ISHS Acta Horticulturae 494: II International Symposium on Chestnut
GENETIC TRANSFORMATION OF EUROPEAN CHESTNUT (CASTANEA SATIVA MILL.) WITH GENES OF INTEREST |
| Authors: | R. da Costa Seabra, M.S. Pais |
| Keywords: | Transformation, chestnut, Castanea sativa Mill., Agrobacterium tumefaciens |
| Abstract:
Agrobacterium-mediated transformation of stem segments of micropropagated plantlets of Castanea sativa was performed with the LBA 4404 strain of Agrobacterium tumefaciens, harbouring the plasmid pGJ40 which contains a T-region with Neomicin Phosphotransferase (nptII), ß-1,3- glucanase (glu) and chitinase (chi) genes. Transgenic plants of Castanea sativa integrating nptII and glu genes were obtained. Different schemes of selection, as well as different aminoglycoside antibiotics, were tested for the screening of the nptII transgenics. Paromomycin proved to be better than kanamycin and geneticin for the selection of nptII transgenics. The transgenic plantlets, regenerated from stem segments, were multiplied in vitro. Although some plantlets kept the glu genes integrated after 2–3 subcultures in vitro, some others lost them. | |
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