A transposon-mediate inactivation of a CYCLOIDEA-like gene originates polysymmetric and androgynous ray flowers in Helianthus annuus

Genetica

Marco Fambrini, Mariangela Salvini, Claudio Pugliesi

Correspondence:

Pugliesi Claudio, Dipartimento di Biologia delle Piante Agrarie, Sezione di Genetica Agraria, Università di Pisa, Via Matteotti 1B, I-56124 Pisa, Italy.

E-mail:

Supplemental Fig. 7Nucleotide sequence alignments of the CYC10/CYC11 (a) and the RAG19/CYC11 (b) PCR amplified products from wild type (TURF), tubular ray flower mutant (turf) of sunflower (Helianthus annuus L.) andindividuals of a progenyobtained from a self-fertilized revertant. a Nucleotide sequences of homozygous (TT)and heterozygous (Tt) individuals of a progeny obtained from a self-fertilized revertant and TURF. bNucleotide sequences of heterozygous (Tt) and homozygous (tt) individuals of a progeny obtained froma self-fertilized revertantand turf. Underlined in bold is a portion of the CACTA transposable element (Tetu1). The three bp (ATA)target site duplication (TSD) are in white letters and highlighted in black. Highlighted in yellow and grey are the primers CYC10 and CYC11 used in a, respectively. Highlighted in light-blue and grey are the primers RAG19 and CYC11 used in b,respectively. The arrows indicate the 5’-3’-primer direction. The start and the stop codons are in bold, underlined and highlighted in green and magenta, respectively. The intron, located in the 3’-UTR region is in italic and underlined. Asterisks indicate identical nucleotides.

