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Biochemical mechanism and molecular basis for ALS-inhibiting herbicide resistance in sugarbeet (Beta vulgaris) somatic cell selections

Published online by Cambridge University Press:  12 June 2017

Terry R. Wright
Affiliation:
Department of Crop and Soil Sciences, Michigan State University, East Lansing, MI 48824-1325
Newell F. Bascomb
Affiliation:
American Cyanamid, Agricultural Products Research Division, Princeton, NJ 08543-0400
Stephen F. Sturner
Affiliation:
American Cyanamid, Agricultural Products Research Division, Princeton, NJ 08543-0400
Donald Penner*
Affiliation:
Department of Crop and Soil Sciences, Michigan State University, East Lansing, MI 48824-1325
*
Corresponding author. [email protected]

Abstract

Three sugarbeet selections differing in cross-resistance to three classes of acetolactate synthase (ALS)-inhibiting herbicides have been developed using somatic cell selection. Sugarbeet selections resistant to imidazolinone herbicides, Sir-13 and 93R30B, do not metabolize [14C]-imazethapyr any faster or differently than sensitive, wild-type sugarbeets or a sulfonylurea-resistant/imidazolinone-sensitive selection, Sur. ALS specific activity from the three herbicide-resistant selections ranged from 73 to 93% of the wild-type enzyme extracts in the absence of herbicide, indicating enzyme overexpression was not a factor in resistance. Acetolactate synthase from Sir-13 plants showed a 40-fold resistance to imazethapyr but no resistance to chlorsulfuron or flumetsulam. Polymerase chain reaction amplification and sequencing of two regions of the ALS gene spanning all known sites for ALS-based herbicide resistance in plants indicated a single nucleotide change in the Sir-13 gene (G337 to A337) resulting in a deduced substitution of threonine for alanine at position 113 in the sugarbeet amino acid sequence. Sur ALS was not significantly resistant to imazethapyr, but was 1,000- and 50-fold resistant to chlorsulfuron and flumetsulam, respectively. Sur gene sequencing indicated a single nucleotide change (C562 to T562) resulting in a serine for proline substitution at position 188 of the ALS primary structure. The 93R30B nucleotide sequence indicated two mutations resulting in two deduced amino acid substitutions: threonine for alanine at position 113 plus serine for proline at position 188. The 93R30B double mutant incorporated the changes observed in each of the single mutants above and correlated with higher resistance levels to imazethapyr (> 1,000-fold), chlorsulfuron (4,300-fold), and flumetsulam (200-fold) at the ALS level than observed in either of the single mutants. 93R30B represents the first double mutant derived by a two-step selection process that incorporates two class-specific ALS-inhibitor resistance mutations to form a single broad cross-resistance trait. The interaction of the two altered amino acids is synergistic with respect to enzyme resistance vs. the resistance afforded by each of the individual mutations.

Type
Physiology, Chemistry, and Biochemistry
Copyright
Copyright © 1998 by the Weed Science Society of America 

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