ISHS Acta Horticulturae 596: VIII International Symposium on Pear
FACTORS AFFECTING AGROBACTERIUM-MEDIATED TRANSFORMATION OF WILD PEAR (PYRUS COMMUNIS, VAR PYRASTER) |
| Authors: | E. Caboni, P. Lauri, B. Watillon |
| Keywords: | Agrobacterium tumefaciens, bacterial strain, gene transfer, in vitro culture, leaf explant, shoot regeneration, shoot vegetative apex. |
| Abstract:
Wounded leaves of in vitro micropropagated shoots of a wild pear (Pyrus communis, var Pyraster) genotypes were infected with two disarmed strains, EHA105 or C58C1 both harbouring the plasmid pBIN19GUS-Int carrying nptII and uidA genes or a plasmid having the uidA sequence substituted with a gene sequence of lactoferricin B with reported anctibacterial activity. Beside the strain used, the effect of the following factors on transformation efficiency (GUS response) and/or regeneration ability was also evaluated: bacterial culture dilution, pre-induction treatment, co-cultivation period length, time and modality of kanamicin application for selection of transgenics. Among these factors, some, such as length of the induction treatment and of the co-cultivation period and time of application of antibiotic selection were showed to be critical in increasing transformation efficiency. Some putative transgenic shoots, obtained with both the bacterial strains carrying sequence of lactoferricin B, were regenerated and multiplied for several sub-cultures on kanamycin. The molecular analyses of these clones are now in progress | |
Download Adobe Acrobat Reader (free software to read PDF files) | |
