US 20180346920A1 ( 19) United States (12 ) Patent Application Publication (10 ) Pub. No. : US 2018 /0346920 A1 Nandakumar et al. (43 ) Pub . Date : Dec . 6 , 2018 ( 54 ) MUTATIONS CONFERRING ACETYL -COA (52 ) U . S . CI. CARBOXYLASE ( ACC ) INHIBITING CPC . .. .. .. C12N 15 / 8274 ( 2013 .01 ) ; C12N 9 / 93 HERBICIDE TOLERANCE IN SORGHUM ( 2013 .01 ) ; C12Y 604 / 01002 ( 2013 .01 ) ; A01H ( 71 ) Applicant: Chromatin , Inc ., Chicago , IL (US ) 6 / 4666 (2018 . 05 ) ; C12N 15 /8206 ( 2013 .01 ) (72 ) Inventors: Rangaraj Nandakumar, Lubbock , TX (US ) ; Song Luo , Chicago , IL (US ) ; (57 ) ABSTRACT Scott A . Staggenborg , Lubbock , TX (US ) The invention provides for sorghum plants and plant parts developed through tissue culture , gene editing or other ( 21 ) Appl. No. : 15 / 993 ,081 methods of mutagenesis in which the plant or plant parts (22 ) Filed : May 30 , 2018 have increased tolerance to one or more acetyl - CoA car boxylase (ACC ) herbicides at levels that would normally Related U . S . Application Data inhibit the growth of wild - type sorghum plants . In this ( 60 ) Provisional application No . 62/ 513 , 074 , filed on May context, the sorghum plantmay be tolerant to any herbicide 31 , 2017 capable of inhibiting acetyl- CoA carboxylase enzyme activ ity . The present invention allows for the screening of ACC Publication Classification herbicide tolerant hybrids with markers or application of (51 ) Int . CI. ACC inhibiting herbicides , and for the removal of unwanted C12N 15 / 82 (2006 .01 ) vegetation with application of ACC inhibiting herbicides C12N 9 / 00 ( 2006 .01 ) from seed and grain production fields. AOIH 6 / 46 ( 2006 .01 ) Specification includes a Sequence Listing . Patent Application Publication Dec. 6 , 2018 Sheet 1 of 22 US 2018 / 0346920 A1 BTX430 control calli . SO FOP - 13 : W1999C mutant calliLT 3214(BTX430 ) Figure 1 . Patent Application Publication Dec . 6 , 2018 Sheet 2 of 22 US 2018 / 0346920 A1 Control BTX430 Control W1999C mutant calli (BTX430 ) Figure 2 . Patent Application Publication Dec . 