Gene expression in response to glyphosate treatment in fleabane (<scp><i>Conyza bonariensis</i></scp>) – glyphosate death response and candidate resistance genes

Hereward, James; Werth, Jeff; Thornby, David; Keenan, Michelle; Chauhan, Bhagirath Singh; Walter, Gimme H.

doi:10.1002/ps.4804

Cited by 17 publications

(31 citation statements)

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“…The mechanism of glyphosate resistance remains to be characterized fully in other NRR A. trifida biotypes as noted above, in several RR A. trifida accessions, 15,19,62 and in other species. 45,63 Future research objectives are to investigate cellular uptake and subcellular distribution of glyphosate 19,64 and transcriptome profiling 65 in Wisconsin A. trifida R and S phenotypes.…”

Section: Resultsmentioning

confidence: 99%

Physiological and molecular analysis of glyphosate resistance in non‐rapid response Ambrosia trifida from Wisconsin

Wilson

Takano

Horn

et al. 2019

Pest Management Science

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BACKGROUND We previously identified a glyphosate‐resistant A. trifida phenotype from Wisconsin USA that showed a non‐rapid response to glyphosate. The mechanism of glyphosate resistance in this phenotype has yet to be elucidated. We conducted experiments to investigate non‐target‐site resistance and target‐site resistance mechanisms. The roles of glyphosate absorption, translocation, and metabolism in resistance of this phenotype have not been reported previously, nor have EPSPS protein abundance or mutations to the full‐length sequence of EPSPS. RESULTS Whole‐plant dose–response results confirmed a 6.5‐level of glyphosate resistance for the resistant (R) phenotype compared to a susceptible (S) phenotype. Absorption and translocation of 14C‐glyphosate were similar between R and S phenotypes over 72 h. Glyphosate and AMPA concentrations in leaf tissue did not differ between R and S phenotypes over 96 h. In vivo shikimate leaf disc assays confirmed that glyphosate EC50 values were 4.6‐ to 5.4‐fold greater for the R than S phenotype. Shikimate accumulation was similar between phenotypes at high glyphosate concentrations (>1000 μM), suggesting that glyphosate entered chloroplasts and inhibited EPSPS. This finding was supported by results showing that EPSPS copy number and EPSPS protein abundance did not differ between R and S phenotypes, nor did EPSPS sequence at Gly101, Thr102, and Pro106 positions. Comparison of full‐length EPSPS sequences found five nonsynonymous polymorphisms that differed between R and S phenotypes. However, their locations were distant from the glyphosate target site and, therefore, not likely to affect enzyme‐glyphosate interaction. CONCLUSION The results suggest that a novel mechanism confers glyphosate resistance in this A. trifida phenotype. © 2019 Society of Chemical Industry

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Section: Resultsmentioning

confidence: 99%

Physiological and molecular analysis of glyphosate resistance in non‐rapid response Ambrosia trifida from Wisconsin

Wilson

Takano

Horn

et al. 2019

Pest Management Science

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“…However, the results of the present work show that until 192 and 288 HAT for GS and GR biotypes, respectively, the glyphosate treatment does not interfere negatively in RIN integrity (Figures 2, 3, 4, 5 and 6). Thus, for further molecular analysis such as RNA-Seq and RT-qPCR, the RNA integrity results indicate that studies can be performed in times higher than 24 and 48 HAT Hereward et al, 2018) to capture as much as possible the process of glyphosate plant detoxification ( Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…Several studies have been conducted in Conyza spp. with the objectives to understand the GR mechanisms (Feng et al, 2004;Ferreira et al, 2008;Peng et al, 2010;Ge et al, 2014;Cardinali et al, 2015;Tani et al, 2015;Moretti et al, 2017;Kleinman and Rubin, 2017;González-Torralva et al, 2017;Hereward et al, 2018;Amaro-Blanco et al, 2018). These studies have found that the GR mechanisms are related to herbicide transport, translocation, uptake, metabolism, and alterations at the 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) enzyme, which is the glyphosate target site.…”

Section: Introductionmentioning

confidence: 99%

“…These studies have found that the GR mechanisms are related to herbicide transport, translocation, uptake, metabolism, and alterations at the 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) enzyme, which is the glyphosate target site. However, the specific GR mechanisms remain unclear in Conyza spp., even after the adoption of the powerful approaches like RNA-Sequencing (RNA-Seq) and real-time quantitative reverse transcriptase (RT-qPCR) Hereward et al, 2018). RNA-Seq and RT-qPCR are molecular approaches used to evaluate changes in gene expression in response to stress, in this case, herbicide treatment.…”

