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QUIK STATS (last updated Jan 20, 2015 )
NOTES ABOUT THIS BIOTYPE
A triple target-site mutation confers high levels of resistance to glyphosate in an Amaranthus quitensis population from Argentina Sarah-Jane Hutchingsa, Eddie McIndoea, Ryan Carlinb, Wenjin Yub, Anushka Howella, Weining Gub, Wenling Wangb, Mike Langforda, Jo Mattocksa and Shiv-Shankar Kaunduna a Syngenta, Jealott’s Hill International Research Centre, RG42 6EY Bracknell, UK b Syngenta, Research Triangle Park, NC USA E:mail: sarah-jane.hutchings@syngenta.com
Resistance to glyphosate has evolved and is quickly spreading in Amaranthus quitensis due to excessive use of the EPSPS-inhibiting herbicide in Argentinian soybean production systems. Here, we confirmed resistance to glyphosate and determined the mechanism involved in an A. quitensis population (AMAQU-R) from Santa Fe Province, Argentina. AMAQU-R plants survived glyphosate rates as high as 48 kg ai/ha, whereas individuals from the sensitive populations (AMAQU-S1 and AMAQU-S2) were killed at 750 g ai/ha or below. Differential glyphosate uptake, movement or metabolism was not associated with resistance in AMAQU-R and only a small average relative increase (1.94) in EPSPS gene copy number/expression was detected in AMAQU-R. Three linked EPSPS mutations were identified around the binding site of glyphosate in AMAQU-R. These comprised T102I and P106S amino acid substitutions documented to confer high levels of glyphosate resistance, as well as a novel A103V target-site mutation. A 3.5-fold more tolerance to glyphosate was observed for the TIAVPS mutant compared to the double TIPS strain following heterologous gene expression and enzyme analysis. The triple glyphosate resistance mutations may explain the rapid spread of A. quitensis in Argentina and could be a real threat for neighbouring countries under similar glyphosate selection pressure.
ACADEMIC ASPECTS
CONTRIBUTING WEED SCIENTISTS
ACKNOWLEDGEMENTS
Background
The evolution of herbicide‐resistant weeds is one of the most important concerns of global agriculture. Amaranthus hybridus L. is a competitive weed for summer crops in South America. In this article, we intend to unravel the molecular mechanisms by which an A. hybridus population from Argentina has become resistant to extraordinarily high levels of glyphosate.
Results
The glyphosate‐resistant population (A) exhibited particularly high parameters of resistance (GR50 = 20 900 g ai ha−1, Rf = 314), with all plants completing a normal life cycle even after 32X dose application. No shikimic acid accumulation was detected in the resistant plants at any of the glyphosate concentrations tested. Molecular and genetic analyses revealed a novel triple substitution (TAP‐IVS: T102I, A103V, and P106S) in the 5‐enol‐pyruvylshikimate‐3‐phosphate synthase (EPSPS) enzyme of population A and an incipient increase on the epsps relative copy number but without effects on the epspstranscription levels. The novel mechanism was prevalent, with 48% and 52% of the individuals being homozygous and heterozygous for the triple substitution, respectively. In silico conformational studies revealed that TAP‐IVS triple substitution would generate an EPSPS with a functional active site but with an increased restriction to glyphosate binding.
Conclusion
The prevalence of the TAP‐IVS triple substitution as the sole mechanism detected in the highly glyphosate resistant population suggests the evolution of a new glyphosate resistance mechanism arising in A. hybridus. This is the first report of a naturally occurring EPSPS triple substitution and the first glyphosate target‐site resistance mechanism described in A. hybridus.