Maize event dp-098140-6 and compositions and mehtods for the identification and/or detection thereof

Abstract

Compositions and methods related to transgenic glyphosate/ALS inhibitor-tolerant maize plants are provided. Specifically, the present invention provides maize plants having a DP-098140-6 event which imparts tolerance to glyphosate and at least one ALS-inhibiting herbicide. The maize plant harboring the DP-098140-6 event at the recited chromosomal location comprises genomic/transgene junctions having at least the polynucleotide sequence of SEQ ID NO:5 and/or 6. The characterization of the genomic insertion site of the DP-098140-6 event provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in the breeding populations and progeny thereof. Various methods and compositions for the identification, detection, and use of the maize DP-098140-6 events are provided.

1 . An isolated polynucleotide comprising SEQ ID NO: 5 or 6. 
     
     
         2 . The isolated polynucleotide of  claim 1 , wherein said polynucleotide is selected from the group consisting of:
 (a) a nucleotide sequence set forth in SEQ ID NO:48, 7, 8, 9, 10, 49, or 50; and,   (b) a nucleotide sequence comprising a fragment of SEQ ID NO:48, 7, 8, 9, 10, 49, or 50.   
     
     
         3 . A kit for identifying event DP-098140-6 in a biological sample, said kit comprising a first and a second primer, wherein said first and said second primer amplify a polynucleotide comprising a DP-098140-6 specific region. 
     
     
         4 . The kit of  claim 3 , wherein said kit further comprises a polynucleotide for the detection of the DP-098140-6 specific region. 
     
     
         5 . The kit of  claim 3 , wherein said first primer comprises a first fragment of SEQ ID NO: 48 and the second primer comprises a second fragment of SEQ ID NO:48, wherein said first and said second primer flank said DP-098140-6 specific region and share sufficient sequence homology or complementarity to said polynucleotide to amplify said DP-098140-6 specific region. 
     
     
         6 . The kit of  claim 5 , wherein
 a) said first primer comprises a fragment of SEQ ID NO:47 and said second primer comprises a fragment of SEQ ID NO:1;   b) said first primer comprises a fragment of SEQ ID NO:47 and said second primer comprises a fragment of SEQ ID NO:46;   c) said first primer comprises a fragment of SEQ ID NO: 46 and said second primer comprises a fragment of SEQ ID NO:1.   
     
     
         7 . The kit of  claim 5 , wherein said first or said second primer comprises at least 8 consecutive polynucleotides of SEQ ID NO: 48. 
     
     
         8 . The kit of  claim 6 , wherein said first or said second primer comprises at least 8 consecutive polynucleotides of SEQ ID NO:1, 46, or 47. 
     
     
         9 . The kit of  claim 5 , wherein said first or said second primer comprises at least 8 consecutive nucleotides of SEQ ID NO:24. 
     
     
         10 . The kit of  claim 3 , wherein said first or said second primer comprise SEQ ID NO:13, 14, 15, 16, 17, 18, 19, 20, 21, 25, 26, 27, 28, 29, 30, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 51, 52, 53, 54, 55, or 56. 
     
     
         11 . A DNA detection kit comprising at least one polynucleotide that can specifically detect a DP-098140-6 specific region, wherein said polynucleotide comprises at least one DNA molecule of a sufficient length of contiguous nucleotides identical or complementary to SEQ ID NO:48. 
     
     
         12 . The DNA detection kit of  claim 11 , wherein said polynucleotide that can specifically detect a DP-098140-6 specific region comprises a polynucleotide having SEQ ID NO:5 or 6. 
     
     
         13 . The DNA detection kit of  claim 11 , wherein said polynucleotide comprises a sequence which hybridizes under stringent conditions with sequences selected from the group consisting of:
 (a) the sequences of SEQ ID NO:1 and SEQ ID NO:47; and,   (b) the sequences of SEQ ID NO:46 and SEQ ID NO: 47.   
     
     
         14 . A method for identifying event DP-098140-6 in a biological sample, comprising
 (a) contacting said sample with a first and a second primer; and,   (b) amplifying a polynucleotide comprising a DP-098140-6 specific region.   
     
     
         15 . The method of  claim 14 , wherein the polynucleotide comprising the DP-098140-6 specific region comprises SEQ ID NO:5. 
     
     
         16 . The method of  claim 14 , wherein the polynucleotide comprising the DP-098140-6 specific region comprises SEQ ID NO:6. 
     
     
         17 . The method of  claim 14 , further comprising detecting the DP-098140-6 specific region. 
     
     
         18 . The method of  claim 14 , said first primer comprises a first fragment of SEQ ID NO: 48 and the second primer comprises a second fragment of SEQ ID NO:48, wherein said first and said second primer flank said DP-098140-6 specific region and share sufficient sequence homology or complementarity to said polynucleotide to amplify said DP-098140-6 specific region. 
     
     
         19 . The method of  claim 18 , wherein
 a) said first primer comprises a fragment of SEQ ID NO:47 and said second primer comprises a fragment of SEQ ID NO:1;   b) said first primer comprises a fragment of SEQ ID NO:47 and said second primer comprises a fragment of SEQ ID NO:46;   c) said first primer comprises a fragment of SEQ ID NO: 1 and said second primer comprises a fragment of SEQ ID NO:46.   
     
     
         20 . The method of  claim 18 , wherein said first and or said second primer comprises at least 8 consecutive polynucleotides of SEQ ID NO: 48. 
     
