ISHS Acta Horticulturae 596: VIII International Symposium on Pear
COMPARISON OF DECONTAMINATING TREATMENTS ON AGROBACTERIUM-INOCULATED INTERNODE EXPLANTS OF ‘BARTLETT’ PEAR |
| Authors: | P. Negri, L. Manzecchi |
| Keywords: | Pyrus communis, Agrobacterium tumefaciens, genetic transformation, lysozyme |
| Abstract:
In Agrobacterium-mediated genetic transformation experiments the antibiotics commonly added to culture media are often unable by themselves to completely overcome bacterial contamination of explants. After co-cultivation with A. tumefaciens C58C1 pGV3850, ‘Bartlett’ internodes were exposed to vacuum-infiltration with different filter-sterilized washes: either lysozyme (2 mg/ml) or plain water were employed at two pH levels (7, or lowered to 3 with HCl) and, only at pH 7, cefotaxime (500 mg/L). Residual contamination of the explants was assessed on samples, transferred to an antibiotic-free substrate for bacterial growth immediately after the washes (day 0) and after 15, 42, 92 and 133 days of incubation on a cefotaxime-containing plant tissue culture medium. Although none of the treatments by itself was able to eradicate the bacteria, all of them, combined with explant culture on the cefotaxime-containing substrate, assured a faster decontamination than the pH 7 water control. The low pH washes were the most effective, but had inhibitory effects on plant cell proliferation; in contrast, lysozyme at pH 7 provided good decontamination without adverse effects on tissue culture. | |
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