a

TURF CAGTGTAAAGCTTAAGAGGGGTGGTGGTCTCTTAAATGCAACTTTACCCTTAGCTTTATT 60

TT CAGTGTAAAGCTTAAGAGGGGTGGTGGTCTCTTAAATGCAACTTTACCCTTAGCTTTATT 60

Tt CAGTGTAAAGCTTAAGAGGGGTGGTGGTCTCTTAAATGCAACTTTACCCTTAGCTTTATT 60

************************************************************

TURF TGGATCAATGAACTTATATCTAAACTTCTTTGTTAACTATTATTGTTATTGTTTCTTTCA 120

TT TGGATCAATGAACTTATATCTAAACTTCTTTGTTAACTATTATTGTTATTGTTTCTTTCA 120

Tt TGGATCAATGAACTTATATCTAAACTTCTTTGTTAACTATTATTGTTATTGTTTCTTTCA 120

************************************************************

TURF TTTCCAAATTACAATGTTTTCCTCAAACCCTTTTCCACAGATTCCATCTCCCATCCATGT 180

TT TTTCCAAATTACAATGTTTTCCTCAAACCCTTTTCCACAGATTCCATCTCCCATCCATGT 180

Tt TTTCCAAATTACAATGTTTTCCTCAAACCCTTTTCCACAGATTCCATCTCCCATCCATGT 180

************************************************************

TURF CTCACATCCTTTCAATTCCTTCTTTGACCTTGAAAGAAACGATGTATATTTCAACCATCA 240

TT CTCACATCCTTTCAATTCCTTCTTTGACCTTGAAAGAAACGATGTATATTTCAACCATCA 240

Tt CTCACATCCTTTCAATTCCTTCTTTGACCTTGAAAGAAACGATGTATATTTCAACCATCA 240

************************************************************

TURF TGAAGACAACAATAACCCTTTTGTTTCCGGTGATTCTTTTTTACATTCCTACAGCAGTTT 300

TT TGAAGACAACAATAACCCTTTTGTTTCCGGTGATTCTTTTTTACATTCCTACAGCAGTTT 300

Tt TGAAGACAACAATAACCCTTTTGTTTCCGGTGATTCTTTTTTACATTCCTACAGCAGTTT 300

************************************************************

TURF TGCTCCACAACCATCAACCCTACCTCCAGTCACAGACTATATACCTTCAAATGCACAAGA 360

TT TGCTCCACAACCATCAACCCTACCTCCAGTCACAGACTATATACCTTCAAATGCACAAGA 360

Tt TGCTCCACAACCATCAACCCTACCTCCAGTCACAGACTATATACCTTCAAATGCACAAGA 360

************************************************************

TURF GTTAGATAGTCAAAACCAGCTGCTTGATCATGAAGGGTCTGGGTTGCAATATTGTGACAA 420

TT GTTAGATAGTCAAAACCAGCTGCTTGATCATGAAGGGTCTGGGTTGCAATATTGTGACAA 420

Tt GTTAGATAGTCAAAACCAGCTGCTTGATCATGAAGGGTCTGGGTTGCAATATTGTGACAA 420

************************************************************

TURF TTATAGTGATCTTTTGGAGTCAGTAGTTTACCCATCAAAGAAAAAAGTGGCGATTTCAAA 480

TT TTATAGTGATCTTTTGGAGTCAGTAGTTTACCCATCAAAGAAAAAAGTGGCGATTTCAAA 480

Tt TTATAGTGATCTTTTGGAGTCAGTAGTTTACCCATCAAAGAAAAAAGTGGCGATTTCAAA 480

************************************************************

TURF AAAAGATGGGCATAGTAAAATCTATACCGCACAAGGCCCGAGAGATCGGAGGGTGAGATT 540

TT AAAAGATGGGCATAGTAAAATCTATACCGCACAAGGCCCGAGAGATCGGAGGGTGAGATT 540

Tt AAAAGATGGGCATAGTAAAATCTATACCGCACAAGGCCCGAGAGATCGGAGGGTGAGATT 540

************************************************************

TURF GTCCATTGACATTGCAAGAAAGTTTTTTGTGCTTCAAGATTTGCTAGGTTTTGACAAAGC 600

TT GTCCATTGACATTGCAAGAAAGTTTTTTGTGCTTCAAGATTTGCTAGGTTTTGACAAAGC 600

Tt GTCCATTGACATTGCAAGAAAGTTTTTTGTGCTTCAAGATTTGCTAGGTTTTGACAAAGC 600

************************************************************

TURF AAGCAAAACCCTTGATTGGCTCTTTACCAAGTCCAAGAAGGCGATTAAGGAGTTGATTGA 660

TT AAGCAAAACCCTTGATTGGCTCTTTACCAAGTCCAAGAAGGCGATTAAGGAGTTGATTGA 660

Tt AAGCAAAACCCTTGATTGGCTCTTTACCAAGTCCAAGAAGGCGATTAAGGAGTTGATTGA 660

************************************************************

TURF AGAAACAAAGCACTCCTCATCTTCCACTGTTAGTACTAATCAGTGTGAAATGGCTTTCCT 720

