Journal: Molecular Breeding Molecular characterization of allelic variation in spontaneous brown midrib mutants of sor

1. Journal: Molecular Breeding • Molecular characterization of allelic variation in spontaneous brown midrib mutants of sorghum (Sorghum bicolor L Moench) • Sunita Gorthy1, Karthikeyan Mayandi1, Dhara Faldu2 and Monika Dalal1, 2 • 1 Directorate of Sorghum Research, Rajendra Nagar, Hyderabad 500 030, Andhra Pradesh, India • 2 National Research Center on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi 110 012, India • Joint First Authors • Corresponding Author: • Email: monika@nrcpb.org • Telephone No: 91-11 25841787 • Fax: +91 +11 25843984

2. Supplementary Fig. S1 Wild type CGGAGCAAGCGGCGCCGCTGCTGTGCGCGGGCGTAACGGTGTACAGCCCGCTAAAG Template IS21549 CGGAGCAAGCGGCGCCGCTGCTGTGCGTTGGCGTAACGGTGTACAGCCCGCTAAAG Template dCAPS primer AGCGGCGCCGCTGCTGAACG Wild type CGGAGCAAGCGGCGCCGCTGCTGAACGCGGGCGTAACGGTGTACAGCCCGCTAAAG Product IS21549 CGGAGCAAGCGGCGCCGCTGCTGAACGTTGGCGTAACGGTGTACAGCCCGCTAAAG Product Wild type CGGAGCAAGCGGCGCCGCTGCTGAACGCGGGCGTAACGGTGTACAGCCCGCTAAAG Product IS21549 CGGAGCAAGCGGCGCCGCTGCTGAA & CGTTGGCGTAACGGTGTACAGCCCGCTAAAG Product Digestion with AclI (AA↓CGTT)

3. IS21549 PEQAAPLLCVGVTVYSPLKAFGLTA 179 Sorghum bicolor PEQAAPLLCAGVTVYSPLKAFGLTA 179 Saccharumofficinarum PEQAAPLLCAGVTVYSPLKAFGLTT 179 Zeamays PEQAAPLLCAGVTVYSPLKHFGLTT 179 Panicumvirgatum PEQAAPLLCAGVTVYSPLKHFGLTA 179 Triticumaestivum PEQAAPLLCAGVTVYSPLKHFGLMT 180 Hordeumvulgare PEQAAPLLCAGVTVYSPLKHFGLMT 180 Festucaarundinacea PEQAAPLLCAGVTVYSPLKHFGLMT 180 Loliumperenne PEQAAPLLCAGVTVYSPLKHFGLMT 180 Brachypodiumdistachyon PEQAAPLLCAGVTVYSPLKHFGLMS 180 Oryza sativa PEQAAPLLCAGLTVYSPLKHFGLMS 175 Oryzaglaberrima PEQAAPLLCAGLTVYSPLKHFGLMS 179 Bambusaglaucescens PEQAAPLLCAGVTVYSPLKHFGLMA 179 Gossypiumhirsutum PEQVAPLLCAGVTVYSPLNHFGLMG 179 Hibiscus cannabinus PEQVAPLLCAGITVYSPLNHFGLTA 188 Linum album PEQAAPLLCAGITVYSPLNHFGLKQ 179 Ocimumbasilicum PEQAAPLLCAGVTVYSPLNHFGLKQ 179 Strigaasiatica PEQAAPLLCAGVTVYSPLNHFGLKQ 174 Heveabrasiliensis AEQAAPLLCAGLTVYSPLSHFGLNK 179 Citrus sinensis LEQAAPLLCAGVTVFSPLSHFGLKQ 179 Ricinuscommunis PEQAAPLLCAGVTVYSPLSHFGLKK 179 Vitisvinifera PEQAAPLLCAGVTVYSPLSHFGLKK 179 Populusnigra PEQAAPLLCAGLTVYSPLKHFGLKQ 179 Arabidopsis lyrata VEQAAPLLCAGVTVYSPLSHFGLMA 179 Arabidopsis thaliana VEQAAPLLCAGVTVYSPLSHFGLMA 180 Artemisia annua PEQVAPLLCAGVTVYSPLNYFGLKK 179 Araliacordata PEQAAPLLCAGVTVYSPLTHFGLKE 179 Nicotianatabacum PEQAAPLLCAGITVYSPLNHFGFKQ 179 Capsicum annuum PEQAAPLLCAGVTVYSPLNHFGFNQ 179 Medicago sativa PEQVAPLLCAGVTVYSPLSHFGLKT 180 Medicagotruncatula PEQVAPLLCAGVTVYSPLSHFGLKT 180 Glycine max PEQVAPLLCAGVTVYSPLVRFGLKE 179 Leucaenaleucocephala PEQVAPLLCAGVTVYSPLSHFGLRE 179 Eucalyptus globulus SEQAAPLMCAGVTVYSPLVRFGLKQ 179 Pinusradiata LEQAAPLLCAGVTVFSPMKHFAMTE 179 Piceaabies LEQAAPLLCAGVTVYSPMKHFGMTE 179 **.***:*.*:**:**: *.: Supplementary Fig. S2

