glufosinate or dicamba or glufosinate and dicamba herbicides with itself or another corn plant, wherein A representative seed sample containing said event has been deposited with the ATCC under Accession Number PTA-120860, said corn event containing a sequence selected from the group consisting of SEQ ID NOs: 1-10; collection of obtained seeds; - 22 040754 growing seeds to obtain progeny plants; treatment of progeny plants with herbicides glufosinate or dicamba or glufosinate and dicamba; and selecting a progeny plant resistant to the herbicides glufosinate and dicamba. 22. A method for growing a transgenic corn plant resistant to glufosinate and dicamba herbicides, comprising a) planting a corn seed containing a transgenic event conferring tolerance to the herbicides glufosinate or dicamba or glufosinate and dicamba, wherein a representative seed sample containing said event is deposited with the ATCC under accession number PTA-120860, said corn event containing a sequence selected from the group consisting of SEQ ID NO: 1-10; b) allowing the plant to grow from said seed; and c) treating the plant with the herbicides glufosinate or dicamba or glufosinate and dicamba. 23. A method for obtaining a plant resistant to the use of herbicides glufosinate and dicamba, including a) providing a DNA construct containing SEQ ID NO: 9; b) introducing the DNA construct into a corn plant cell; And c) regenerating a cell of a corn plant to produce a plant, such as a corn plant, resistant to the use of the herbicides glufosinate and dicamba. 24. The method of claim 23, wherein inserting the construct DNA into a corn plant cell results in a sequence selected from SEQ ID NOs: 1-8 and 10 as part of the corn cell genome. 25. A corn plant obtained by the method of claim 23. 26. A plant that is a descendant of the corn plant according to claim 25. 27. Seed of a corn plant according to claim 25. 28. A part of a corn plant according to claim 25.