Production of transgenic Eucalyptus grandis x E. urophylla using the sonication-assisted Agrobacterium transformation (SAAT) system

Functional Plant Biology : FPB
Esteban Roberto GonzálezCarlos Alberto Labate

Abstract

A method for genetic transformation of germinating seeds and seedlings of Eucalyptus grandis × E. urophyllais described using the sonication-assisted Agrobacterium-mediated transformation (SAAT) system. Seeds germinated for 2 d, and 15-d-old seedlings, sonicated for 30 s, had the highest percentage of β-glucuronidase (GUS) transient expression (21.7 and 37.4%, respectively). Pre-sonication greatly enhanced the efficiency of transformation. The differential transformation of tissues was also investigated, with seeds imbibed for 2 d having over 90% of the blue sectors localised in cotyledons and in the intersection of the hypocotyls and roots, whereas in 5-d-old seedlings, 70% of GUS activity was detected in cotyledons. However, 15-17-d-old seedlings had around 60% of transformed sectors localised in the first pair of leaves. The efficiency of the method was also assessed using a chimeric construct containing the Lhcb1*2 gene of the 28 kDa chlorophyll a/b binding pea protein from the LHCII antenna. Four stable transformants were confirmed by genomic blotting.

Citations

Aug 24, 2004·Plant Cell Reports·Antanas V SpokeviciusGerd Bossinger
Mar 1, 2011·Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology·V Girijashankar
Jan 9, 2008·Antonie van Leeuwenhoek·Fernando D AndreoteWelington L Araújo
Sep 22, 2005·Genetica·Fiona S PokeJames B Reid

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