Transformation protocol.  Transformation was achieved via direct gene transfer, therefore no vector agent was used .  AgrEvo has requested that the details of the methodology be maintained by APHIS as confidential business information. Tissues were transformed with a fragment of DNA which contained the bar gene and from the plasmid, then used this fragment to transform the rice callus tissue.<br>Genes introduced to develop LLRICE06 and LLRICE62.  Transformation events LLRICE06 and LLRICE62 were developed by introducing a single gene,  the bar gene, into the varieties M202 and Bengal.  The bar gene was derived from the soil-borne bacterium Streptomyces hygroscopicus, an organism which is not a plant pest.  The bar gene encodes  phosphinothricin-N-acetyltransferase (PAT), an enzyme which inactivates the herbicide glufosinate ammonium.  The bar gene was engineered into the rice with noncoding DNA regulatory sequences attached to the coding region of bar.  The regulatory sequences are the 35S promoter and terminator sequences derived from cauliflower mosaic virus.   Neither the bar gene nor its associated noncoding DNA regulatory sequences cause plant disease or pose any plant pest risk.<br>Expression of genes introduced into lines LLRICE06 and LLRICE62.  The rice transformation events LLRICE06 and LLRICE62 expressed the PAT protein encoded by the bar gene cassette.  The PAT protein expressed in these plants was characterized and found to be equivalent to the previously characterized reference standards.