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A rapid in vivo shikimate accumulation assay with excised leaf discs

Published online by Cambridge University Press:  20 January 2017

Dale L. Shaner ,
Talia Nadler-Hassar ,
W. Brien Henry  and
Clifford H. Koger
Talia Nadler-Hassar
Affiliation:
Colorado State University, Biological Science and Pest Management, Weed Science Lab, Fort Collins, CO 80527
W. Brien Henry
Affiliation:
USDA-ARS, Central Plains Resources Management Research Unit, 40335 County Road GG, Akron, CO 80720
Clifford H. Koger
Affiliation:
USDA-ARS, Southern Weed Science Research Unit, P.O. Box 350, Stoneville, MS 38776
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Abstract

An in vivo shikimate accumulation assay with excised leaf tissue was developed to provide a fast and reliable method for identifying glyphosate-resistant plants. The assay is based on glyphosate-induced accumulation of shikimate. There was a linear accumulation of shikimate in excised leaf discs of soybean and canola treated with 250 μM glyphosate for 48 h. The IC50 for the accumulation of shikimate in soybean and corn leaf discs was 34 and 87 μM, respectively. Leaf discs excised from glyphosate-resistant corn or soybean did not accumulate shikimate when treated with 500 μM glyphosate. Leaf discs taken from a number of field-grown plants accumulated shikimate in a glyphosate dose–dependent manner. The accumulation of shikimate was dependent on light and the age of the leaf from which the disc was taken. The assay worked either in 96-well microtiter plates or in vials, and it clearly differentiated between glyphosate-resistant and -susceptible crops in which the resistance is due to an alteration of the target site for glyphosate. The assay was simple and robust and has the potential to be used as a high throughput assay to detect glyphosate resistance in weeds.

Keywords

Glyphosatecanola, Brassica napus L. ‘Hayola 420’corn, Zea mays L. ‘Pioneer 37M34’, ‘Dekalb DK493RR/BTY’soybean, Glycine max (L.) Merr., ‘Asgrow A2869’, ‘Asgrow AG3003’Glyphosate resistanceresistance detection
Type
Physiology, Chemistry, and Biochemistry
Information
Weed Science , Volume 53 , Issue 6 , December 2005 , pp. 769 - 774
Copyright
Copyright © Weed Science Society of America 

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