Section: Introductionmentioning

confidence: 99%

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Does the Glyphosate Treatment Interfere Negatively on RNA Integrity in Glyphosate-Resistant and -Sensitive Conyza bonariensis?

et al. 2019

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The hairy fleabane (Conyza bonariensis (L.) Cronq.) is among the most problematic glyphosate-resistant weeds to manage around the world. In weed science, molecular approaches such as RNA sequencing (RNA-Seq) and quantitative reverse-transcription polymerase chain reaction (RT-qPCR) have been employed to study molecular responses to glyphosate treatment in Conyza species. Glyphosate treatment leads to reactive oxygen species (ROS) production in plants which could damage the RNA. Degraded RNA is an issue and can compromise further molecular analysis. Thus, the objective of this study was to evaluate whether glyphosate treatment interferes negatively on RNA integrity of glyphosate-resistant andsensitive hairy fleabane biotypes. Two experiments were performed using glyphosate doses from 0 to 11,840 g a.e. ha -1 and evaluated in a time-course until 288 hours after treatment. The total of 86 RNA samples were evaluated. The RNA integrity was evaluated in a Bioanalyzer 2100 equipment according to RNA integrity number (RIN) scores and electrophoresis gel. The RIN scores ranged from 5.1 to 9.0. Glyphosate doses do not reduce the RIN scores in both glyphosate-resistant and -sensitive biotypes of hairy fleabane. Visual and automatic analysis of electrophoresis gel show suitable results for all RNA samples, with well-defined bands at 28S and 18S positions and no degradation. The results of the analysis indicate that glyphosate treatment does not affect the RNA integrity of glyphosate-resistant and -sensitive biotypes of hairy fleabane until 288 and 192 hours after glyphosate treatment, respectively. The RNA integrity analysis provides useful results to evaluate the RNA condition for further analysis. However, the costs were around US$ 14.25 per sample, considering only reagents. These results are useful for planning future timecourse experiments in Conyza spp. after glyphosate treatment.Keywords: hairy fleabane, molecular analysis, RNA integrity number (RIN). (Conyza bonariensis (L.) Cronq.) está entre as plantas daninhas resistentes ao glifosato mais difíceis de serem manejadas em todo o mundo. Na ciência das plantas daninhas, abordagens moleculares, como o sequenciamento de RNA (RNA-Seq) e a reação da transcriptase reversa da polimerase em tempo real (RT-qPCR), têm sido empregadas para estudar as respostas moleculares ao tratamento com glifosato em espécies de Conyza. No entanto, o tratamento com glifosato leva à produção de espécies reativas de oxigênio (ROS) em plantas, que podem danificar o RNA. A degradação do RNA é um problema e pode comprometer futuras análises moleculares. Assim, o objetivo deste estudo foi avaliar se o tratamento com glifosato interfere negativamente na integridade do RNA de biótipos de buva resistente e sensível ao glifosato. Dois experimentos foram realizados I D I D I D Planta Daninha 2019; v37:e019217909 PIASECKI, C. et al. Does the glyphosate treatment interfere negatively on RNA integrity in glyphosate-resistant and -sensitive ... 2 utilizando doses de glifosato de 0 a 11.840 g e.a. ha -1 e av...

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“…Reduced dicamba translocation was found in dicamba‐resistant Kochia scoparia with no differences in dicamba metabolism, and a transcriptomics study suggested that increased flavonol pathway transcription was associated with dicamba resistance . Glyphosate resistance was investigated in Conyza bonariensis using transcriptomics, showing the lack of a resistance conferring mutation in the three EPSPS copies carried on each of three genomes of this hexaploid species, and producing a list of candidate genes that could be involved in non‐target‐site glyphosate resistance . The evolution of plant protoporphyrinogen oxidase genes is reviewed, along with implications for herbicide resistance .…”

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confidence: 99%

Introduction to Pest Management Science special issue for GHRC 2017

Gaines¹,

Shaner²,

Dayan³

2018

Pest Management Science

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Gene expression in response to glyphosate treatment in fleabane (Conyza bonariensis) – glyphosate death response and candidate resistance genes

Cited by 17 publications

References 58 publications

Physiological and molecular analysis of glyphosate resistance in non‐rapid response Ambrosia trifida from Wisconsin

Physiological and molecular analysis of glyphosate resistance in non‐rapid response Ambrosia trifida from Wisconsin

Does the Glyphosate Treatment Interfere Negatively on RNA Integrity in Glyphosate-Resistant and -Sensitive Conyza bonariensis?

Introduction to Pest Management Science special issue for GHRC 2017

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