# US 2014/0196174 A1 · Development of Phytophthora Resistant Potato with Increased Yield > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0196174 A1 - Document ID: US20140196174A1 - Status: Published application - Kind: A1 - Application number: 14/115,711 - Filed: 2012-05-24 - Published: 2014-07-10 - Earliest claimed: 2011-05-24 - Assignees: BASF Plant Science GmbH - Inventors: Holger Schultheiss; Timo Böhme - Web page: https://patentgrep.com/patent/US20140196174A1 ## CPC - C12N 15/8279: {for biotic stress resistance, pathogen resistance, disease resistance} - C12N 15/8282: {for fungal resistance} - C12Q 1/6895: for plants, fungi or algae - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract The present invention relates to transgenic potato plants having an increased resistance against Phytophthora infestans and a comparable yield of potato tubers compared with the wildtype potato plants, wherein the blb1-gen and blb2-gen are integrated within a specific genetic background into the potato plant. ## Claims ### Claim 1 A Phythophthora-resistant transgenic potato plant, seed, tuber, plant cell or tissue thereof comprising a nucleic acid molecule comprising a nucleotide sequence having at least 80% identity with SEQ ID NO: 1. ### Claim 2 A Phythophthora-resistant transgenic potato plant, seed, tuber, plant cell or tissue thereof comprising: a) a recombinant construct comprising a nucleic acid sequence having at least 80% identity with SEQ ID NO: 2 or SEQ ID NO: 3; and b) further comprising a junction comprising a nucleic acid sequence selected from the group consisting of: i) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 126 and 136 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 4 and SEQ ID NO: 5; ii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 505 and 515 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 6 and SEQ ID NO: 7; iii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 625 and 635 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 8 and SEQ ID NO: 9; and/or iv) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 4752 and 4762 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 10 and SEQ ID NO: 11; and/or further comprising a junction comprising a nucleic acid sequence selected from the group consisting of: v) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 282 and 292 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 12 and SEQ ID NO: 13; vi) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 877 and 887 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 14 and SEQ ID NO: 15; vii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 827 and 837 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 16 and SEQ ID NO: 17; and/or viii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 9905 and 9915 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 18 and SEQ ID NO: 19. ### Claim 3 (depends on 1) The Phythophthora-resistant transgenic potato, seed, tuber, plant cell or tissue of claim 1 comprising: a) a recombinant construct comprising a nucleic acid sequence having at least 80% identity with SEQ ID NO: 2 or SEQ ID NO: 3; and b) further comprising a junction comprising a nucleic acid sequence selected from the group consisting of: i) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 131 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 4 and SEQ ID NO: 5; ii) a nucleic acid sequence that can be used to amplify a nucleotide fragment 510 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 6 and SEQ ID NO: 7; iii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 630 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 8 and SEQ ID NO: 9; and/or iv) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 4757 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 10 and SEQ ID NO: 11; and/or further comprising a junction comprising a nucleic acid sequence selected from the group consisting of: v) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 287 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 12 and SEQ ID NO: 13; vi) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 882 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 14 and SEQ ID NO: 15; vii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 832 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 16 and SEQ ID NO: 17; and/or viii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 9910 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 18 and SEQ ID NO: 19. ### Claim 4 A method for providing a Phythophthora-resistant transgenic potato plant comprising the following steps: a) introducing a recombinant nucleic acid comprising a nucleic acid sequence having at least 80% identity with SEQ ID NO: 2 or SEQ ID NO: 3 into the genome of potato plant cells; b) integrating said recombinant nucleic acid into the genome; c) regenerating a plant from said plant cells; d) selecting a plant comprising a nucleic acid comprising a nucleic acid sequence having at least 80% identity with SEQ ID NO: 2 or SEQ ID NO: 3 and a junction comprising a nucleic acid sequence selected from the group consisting of: i) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 126 and 136 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 4 and SEQ ID NO: 5; ii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 505 and 515 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 6 and SEQ ID NO: 7; iii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 625 and 635 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 8 and SEQ ID NO: 9; and/or iv) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 4752 and 4762 basepairs, using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 10 and SEQ ID NO: 11; and/or further comprising a junction comprising a nucleic acid sequence selected from the group consisting of: v) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 282 and 292 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 12 and SEQ ID NO: 13; vi) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 877 and 887 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 14 and SEQ ID NO: 15; vii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 827 and 837 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 16 and SEQ ID NO: 17; and/or viii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of between 9905 and 9915 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 18 and SEQ ID NO: 19. ### Claim 5 (depends on 4) The method for providing a Phythophthora-resistant transgenic potato plant of claim 4, wherein in step d) a plant is selected comprising a nucleic acid comprising a nucleic acid sequence having at least 80% identity with SEQ ID NO: 2 or SEQ ID NO: 3 and a junction comprising a nucleic acid sequence selected from the group consisting of: i) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 131 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 4 and SEQ ID NO: 5; ii) a nucleic acid sequence that can be used to amplify a nucleotide fragment 510 basepairs, using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 6 and SEQ ID NO: 7; iii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 630 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 8 and SEQ ID NO: 9; and/or iv) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 4757 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 10 and SEQ ID NO: 11; and/or further comprising