Onion Discrimination Method

12:

• A method of discriminating traits of an onion, comprising:
• a first determination step of determining presence of the nucleotide sequence of SEQ ID NO:1 in a nucleic acid derived from the onion;
• and
• a second determination step of determining presence of the nucleotide sequence of SEQ ID NO:2 in the nucleic acid derived from the onion, wherein
• the onion is discriminated to be an onion with no pungent taste and/or tear-inducing property if the presence of the nucleotide sequence of SEQ ID NO:1 is not determined in the first determination step and the presence of the nucleotide sequence of SEQ ID NO:2 is determined in the second determination step, and
• the onion is discriminated to be an onion with a pungent taste and/or a tear-inducing property if the presence of the nucleotide sequence of SEQ ID NO:1 is determined in the first determination step, and the presence of the nucleotide sequence of SEQ ID NO:2 is determined in the second determination step.

14:

• The method according to claim 12, wherein
• the first determination step comprises:
• performing a first nucleic acid amplification reaction on a genomic DNA or a cDNA of the onion as a template using a first primer set comprising:
• a first primer comprising a polynucleotide comprising a first nucleotide sequence at the 3′ end, wherein the first nucleotide sequence is identical to a first partial nucleotide sequence of 17 or more consecutive nucleotides comprised in the nucleotide sequence of SEQ ID NO:1, the first partial nucleotide sequence comprising one or more first nucleotides selected from nucleotides at positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 within 2 nucleotides from the 3′ end, or wherein the first nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the first partial nucleotide sequence;
• and/or a second primer comprising a polynucleotide comprising a second nucleotide sequence at the 3′ end, wherein the second nucleotide sequence is identical to a second partial nucleotide sequence of 17 or more consecutive nucleotides comprised in a complementary nucleotide sequence to the nucleotide sequence of SEQ ID NO:1, the second partial nucleotide sequence comprising one or more nucleotides complementary to one or more second nucleotides selected from nucleotides at positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 in the nucleotide sequence of SEQ ID NO:1 within 2 nucleotides from the 3′ end, or wherein the second nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the second partial nucleotide sequence;
• and
• confirming an amplification product of the first nucleic acid amplification reaction, and
• the second determination step comprises:
• performing a second nucleic acid amplification reaction on a genomic DNA of the onion as a template using a second primer set comprising:
• a third primer comprising a polynucleotide comprising a third nucleotide sequence at the 3′ end, wherein the third nucleotide sequence is identical to a third partial nucleotide sequence of 17 or more consecutive nucleotides comprised in the nucleotide sequence of SEQ ID NO:2, the third partial nucleotide sequence comprising one or more third nucleotides selected from nucleotides at positions 34, 70, 1667, 1703, 127, 409, 943, and 1822 within 2 nucleotides from the 3′ end, or wherein the third nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the third partial nucleotide sequence;
• and/or a fourth primer comprising a polynucleotide comprising a fourth nucleotide sequence at the 3′ end, wherein the fourth nucleotide sequence is identical to a fourth partial nucleotide sequence of 17 or more consecutive nucleotides comprised in a complementary nucleotide sequence to the nucleotide sequence of SEQ ID NO:2, the fourth partial nucleotide sequence comprising one or more nucleotides complementary to one or more fourth nucleotides selected from nucleotides at positions 34, 70, 1667, 1703, 127, 409, 943, and 1822 in the nucleotide sequence of SEQ ID NO:2 within 2 nucleotides from the 3′ end, or wherein the fourth nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the fourth partial nucleotide sequence;
• and
• confirming an amplification product of the second nucleic acid amplification reaction.

15:

