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Improved genetic transformation protocol for cork oak (Quercus suber L.)

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Abstract

An optimized protocol for Agrobacterium tumefaciens-mediated transformation of mature Quercus suber L. embryogenic masses is reported. In this work several variables were tested. Plant genotype, explant type and time elapsed between the last subculture and inoculation, i.e. the explant preculture period, were found to be very important. Interaction between inoculum density and cocultivation period influenced the transformation efficiency as well. A transformation efficiency (i.e. percentage of the inoculated explants that yielded independent transgenic embryogenic lines) of up to 43% was obtained, greatly improving the previously described method for plant transformation of adult-selected cork oak. It was also shown that this protocol could be applied to various genotypes.

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Abbreviations

GE:

number of GUS spots per inoculated explant three weeks after the inoculation

GUS:

β-glucuronidase enzyme

GUS+ :

percentage of Kan+ lines positive for β-glucuronidase

IG:

percentage of inoculated explants showing GUS spots three weeks after the inoculation

Kan+ :

percentage of inoculated explants producing kanamycin-resistant embryos after 4 months of culture on selective medium

MSSH:

expression-proliferation medium

NptII:

neomycin phosphotransferase gene II

OD600 nm :

optical density of the bacterial cultures at 600 nm

GusA:

β-glucuronidase gene

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Acknowledgements

Authors thank Dr. M. Toribio for kindly providing the cork oak embryogenic lines and M. Cortizo and M. González for revising the English language. R. Álvarez was supported by a FICYT research fellowship funded by “Gobierno del Principado de Asturias, Spain”.

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  1. Departamento de Biología de Organismos y Sistemas, Universidad de Oviedo, C/. Catedrático Rodrigo Uría s/n, Oviedo, 33011, Spain

    Rubén Álvarez & Ricardo Javier Ordás

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  1. Rubén Álvarez
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  2. Ricardo Javier Ordás
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Correspondence to Ricardo Javier Ordás.

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Álvarez, R., Ordás, R.J. Improved genetic transformation protocol for cork oak (Quercus suber L.). Plant Cell Tiss Organ Cult 91, 45–52 (2007). https://doi.org/10.1007/s11240-007-9276-6

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