Stacked herbicide tolerance event 8264.42.32.1, related transgenic soybean lines, and detection thereof

A plant cell comprising an expression cassette, inserted transgenically into chromosome 15 at a locus flanked by or including SEQ ID NO:1 and SEQ ID NO:2, said expression cassette comprising:

• a. a first plant transcription unit which expresses a glyphosate herbicide tolerance gene;
• b. a second plant transcription unit which expresses an aryloxyalkanoate herbicide tolerance gene;
• and
• c. a third plant transcription unit which expresses a glufosinate herbicide tolerance gene.

A method for determining event zygosity of a soybean plant comprising soybean event pDAB8264.42.32.1, as present in seed deposited under ATCC Accession No. PTA-11993, said event comprising a transgene construct, said transgene construct being flanked by a 5′ flanking soybean genomic DNA and a 3′ flanking soybean genomic DNA, said method comprising:

• obtaining a DNA sample of genomic DNA from said soybean plant;
• producing a contacted sample by contacting said DNA sample with
• a. a first event primer and a second event primer, wherein said first event primer specifically binds said transgene construct, said second event primer specifically binds said 5′ soybean genomic flanking DNA or said 3′ soybean genomic flanking DNA, and wherein said first event primer and said second event primer produce an event amplicon when subjected to TAQMAN PCR conditions
• b. a reference forward primer and a reference reverse primer that produce a reference amplicon from an endogenous soybean reference gene when subjected to TAQMAN PCR conditions
• c. a fluorescent event probe that hybridizes with said event amplicon
• d. a fluorescent reference probe that hybridizes with said reference amplicon;
• subjecting said contacted sample to fluorescence-based endpoint TAQMAN PCR conditions;
• quantitating said fluorescent event probe that hybridized to said event amplicon;
• quantitating said fluorescent reference probe that hybridized to said reference amplicon;
• comparing amounts of hybridized fluorescent event probe to hybridized fluorescent reference probe;
• and
• determining zygosity of pDAB8264.42.32.1 by comparing fluorescence ratios of hybridized fluorescent event probe and hybridized fluorescent reference probe.