Date of Graduation

5-2012

Document Type

Thesis

Degree Name

Master of Science in Crop, Soil & Environmental Sciences (MS)

Degree Level

Graduate

Department

Crop, Soil & Environmental Sciences

Advisor

Nilda R. Burgos

Committee Member

Robert C. Scott

Second Committee Member

Mary C. Savin

Third Committee Member

Andronikos Mauromoustakos

Fourth Committee Member

Nicholas Polge

Keywords

Biological sciences, Epsps gene, Glyphosate, Herbicide resistance, Herbicide resistance mechanisms, Italian ryegrass, Weed science

Abstract

Italian ryegrass is a principal weed problem in wheat production fields in the Southern US. Resistance to herbicides diclofop, mesosulfuron, and pinoxaden among ryegrass populations has been reported. Glyphosate-resistant Italian ryegrass populations were identified in Desha County, Arkansas. This research aimed to 1) determine resistance patterns to ACCase (diclofop and pinoxaden) and ALS (imazamox, mesosulfuron, and pyroxsulam) herbicides among Italian ryegrass populations from the southern US; 2) determine if cytochrome P450-mediated enhanced herbicide metabolism contributed to resistance; and 3) elucidate the resistance mechanism to glyphosate in four Arkansas populations (Des03, Des05, Des14, and D8). For objective 1, 30 accessions from problematic fields in the southern US between 2008 and 2010 were subjected to dose-response bioassays. Among the 30 accessions, 27 were resistant to both diclofop and mesosulfuron, 25 of which were also resistant to pyroxsulam. Ten Arkansas accessions collected in 2008 were resistant to diclofop, mesosulfuron, pyroxsulam, and imazamox. One accession from Georgia and three accessions from North Carolina were resistant to diclofop, mesosulfuron, pyroxsulam, and pinoxaden. For objective 2, six ryegrass populations with different resistance patterns to glyphosate, ALS- and ACCase herbicides, were treated with P450 inhibitors malathion (1000 g ai ha-1) and 1-aminobenzotriazole (100 µM ABT) before herbicide application. Malathion improved herbicide activity in some populations; but did not completely overcome resistance to any herbicide. This indicates that P450-mediated metabolism is only partially responsible for resistance in these populations. For objective 3, plants from Des03 population were analyzed for resistance level, EPSPS genetic mutation(s), EPSPS enzyme activity, and EPSPS gene copy number. The absorption and translocation of 14C-glyphosate were similar in R and S plants. The EPSPS gene in the R plants did not contain any point mutation(s) associated with glyphosate resistance. Resistance to glyphosate in Des03 is due to increased basal EPSPS enzyme activity resulting from amplification of the EPSPS gene. Follow-up experiments conducted on other glyphosate-R populations Des05, Des14, and D8 showed 11-fold to 516-fold more copies of the EPSPS gene in resistant plants than their susceptible counterparts indicating that EPSPS gene amplification also confers resistance to glyphosate in these populations.

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