Multiple resistant Eleusine indica from Malaysia

International Survey of Herbicide-Resistant Weeds

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Monday, August 19, 2019

MULTIPLE RESISTANT GOOSEGRASS
(Eleusine indica)

Multiple Resistance: 4 Sites of Action
ACCase inhibitors (A/1)
PSI Electron Diverter (D/22)
EPSP synthase inhibitors (G/9)
Glutamine synthase inhibitors (H/10)

Malaysia, Jerantut

INTRODUCTION		GOOSEGRASS
Goosegrass (Eleusine indica) is a monocot weed in the Poaceae family. In Malaysia this weed first evolved multiple resistance (to 4 herbicide sites of action) in 2009 and infests Oil Palm Nursery. Multiple resistance has evolved to herbicides in the Groups A/1, D/22, G/9, and H/10. These particular biotypes are known to have resistance to butroxydim, fluazifop-P-butyl, glufosinate-ammonium, glyphosate, haloxyfop-methyl, and paraquat and they may be cross-resistant to other herbicides in the Groups A/1, D/22, G/9, and H/10. The 'Group' letters/numbers that you see throughout this web site refer to the classification of herbicides by their site of action. To see a full list of herbicides and HRAC herbicide classifications click here.

QUIK STATS (last updated Jan 20, 2015 )
Common Name	Goosegrass
Species	Eleusine indica
Group	ACCase inhibitors (A/1) PSI Electron Diverter (D/22) EPSP synthase inhibitors (G/9) Glutamine synthase inhibitors (H/10)
Herbicides	butroxydim, fluazifop-P-butyl, glufosinate-ammonium, glyphosate, haloxyfop-methyl, and paraquat
Location	Malaysia, Jerantut
Year	2009
Situation(s)	Oil Palm Nursery
Contributors - (Alphabetically)	Baki Bakar, and Stephen Powles
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NOTES ABOUT THIS BIOTYPE

GENERAL

JALALUDIN A, YU Q & POWLES SB (2015). Multiple resistance across glufosinate, glyphosate, paraquat and ACCase-inhibiting herbicides in an Eleusine indica population. Weed Research 55, 82–89.

An Eleusine indica population was previously reported as the first global case of field-evolved glufosinate resistance. This study re-examines glufosinate resistance and investigates multiple resistance to other herbicides in the population. Dose–response experiments with glufosinate showed that the resistant population is 5-fold and 14-fold resistant relative to the susceptible population, based on GR50 and LD50 R/S ratio respectively. The selected glufosinate-resistant subpopulation also displayed a high-level resistance to glyphosate, with the respective GR50 and LD50 R/S ratios being 12- and 144-fold. In addition, the subpopulation also displayed a level of resistance to paraquat and ACCase-inhibiting herbicides fluazifop-P-butyl, haloxyfop-P-methyl and butroxydim. ACCase gene sequencing revealed that the Trp-2027-Cys mutation is likely responsible for resistance to the ACCase inhibitors examined. Here, we confirm glufosinate resistance and importantly, we find very high-level glyphosate resistance, as well as resistance to paraquat and ACCase-inhibiting herbicides. This is the first confirmed report of a weed species that evolved multiple resistance across all the three non-selective global herbicides, glufosinate, glyphosate and paraquat.

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ACADEMIC ASPECTS

Confirmation Tests

Greenhouse, and Laboratory trials comparing a known susceptible Goosegrass biotype with this Goosegrass biotype have been used to confirm resistance. For further information on the tests conducted please contact the local weed scientists that provided this information.

Genetics

Genetic studies on Group A, D, G, H/1, 22, 9, 10 resistant Goosegrass have not been reported to the site. There may be a note below or an article discussing the genetics of this biotype in the Fact Sheets and Other Literature

Mechanism of Resistance

Studies on the mechanism of resistance of multiple resistant Goosegrass from Malaysia indicate that resistance is due to unknown, and an altered target site. There may be a note below or an article discussing the mechanism of resistance in the Fact Sheets and Other Literature

Relative Fitness

There is no record of differences in fitness or competitiveness of these resistant biotypes when compared to that of normal susceptible biotypes. If you have any information pertaining to the fitness of multiple resistant Goosegrass from Malaysia please update the database.

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CONTRIBUTING WEED SCIENTISTS

BAKI BAKAR

Senior Research Fellow Institute of Biological Sciences, University Of Malaya Dept. Of Botany Faculty of Science Building Kuala Lumpur, 50603 , Kuala Lumpur Malaysia Email Baki Bakar Web : Web Site Link

STEPHEN POWLES

Professor And Research Director University Of Western Australia School Of Plant Biology 2276 Dahlk Circle Crawley, 6009, Western Australia Australia Email Stephen Powles Web : Web Site Link

ACKNOWLEDGEMENTS
The Herbicide Resistance Action Committee, The Weed Science Society of America, and weed scientists in Malaysia have been instrumental in providing you this information. Particular thanks is given to Baki Bakar, and Stephen Powles for providing detailed information.

