Transfer of a mutant plant glutamate 1-semialdehyde aminotransferase gene from the nuclear to the plastid genome confers gabaculine resistance in tobacco
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New selection systems are required to extend plastid transformation to a more significant number of plant species. After demonstrating that a bacterial mutant glutamate 1-semialdehyde aminotransferase (GSA) gene is functional in tobacco plastids and confers resistance to the phytotoxin gabaculine, in this work, we transformed the tobacco plastome with a mutated GSA gene derived from alfalfa, along with the aadA selectable marker gene conferring spectinomycin resistance. Selection exploiting gabaculine was not effective to directly regenerate transplastomic events, but some events regenerated using conventional spectinomycin selection acquired gabaculine resistance during the second or third regeneration rounds with gabaculine selection. Gabaculine selection with our plant mutant GSA gene can be further investigated for plastid transformation of species that are not transformable with the aadA-spectinomycin selection system.
Resistance to the phytotoxin gabaculine was obtained in tobacco by introducing a mutant plant gene in the plastid genome; it may be implemented as a tool in plastid genetic transformation.
KeywordsChloroplast In vitro selection Gabaculine Transgenic plants Transplastomic plants
This work was funded by the Department of Agricultural Food and Environmental Sciences of the University of Perugia, project: “Ricerca di base 2015: Sviluppo di un nuovo gene marcatore selezionabile di origine vegetale per la trasformazione del genoma plastidiale”. This work has been conducted under the auspices of Consorzio Interuniversitario per le Biotecnologie (CIB).
DR and MB designed the study, MB, FDM, AP, MM, TG carried out the experiments; DR and MB wrote the manuscript; all authors revised the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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