6 , 2018 Sheet 3 of 22 US 2018 / 0346920 A1 Control plants (BTX430 ) W1999C mutant plants Figure 3 . Patent Application Publication Dec. 6 , 2018 Sheet 4 of 22 US 2018 / 0346920 A1 Figure 4 . DNA and protein sequence data SEQ ID No. 1 . ACC Carboxyl Transferase domain sequence (Wild Type BTX430 ) . GCAAACTCTGGTGCTAGGATTGGCATAGCTGATGAAGTAAAATCTTGCTTC CGTGTTGGGTGGTCTGACGAAGGCAGCCCTGAGCGAGGGTTTCAGTACAT CTATCTGACTGAAGAAGACTATGCCCGTATTAGCTCTTCTGTTATAGCACAT AAGCTGCAGCTAGATAGCGGTGAAATTAGGTGGATTATTGACTCTGTTGTGTI LAYA1A 1 GGCAAGGAGGATGGGCTTGGTGTTGAGAACATACATGGAAGTGCTGCTAT CGCCAGTGCTTATTCTAGGGCATATGAGGAGACATTTACACTTACATTTGT GACCGGACGGACTGTAGGAATAGGAGCTTATCTTGCTAGACTTGGTATACG GTGCATACAGCGTCTTGACCAGCCGATTATTTTAACAGGGTTTTCTGCCCT GAACAAGCTCCTTGGGCGGGAAGTGTACAGCTCCCACATGCAGCTTGGTGLY GTCCTAAGATCATGGCGACCAATGGTGTTGTCCACCTGACTGTTCCAGATG ACCTTGAAGGTGTTTCCAATATATTGAGGTGGCTCAGCTATGTTCCTGCAA ACATTGGTGGACCTCTTCCTATTACCAAACCTTTGGACCCTCCAGACAGAC CTGTTGCATACATCCCTGAGAACACATGCGATCCACGTGCAGCCATCCGTG GTGTAGATGACAGCCAAGGGAAATGGTTGGGTGGTATGTTTGACAAAGACA GCTTTGTGGAGACATTTGAAGGATGGGCAAAAACAGTGGTTACTGGCAGAG CAAAGCTTGGAGGAATTCCTGTGGGTGTCATAGCTGTGGAGACACAGACCA TGATGCAGCTTGTCCCTGCTGATCCAGGTCAGCTTGATTCCCATGAGCGAT CCGTTCCTCGGGCTGGACAAGTGTGGTTCCCAGATTCTGCAACCAAGACAG CTCAGGCATTATTAGACTTCAACCGTGAAGGATTGCCTCTGTTTATCCTGG CTAACTGGAGAGGTTTCTCTGGTGGACAGAGAGATCTCTTTGAAGGAATTC TTCAGGCTGGGTCAACAATTGTCGAGAACCTTAGGACATATAATCAGCCTG CGTTTGTCTACATTCCTATGGCTGGAGAGCTTCGTGGAGGAGCTTGGGTTG TGGTCGATAGCAAAATAAATCCAGACCGCATTGAGTGTTATGCTGAGAGGA CTGCCAAAGGTAATGTTCTCGAACCTCAAGGGTTAATTGAAATCAAGTTCA GGTCAGAGGAACTCCAAGACTGTATGGGTAGGCTTGACCCCGAGTTGATAA ATCTGAAAGCAAAACTCCAAGATGTAAAGCATGGAAATGGAAGTCTACCAG ACATAGAATCCCTTCAGAAGAGTATAGAAGCACGTACGAAACAGTTGCTGC CITTATATACCCAGATTGCAATACGGTITGCTGAATTGCATGATACTTCCCT AAGAATGGCAGCTAAAGGCGTGATTAAGAAAGTTGTAGACTGGGAAGAATC Patent Application Publication Dec . 6 , 2018 Sheet 5 of 22 US 2018 / 0346920 A1 Figure 4 Cont. ACGCTCTTTCTTCTATAAAAGGCTACGGAGAAGGATCTCTGAAGATGTTCT TGCAAAAGAAATAAGACATATAGTCGGTGACAACTTCACTCACCAATCAGC AATGGAGCTCATCAAGGAATGGTACCTGGCTTCTCCAGCCACAGCAGGAAG CACTGGATGGGATGACGATGATGCATTTGTTGCCTGGAAGGACAGTCCTGA AAACTACAATGGATATATCCAAGAGCTAAGGGCTCAAAAAGTGTCTCAGTC GCTCTCTGATCTCACTGACTCCAGTTCAGATCTACAAGCATTCTCGCAGGG TCTTTCTACGCTATTAGATAAGATGGATCCCTCTCAAAGAGCGAAGTTTGTT CAGGAAGTCAAGAAGGTCCTTGGTTGA Chromatin mutant sequences SEQ ID No . 2 . ACCI ( TGG to TGC ) GCAAACTCTGGTGCTAGGATTGGCATAGCTGATGAAGTAAAATCTTGCTTC CGTGTTGGGTGGTCTGACGAAGGCAGCCCTGAGCGAGGGTTTCAGTACAT CTATCTGACTGAAGAAGACTATGCCCGTATTAGCTCTTCTGTTATAGCACAT AAGCTGCAGCTAGATAGCGGTGAAATTAGGTGGATTATTGACTCTGTTGTGf GGCAAGGAGGATGGGCTTGGTGTTGAGAACATACATGGAAGTGCTGCTAT CGCCAGTGCTTATTCTAGGGCATATGAGGAGACATTTACACTTACATTTGT GACCGGACGGACTGTAGGAATAGGAGCTTATCTTGCTAGACTTGGTATACG