Evolution of target and non-target based multiple herbicide resistance in a single Palmer amaranth (Amaranthus palmeri) population from Kansas

The evolution of resistance to multiple herbicides in Palmer amaranth is a major challenge for its management. In this study, a Palmer amaranth population from Hutchinson, Kansas (HMR), was characterized for resistance to inhibitors of photosystem II (PSII) (e.g., atrazine), acetolactate synthase (ALS) (e.g., chlorsulfuron), and EPSP synthase (EPSPS) (e.g., glyphosate), and this resistance was investigated. About 100 HMR plants were treated with field-recommended doses (1x) of atrazine, chlorsulfuron, and glyphosate, separately along with Hutchinson multipleherbicide (atrazine, chlorsulfuron, and glyphosate)-susceptible (HMS) Palmer amaranth as control. The mechanism of resistance to these herbicides was investigated by sequencing or amplifying the psbA, ALS, and EPSPS genes, the molecular targets of atrazine, chlorsulfuron, and glyphosate, respectively. Fifty-two percent of plants survived a 1x (2,240 g ai ha(-1)) atrazine application with no known psbA gene mutation, indicating the predominance of a non-target site resistance mechanism to this herbicide. Forty-two percent of plants survived a 1x (18 g ai ha(-1)) dose of chlorsulfuron with proline(197)serine, proline(197)threonine, proline(197)alanine, and proline(197)asparagine, or tryptophan 574 leucine mutations in the ALS gene. About 40% of the plants survived a 1x (840 g ae ha(-1)) dose of glyphosate with no known mutations in the EPSPS gene. Quantitative PCR results revealed increased EPSPS copy number (50 to 140) as the mechanism of glyphosate resistance in the survivors. The most important finding of this study was the evolution of resistance to at least two sites of action (SOAs) (similar to 50% of plants) and to all three herbicides due to target site as well as non-target site mechanisms. The high incidence of individual plants with resistance to multiple SOAs poses a challenge for effective management of this weed.