(277) Behavior of Glomus intraradices in Agave Roots Transformed with Agrobacterium rhizogenes

Abstract

The present work is the first report in vitro on root induction of Agave salmiana Otto, using Agrobacterium rhizogenes. Several concentrations of bacteria and acetosyringone were used, and different inoculation sites were tested, such as leaves, shaft, and root. Incubation time in darkness was 6 days. The transformed adventitious roots appeared 25 days after inoculation. The best treatment was when the shaft was inoculated with: 1 × 108 bacteria/mL and 100 μm acetosyringone; in this treatment, induction of transformed roots was 57.5% in the inoculated sites. The activity and presence of the foreign genes in the transformed roots of A. salmianawere verified as follows: 1) histochemical staining for GUS activity was determined in 80% of the tested root; and 2) molecular analysis via PCR was made to verify the presence of nptII gene and rol B gene (both were present in 60% of the tested root). This is the first report of the arbuscular mycorrhizal colonization on wild roots and transformed roots of Agavewith Glomus intraradicesSchenck and Smith. The result of the monoxenic culture was as follows: mother spore germinated 5 days; the colonization of the transformed roots was 70%. Then we proceeded to the recovery of daughter spores, in which we obtained an average 300 daughter spores per petri dish, 6 months after inoculation.