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Transgenic broccoli plants were obtained by transformation with Agrobacterium tumefaciens carrying endochitinase, gus and nptII genes. The effects of the type and age of explant, Agrobacterium strain, and silver nitrate on transformation efficiency were examined. Cotyledonary-petioles from 5-day-old seedlings were successfully transformed with octopine strain LBA4404 or nopaline strain GV3101. No transformed shoots were obtained from 15-day-old cotyledonary-petioles or from hypocotyls at any developmental stage. Higher transformation rates were obtained with GV3101 (11-16%), compared to LBA4404 (1-6%). Improvement in transformation rates was also observed when explants were exposed to silver nitrate during the first 7 days of selection, but only when the LBA4404 strain was used. Use of silver nitrate on explants inoculated with GV3101 did not increase shoot differentiation. Selection for kanamycin resistance allowed differentiation of many escapes, but transgenic shoots were also recovered. Transgenic status of green regenerated plants was confirmed by NptII and PCR assays which showed a high correlation with root formation on selective medium. The efficient protocol developed for broccoli transformation is suggested for use with other Brassica oleracea varieties.

Palabras clave: Brassica oleracea var. italica, cotyledonary-petioles, nopaline, octopine, silver nitrate, transformation efficiency.
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Universidad Autónoma del Estado de México
Sistema de Información Científica Redalyc ®
Versión 3.0 | 2018
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