Biochemical characterization of metabolism-based atrazine resistance in Amaranthus tuberculatus and identification of an expressed GST associated with resistance

Anton F. Evans, Sarah R. O'Brien, Rong Ma, Aaron G. Hager, Chance W. Riggins, Kris N. Lambert, Dean E. Riechers

Research output: Contribution to journalArticle

Abstract

Rapid detoxification of atrazine in naturally tolerant crops such as maize (Zea mays) and grain sorghum (Sorghum bicolor) results from glutathione S-transferase (GST) activity. In previous research, two atrazine-resistant waterhemp (Amaranthus tuberculatus) populations from Illinois, U.S.A. (designated ACR and MCR), displayed rapid formation of atrazine-glutathione (GSH) conjugates, implicating elevated rates of metabolism as the resistance mechanism. Our main objective was to utilize protein purification combined with qualitative proteomics to investigate the hypothesis that enhanced atrazine detoxification, catalysed by distinct GSTs, confers resistance in ACR and MCR. Additionally, candidate AtuGST expression was analysed in an F2 population segregating for atrazine resistance. ACR and MCR showed higher specific activities towards atrazine in partially purified ammonium sulphate and GSH affinity-purified fractions compared to an atrazine-sensitive population (WCS). One-dimensional electrophoresis of these fractions displayed an approximate 26-kDa band, typical of GST subunits. Several phi- and tau-class GSTs were identified by LC-MS/MS from each population, based on peptide similarity with GSTs from Arabidopsis. Elevated constitutive expression of one phi-class GST, named AtuGSTF2, correlated strongly with atrazine resistance in ACR and MCR and segregating F2 population. These results indicate that AtuGSTF2 may be linked to a metabolic mechanism that confers atrazine resistance in ACR and MCR.

Original languageEnglish (US)
Pages (from-to)1238-1249
Number of pages12
JournalPlant Biotechnology Journal
Volume15
Issue number10
DOIs
StatePublished - Oct 2017

Fingerprint

Amaranthus tuberculatus
Amaranthus
Atrazine
atrazine
Glutathione Transferase
glutathione transferase
metabolism
Sorghum
Population
Zea mays
Glutathione S-Transferase pi
grain sorghum
Ammonium Sulfate
Vulnerable Populations
resistance mechanisms
Sorghum bicolor
Arabidopsis
ammonium sulfate
Proteomics
proteomics

Keywords

  • Amaranthus tuberculatus
  • detoxification
  • glutathione S-transferase
  • herbicide metabolism
  • oxidative stress
  • proteome
  • weed resistance

ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Plant Science

Cite this

Biochemical characterization of metabolism-based atrazine resistance in Amaranthus tuberculatus and identification of an expressed GST associated with resistance. / Evans, Anton F.; O'Brien, Sarah R.; Ma, Rong; Hager, Aaron G.; Riggins, Chance W.; Lambert, Kris N.; Riechers, Dean E.

In: Plant Biotechnology Journal, Vol. 15, No. 10, 10.2017, p. 1238-1249.

Research output: Contribution to journalArticle

@article{56628a7cf07a4d0aab03260a32694102,
title = "Biochemical characterization of metabolism-based atrazine resistance in Amaranthus tuberculatus and identification of an expressed GST associated with resistance",
abstract = "Rapid detoxification of atrazine in naturally tolerant crops such as maize (Zea mays) and grain sorghum (Sorghum bicolor) results from glutathione S-transferase (GST) activity. In previous research, two atrazine-resistant waterhemp (Amaranthus tuberculatus) populations from Illinois, U.S.A. (designated ACR and MCR), displayed rapid formation of atrazine-glutathione (GSH) conjugates, implicating elevated rates of metabolism as the resistance mechanism. Our main objective was to utilize protein purification combined with qualitative proteomics to investigate the hypothesis that enhanced atrazine detoxification, catalysed by distinct GSTs, confers resistance in ACR and MCR. Additionally, candidate AtuGST expression was analysed in an F2 population segregating for atrazine resistance. ACR and MCR showed higher specific activities towards atrazine in partially purified ammonium sulphate and GSH affinity-purified fractions compared to an atrazine-sensitive population (WCS). One-dimensional electrophoresis of these fractions displayed an approximate 26-kDa band, typical of GST subunits. Several phi- and tau-class GSTs were identified by LC-MS/MS from each population, based on peptide similarity with GSTs from Arabidopsis. Elevated constitutive expression of one phi-class GST, named AtuGSTF2, correlated strongly with atrazine resistance in ACR and MCR and segregating F2 population. These results indicate that AtuGSTF2 may be linked to a metabolic mechanism that confers atrazine resistance in ACR and MCR.",
keywords = "Amaranthus tuberculatus, detoxification, glutathione S-transferase, herbicide metabolism, oxidative stress, proteome, weed resistance",
author = "Evans, {Anton F.} and O'Brien, {Sarah R.} and Rong Ma and Hager, {Aaron G.} and Riggins, {Chance W.} and Lambert, {Kris N.} and Riechers, {Dean E.}",
year = "2017",
month = "10",
doi = "10.1111/pbi.12711",
language = "English (US)",
volume = "15",
pages = "1238--1249",
journal = "Plant Biotechnology Journal",
issn = "1467-7644",
publisher = "Wiley-Blackwell",
number = "10",

