ABSTRACT
Although it is generally accepted that plant in vitro culture response is influenced by the donor genotype, the genetic and molecular bases of this phenomenon are barely known. As a consequence, the optimization of tissue culture protocols is mainly empirically done. Researchers of the IGEAF studied the genetic basis of the in vitro regeneration of various plant species, including the tissue culture response of artificially induced barley mutants. One barley mutant, MC 169, carries a nuclear gene, recently described controlling the root growth in hydroponic cultivation. Under this condition, the roots of MC 169 mutant plants were longer than those of the original wild type line MC 182, a fact that was associated with a reduced ethylene biosynthesis. On the other hand, it is known that ethylene accumulation is inhibitory for in vitro regeneration of several plant species. In this study, we compared the in vitro culture response of mutant MC 169 with that of its mother line MC 182. The data about induction and regeneration of calli as well as those of habituated calli formation demonstrated that mutant MC 169 and its mother line MC 182 show a similar in vitro behaviour.
ABSTRACT
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 percent 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 expresion of the beta-glucuronidase enzyme, 42 percent 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.