INVESTIGADORES
LUNA claudia veronica
artículos
Título:
Identification and control of bacterial contaminants from Ilex dumosa nodal segments culture in a temporal immersion bioreactor system using 16S rDNA analysis
Autor/es:
LUNA C,; COLLAVINO M.; MROGINSKI L.; SANSBERRO P.
Revista:
PLANT CELL TISSUE AND ORGAN CULTURE
Editorial:
Springer
Referencias:
Lugar: The Netherlands; Año: 2008 vol. 95 p. 13 - 19
ISSN:
0167-6857
Resumen:
A protocol for the production of transgenic plants has been developed for Lotus tenuis via Agrobacterium  mediated transformation of leaf segments. The explants were co-cultivated (for three days) with A. tumefaciens strain harboured the binary vector either pBi RD29A: oat arginine decarboxylase (ADC) or pBi RD29A:GUS, carrying the neomycin phosphotransferase II (nptII) gene in the T-DNA region. Following co-cultivation, the explants were cultured in the regeneration medium (MS plus naphtalenacetic acid and benzyladenine) containing kanamycin (30 ìg ml-1) and cefotaxime (400 ìg ml-1) for 45 days. The explants were subcultured several times (each 2 weeks) in order to maintain the selection pressure during the entire period.  Forty percent of the inoculated explants with pBiRD29:ADC strain  showed 8-10 adventitious shoots per responsive explants through a direct system of regeneration.  Likewise, sixty nine percent of inoculated explants with pBi RD29A:GUS strain produced 13-15 adventitious shoots per responsive explants. Selected transgenic lines were identified by polymerase chain reaction and Southern blot analysis. Three ADC transgenic lines were obtained from 30 infected explants. In the case of GUS, 29 transgenic lines were obtained from 160 analyzed explants. These results correspond to a transformation efficiency of 10 and 18.1%, respectively. More than 90% of the in vitro plantlets could be successfully transferred to soil. All of the transgenic plants obtained in the present study exhibited the same phenotype as the untransformed control under non-stressed condition and the stability of the introduced gene in the cloned materials was established.