• A method of discriminating traits of an onion, comprising
• a first determination step of determining presence of the nucleotide sequence of SEQ ID NO:1 in a nucleic acid derived from the onion,
• the first determination step comprising determining presence of a first mutation site comprising one or more nucleotides selected from nucleotides at positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 in the nucleotide sequence of SEQ ID NO:1 in the nucleic acid, wherein
• the onion is discriminated to be an onion with no pungent taste and/or tear-inducing property if the presence of the nucleotide sequence of SEQ ID NO:1 is not determined in the first determination step,
• the onion is discriminated to be an onion with a pungent taste and/or a tear-inducing property if the presence of the nucleotide sequence of SEQ ID NO:1 is determined in the first determination step, and
• the first determination step comprises:
• performing a first nucleic acid amplification reaction on a genomic DNA or a cDNA of the onion as a template using a first primer set comprising:
• a first primer comprising a first polynucleotide comprising a first nucleotide sequence at the 3′ end, wherein the first nucleotide sequence is identical to a first partial nucleotide sequence of 17 or more consecutive nucleotides comprised in the nucleotide sequence of SEQ ID NO:1, the first partial nucleotide sequence comprising a first nucleotide selected from positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 in the nucleotide sequence of SEQ ID NO:1 within 2 nucleotides from the 3′ end, or wherein the first nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the first partial nucleotide sequence;
• and a second primer comprising a second polynucleotide comprising a second nucleotide sequence at the 3′ end, wherein the second nucleotide sequence is identical to a second partial nucleotide sequence of 17 or more consecutive nucleotides comprised in a complementary nucleotide sequence to the nucleotide sequence of SEQ ID NO:1, the second partial nucleotide sequence comprising a nucleotide complementary to a second nucleotide selected from positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 in the nucleotide sequence of SEQ ID NO:1 within 2 nucleotides from the 3′ end, or wherein the second nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the second partial nucleotide sequence, wherein the first nucleotide is positioned more closely to the 5′ end than the second nucleotide in the nucleotide sequence of SEQ ID NO:1;
• and
• confirming an amplification product of the first nucleic acid amplification reaction.

16:

• A method of discriminating traits of an onion, comprising:
• discriminating whether the onion is an onion with no pungent taste and/or tear-inducing property by a method according to claim 12;
• and
• discriminating whether the onion is an onion with no pungent taste and/or tear-inducing property based on one or more of the following traits as indicator(s):
• a) production of pyruvic acid at disruption of onion cells is reduced compared with conventional varieties;
• b) the quantity of PRENCSO remaining after disruption of onion cells is greater compared with conventional varieties;
• and
• c) production of lachrymatory factor (LF) at disruption of onion cells is reduced compared with conventional varieties.

17:

• A method of discriminating traits of an onion, comprising:
• discriminating whether the onion is an onion with no pungent taste and/or tear-inducing property by a method according to claim 15;
• and
• discriminating whether the onion is an onion with no pungent taste and/or tear-inducing property based on one or more of the following traits as indicator(s):
• a) production of pyruvic acid at disruption of onion cells is reduced compared with conventional varieties;
• b) the quantity of PRENCSO remaining after disruption of onion cells is greater compared with conventional varieties;
• and
• c) production of lachrymatory factor (LF) at disruption of onion cells is reduced compared with conventional varieties.

18:

• A method of breeding an onion with no pungent taste and/or tear-inducing property, comprising:
• discriminating whether an onion is an onion with no pungent taste and/or tear-inducing property by a method according to claim 12;
• and
• using the onion discriminated to be an onion with no pungent taste and/or tear-inducing property to breed an onion.

19:

• A method of breeding an onion with no pungent taste and/or tear-inducing property, comprising:
• discriminating whether an onion is an onion with no pungent taste and/or tear-inducing property by a method according to claim 15;
• and
• using the onion discriminated to be an onion with no pungent taste and/or tear-inducing property to breed an onion.

20:

• A primer set comprising:
• a primer comprising a first polynucleotide comprising a first nucleotide sequence at the 3′ end, wherein the first nucleotide sequence is identical to a first partial nucleotide sequence of 17 or more consecutive nucleotides comprised in the nucleotide sequence of SEQ ID NO:1, the first partial nucleotide sequence comprising a first nucleotide selected from positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 in the nucleotide sequence of SEQ ID NO:1 within 2 nucleotides from the 3′ end, or wherein the first nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the first partial nucleotide sequence;
• and
• a primer comprising a second polynucleotide comprising a second nucleotide sequence at the 3′ end, wherein the second nucleotide sequence is identical to a second partial nucleotide sequence of 17 or more consecutive nucleotides comprised in a complementary nucleotide sequence to the nucleotide sequence of SEQ ID NO:1, the second partial nucleotide sequence comprising a nucleotide complementary to a second nucleotide selected from positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 in the nucleotide sequence of SEQ ID NO:1 within 2 nucleotides from the 3′ end, or wherein the second nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the second partial nucleotide sequence, wherein
• the first nucleotide is positioned more closely to the 5′ end than the second nucleotide in the nucleotide sequence of SEQ ID NO:1.