a junction comprising a nucleic sequence selected from the group consisting of: v) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 287 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 12 and SEQ ID NO: 13; vi) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 882 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 14 and SEQ ID NO: 15; vii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 832 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 16 and SEQ ID NO: 17; and/or viii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 9910 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 18 and SEQ ID NO: 19. ### Claim 6 A kit comprising the primer pairs for the detection of the specific integration place, selected from the group consisting of SEQ ID NO: 4 and 5; SEQ ID NO: 6 and 7; SEQ ID NO: 8 and 9; SEQ ID NO: 10 and 14; SEQ ID NO: 12 and 13; SEQ ID NO: 14 and 15; SEQ ID NO: 16 and and/or SEQ ID NO: 18 and 19. ### Claim 7 (depends on 6) A detection method for the detection of the specific integration place comprising: a) isolating a nucleic acid sequence from a potato plant, seed, tuber, plant cell or tissue thereof as a test sample; b) exposing said test sample, a positive and a negative sample with nucleotide sequence selected from at least one set of primer pairs defined in claim 6 under PCR-conditions; and c) evaluating the amplification of a nucleotide fragment selected from the group consisting of a nucleotide fragment i) of between 126 and 136 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 4 and SEQ ID NO: 5; ii) of between 505 and 515 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 6 and SEQ ID NO: 7, of 625 and 635 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 8 and SEQ ID NO: 9; and/or iii) of 4752 and 4762 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 10 and SEQ ID NO: 11 and/or selected from the group consisting of a nucleotide fragment; iv) of between 282 and 292 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 12 and SEQ ID NO: 13; v) of 877 and 292 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 14 and SEQ ID NO: 15; vi) of 827 and 837 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequence of SEQ ID NO: 16 and SEQ ID NO: 17; and/or vii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 9905 and 9915 basepairs, using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 18 and SEQ ID NO: 19 compared with said positive and negative control. ### Claim 8 (depends on 7) The detection method of claim 7, wherein step c) comprises evaluating the amplification of a nucleotide fragment selected from the group consisting of a nucleotide fragment: i) of 131 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 4 and SEQ ID NO: 5; ii) of 510 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 6 and SEQ ID NO: 7; iii) of 630 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 8 and SEQ ID NO: 9; and/or iv) of 4757 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 10 and SEQ ID NO: 1 and/or evaluating the amplification of a nucleotide fragment selected from the group consisting of a nucleotide fragment selected from the group consisting of a nucleotide fragment; v) of 287 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 12 and SEQ ID NO: 13; vi) of 882 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 14 and SEQ ID NO: 15; vii) of 832 basepairs when using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 16 and SEQ ID NO: 17; and/or viii) a nucleic acid sequences that can be used to amplify a nucleotide fragment of 9910 basepairs, using a polymerase chain reaction with two primers having the nucleotide sequence of SEQ ID NO: 18 and SEQ ID NO: 19 compared with said positive and negative control. ### Claim 9 (depends on 6) A plant, seed, tuber, plant cell or tissue thereof detectable by the kit of claim 6. ### Claim 10 A polynucleotide comprising: a) a recombinant nucleic acid having at least 80% identity with SEQ ID NO: 2 or SEQ ID NO: 3; and b) further comprising a junction comprising a nucleic acid sequence selected from the group consisting of: i) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 131 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 4 and SEQ ID NO: 5; ii) a nucleic acid sequence that can be used to amplify a nucleotide fragment 510 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 6 and SEQ ID NO: 7; iii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 630 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 8 and SEQ ID NO: 9; and/or iv) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 4757 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 10 and SEQ ID NO: 11; and/or iv) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 287 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 12 and SEQ ID NO: 13; v) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 882 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 14 and SEQ ID NO: 15; vi) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 832 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 16 and SEQ ID NO: 17; and/or vii) a nucleic acid sequence that can be used to amplify a nucleotide fragment of 13234 basepairs using a polymerase chain reaction with two primers having the nucleotide sequences of SEQ ID NO: 18 and SEQ ID NO: 19 stably integrated into a potato plant cell nucleus. ### Claim 11 A polynucleotide comprising a nucleotide sequence having at least 80% identity SEQ ID NO: 1 stably integrated into a potato plant cell nucleus. ### Claim 12 (depends on 10) The polynucleotide of claim 10, wherein the nucleotide sequence having at least 80% identity SEQ ID NO: 1 comprises the blbl gene and the blb2 gene. ### Claim 13 (depends on 12) The polynucleotide of claim 12, wherein the polynucleotide sequence further comprises one or more of SEQ ID NOS: 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, and/or 19. ### Claim 14 (depends on 10) A Phythophthora-resistant transgenic potato plant, seed, tuber, plant cell or tissue thereof comprising the polynucleotide of claim 10. ### Claim 15 (depends on 1) The Phythophthora-resistant transgenic potato plant, seed, tuber, plant cell or tissue thereof of claim 1, wherein the nucleic acid molecule or recombinant construct comprises the blb1 gene and the blb2 gene. ### Claim 16 (depends on 7) A plant, seed, tuber, plant cell or tissue thereof detectable by the method of claim 7. ### Claim 17 (depends on 11) The Phythophthora-resistant transgenic potato plant, seed, tuber, plant cell or tissue thereof comprising the polynucleotide of claim 11. ### Claim 18 (depends on 12) The Phythophthora-resistant transgenic potato plant, seed, tuber, plant cell or tissue thereof comprising the polynucleotide of claim 12. ### Claim 19 (depends on 13) The Phythophthora-resistant transgenic potato plant, seed, tuber, plant cell or tissue thereof comprising the polynucleotide of claim 13. ### Claim 20 (depends on 2) The Phythophthora-resistant transgenic potato plant, seed, tuber. plant cell or tissue thereof of claim 2, wherein the nucleic acid molecule or recombinant construct comprises the blb1 gene and the blb2 gene. ### Claim 21 (depends on 3) The Phythophthora-resistant transgenic potato plant, seed, tuber, plant cell or tissue thereof of claim 3, wherein the nucleic acid molecule or recombinant construct comprises the blb1 gene and the blb2 gene.