     
         21 . The method of  claim 19 , wherein said first and said second primer comprises at least 8 consecutive polynucleotides of SEQ ID NO:1, 46 or 47. 
     
     
         22 . The method of  claim 14 , wherein said first and said second primer comprise SEQ ID NO:13, 14, 15, 16, 17, 18, 19, 20, 21, 25, 26, 27, 28, 29, 30, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 51, 52, 53, 54, 55, or 56. 
     
     
         23 . A method for identifying event DP-098140-6 in a biological sample, comprising
 (a) contacting said sample with a first and a second primer;   (b) performing a DNA amplification reaction, thereby producing a DNA amplicon molecule; and   (c) detecting said DNA amplicon molecule, wherein the detection of said DNA amplicon molecule in said DNA amplification reaction indicates the presence of maize event DP-098140-6.   
     
     
         24 . The method of  claim 23 , wherein said first primer comprises a first fragment of SEQ ID NO: 48 and the second primer comprises a second fragment of SEQ ID NO:48, wherein said first and said second primer flank said DP-098140-6 specific region and share sufficient sequence homology or complementarity to said polynucleotide to amplify said DP-098140-6 specific region. 
     
     
         25 . The method of  claim 23 , wherein said first and second primer are selected from the group consisting of:
 (a) the sequences comprising SEQ ID NO:13 and SEQ ID NO:14;   (b) the sequences comprising SEQ ID NO:15 and SEQ ID NO:16;   (c) the sequences comprising SEQ ID NO:17 and SEQ ID NO:18;   (d) the sequences comprising SEQ ID NO:20 and SEQ ID NO:21;   (e) the sequences comprising SEQ ID NO:51 and SEQ ID NO:52; and,   (f) the sequence comprising SEQ ID NO: 54 and 55.   
     
     
         26 . A method of detecting the presence of DNA corresponding to a DP-098140-6 event in a sample, the method comprising:
 (a) contacting the sample with a polynucleotide probe that hybridizes under stringent hybridization conditions with DNA from maize event DP-098140-6 and specifically detects the DP-098140-6 event;   (b) subjecting the sample and probe to stringent hybridization conditions; and   (c) detecting hybridization of the probe to the DNA, wherein detection of hybridization indicates the presence of the DP-098140-6 event.   
     
     
         27 . The method of  claim 26 , wherein said sample comprises maize tissue. 
     
     
         28 . A polynucleotide comprising a sequence selected from the group consisting of SEQ ID NO:13, 14, 15, 16, 17, 18, 19, 20, 21, 24, 25, 26, 27, 28, 29, 30, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 51, 52, 53, 54, 55, 56 or its complement. 
     
     
         29 . A pair of DNA molecules comprising a first DNA molecule and a second DNA molecule, said first DNA molecule comprising a first fragment of SEQ ID NO: 48 and the second DNA molecule comprising a second fragment of SEQ ID NO:48, wherein said first and said second DNA molecule flank said DP-098140-6 specific region of SEQ ID NO:51 and share sufficient sequence homology or complementarity to said polynucleotide to amplify said DP-098140-6 specific region. 
     
     
         30 . A method for confirming seed purity or a method for screening seeds in a seed lot for a DP-098140-6 event comprising
 (a) contacting said sample with a first and a second primer;   (b) performing a DNA amplification reaction, thereby producing a DNA amplicon molecule; and   (c) detecting said DNA amplicon molecule, wherein the detection of said DNA amplicon molecule in said DNA amplification reaction indicates the presence of maize event DP-098140-6.   
     
     
         31 . The method of  claim 30 , wherein said first primer comprises a first fragment of SEQ ID NO: 48 and the second primer comprises a second fragment of SEQ ID NO:48, wherein said first and said second primer flank said DP-098140-6 specific region and share sufficient sequence homology or complementarity to said polynucleotide to amplify said DP-098140-6 specific region. 
     
     
         32 . The method of  claim 31 , wherein
 a) said first primer comprises a fragment of SEQ ID NO:47 and said second primer comprises a fragment of SEQ ID NO:1;   b) said first primer comprises a fragment of SEQ ID NO:47 and said second primer comprises a fragment of SEQ ID NO:46;   c) said first primer comprises a fragment of SEQ ID NO:1 and said second primer comprises a fragment of SEQ ID NO:46.   
     
     
         33 . The method of  claim 31 , wherein said first or said second primer comprises at least 8 consecutive polynucleotides of SEQ ID NO:48. 
     
     
         34 . The method of  claim 32 , wherein said second primer and said first primer comprise at least 8 consecutive polynucleotides of SEQ ID NO:1, 46 or 47. 
     
     
         35 . The method of  claim 34 , wherein said first or said second primer comprise SEQ ID NO:13, 14, 15, 16, 17, 18, 19, 20, 21, 25, 26, 27, 28, 29, 30, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 51, 52, 53, 54, 55, or 56. 
     
     
         36 . A method for confirming seed purity or for screening for the presence of a DP-098140-6 event in a seed lot comprising
 (a) contacting a sample comprising maize DNA with a polynucleotide probe that hybridizes under stringent hybridization conditions with DNA from maize event DP-098140-6 and specifically detects the DP-098140-6 event;   (b) subjecting the sample and probe to stringent hybridization conditions; and   (c) detecting hybridization of the probe to the DNA, wherein detection of hybridization indicates the presence of the DP-098140-6 event.