Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat Varieties
The development of a robust Agrobacterium-mediated transformation protocol for a recalcitrant species like durum wheat requires the identification and optimization of factors affecting T-DNA delivery and plant regeneration. The purpose of this research was to compare the behavior of diverse durum wh...
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MDPI AG
2016-10-01
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| Series: | Molecules |
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| Online Access: | http://www.mdpi.com/1420-3049/21/11/1440 |
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| author | Wilma Sabetta Cristina Crosatti Alexandra Soltész Valentina Di Rienzo Cinzia Montemurro |
| author_facet | Wilma Sabetta Cristina Crosatti Alexandra Soltész Valentina Di Rienzo Cinzia Montemurro |
| author_sort | Wilma Sabetta |
| collection | DOAJ |
| description | The development of a robust Agrobacterium-mediated transformation protocol for a recalcitrant species like durum wheat requires the identification and optimization of factors affecting T-DNA delivery and plant regeneration. The purpose of this research was to compare the behavior of diverse durum wheat genotypes during in vitro culture and Agrobacterium tumefaciens-mediated transformation, using immature embryos as explants. Apart from plant genotype, two of the main influencing factors for a successful genetic transformation have been examined here, i.e., auxin source (Dicamba and 2,4-D) and duration of the pre-culture period (one, seven and 21 days). The addition of Dicamba to the media in combination with seven days pre-cultivation resulted in a general enhancement of T-DNA delivery for most of the analyzed cultivars, as revealed by β-glucuronidase (GUS) histochemical assay. Although all genotypes were able to produce calli, significant differences were detected in regeneration and transformation efficiencies, since only two (Karalis and Neolatino) out of 14 cultivars produced fertile transgenic plants. The estimated transformation efficiencies were 6.25% and 1.66% for Karalis and Neolatino, respectively, and χ2 analysis revealed the stable integration and segregation of the gus transgene in T1 and T2 progenies. This research has demonstrated that, among the influencing factors, genotype and auxin type play the most important role in the success of durum wheat transformation. |
| first_indexed | 2024-04-12T12:36:51Z |
| format | Article |
| id | doaj.art-eef8cb601f17488d8ade73821f00d52a |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-04-12T12:36:51Z |
| publishDate | 2016-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-eef8cb601f17488d8ade73821f00d52a2022-12-22T03:32:52ZengMDPI AGMolecules1420-30492016-10-012111144010.3390/molecules21111440molecules21111440Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat VarietiesWilma Sabetta0Cristina Crosatti1Alexandra Soltész2Valentina Di Rienzo3Cinzia Montemurro4Sinagri S.r.l., Spin-off, University of Bari, Via Amendola 165/A, Bari 70126, ItalyCouncil for Agricultural Research and Economics, Genomics Research Centre, Via S.Protaso 302, Fiorenzuola d’Arda (PC) 29017, ItalyAgricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, Brunszvik u. 2., Martonvásár H-2462, HungarySinagri S.r.l., Spin-off, University of Bari, Via Amendola 165/A, Bari 70126, ItalySinagri S.r.l., Spin-off, University of Bari, Via Amendola 165/A, Bari 70126, ItalyThe development of a robust Agrobacterium-mediated transformation protocol for a recalcitrant species like durum wheat requires the identification and optimization of factors affecting T-DNA delivery and plant regeneration. The purpose of this research was to compare the behavior of diverse durum wheat genotypes during in vitro culture and Agrobacterium tumefaciens-mediated transformation, using immature embryos as explants. Apart from plant genotype, two of the main influencing factors for a successful genetic transformation have been examined here, i.e., auxin source (Dicamba and 2,4-D) and duration of the pre-culture period (one, seven and 21 days). The addition of Dicamba to the media in combination with seven days pre-cultivation resulted in a general enhancement of T-DNA delivery for most of the analyzed cultivars, as revealed by β-glucuronidase (GUS) histochemical assay. Although all genotypes were able to produce calli, significant differences were detected in regeneration and transformation efficiencies, since only two (Karalis and Neolatino) out of 14 cultivars produced fertile transgenic plants. The estimated transformation efficiencies were 6.25% and 1.66% for Karalis and Neolatino, respectively, and χ2 analysis revealed the stable integration and segregation of the gus transgene in T1 and T2 progenies. This research has demonstrated that, among the influencing factors, genotype and auxin type play the most important role in the success of durum wheat transformation.http://www.mdpi.com/1420-3049/21/11/1440durum wheatAgrobacteriumDicamba2,4-Dpre-cultureimmature embryos |
| spellingShingle | Wilma Sabetta Cristina Crosatti Alexandra Soltész Valentina Di Rienzo Cinzia Montemurro Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat Varieties Molecules durum wheat Agrobacterium Dicamba 2,4-D pre-culture immature embryos |
| title | Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat Varieties |
| title_full | Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat Varieties |
| title_fullStr | Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat Varieties |
| title_full_unstemmed | Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat Varieties |
| title_short | Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat Varieties |
| title_sort | screening auxin response in vitro culture aptitude and susceptibility to agrobacterium mediated transformation of italian commercial durum wheat varieties |
| topic | durum wheat Agrobacterium Dicamba 2,4-D pre-culture immature embryos |
| url | http://www.mdpi.com/1420-3049/21/11/1440 |
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