NDLI: Studies on genetic transformation of olive (Olea europaea L.) somatic embryos: I. Evaluation of different aminoglycoside antibiotics for nptII selection; II. Transient transformation via particle bombardment

Content Provider	Springer Nature Link
Author	Pérez Barranco, Gema Torreblanca, Rocío Padilla, Isabel M. G. Sánchez Romero, Carolina Pliego Alfaro, Fernando Mercado, José A.
Copyright Year	2009
Abstract	As a first step to the establishment of a genetic transformation protocol for olive somatic embryos obtained from the seeds of cv. ‘Picual’, the efficiencies of different aminoglycoside antibiotics as selective agents to be used with the nptII marker gene, and the particle bombardment technique for transient transformation have been evaluated. Among the three antibiotics tested, paromomycin and kanamycin showed a similar inhibitory effect and, at 200 mg l$^{−1}$, both of them impaired callus growth after 8 weeks of culture. However, when isolated embryos were cultured in the presence of these antibiotics, a 20% of the embryos still remained viable at 400 mg l$^{−1}$. Neomycin was discarded as a selective agent since it showed only a moderate toxic effect. Contrary to solid medium, when olive callus was cultured in liquid medium supplemented with different paromomycin concentrations for 3 weeks, the callus growth was impaired at the lowest antibiotic concentration, 3 mg l$^{−1}$. Best conditions for transient transformation of olive callus using PDS-1000/He system were a 6 cm target distance and a 900 psi bombardment pressure. pCGU∆1 plasmid, containing the gus gene under the control of sunflower ubiquitin promoter yielded a significantly higher number of gus expression areas per bombarded explant than pGUSINT or pJGUS5 plasmids, where the gus gene is driven by CaMV35S promoter or CaMV35S with enhancer, respectively. Almost 45% of bombarded explants showed gus expression 12 weeks after bombardment.
Starting Page	243
Ending Page	251
Page Count	9
File Format	PDF
ISSN	01676857
Journal	Plant Cell, Tissue and Organ Culture (PCTOC)
Volume Number	97
Issue Number	3
e-ISSN	15735044
Language	English
Publisher	Springer Netherlands
Publisher Date	2009-03-18
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Biolistics Genetic transformation Olive Particle bombardment Selectable marker Somatic embryos Plant Pathology Plant Genetics & Genomics Plant Physiology Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Horticulture

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