Potential use of theaux2 gene fromAgrobacterium rhizogenes as a conditional negative marker in transgenic cabbage

An originalAgrobacterium tumefaciens-mediated transformation procedure, based on the actions of both wild type and disarmed bacterial strains, was developed. Theaux2 gene ofA. rhizogenes was introduced into a rapid-cycling genotype of cabbage (Brassica oleracea L.). Theaux2 gene product converts naphthalene acetamide into the auxin naphthalene acetic acid. Expression of this gene in the transgenic progeny grownin vitro led to an altered root phenotype. On a medium supplemented with napthalene acetamide (NAM), two of the three analysed progenies were characterized by the formation of callus instead of roots, whereas on a NAM-free medium all the plantlets from these progenies presented a normal phenotype. Expression of theaux2 gene was also assessed under horticultural conditions by sowing seeds in sand and watering them with a nutritive solution supplemented with NAM. Under these conditions, NAM inhibited the formation of a root system in transgenic plantlets and induced the death of the transgenic plantlets three to four weeks after germination. Thus,aux2 acts as a lethal conditional marker which could be used in negative selection of cabbage. Potential utilization of theaux2 gene to screen spontaneous androgenetic plants in order to transfer cytoplasmic male sterility in a single generation is discussed.