GTGCATACAGCGTCTTGACCAGCCAATTATTTTAACAGGGTTTTCTGCCCTYTTET GAACAAGCTCCTTGGGCGGGAAGTGTACAGCTCCCACATGCAGCTTGGTG GTCCTAAGATCATGGCGACCAATGGTGTTGTCCACCTGACTGTTCCAGATG ACCTTGAAGGTGTTTCCAATATATTGAGGTGGCTCAGCTATGTTCCTGCAA ACATTGGTGGACCTCTTCCTATTACCAAACCTTTGGACCCTCCAGACAGAC CTGTTGCATACATCCCTGAGAACACATGCGATCCACGTGCAGCCATCCGTG GTGTAGATGACAGCCAAGGGAAATGGTTGGGTGGTATGTTTGACAAAGACA GCTTTGTGGAGACATTTGAAGGATGGGCAAAAACAGTGGTTACTGGCAGAGYEN ry CAAAGCTTGGAGGAATTCCTGTGGGTGTCATAGCTGTGGAGACACAGACCA TGATGCAGCTTGTCCCTGCTGATCCAGGTCAGCTTGATTCCCATGAGCGATTI CCGTTCCTCGGGCTGGACAAGTGTGOTTCCCAGATTCTGCAACC CTCAGGCATTATTAGACTTCAACCGTGAAGGATTGCCTCTGTTTATCCTGG 19 CTAACTGGAGAGGTTTCTCTGGTGGACAGAGAGATCTCTTTGAAGGAATTC1 1 l Patent Application Publication Dec . 6 , 2018 Sheet 6 of 22 US 2018 / 0346920 A1 Figure 4 Cont. TTCAGGCTGGGTCAACAATTGTCGAGAACCTTAGGACATATAATCAGCCTG1 . TE W CGTTTGTCTACATTCCTATGGCTGGAGAGCTTCGTGGAGGAGCTTGGGTTG TGGTCGATAGCAAAATAAATCCAGACCGCATTGAGTGTTATGCTGAGAGGA CTGCCAAAGGTAATGTTCTCGAACCTCAAGGGTTAATTGAAATCAAGTTCA GGTCAGAGGAACTCCAAGACTGTATGGGTAGGCTTGACCCCGAGTTGATAA ATCTGAAAGCAAAACTCCAAGATGTAAAGCATGGAAATGGAAGTCTACCAG ACATAGAATCCCTTCAGAAGAGTATAGAAGCACGTACGAAACAGTTGCTGC CTTTATATACCCAGATTGCAATACGGTTTGCTGAATTGCATGATACTTCCCT AAGAATGGCAGCTAAAGGCGTGATTAAGAAAGTTGTAGACTGGGAAGAATC ACGCTCTTTCTTCTATAAAAGGCTACGGAGAAGGATCTCTGAAGATGTTCT TGCAAAAGAAATAAGACATATAGTCGGTGACAACTTCACTCACCAATCAGC AATGGAGCTCATCAAGGAATGGTACCTGGCTTCTCCAGCCACAGCAGGAAG CACTGGATGGGATGACGATGATGCATTTGTTGCCTGGAAGGACAGTCCTGA AAACTACAATGGATATATCCAAGAGCTAAGGGCTCAAAAAGTGTCTCAGTC GCTCTCTGATCTCACTGACTCCAGTTCAGATCTACAAGCATTCTCGCAGGG TCTTTCTACGCTATTAGATAAGATGGATCCCTCTCAAAGAGCGAAGTTTGTT CAGGAAGTCAAGAAGGTCCTTGGTTGA SEQ ID No. 3 . ACC2 ( TGG to TCG ) GCAAACTCTGGTGCTAGGATTGGCATAGCTGATGAAGTAAAATCTTGCTTC CGTGTTGGGTGGTCTGACGAAGGCAGCCCTGAGCGAGGGTTTCAGTACAT CTATCTGACTGAAGAAGACTATGCCCGTATTAGCTCTTCTGTTATAGCACATAIR AT mm AAGCTGCAGCTAGATAGCGGTGAAATTAGGTGGATTATTGACTCTGTTGTGTA I JA GGCAAGGAGGATGGGCTTGGTGTTGAGAACATACATGGAAGTGCTGCTAT CGCCAGTGCTTATTCTAGGGCATATGAGGAGACATTTACACTTACATTTGT GACCGGACGGACTGTAGGAATAGGAGCTTATCTTGCTAGACTTGGTATACG GTGCATACAGCGTCTTGACCAGCCAATTATTTTAACAGGGTTTTCTGCCCT GAACAAGCTCCTTGGGCGGGAAGTGTACAGCTCCCACATGCAGCTTGGTG GTCCTAAGATCATGGCGACCAATGGTGTTGTCCACCTGACTGTTCCAGATG ACCTTGAAGGTGTTTCCAATATATTGAGGTGGCTCAGCTATGTTCCTGCAALIIGA WIN! Patent Application Publication Dec. 6 , 2018 Sheet 7 of 22 US 2018 / 0346920 A1 Figure 4 Cont. ACATTGGTGGACCTCTTCCTATTACCAAACCTTTGGACCCTCCAGACAGAC