<p style="text-align: justify;"><b>The Introduced DNA</b></p>

<p style="text-align: justify;">Southern Blot and segregation analyses demonstrated the stable integration of the inserted DNA at a single genetic locus in both MS1 and RF2. The <i>barnase</i>, <i>bar</i> and <i>neo</i> genes were integrated into MS1 and similarly, the <i>barstar</i>, <i>bar</i> and <i>neo</i> genes were integrated in RF2. Additional Souther blot analyses demonstrated that there was no incorporation of any plasmid sequences outside of the T-DNA region.</p>

<p style="text-align: justify;"><b>Genetic Stability of the Introduced Trait</b></p>

<p style="text-align: justify;">MS1 and RF2 are several generations removed from the original transformants. Comparisons to the original transformants, in addition to data from several generations of backcrossing demonstrated the stable inheritance of the novel traits in lines MS1 and RF2. Segregation analysis confirmed the data from Southern blots and indicated that the novel traits in both transformation events segregated as a single locus according to the rules of Mendelian inheritance.</p>

<p style="text-align: justify;"><b>Expressed Material</b></p>

<p style="text-align: justify;">Transcription of the <i>bar</i> gene was detected in both leaf and flower bud tissue by Northern blot analysis and was estimated to be 0.8-1.6 pg/mg total RNA and 0.1-0.2 pg/mg total RNA, respectively. Similar analyses performed on canola line RF2 demonstrated that the <i>barstar</i> gene was only transcribed in flower bud tissue at a level of 0.4-0.8 pg/mg total RNA. Both genes were expressed only during the early stages of anther development in the tapetum cell layer of the anther.</p>

<p style="text-align: justify;">The NPTII encoding gene was linked to a weak constitutive promoter and expression of this enzyme was not detected in pollen samples or unprocessed honey.</p>