21:

• A primer set comprising:
• a primer comprising a third polynucleotide comprising a third nucleotide sequence at the 3′ end, wherein the third nucleotide sequence is identical to a third partial nucleotide sequence of 17 or more consecutive nucleotides comprised in the nucleotide sequence of SEQ ID NO:2, the third partial nucleotide sequence comprising a third nucleotide selected from positions 34, 70, 1667, 1703, 127, 409, 943, and 1822 in the nucleotide sequence of SEQ ID NO:2 within 2 nucleotides from the 3′ end, or wherein the third nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the third partial nucleotide sequence;
• and
• a primer comprising a fourth polynucleotide comprising a fourth nucleotide sequence at the 3′ end, wherein the fourth nucleotide sequence is identical to a fourth partial nucleotide sequence of 17 or more consecutive nucleotides comprised in a complementary nucleotide sequence to the nucleotide sequence of SEQ ID NO:2, the fourth partial nucleotide sequence comprising a nucleotide complementary to a fourth nucleotide selected from positions 34, 70, 1667, 1703, 127, 409, 943, and 1822 in the nucleotide sequence of SEQ ID NO:2 within 2 nucleotides from the 3′ end, or wherein the fourth nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the fourth partial nucleotide sequence, wherein
• the third nucleotide is positioned more closely to the 5′ end than the fourth nucleotide in the nucleotide sequence of SEQ ID NO:2.

22:

• A kit for discriminating traits of an onion comprising:
• one selected from
• a first primer comprising a first polynucleotide comprising a first nucleotide sequence at the 3′ end, wherein the first nucleotide sequence is identical to a first partial nucleotide sequence of 17 or more consecutive nucleotides comprised in the nucleotide sequence of SEQ ID NO:1, the first nucleotide sequence comprising a first nucleotide selected from positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 in the nucleotide sequence of SEQ ID NO:1 within 2 nucleotides from the 3′ end, or wherein the first nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the first partial nucleotide sequence,
• a second primer comprising a second polynucleotide comprising a second nucleotide sequence at the 3′ end, wherein the second nucleotide sequence is identical to a second partial nucleotide sequence of 17 or more consecutive nucleotides comprised in a complementary nucleotide sequence to the nucleotide sequence of SEQ ID NO:1, the second partial nucleotide sequence comprising a nucleotide complementary to a second nucleotide selected from positions 94, 130, 1312, 1348, 187, 358, 791, and 1467 in the nucleotide sequence of SEQ ID NO:1 within 2 nucleotides from the 3′ end, or wherein the second nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the second partial nucleotide sequence, and
• a primer set comprising:
• the first primer and
• the second primer, wherein
• the first nucleotide is positioned more closely to the 5′ end than the second nucleotide in the nucleotide sequence of SEQ ID NO:1;
• and
• one selected from
• a third primer comprising a third polynucleotide comprising a third nucleotide sequence at the 3′ end, wherein the third nucleotide sequence is identical to a third partial nucleotide sequence of 17 or more consecutive nucleotides comprised in the nucleotide sequence of SEQ ID NO:2, the third partial nucleotide sequence comprising a third nucleotide selected from positions 34, 70, 1667, 1703, 127, 409, 943, and 1822 in the nucleotide sequence of SEQ ID NO:2 within 2 nucleotides from the 3′ end, or wherein the third nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the third partial nucleotide sequence;
• a fourth primer comprising a fourth polynucleotide comprising a fourth nucleotide sequence at the 3′ end, wherein the fourth nucleotide sequence is identical to a fourth partial nucleotide sequence of 17 or more consecutive nucleotides comprised in a complementary nucleotide sequence to the nucleotide sequence of SEQ ID NO:2, the fourth partial nucleotide sequence comprising a nucleotide complementary to a fourth nucleotide selected from positions 34, 70, 1667, 1703, 127, 409, 943, and 1822 in the nucleotide sequence of SEQ ID NO:2 within 2 nucleotides from the 3′ end, or wherein the fourth nucleotide sequence is identical in a region of 3 nucleotides at the 3′ end and homologous in the remaining region on the 5′ end side to the fourth partial nucleotide sequence;
• and
• a primer set comprising:
• the third primer and
• the fourth primer, wherein
• the third nucleotide is positioned more closely to the 5′ end than the fourth nucleotide in the nucleotide sequence of SEQ ID NO:2.