TT AGAAACAAAGCACTCCTCATCTTCCACTGTTAGTACTAATCAGTGTGAAATGGCTTTCCT 720

Tt AGAAACAAAGCACTCCTCATCTTCCACTGTTAGTACTAATCAGTGTGAAATGGCTTTCCT 720

************************************************************

TURF AGAGACCATCAAGGGAGGATCAGGATCAGATGAAGATAAAGGGCAAAAAATGTCAGCTCT 780

TT AGAGACCATCAAGGGAGGATCAGGATCAGATGAAGATAAAGGGCAAAAAATGTCAGCTCT 780

Tt AGAGACCATCAAGGGAGGATCAGGATCAGATGAAGATAAAGGGCAAAAAATGTCAGCTCT 780

************************************************************

TURF CAGGTTTCTTGATGGAGAAAAGAAGAAAATGACACAAAAATGCAAATCTGGATTTCATGC 840

TT CAGGTTTCTTGATGGAGAAAAGAAGAAAATGACACAAAAATGCAAATCTGGATTTCATGC 840

Tt CAGGTTTCTTGATGGAGAAAAGAAGAAAATGACACAAAAATGCAAATCTGGATTTCATGC 840

************************************************************

TURF TAGTCTTGCAAGAGGCCAGTCAAGGGCAGAGGCAAGAGCAAGGGCTAGGGAGAGAACAAA 900

TT TAGTCTTGCAAGAGGCCAGTCAAGGGCAGAGGCAAGAGCAAGGGCTAGGGAGAGAACAAA 900

Tt TAGTCTTGCAAGAGGCCAGTCAAGGGCAGAGGCAAGAGCAAGGGCTAGGGAGAGAACAAA 900

************************************************************

TURF ACAAAAATTGCGCATCAAGGAGCTAGACAACGACTTGAAGAAAATTCCTGATGATTACCC 960

TT ACAAAAATTGCGCATCAAGGAGCTAGACAACGACTTGAAGAAAATTCCTGATGATTACCC 960

Tt ACAAAAATTGCGCATCAAGGAGCTAGACAACGACTTGAAGAAAATTCCTGATGATTACCC 960

************************************************************

TURF TTGTCATGCATTAAGCCCCTCAAACACAACACTCCAATCAAACTCTTGGGGTCAATTTGA 1020

TT TTGTCATGCATTAAGCCCCTCAAACACAACACTCCAATCAAACTCTTGGGGTCAATTTGA 1020

Tt TTGTCATGCATTAAGCCCCTCAAACACAACACTCCAATCAAACTCTTGGGGTCAATTTGA 1020

************************************************************

TURF ATCACAAAGTGACTATAATGACATCCTTCATGGATCAATGCTGGAACAAAGGTTTTCAGT 1080

TT ATCACAAAGTGACTATAATGACATCCTTCATGGATCAATGCTGGAACAAAGGTTTTCAGT 1080

Tt ATCACAAAGTGACTATAATGACATCCTTCATGGATCAATGCTGGAACAAAGGTTTTCAGT 1080

************************************************************

TURF ATCATCTTCAACTTTGTATACTTACAATCACAACTTTGTTGTATCAGATGAGTGCATCTC 1140

TT ATCATCTTCAACTTTGTATACTTACAATCACAACTTTGTTGTATCAGATGAGTGCATCTC 1140

Tt ATCATCTTCAACTTTGTATACTTACAATCACAACTTTGTTGTATCAGATGAGTGCATCTC 1140

************************************************************

TURF CCAAATTTCATCCAAAGGTATGCCTAAATTTAAAGTAATTCACGAGGAGCAGAGTCACCG 1200

TT CCAAATTTCATCCAAAGGTATGCCTAAATTTAAAGTAATTCACGAGGAGCAGAGTCACCG 1200

Tt CCAAATTTCATCCAAAGGTATGCCTAAATTTAAAGTAATTCACGAGGAGCAGAGTCACCG 1200

************************************************************

TURF TGCAATGATATAGATCTACAAAAGGTAGATTAATTAATCAAATACAAAACACGGTAGTTA 1260

TT TGCAATGATATAGATCTACAAAAGGTAGATTAATTAATCAAATACAAAACACGGTAGTTA 1260

Tt TGCAATGATATAGATCTACAAAAGGTAGATTAATTAATCAAATACAAAACACGGTAGTTA 1260

************************************************************

TURF CTGAATATAGTTTTTAAGATTTTATATGCCTTTGTTTTGTCAACAGAACTCTGAAAGGGG 1320

TT CTGAATATAGTTTTTAAGATTTTATATGCCTTTGTTTTGTCAACAGAACTCTGAAAGGGG 1320

Tt CTGAATATAGTTTTTAAGATTTTATATGCCTTTGTTTTGTCAACAGAACTCTGAAAGGGG 1320

************************************************************

TURF TGCTGGAGAAAATCTAGTCTTCATCAGCCGGCAAATAGATCTAGTAACCACTTCTCGGCA 1380

