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An in vivo Acetolactate Synthase Assay

Published online by Cambridge University Press:  12 June 2017

David M. Simpson ,
Edward W. Stoller  and
Loyd M. Wax
David M. Simpson
Affiliation:
Dept. of Agronomy, Univ. of Illinois
Edward W. Stoller
Affiliation:
USDA-ARS, Urbana, IL
Loyd M. Wax
Affiliation:
USDA-ARS, Urbana, IL
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Abstract

A method was developed and tested for in vivo assay of acetolactate synthase (ALS). The method used foliar application of 1,1-cyclopropanedicarboxylic acid (CPCA) to inhibit ketol-acid reductoisomerase, the enzyme immediately following ALS in biosynthesis of branched-chain amino acids, thereby causing accumulation of acetolactate. Since the amount of acetolactate accumulation is a function of carbon flux through ALS, quantification of acetolactate accumulation determined ALS activity. Accumulation of acetolactate in soybean leaves resulted from CPCA rates as low as 15 g/ha and occurred within 1.5 h. Accumulation rates in soybean leaflets declined with leaf age from 84 μg/h/g tissue at 3 d to 17 μg/h/g tissue at 7 d. Foliar application of CPCA also caused acetolactate accumulation in corn, grain sorghum, velvetleaf, common cocklebur, and smooth pigweed. The ability of the in vivo assay to quantify the reduction in ALS activity following applications of ALS-inhibiting herbicides was validated by comparing ALS activity following thifensulfuron application to ‘Williams 82’ soybean, which has a sulfonylurea-sensitive ALS, and ‘Asgrow 3200’ soybean, which has a sulfonylurea-insensitive ALS. Thifensulfuron reduced ALS activity in Williams 82 soybean to 0, 0.8, 3.3, and 15.6% of the CPCA control at 6, 12, 24, and 48 HAT, but ALS activity in Asgrow 3200 soybean was reduced only to 34, 40, 57, and 88% of the CPCA control.

Keywords

Thifensulfuron, 3-[[[[(4-methoxy-6-methyl-1,3,5-triazin-2-yl)amino]carbonyl]amino]sulfonyl]-2-thiophenecarboxylic acidcommon cocklebur, Xanthium strumarium L. # XANSTsmooth pigweed, Amaranthus hybridus L. # AMACHvelvetleaf, Abutilon theophrasti Medicus # ABUTHcorn, Zea mays L.grain sorghum, Sorghum bicolor (L.) Moenchsoybean, Glycine max (L.) Merr.Ketol-acid reductoisomeraseAcetolactate reductoisomeraseEC 4.1.3.18EC 1.1.186
Type
Research
Information
Weed Technology , Volume 9 , Issue 1 , March 1995 , pp. 17 - 22
Copyright
Copyright © 1995 by the Weed Science Society of America 

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