CTGTTGCATACATCCCTGAGAACACATGCGATCCACGTGCAGCCATCCGTG GTGTAGATGACAGCCAAGGGAAATGGTTGGGTGGTATGTTTGACAAAGACA GCTTTGTGGAGACATTTGAAGGATGGGCAAAAACAGTGGTTACTGGCAGAG CAAAGCTTGGAGGAATTCCTGTGGGTGTCATAGCTGTGGAGACACAGACCA TGATGCAGCTTGTCCCTGCTGATCCAGGTCAGCTTGATTCCCATGAGCGAT CCGTTCCTCGGGCTGGACAAGTGICOTTCCCAGATTCTGCAACCAAGACAG CTCAGGCATTATTAGACTTCAACCGTGAAGGATTGCCTCTGTTTATCCTGG CTAACTGGAGAGGTTTCTCTGGTGGACAGAGAGATCTCTTTGAAGGAATTC TTCAGGCTGGGTCAACAATTGTCGAGAACCTTAGGACATATAATCAGCCTG CGTTTGTCTACATTCCTATGGCTGGAGAGCTTCGTGGAGGAGCTTGGGTTG TGGTCGATAGCAAAATAAATCCAGACCGCATTGAGTGTTATGCTGAGAGGA CTGCCAAAGGTAATGTTCTCGAACCTCAAGGGTTAATTGAAATCAAGTTCA GGTCAGAGGAACTCCAAGACTGTATGGGTAGGCTTGACCCCGAGTTGATAA ATCTGAAAGCAAAACTCCAAGATGTAAAGCATGGAAATGGAAGTCTACCAG ACATAGAATCCCTTCAGAAGAGTATAGAAGCACGTACGAAACAGTTGCTGC CTTTATATACCCAGATTGCAATACGGTTTGCTGAATTGCATGATACTTCCCTOLU AAGAATGGCAGCTAAAGGCGTGATTAAGAAAGTTGTAGACTGGGAAGAATC ACGCTCTTTCTTCTATAAAAGGCTACGGAGAAGGATCTCTGAAGATGTTCT TGCAAAAGAAATAAGACATATAGTCGGTGACAACTTCACTCACCAATCAGC AATGGAGCTCATCAAGGAATGGTACCTGGCTTCTCCAGCCACAGCAGGAAG CACTGGATGGGATGACGATGATGCATTTGTTGCCTGGAAGGACAGTCCTGA AAACTACAATGGATATATCCAAGAGCTAAGGGCTCAAAAAGTGTCTCAGTC GCTCTCTGATCTCACTGACTCCAGTTCAGATCTACAAGCATTCTCGCAGGG TCTTTCTACGCTATTAGATAAGATGGATCCCTCTCAAAGAGCGAAGTTTGTT CAGGAAGTCAAGAAGGTCCTTGGTTGA SEQ ID No. 4 ACC3 (GCA to GTA ) GCAAACTCTGGTGCTAGGATTGGCATAGCTGATGAAGTAAAATCTTGCTTC CGTGTTGGGTGGTCTGACGAAGGCAGCCCTGAGCGAGGGTTTCAGTACAT CTATCTGACTGAAGAAGACTATGCCCGTATTAGCTCTTCTGTTATAGCACAT. 12 . Y L L . + Patent Application Publication Dec . 6 , 2018 Sheet 8 of 22 US 2018 / 0346920 A1 Figure 4 Cont. AAGCTGCAGCTAGATAGCGGTGAAATTAGGTGGATTATTGACTCTGTTGTG GAGGATGGGCTTGGTGTTGAGAACATACATGGAAGTGCTGCTAT2 t CGCCAGTGCTTATTCTAGGGCATATGAGGAGACATTTACACTTACATTTGT GACCGGACGGACTGTAGGAATAGGAGCTTATCTTGCTAGACTTGGTATACG GTGCATACAGCGTCTTGACCAGCCAATTATTTTAACAGGGTTTTCTGCCCT GAACAAGCTCCTTGGGCGGGAAGTGTACAGCTCCCACATGCAGCTTGGTG GTCCTAAGATCATGGCGACCAATGGTGTTGTCCACCTGACTGTTCCAGATG ACCTTGAAGGTGTTTCCAATATATTGAGGTGGCTCAGCTATGTTCCTGCAA ACATTGGTGGACCTCTTCCTATTACCAAACCTTTGGACCCTCCAGACAGAC CTGTTGCATACATCCCTGAGAACACATGCGATCCACGTGCAGCCATCCGTG GTGTAGATGACAGCCAAGGGAAATGGTTGGGTGGTATGTTTGACAAAGACA GCTTTGTGGAGACATTTGAAGGATGGGCAAAAACAGTGGTTACTGGCAGAG CAAAGCTTGGAGGAATTCCTGTGGGTGTCATAGCTGTGGAGACACAGACCA TGATGCAGCTTGTCCCTGCTGATCCAGGTCAGCTTGATTCCCATGAGCGAT CCGTTCCTCGGGCTGGACAAGTGTCGTTCCCAGATTCTGRAACCAAGACAG CTCAGGCATTATTAGACTTCAACCGTGAAGGATTGCCTCTGTTTATCCTGG