TT TGCTGGAGAAAATCTAGTCTTCATCAGCCGGCAAATAGATCTAGTAACCACTTCTCGGCA 1380

Tt TGCTGGAGAAAATCTAGTCTTCATCAGCCGGCAAATAGATCTAGTAACCACTTCTCGGCA 1380

************************************************************

TURF CTAGTTGATTGG 1392

TT CTAGTTGATTGG 1392

Tt CTAGTTGATTGG 1392

************

b

turf GGTTGCATTTGAGTGTAACAAAGGTTGCATTTTGGTGTAGAAAAGGTTGCGTTTGAGGTG 60

Tt GGTTGCATTTGAGTGTAACAAAGGTTGCATTTTGGTGTAGAAAAGGTTGCGTTTGAGGTG 60

tt GGTTGCATTTGAGTGTAACAAAGGTTGCATTTTGGTGTAGAAAAGGTTGCGTTTGAGGTG 60

************************************************************

turf GCAAAACTTGCATTTGAGATAACAAAGGTTACATTTGAGTGCTAGAAAAGTTGCACTGAA 120

Tt GCAAAACTTGCATTTGAGATAACAAAGGTTACATTTGAGTGCTAGAAAAGTTGCACTGAA 120

tt GCAAAACTTGCATTTGAGATAACAAAGGTTACATTTGAGTGCTAGAAAAGTTGCACTGAA 120

************************************************************

turf AGTAAAAAGAGTTGCGTTAGGATATAAAAAAGGTTGCATTATGAAAAGGTTGCATTTGAA 180

Tt AGTAAAAAGAGTTGCGTTAGGATATAAAAAAGGTTGCATTATGAAAAGGTTGCATTTGAA 180

tt AGTAAAAAGAGTTGCGTTAGGATATAAAAAAGGTTGCATTATGAAAAGGTTGCATTTGAA 180

************************************************************

turf TAGTAAATGCAACTCTTTAAAAAAAGGTTGCATTTTCTAACAAAAAGGTTGCATTAGGTC 240

Tt TAGTAAATGCAACTCTTTAAAAAAAGGTTGCATTTTCTAACAAAAAGGTTGCATTAGGTC 240

tt TAGTAAATGCAACTCTTTAAAAAAAGGTTGCATTTTCTAACAAAAAGGTTGCATTAGGTC 240

************************************************************

turf TAATGCAACTTCATCTAATGCAGCCCTTGATAAAAGGTTGCATTAGGCCTAATGCAACCT 300

Tt TAATGCAACTTCATCTAATGCAGCCCTTGATAAAAGGTTGCATTAGGCCTAATGCAACCT 300

tt TAATGCAACTTCATCTAATGCAGCCCTTGATAAAAGGTTGCATTAGGCCTAATGCAACCT 300

************************************************************

turf TTTTAACCCCTAATGCAACATTTTATAAGGGTTGCATTAGCATCATTTTCTTGTAGTGAT 360

Tt TTTTAACCCCTAATGCAACATTTTATAAGGGTTGCATTAGCATCATTTTCTTGTAGTGAT 360

tt TTTTAACCCCTAATGCAACATTTTATAAGGGTTGCATTAGCATCATTTTCTTGTAGTGAT 360

************************************************************

turf AGTAAAATCTATACCGCACAAGGCCCGAGAGATCGGAGGGTGAGATTGTCCATTGACATT 420

Tt AGTAAAATCTATACCGCACAAGGCCCGAGAGATCGGAGGGTGAGATTGTCCATTGACATT 420

tt AGTAAAATCTATACCGCACAAGGCCCGAGAGATCGGAGGGTGAGATTGTCCATTGACATT 420

************************************************************

turf GCAAGAAAGTTTTTTGTGCTTCAAGATTTGCTAGGTTTTGACAAAGCAAGCAAAACCCTT 480

Tt GCAAGAAAGTTTTTTGTGCTTCAAGATTTGCTAGGTTTTGACAAAGCAAGCAAAACCCTT 480

tt GCAAGAAAGTTTTTTGTGCTTCAAGATTTGCTAGGTTTTGACAAAGCAAGCAAAACCCTT 480

************************************************************

turf GATTGGCTCTTTACCAAGTCCAAGAAGGCGATTAAGGAGTTGATTGAAGAAACAAAGCAC 540

Tt GATTGGCTCTTTACCAAGTCCAAGAAGGCGATTAAGGAGTTGATTGAAGAAACAAAGCAC 540

tt GATTGGCTCTTTACCAAGTCCAAGAAGGCGATTAAGGAGTTGATTGAAGAAACAAAGCAC 540

************************************************************

turf TCCTCATCTTCCACTGTTAGTACTAATCAGTGTGAAATGGCTTTCCTAGAGACCATCAAG 600

Tt TCCTCATCTTCCACTGTTAGTACTAATCAGTGTGAAATGGCTTTCCTAGAGACCATCAAG 600

tt TCCTCATCTTCCACTGTTAGTACTAATCAGTGTGAAATGGCTTTCCTAGAGACCATCAAG 600

