title: Screening Auxin Response, In Vitro Culture Aptitude and Susceptibility to Agrobacterium-Mediated Transformation of Italian Commercial Durum Wheat Varieties.
abstract: The development of a robustAgrobacterium-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 andAgrobacterium 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 thegustransgene 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.
authors: Sinagri S.r.l., Spin-off, University of Bari, Via Amendola 165/A, Bari 70126, Italy. wilma.sabetta@libero.it.
authors: Wilma
authors: W
authors: Sabetta
authors: Council for Agricultural Research and Economics, Genomics Research Centre, Via S.Protaso 302, Fiorenzuola d'Arda (PC) 29017, Italy. cristina.crosatti@crea.gov.it.
authors: Cristina
authors: C
authors: Crosatti
authors: Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, Brunszvik u. 2., Martonvásár H-2462, Hungary. soltesz.alexandra@agrar.mta.hu.
authors: Alexandra
authors: A
authors: Soltész
authors: Sinagri S.r.l., Spin-off, University of Bari, Via Amendola 165/A, Bari 70126, Italy. valentina.dirienzo@gmail.com.
authors: Valentina
authors: V
authors: Di Rienzo
authors: Sinagri S.r.l., Spin-off, University of Bari, Via Amendola 165/A, Bari 70126, Italy. cinzia.montemurro@uniba.it.|Department of Soil, Plant and Food Sciences, University of Bari, Via Amendola 165/A, Bari 70126, Italy. cinzia.montemurro@uniba.it.
authors: Cinzia
authors: C
authors: Montemurro
mesh_terms: D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology; D016960:Agrobacterium tumefaciens / Q000502:physiology*; D004269:DNA, Bacterial / Q000235:genetics*; D003996:Dicamba / Q000494:pharmacology; D005838:Genotype; D007210:Indoleacetic Acids / Q000494:pharmacology*; D030821:Plants, Genetically Modified / Q000254:growth & development / Q000382:microbiology; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic; D014908:Triticum / Q000235:genetics* / Q000254:growth & development / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial; D007210:Indoleacetic Acids; C036483:T-DNA; D015084:2,4-Dichlorophenoxyacetic Acid; D003996:Dicamba
keywords: 2,4-D; Agrobacterium; Dicamba; durum wheat; immature embryos; pre-culture