}

TY - JOUR

T1 - Biochemical characterization of metabolism-based atrazine resistance in Amaranthus tuberculatus and identification of an expressed GST associated with resistance

AU - Evans, Anton F.

AU - O'Brien, Sarah R.

AU - Ma, Rong

AU - Hager, Aaron G.

AU - Riggins, Chance W.

AU - Lambert, Kris N.

AU - Riechers, Dean E.

PY - 2017/10

Y1 - 2017/10

N2 - Rapid detoxification of atrazine in naturally tolerant crops such as maize (Zea mays) and grain sorghum (Sorghum bicolor) results from glutathione S-transferase (GST) activity. In previous research, two atrazine-resistant waterhemp (Amaranthus tuberculatus) populations from Illinois, U.S.A. (designated ACR and MCR), displayed rapid formation of atrazine-glutathione (GSH) conjugates, implicating elevated rates of metabolism as the resistance mechanism. Our main objective was to utilize protein purification combined with qualitative proteomics to investigate the hypothesis that enhanced atrazine detoxification, catalysed by distinct GSTs, confers resistance in ACR and MCR. Additionally, candidate AtuGST expression was analysed in an F2 population segregating for atrazine resistance. ACR and MCR showed higher specific activities towards atrazine in partially purified ammonium sulphate and GSH affinity-purified fractions compared to an atrazine-sensitive population (WCS). One-dimensional electrophoresis of these fractions displayed an approximate 26-kDa band, typical of GST subunits. Several phi- and tau-class GSTs were identified by LC-MS/MS from each population, based on peptide similarity with GSTs from Arabidopsis. Elevated constitutive expression of one phi-class GST, named AtuGSTF2, correlated strongly with atrazine resistance in ACR and MCR and segregating F2 population. These results indicate that AtuGSTF2 may be linked to a metabolic mechanism that confers atrazine resistance in ACR and MCR.

AB - Rapid detoxification of atrazine in naturally tolerant crops such as maize (Zea mays) and grain sorghum (Sorghum bicolor) results from glutathione S-transferase (GST) activity. In previous research, two atrazine-resistant waterhemp (Amaranthus tuberculatus) populations from Illinois, U.S.A. (designated ACR and MCR), displayed rapid formation of atrazine-glutathione (GSH) conjugates, implicating elevated rates of metabolism as the resistance mechanism. Our main objective was to utilize protein purification combined with qualitative proteomics to investigate the hypothesis that enhanced atrazine detoxification, catalysed by distinct GSTs, confers resistance in ACR and MCR. Additionally, candidate AtuGST expression was analysed in an F2 population segregating for atrazine resistance. ACR and MCR showed higher specific activities towards atrazine in partially purified ammonium sulphate and GSH affinity-purified fractions compared to an atrazine-sensitive population (WCS). One-dimensional electrophoresis of these fractions displayed an approximate 26-kDa band, typical of GST subunits. Several phi- and tau-class GSTs were identified by LC-MS/MS from each population, based on peptide similarity with GSTs from Arabidopsis. Elevated constitutive expression of one phi-class GST, named AtuGSTF2, correlated strongly with atrazine resistance in ACR and MCR and segregating F2 population. These results indicate that AtuGSTF2 may be linked to a metabolic mechanism that confers atrazine resistance in ACR and MCR.

KW - Amaranthus tuberculatus

KW - detoxification

KW - glutathione S-transferase

KW - herbicide metabolism

KW - oxidative stress

KW - proteome

KW - weed resistance

UR - http://www.scopus.com/inward/record.url?scp=85016713186&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85016713186&partnerID=8YFLogxK

U2 - 10.1111/pbi.12711

DO - 10.1111/pbi.12711

M3 - Article

C2 - 28218978

AN - SCOPUS:85016713186

VL - 15

SP - 1238

EP - 1249

JO - Plant Biotechnology Journal

JF - Plant Biotechnology Journal

SN - 1467-7644

IS - 10

ER -