An Efficient Agrobacterium -Mediated Genetic Transformation Method for Solanum betaceum Cav. Embryogenic Callus

Somatic embryogenesis in (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 syste...

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Bibliographic Details
Published in:Plants (Basel) 2023-03, Vol.12 (5), p.1202
Main Authors: Cordeiro, Daniela, Alves, Ana, Ferraz, Ricardo, Casimiro, Bruno, Canhoto, Jorge, Correia, Sandra
Format: Article
Language:English
Subjects:
Agrobacterium
Agrobacterium tumefaciens
Antibiotic resistance
Antibiotics
Bacteria
Biotechnology
Callus
Cell lines
Clumps
Cold shock
Cold treatment
Comparative analysis
Embryonic growth stage
Environmental aspects
Explants
functional genomics
Genes
Genetic engineering
Genetic transformation
Genomes
Growth
Kanamycin
Morphogenesis
Neomycin
Neomycin phosphotransferase
Phosphotransferase
Physiological aspects
plant cell culture
Plant tissue culture
Polyvinylpyrrolidone
Protocol
Reporter gene
Solanum
Solanum betaceum
Somatic embryogenesis
Transformations
Transgenic plants
tree tomato
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Summary:Somatic embryogenesis in (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 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 strains, EHA105 and LBA4404, both harboring the p35SGUSINT plasmid, carrying the reporter gene for β-glucuronidase ( ) and the marker gene neomycin phosphotransferase ( ), 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 insertion in the genome. The protocol presented provides a useful tool for functional gene analysis and biotechnology approaches.
ISSN:2223-7747
2223-7747
DOI:10.3390/plants12051202