Abstract：
:An efficient in vitro regeneration system using epicotyl segments was developed and then used for optimizing genetic transformation of the Tunisian 'Maltese half-blood' (Citrus sinensis) variety using phosphinothricin (PPT) resistance as a selectable marker. The maximum regeneration efficiency was achieved after incubating epicotyl explants (excised in an oblique manner) in MT culture media containing BAP (4 mg/l) and IAA (0.3 mg/l) hormonal combination in the dark for 3 weeks before their transfer to light. Data from the genetic transformation assays indicated that the highest number of regenerated-transformants was reached when the selection phase was conducted in MT culture media containing PPT (0.25 mg/l) and Carbenicillin (500 mg/l) for 3 weeks in the dark followed by 8 weeks of light. After that, transformed buds were maintained for eight additional weeks in the same culture media but with reduced PPT concentration (0.125 mg/l) before decreasing Carbenicillin dose (250 mg/l) at the second half of this last incubation period which allowed both a good shoot proliferation and an optimal rooting efficiency. Based on molecular analyses, the transgenicity of 21.42% of the regenerated vitroplants was confirmed. The developed regeneration and transformation procedures of the elite 'Maltese half-blood' variety can be used for orchard renewal as well as for functional studies and genome editing purposes to develop new cultivars with the desired genetic traits.
journal_name
3 Biotechjournal_title
3 Biotechauthors
Jardak R,Boubakri H,Zemni H,Gandoura S,Mejri S,Mliki A,Ghorbel Adoi
10.1007/s13205-020-2097-6subject
Has Abstractpub_date
2020-03-01 00:00:00pages
99issue
3eissn
2190-572Xissn
2190-5738pii
2097journal_volume
10pub_type
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