2 research outputs found
Towards genetic transformation of local onion varieties
The aim of this work was to explore the possibility of obtaining
transgenic plants of onion varieties cultivated in Argentina , starting
from calli induced from mature zygotic embryos, using two strains of
Agrobacterium tumefaciens as transfection vectors. Mature embryos
from three varieties of 'Valenciana' onion, Torrentina, Cobriza INTA
and Grano de oro were in vitro cultivated for callus induction. After
three to four months an average of 57.4% success for the three
varieties was reached. Transformation was carried out with Agl1 or LBA
4404 A . tumefaciens strains, both carrying a binary vector containing
the marker gene gus a and the selection gene nptII. Selection was done
in callus induction medium containing 10 mgL-1 geneticin during three
subcultures. At the end of the selection period, 342 portions of calli
were recovered and transferred to regeneration medium. Of the selected
calli evaluated by the expresi\uf3n of the \u3b2-glucuronidase
enzyme, 42% presented extensive blue areas or were completely blue. At
the end of the first subculture in the regeneration medium, 54 calli
were considered potentially organogenic because of the green areas
observed. At the end of the wole regeneration period, just one normal
plant was obtained, that was negative to PCR analysis using specific
primers for gus a and nptII. All selected calli came from the
Torrentina variety and the highest quantity of them were transformed
with the strain LBA 4404
Towards genetic transformation of local onion varieties
The aim of this work was to explore the possibility of obtaining
transgenic plants of onion varieties cultivated in Argentina, starting
from calli induced from mature zygotic embryos, using two strains of
Agrobacterium tumefaciens as transfection vectors. Mature embryos
from three varieties of 'Valenciana' onion, Torrentina, Cobriza INTA
and Grano de oro were in vitro cultivated for callus induction. After
three to four months an average of 57.4% success for the three
varieties was reached. Transformation was carried out with Agl1 or LBA
4404 A. tumefaciens strains, both carrying a binary vector containing
the marker gene gus a and the selection gene nptII. Selection was done
in callus induction medium containing 10 mgL-1 geneticin during three
subcultures. At the end of the selection period, 342 portions of calli
were recovered and transferred to regeneration medium. Of the selected
calli evaluated by the expresi\uf3n of the \u3b2-glucuronidase
enzyme, 42% presented extensive blue areas or were completely blue. At
the end of the first subculture in the regeneration medium, 54 calli
were considered potentially organogenic because of the green areas
observed. At the end of the wole regeneration period, just one normal
plant was obtained, that was negative to PCR analysis using specific
primers for gus a and nptII. All selected calli came from the
Torrentina variety and the highest quantity of them were transformed
with the strain LBA 4404