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sunTILL: a TILLING resource for gene function analysis in sunflower

BACKGROUND: Cultivated sunflower (Helianthus annus L.) is a globally important oilseed crop, subjected to intensive genetic and genomic studies. Although classical mutagenesis has successfully been applied to Helianthus genus in the past, we have developed the first sunflower TILLING resource. RESUL...

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Autores principales: Sabetta, Wilma, Alba, Vittorio, Blanco, Antonio, Montemurro, Cinzia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3169506/
https://www.ncbi.nlm.nih.gov/pubmed/21718494
http://dx.doi.org/10.1186/1746-4811-7-20
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author Sabetta, Wilma
Alba, Vittorio
Blanco, Antonio
Montemurro, Cinzia
author_facet Sabetta, Wilma
Alba, Vittorio
Blanco, Antonio
Montemurro, Cinzia
author_sort Sabetta, Wilma
collection PubMed
description BACKGROUND: Cultivated sunflower (Helianthus annus L.) is a globally important oilseed crop, subjected to intensive genetic and genomic studies. Although classical mutagenesis has successfully been applied to Helianthus genus in the past, we have developed the first sunflower TILLING resource. RESULTS: To balance the maximum mutation density with an acceptable plant survival rate, a 'kill curve' analysis was first conducted with different ethylmethanesulfonate (EMS) dosages and different exposure times. According to the germination rate, a treatment with 0.7% EMS for 6 h was chosen. An M(2 )progeny of 3,651 fertile plants was obtained. Totally, 4.79% of the whole population showed clear aberrant phenotypes. A microsatellite analysis on a representative sample of the original seed stock and mutant lines confirmed the uniformity of the genetic background of plant material. The TILLING procedure was successfully applied to sunflower genome, initially by a CelI-nuclease mismatch cleavage assay coupled with a DNA-pooling level test. To investigate the efficiency of the mutagenic treatment, a pilot screening was carried out on 1,152 M(2) lines focusing on four genes, three involved in the fatty acid biosynthetic pathway and one for downy mildew resistance. A total of 9 mutant lines were identified and confirmed by sequencing; thereby, the estimated overall mutation frequency for the pilot assay resulted to be 1/475 kb. CONCLUSION: A first TILLING population for a high throughput identification of EMS-induced point mutations in sunflower genome has been successfully obtained. This represents a powerful tool to a better understanding of gene function in sunflower.
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spelling pubmed-31695062011-09-09 sunTILL: a TILLING resource for gene function analysis in sunflower Sabetta, Wilma Alba, Vittorio Blanco, Antonio Montemurro, Cinzia Plant Methods Research BACKGROUND: Cultivated sunflower (Helianthus annus L.) is a globally important oilseed crop, subjected to intensive genetic and genomic studies. Although classical mutagenesis has successfully been applied to Helianthus genus in the past, we have developed the first sunflower TILLING resource. RESULTS: To balance the maximum mutation density with an acceptable plant survival rate, a 'kill curve' analysis was first conducted with different ethylmethanesulfonate (EMS) dosages and different exposure times. According to the germination rate, a treatment with 0.7% EMS for 6 h was chosen. An M(2 )progeny of 3,651 fertile plants was obtained. Totally, 4.79% of the whole population showed clear aberrant phenotypes. A microsatellite analysis on a representative sample of the original seed stock and mutant lines confirmed the uniformity of the genetic background of plant material. The TILLING procedure was successfully applied to sunflower genome, initially by a CelI-nuclease mismatch cleavage assay coupled with a DNA-pooling level test. To investigate the efficiency of the mutagenic treatment, a pilot screening was carried out on 1,152 M(2) lines focusing on four genes, three involved in the fatty acid biosynthetic pathway and one for downy mildew resistance. A total of 9 mutant lines were identified and confirmed by sequencing; thereby, the estimated overall mutation frequency for the pilot assay resulted to be 1/475 kb. CONCLUSION: A first TILLING population for a high throughput identification of EMS-induced point mutations in sunflower genome has been successfully obtained. This represents a powerful tool to a better understanding of gene function in sunflower. BioMed Central 2011-06-30 /pmc/articles/PMC3169506/ /pubmed/21718494 http://dx.doi.org/10.1186/1746-4811-7-20 Text en Copyright ©2011 Sabetta et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Sabetta, Wilma
Alba, Vittorio
Blanco, Antonio
Montemurro, Cinzia
sunTILL: a TILLING resource for gene function analysis in sunflower
title sunTILL: a TILLING resource for gene function analysis in sunflower
title_full sunTILL: a TILLING resource for gene function analysis in sunflower
title_fullStr sunTILL: a TILLING resource for gene function analysis in sunflower
title_full_unstemmed sunTILL: a TILLING resource for gene function analysis in sunflower
title_short sunTILL: a TILLING resource for gene function analysis in sunflower
title_sort suntill: a tilling resource for gene function analysis in sunflower
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3169506/
https://www.ncbi.nlm.nih.gov/pubmed/21718494
http://dx.doi.org/10.1186/1746-4811-7-20
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