INBRED TRANSGENIC CANOLA LINE NS-B50027-4 AND SEEDS THEREOF

1 . A method of detecting the presence of an elite event NS-B50027-4, representative seed of an inbred NS-B50027-4 canola line having been deposited under ATCC Accession Number PTA-123186, in a sample comprising a plant DNA, said method comprises the steps of:

(a) contacting said sample with:

(i) a first nucleic acid primer that binds to a flanking junction region of Brassica genome of NS-B50027-4, and

(ii) a second nucleic acid primer that binds to a transgene of NS-B50027-4;

(b) subjecting said sample to a polymerase chain reaction assay; and

(c) assaying the amplicons generated between said first and second nucleic acid primers, wherein the presence of said generated amplicons is diagnostic of elite event NS-B50027-4 in said sample, and

wherein said inbred canola line NS-B50027-4 contains within its genome:

a first transgenic locus, located on a first chromosome, comprising one copy of each of a M pusilla Δ 6-desaturase, a P. cordata Δ 5-elongase, a P. salina Δ 5-desaturase, and a P. pastoris Δ 15/ω3-desaturase gene; and

a second transgenic locus, located on a second chromosome, comprising two copies of each of a Micromonas pusilla Δ 6-desaturase, a Pyramimonas cordata Δ 5-elongase, a Pavlova salina Δ5-desaturase, a Pichia pastoris Δ 15/ω3-desaturase, a Pavlova salina Δ4-desaturase, a Lachancea kluyveri Δ 12-desaturase, and a P. cordata Δ 6-elongase gene.

2 . The method of claim 2 , wherein said first and second nucleic acid primers are the primers as shown in SEQ. ID NO:1 to SEQ ID NO:90.

3 . A kit comprising components to carry out the method of claim 1 or claim 2 .

4 . A method of detecting the presence of at least one transgenic locus in a sample, wherein said method comprises the steps of:

(a) contacting said sample that comprises a plant DNA with:

(i) at least one first nucleic acid primer that binds to a first transgene-flanking junction region of a first transgenic locus, said at least one first nucleic acid primer selected from the group of primers consisting of SEQ ID NO:17-SEQ ID NO:31 and SEQ ID NO:83-SEQ ID NO:86; or

(ii) at least one second nucleic acid primer that binds to a second transgene-flanking junction region of said first transgenic locus, said at least one second nucleic acid primer selected from the group of primers consisting of SEQ ID NO:32-SEQ ID NO:41; or

(iii) at least one third nucleic acid primer that binds to a first transgene-flanking junction region of a second transgenic locus, said at least one third nucleic acid primer selected from the group of primers consisting of SEQ ID NO:42-SEQ ID NO:62; or

(iv) at least one fourth nucleic acid primer that binds to a second transgene-flanking junction region of said second transgenic locus, said at least one fourth nucleic acid primer selected from the group of primers consisting of SEQ ID NO:63-SEQ ID NO:90; or

(v) at least ten contiguous nucleotides of any preceding nucleic acid primers of (i), (ii), (iii) and (iv) or complements thereof;

(b) subjecting said sample to polymerase chain reaction assay; and

(c) assaying for amplicons generated between said primers, wherein the presence of said amplicons is diagnostic of the presence of at least one transgenic locus from a segregant of an inbred canola line NS-B50027-4,

wherein said inbred canola line NS-B50027-4, representative seed of an inbred NS-B50027-4 canola line having been deposited under ATCC Accession Number PTA-123186.

5 . The method of claim 4 , wherein said polymerase chain reaction assay is a KASP™ genotyping assay.

6 . The method of claim 4 , further comprising the step of contacting said sample with a nucleic acid primer that binds to a transgene sequence, said nucleic acid primer is selected from the group of primers consisting of those depicted in SEQ ID NO:1-SEQ ID NO:8, at least ten contiguous nucleotides thereof, or complements thereof.

7 . The method of claim 4 , further comprising the step of contacting said sample with a nucleic acid primer that binds to a transgene sequence, said nucleic acid primer is selected from the group consisting of those depicted in SEQ ID NO:1-SEQ ID NO:16, at least ten contiguous nucleotides thereof, or complements thereof.

8 . A kit comprising components to carry out the method of claim 4 .

9 . A recombinant nucleic acid molecule extracted from an inbred canola line NS-B50027-4, or a progeny or segregant thereof, wherein a representative seed of said inbred canola line NS-B50027-4 has been deposited under ATCC Accession Number PTA-123186, wherein said NS-B50027-4 contains within its genome:

a first transgenic locus comprising one copy of each of a M. pusilla Δ 6-desaturase, a P. cordata Δ 5-elongase, a P. salina Δ 5-desaturase, and a P. pastoris Δ 15/ω3-desaturase gene,

a second transgenic locus comprising two copies of each of a Micromonas pusilla Δ 6-desaturase, a Pyramimonas cordata Δ 5-elongase, a Pavlova salina Δ5-desaturase, a Pichia pastoris Δ 15/ω3-desaturase, a Pavlova salina Δ4-desaturase, a Lachancea kluyveri Δ 12-desaturase, and a P. cordata Δ 6-elongase gene;

wherein said first transgenic locus is located on a first chromosome, and said second transgenic locus is located on a second chromosome;

wherein said inbred canola line NS-B50027-4, or a progeny or segregant thereof, comprises either said first transgenic locus, said second transgenic locus, or both said transgenic loci.

10 . The recombinant nucleic acid molecule of claim 9 , wherein said recombinant nucleic acid molecule has the nucleic acid sequence of (a) nucleotides 1160 to 47773 of SEQ ID NO:41 or a complement thereof; (b) nucleotides 2090 to 14201 of SEQ ID NO:40 or a complement thereof; or (c) both (a) and (b).

11 . The recombinant nucleic acid molecule of claim 10 , wherein said nucleic acid molecule of (a) is at least 80%, 95%, 97%, 98%, 99%, or 99.5% identical to the nucleotide sequence of SEQ ID NO:41 from nucleotide position 1268 to nucleotide position 47662 or a complement thereof.

12 . The recombinant nucleic acid molecule of claim 10 , wherein said nucleic acid molecule of (b) is at least 80%, 95%, 97%, 98%, 99%, or 99.5% identical to the nucleotide sequence of SEQ ID NO:40 from nucleotide position 2090 to nucleotide position 14201 or a complement thereof.