Basta tolerance as a selectable and screening marker for transgenic plants of Norway spruce |
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Authors: | V Brukhin D Clapham M Elfstrand S von Arnold |
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Institution: | (1) Swedish University of Agricultural Sciences, Department of Forest Genetics, Box 7027, S-750 07 Uppsala, Sweden e-mail: David.Clapham@sgen.slu.se Fax: +46-18-673279, SE |
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Abstract: | 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.
Received: 21 October 1999 / Revision received: 24 January 2000 / Accepted: 24 January 2000 |
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Keywords: | Basta resistance bar gene Gold particle bombardment Picae abies Norway spruce Transformation |
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