Monitoring field releases of genetically modified sugar beets for persistence of transgenic plant DNA and horizontal gene transfer

Abstract

Field releases of transgenic rizomania-resistant sugar beet (<it>Beta vulgaris</it>) plants were accompanied by a study of the persistence of DNA from transgenic sugar beet litter in soil and of horizontal gene transfer of plant DNA to bacteria. The transgenic sugar beets contained the marker genes <it>npt</it>II and <it>bar</it> under the control of the bidirectional TR1/2 promoter conferring kanamycin (Km) and glufosinate ammonium resistance to the plant. Primer systems targeting the construct allowed the specific and sensitive detection of the transgenic DNA in soil. Soil samples were analyzed by cultivation of bacteria on nonselective and Km-selective media to determine the proportion of Km-resistant bacteria and to monitor the culturable fraction for incorporation of transgenic plant DNA. To detect the presence of transgenic DNA independently from cultivation, total soil DNA was extracted and amplified by PCR with three different primer sets specific for the transgenic DNA. Long-term persistence of transgenic DNA could be shown under field conditions (up to 2 years) and also in soil microcosms with introduced transgenic plant DNA. No construct-specific sequences were detected by dot blot hybridizations of bacterial isolates. The experimental limitations of detecting horizontal gene transfer from plants to bacteria under field conditions are discussed.