Basta tolerance as a selectable and screening marker for transgenic plants of Norway spruce

The bar gene conferring resistance to the herbicide Basta (containing phosphinothricin) was transferred to embryogenic cultures of Picea abies by particle bombardment and transformants were selected on Basta medium. In total, 83 9-month-old transgenic plants of Picea abies from six transformed sublines were analysed for continued tolerance to Basta. PCR analysis showed that the bar gene was present in all transformed plants but not in the control plants. Northern blot analysis showed differences in expression level among plants from the same subline as well as among sublines. A simple biotest for screening for Basta tolerance based on the colour change of detached needles induced by Basta was developed. The tolerance to Basta varied among the plants from different sublines. Needles from four of the sublines were resistant to 100 mg l^-1 phosphinothricin, a concentration inducing yellowing in control needles, while plants from the other two sublines were on average two to four times as resistant as untransformed control plants. The biotest enables rapid semi-quantitative monitoring for continued transgene expression in long-lived tree species.