************************************************************

turf GGAGGATCAGGATCAGATGAAGATAAAGGGCAAAAAATGTCAGCTCTCAGGTTTCTTGAT 660

Tt GGAGGATCAGGATCAGATGAAGATAAAGGGCAAAAAATGTCAGCTCTCAGGTTTCTTGAT 660

tt GGAGGATCAGGATCAGATGAAGATAAAGGGCAAAAAATGTCAGCTCTCAGGTTTCTTGAT 660

************************************************************

turf GGAGAAAAGAAGAAAATGACACAAAAATGCAAATCTGGATTTCATGCTAGTCTTGCAAGA 720

Tt GGAGAAAAGAAGAAAATGACACAAAAATGCAAATCTGGATTTCATGCTAGTCTTGCAAGA 720

tt GGAGAAAAGAAGAAAATGACACAAAAATGCAAATCTGGATTTCATGCTAGTCTTGCAAGA 720

************************************************************

turf GGCCAGTCAAGGGCAGAGGCAAGAGCAAGGGCTAGGGAGAGAACAAAACAAAAATTGCGC 780

Tt GGCCAGTCAAGGGCAGAGGCAAGAGCAAGGGCTAGGGAGAGAACAAAACAAAAATTGCGC 780

tt GGCCAGTCAAGGGCAGAGGCAAGAGCAAGGGCTAGGGAGAGAACAAAACAAAAATTGCGC 780

************************************************************

turf ATCAAGGAGCTAGACAACGACTTGAAGAAAATTCCTGATGATTACCCTTGTCATGCATTA 840

Tt ATCAAGGAGCTAGACAACGACTTGAAGAAAATTCCTGATGATTACCCTTGTCATGCATTA 840

tt ATCAAGGAGCTAGACAACGACTTGAAGAAAATTCCTGATGATTACCCTTGTCATGCATTA 840

************************************************************

turf AGCCCCTCAAACACAACACTCCAATCAAACTCTTGGGGTCAATTTGAATCACAAAGTGAC 900

Tt AGCCCCTCAAACACAACACTCCAATCAAACTCTTGGGGTCAATTTGAATCACAAAGTGAC 900

tt AGCCCCTCAAACACAACACTCCAATCAAACTCTTGGGGTCAATTTGAATCACAAAGTGAC 900

************************************************************

turf TATAATGACATCCTTCATGGATCAATGCTGGAACAAAGGTTTTCAGTATCATCTTCAACT 960

Tt TATAATGACATCCTTCATGGATCAATGCTGGAACAAAGGTTTTCAGTATCATCTTCAACT 960

tt TATAATGACATCCTTCATGGATCAATGCTGGAACAAAGGTTTTCAGTATCATCTTCAACT 960

************************************************************

turf TTGTATACTTACAATCACAACTTTGTTGTATCAGATGAGTGCATCTCCCAAATTTCATCC 1020

Tt TTGTATACTTACAATCACAACTTTGTTGTATCAGATGAGTGCATCTCCCAAATTTCATCC 1020

tt TTGTATACTTACAATCACAACTTTGTTGTATCAGATGAGTGCATCTCCCAAATTTCATCC 1020

************************************************************

turf AAAGGTATGCCTAAATTTAAAGTAATTCACGAGGAGCAGAGTCACCGTGCAATGATATAG 1080

Tt AAAGGTATGCCTAAATTTAAAGTAATTCACGAGGAGCAGAGTCACCGTGCAATGATATAG 1080

tt AAAGGTATGCCTAAATTTAAAGTAATTCACGAGGAGCAGAGTCACCGTGCAATGATATAG 1080

************************************************************

turf ATCTACAAAAGGTAGATTAATTAATCAAATACAAAACACGGTAGTTACTGAATATAGTTT 1140

Tt ATCTACAAAAGGTAGATTAATTAATCAAATACAAAACACGGTAGTTACTGAATATAGTTT 1140

tt ATCTACAAAAGGTAGATTAATTAATCAAATACAAAACACGGTAGTTACTGAATATAGTTT 1140

************************************************************

turf TTAAGATTTTATATGCCTTTGTTTTGTCAACAGAACTCTGAAAGGGGTGCTGGAGAAAAT 1200

Tt TTAAGATTTTATATGCCTTTGTTTTGTCAACAGAACTCTGAAAGGGGTGCTGGAGAAAAT 1200

tt TTAAGATTTTATATGCCTTTGTTTTGTCAACAGAACTCTGAAAGGGGTGCTGGAGAAAAT 1200

************************************************************

turf CTAGTCTTCATCAGCCGGCAAATAGATCTAGTAACCACTTCTCGGCACTAGTTGATTGG 1259

Tt CTAGTCTTCATCAGCCGGCAAATAGATCTAGTAACCACTTCTCGGCACTAGTTGATTGG 1259

tt CTAGTCTTCATCAGCCGGCAAATAGATCTAGTAACCACTTCTCGGCACTAGTTGATTGG 1259

***********************************************************