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Plant Physiol, January 2003, Vol. 131, pp. 139-146
Ethylmethanesulfonate Saturation Mutagenesis in Arabidopsis to
Determine Frequency of Herbicide Resistance
Georg
Jander,1*
Scott R.
Baerson,2
Jebecka A.
Hudak,3
Kathleen A.
Gonzalez,
Kenneth
J.
Gruys, and
Robert L.
Last4
Cereon Genomics, 45 Sidney Street, Cambridge, Massachusetts 02139 (G.J., J.A.H., R.L.L.); and Monsanto Company, 700 Chesterfield Parkway,
St. Louis, Missouri 63198 (S.R.B., K.A.G., K.J.G.)
Plant resistance to glyphosate has been reported far less
frequently than resistance to sulfonylurea and imidazolinone
herbicides. However, these studies tend to be anecdotal, without side
by side comparisons for a single species or natural isolate. In this
study, we tested the frequencies of resistance of three herbicides in a
controlled ethylmethanesulfonate (EMS) saturation mutagenesis experiment, allowing a direct comparison of the frequencies at which
resistant mutant plants arise. The 100% growth inhibition dose rates
of glyphosate, chlorsulfuron (a sulfonylurea herbicide), and
imazethapyr (an imidazolinone herbicide) were determined for Arabidopsis. Populations of EMS-mutagenized M2 seedlings
were sprayed with twice the 100% growth inhibition dose of glyphosate, chlorsulfuron, or imazethapyr, and herbicide-resistant mutants were
identified. Although there were no glyphosate-resistant mutants among
M2 progeny of 125,000 Columbia and 125,000 Landsberg
erecta M1 lines, chlorsulfuron resistance
and imazethapyr resistance each appeared at frequencies of 3.2 × 10 5. Given the observed frequency of herbicide resistance
mutations, we calculate that there are at least 700 mutations in each
EMS-mutagenized Arabidopsis line and that fewer than 50,000 M1 lines are needed to have a 95% chance of finding a
mutation in any given G:C base pair in the genome. As part of this
study, two previously unreported Arabidopsis mutations conferring
resistance to imidazolinone herbicides, csr1-5
(Ala-122-Thr) and csr1-6 (Ala-205-Val), were discovered. Neither of these mutations caused enhanced resistance to chlorsulfuron in Arabidopsis.
1
Present address: Boyce Thompson Institute for
Plant Research, Tower Road, Ithaca, NY 14853.
2
Present address: U.S. Department of Agriculture,
Agricultural Research Service, Natural Products Utilization Research
Unit, P.O. Box 8048, University, MS 38677.
3
Present address: Infinity Pharmaceuticals, 650 Albany
Street, Boston, MA 02118.
4
Present address: Max Planck Institut for Chemical
Ecology, Beutenberg Campus, Winzerlaer Strasse 10, 07745, Jena, Germany.
*
Corresponding author; e-mail gj32{at}cornell.edu;
fax 607-254-2958.
© 2003 American Society of Plant Biologists
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