4. bmr6-ref MGSLASERKVVGWAARDATGHLSPYTYTLRNTGPEDVVVKVLYCGICHTDIHQAKNHLGA 60 bmr6-3 MGSLASERKVVGWAARDATGHLSPYTYTLRNTGPEDVVVKVLYCGICHTDIHQAKNHLGA 60 BTx623 MGSLASERKVVGWAARDATGHLSPYTYTLRNTGPEDVVVKVLYCGICHTDIHQAKNHLGA 60 IS23253 MGSLASERKVVGWAARDATGHLSPYTYTLRNTGPEDVVVKVLYCGICHTDIHQAKNHLGA 60 IS23789 MGSLASERKVVGWAARDATGHLSPYTYTLRNTGPEDVVVKVLYCGICHTDIHQAKNHLGA 60 IS21549 MGSLASERKVVGWAARDATGHLSPYTYTLRNTGPEDVVVKVLYCGICHTDIHQAKNHLGA 60 ************************************************************ bmr6-ref SKYPMVPGHEVVGEVVEVGPEVSKYGVGDVVGVGVIVGCCRECSPCKANVEQYCNKKIWS 120 bmr6-3 SKYPMVPGHEVVGEVVEVGPEVSKYGVGDVVGVGVIVGCCRECSPCKANVEQYCNKKIWS 120 BTx623 SKYPMVPGHEVVGEVVEVGPEVSKYGVGDVVGVGVIVGCCRECSPCKANVEQYCNKKIWS 120 IS23253 SKYPMVPGHEVVGEVVEVGPEVSKYGVGDVVGVGVIVGCCRECSPCKANVEQYCNKKIWS 120 IS23789 SKYPMVPGHEVVGEVVEVGPEVSKYGVGDVVGVGVIVGCCRECSPCKANVEQYCNKKIWS 120 IS21549 SKYPMVPGHEVVGEVVEVGPEVSKYGVGDVVGVGVIVGCCRECSPCKANVEQYCNKKIWS 120 ************************************************************ bmr6-ref YNDVYTDGRPT-GGFASTMVVDQKFVVKIPAGLAPEQAAPLLCAGVTVYSPLKAFGLTAP 179 bmr6-3 YNDVYTDGRPTQGGFASTMVVDQKFVVKIPAGLAPEQAAPLLCAGVTVYSPLKAFGLTAP 180 BTx623 YNDVYTDGRPTQGGFASTMVVDQKFVVKIPAGLAPEQAAPLLCAGVTVYSPLKAFGLTAP 180 IS23253 YNDVYTDGRPTQGGFASTMVVDQKFVVKIPAGLAPEQAAPLLCAGVTVYSPLKAFGLTAP 180 IS23789 YNDVYTDGRPTQGGFASTMVVDQKFVVKIPAGLAPEQAAPLLCAGVTVYSPLKAFGLTAP 180 IS21549 YNDVYTDGRPTQGGFASTMVVDQKFVVKIPAGLAPEQAAPLLCVGVTVYSPLKAFGLTAP 180 *********** *******************************.**************** bmr6-ref GLRGGIVGLGGVGHMGVKVAKAMGHHVTVISSSSKKRAEAMDHLGADAYLVSTDAAAMAA 239 bmr6-3 GLRGGIVGLGSVGHMGVKVAKAMGHHVTVISSSSKKRAEAMDHLGADAYLVSTDAAAMAA 240 BTx623 GLRGGIVGLGGVGHMGVKVAKAMGHHVTVISSSSKKRAEAMDHLGADAYLVSTDAAAMAA 240 IS23253 GLRGGIVGLGRVGHMGVKVAKAMGHHVTVISSSSKKRAEAMDHLGADAYLVSTDAAAMAA 240 IS23789 GLRGGIVGLGRVGHMGVKVAKAMGHHVTVISSSSKKRAEAMDHLGADAYLVSTDAAAMAA 240 IS21549 GLRGGIVGLGGVGHMGVKVAKAMGHHVTVISSSSKKRAEAMDHLGADAYLVSTDAAAMAA 240 ********** ************************************************* bmr6-ref AADSLDYIIDTVPVHHPLEPYLSLLRLDGKHVLLGVIGEPLSFVSPMVMLGRKAITGSFI 299 bmr6-3 AADSLDYIIDTVPVHHPLEPYLSLLRLDGKHVLLGVIGEPLSFVSPMVMLGRKAITGSFI 300 BTx623 AADSLDYIIDTVPVHHPLEPYLSLLRLDGKHVLLGVIGEPLSFVSPMVMLGRKAITGSFI 300 IS23253 AADSLDYIIDTVPVHHPLEPYLSLLRLDGKHVLLGVIGEPLSFVSPMVMLGRKAITGSFI 300 IS23789 AADSLDYIIDTVPVHHPLEPYLSLLRLDGKHVLLGVIGEPLSFVSPMVMLGRKAITGSFI 300 IS21549 AADSLDYIIDTVPVHHPLEPYLSLLRLDGKHVLLGVIGEPLSFVSPMVMLGRKAITGSFI 300 ************************************************************ bmr6-ref GSIDETAEVLQFCVDKGLTSQIEVVKMGYVNEALERLERNDVRYRFVVDVAGSNVEEDAA 359 bmr6-3 GSIDETAEVLQFCVDKGLTSQIEVVKMGYVNEALERLERNDVRYRFVVDVAGSNVEEDAA 360 BTx623 GSIDETAEVLQFCVDKGLTSQIEVVKMGYVNEALERLERNDVRYRFVVDVAGSNVEEDAA 360 IS23253 GSIDETAEVLQFCVDKGLTSQIEVVKMGYVNEALERLERNDVRYRFVVDVAGSNVEEDAA 360 IS23789 GSIDETAEVLQFCVDKGLTSQIEVVKMGYVNEALERLERNDVRYRFVVDVAGSNVEEDAA 360 IS21549 GSIDETAEVLQFCVDKGLTSQIEVVKMGYVNEALERLERNDVRYRFVVDVAGSNVEEDAA 360 ************************************************************ bmr6-ref DAPSN- 364 bmr6-3 DAPSN- 365 BTx623 DAPSN- 365 IS23253 DAPSN- 365 IS23789 DAPSN- 365 IS21549 DAPSN- 365 ***** Supplementary Fig. S3