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Title: | An Efficient Agrobacterium-Mediated Genetic Transformation Method for Solanum betaceum Cav. Embryogenic Callus | Authors: | Cordeiro, Daniela Alves, Ana Ferraz, Ricardo Casimiro, Bruno Canhoto, Jorge Correia, Sandra |
Keywords: | antibiotic resistance; functional genomics; plant cell culture; somatic embryogenesis; tree tomato | Issue Date: | 2023 | Publisher: | MDPI | Project: | SFRH/BD/136925/2018 SFRH/BD/146485/2019 2021.04958.BD CFE-Centre for Functional Ecology-Science for People & the Planet’s Strategic Plan (UIDB/04004/2020) |
Serial title, monograph or event: | Plants | Volume: | 12 | Issue: | 5 | Abstract: | Somatic embryogenesis in Solanum betaceum (tamarillo) has proven to be an effective model system for studying morphogenesis, since optimized plant regeneration protocols are available, and embryogenic competent cell lines can be induced from different explants. Nevertheless, an efficient genetic transformation system for embryogenic callus (EC) has not yet been implemented for this species. Here, an optimized faster protocol of genetic transformation using Agrobacterium tumefaciens is described for EC. The sensitivity of EC to three antibiotics was determined, and kanamycin proved to be the best selective agent for tamarillo callus. Two Agrobacterium strains, EHA105 and LBA4404, both harboring the p35SGUSINT plasmid, carrying the reporter gene for -glucuronidase (gus) and the marker gene neomycin phosphotransferase (nptII), were used to test the efficiency of the process. To increase the success of the genetic transformation, a cold-shock treatment, coconut water, polyvinylpyrrolidone and an appropriate selection schedule based on antibiotic resistance were employed. The genetic transformation was evaluated by GUS assay and PCR-based techniques, and a 100% efficiency rate was confirmed in the kanamycin-resistant EC clumps. Genetic transformation with the EHA105 strain resulted in higher values for gus insertion in the genome. The protocol presented provides a useful tool for functional gene analysis and biotechnology approaches. | URI: | https://hdl.handle.net/10316/113791 | ISSN: | 2223-7747 | DOI: | 10.3390/plants12051202 | Rights: | openAccess |
Appears in Collections: | FCTUC Ciências da Vida - Artigos em Revistas Internacionais I&D CFE - Artigos em Revistas Internacionais |
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