caa a22 4500 606209638 CHVBK 20210128101029.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00425-014-2197-9 doi (NATIONALLICENCE)springer-10.1007/s00425-014-2197-9 Gene amplification of 5-enol-pyruvylshikimate-3-phosphate synthase in glyphosate-resistant Kochia scoparia [Elektronische Daten] [Andrew Wiersma, Todd Gaines, Christopher Preston, John Hamilton, Darci Giacomini, C. Robin Buell, Jan Leach, Philip Westra] Main conclusion : Field-evolved resistance to the herbicide glyphosate is due to amplification of one of two EPSPS alleles, increasing transcription and protein with no splice variants or effects on other pathway genes. The widely used herbicide glyphosate inhibits the shikimate pathway enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Globally, the intensive use of glyphosate for weed control has selected for glyphosate resistance in 31 weed species. Populations of suspected glyphosate-resistant Kochia scoparia were collected from fields located in the US central Great Plains. Glyphosate dose response verified glyphosate resistance in nine populations. The mechanism of resistance to glyphosate was investigated using targeted sequencing, quantitative PCR, immunoblotting, and whole transcriptome de novo sequencing to characterize the sequence and expression of EPSPS. Sequence analysis showed no mutation of the EPSPS Pro106 codon in glyphosate-resistant K. scoparia, whereas EPSPS genomic copy number and transcript abundance were elevated three- to ten-fold in resistant individuals relative to susceptible individuals. Glyphosate-resistant individuals with increased relative EPSPS copy numbers had consistently lower shikimate accumulation in leaf disks treated with 100μM glyphosate and EPSPS protein levels were higher in glyphosate-resistant individuals with increased gene copy number compared to glyphosate-susceptible individuals. RNA sequence analysis revealed seven nucleotide positions with two different expressed alleles in glyphosate-susceptible reads. However, one nucleotide at the seven positions was predominant in glyphosate-resistant sequences, suggesting that only one of two EPSPS alleles was amplified in glyphosate-resistant individuals. No alternatively spliced EPSPS transcripts were detected. Expression of five other genes in the chorismate pathway was unaffected in glyphosate-resistant individuals with increased EPSPS expression. These results indicate increased EPSPS expression is a mechanism for glyphosate resistance in these K. scoparia populations. Springer-Verlag Berlin Heidelberg, 2014 De novo transcriptome nationallicence EPSPS nationallicence Gene duplication nationallicence Herbicide resistance nationallicence ALS : Acetolactate synthase nationallicence EPSPS : 5-enol-pyruvylshikimate-3-phosphate synthase nationallicence Wiersma Andrew Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut Gaines Todd Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut Preston Christopher School of Agriculture, Food and Wine, University of Adelaide, Adelaide, South Australia, Australia aut Hamilton John Department of Plant Biology, Michigan State University, East Lansing, MI, USA aut Giacomini Darci Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut Robin Buell C. Department of Plant Biology, Michigan State University, East Lansing, MI, USA aut Leach Jan Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut Westra Philip Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut Planta Springer Berlin Heidelberg 241/2(2015-02-01), 463-474 0032-0935 241:2<463 2015 241 425 https://doi.org/10.1007/s00425-014-2197-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00425-014-2197-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wiersma Andrew Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut NATIONALLICENCE 700 1- Gaines Todd Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut NATIONALLICENCE 700 1- Preston Christopher School of Agriculture, Food and Wine, University of Adelaide, Adelaide, South Australia, Australia aut NATIONALLICENCE 700 1- Hamilton John Department of Plant Biology, Michigan State University, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Giacomini Darci Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut NATIONALLICENCE 700 1- Robin Buell C. Department of Plant Biology, Michigan State University, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Leach Jan Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut NATIONALLICENCE 700 1- Westra Philip Department of Bioagricultural Science and Pest Management, Colorado State University, 1177 Campus Delivery, 80523, Fort Collins, CO, USA aut NATIONALLICENCE 773 0- Planta Springer Berlin Heidelberg 241/2(2015-02-01), 463-474 0032-0935 241:2<463 2015 241 425