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Herbicide Resistant Goosegrass Globally
(Eleusine indica)

Country

FirstYear

Situation

Active Ingredients

Site of Action

Argentina

2012

Corn (maize), Fallow, and Soybean

glyphosate

EPSP synthase inhibitors (G/9)

Australia (Queensland)

2015

Sugarcane for sugar

paraquat

PSI Electron Diverter (D/22)

Bolivia

2005

Rice, Soybean, and Wheat

clethodim, cyhalofop-butyl, fluazifop-P-butyl, and haloxyfop-methyl

ACCase inhibitors (A/1)

Bolivia

2007

Soybean

glyphosate

EPSP synthase inhibitors (G/9)

Brazil

2003

Soybean

cyhalofop-butyl, fenoxaprop-P-ethyl, and sethoxydim

ACCase inhibitors (A/1)

Brazil

2016

Corn (maize), Soybean, and Wheat

glyphosate

EPSP synthase inhibitors (G/9)

Brazil

2017

Beans, Corn (maize), Cotton, and Soybean

fenoxaprop-P-ethyl, glyphosate, and haloxyfop-methyl

Multiple Resistance: 2 Sites of Action
ACCase inhibitors (A/1)
EPSP synthase inhibitors (G/9)

China

2010

Unspecified

paraquat

PSI Electron Diverter (D/22)

China

2010

Orchards

glyphosate

EPSP synthase inhibitors (G/9)

Colombia

2006

Coffee

glyphosate

EPSP synthase inhibitors (G/9)

Costa Rica

1989

Industrial sites

imazapyr

ALS inhibitors (B/2)

Costa Rica

2010

Pejibaye palm

glyphosate

EPSP synthase inhibitors (G/9)

Indonesia

2012

Oil Palm Nursery

glyphosate, and paraquat

Multiple Resistance: 2 Sites of Action
PSI Electron Diverter (D/22)
EPSP synthase inhibitors (G/9)

Indonesia

2012

Corn (maize)

paraquat

PSI Electron Diverter (D/22)

Japan

2013

Rice Paddy Levee

glyphosate

EPSP synthase inhibitors (G/9)

Malaysia

1990

Cropland, and Vegetables

fluazifop-P-butyl, and propaquizafop

ACCase inhibitors (A/1)

Malaysia

1990

Vegetables

paraquat

PSI Electron Diverter (D/22)

Malaysia

1997

Orchards

fluazifop-P-butyl, and glyphosate

Multiple Resistance: 2 Sites of Action
ACCase inhibitors (A/1)
EPSP synthase inhibitors (G/9)

Malaysia

2009

Vegetables

glufosinate-ammonium, and paraquat

Multiple Resistance: 2 Sites of Action
PSI Electron Diverter (D/22)
Glutamine synthase inhibitors (H/10)

Malaysia

2009

Oil Palm Nursery

butroxydim, fluazifop-P-butyl, glufosinate-ammonium, glyphosate, haloxyfop-methyl, and paraquat

Multiple Resistance: 4 Sites of Action
ACCase inhibitors (A/1)
PSI Electron Diverter (D/22)
EPSP synthase inhibitors (G/9)
Glutamine synthase inhibitors (H/10)

United States (North Carolina)

1973

Cotton

trifluralin

Microtubule inhibitors (K1/3)

United States (South Carolina)

1974

Cotton, and Soybean

trifluralin

Microtubule inhibitors (K1/3)

United States (Alabama)

1987

Cotton

trifluralin

Microtubule inhibitors (K1/3)

United States (Tennessee)

1988

Cotton, Golf courses, and Turf

pendimethalin, prodiamine, and trifluralin

Microtubule inhibitors (K1/3)

United States (Arkansas)

1989

Cotton, and Cropland

trifluralin

Microtubule inhibitors (K1/3)

United States (Georgia)

1992

Cotton, Golf courses, and Turf

trifluralin

Microtubule inhibitors (K1/3)

United States (Mississippi)

1994

Cotton

pendimethalin, and trifluralin

Microtubule inhibitors (K1/3)

United States (Florida)

1996

Tomatoes

paraquat

PSI Electron Diverter (D/22)

United States (Hawaii)

2003

Turf

metribuzin

Photosystem II inhibitors (C1/5)

United States (Mississippi)

2010

Cotton

glyphosate

EPSP synthase inhibitors (G/9)

United States (Tennessee)

2011

Soybean

glyphosate

EPSP synthase inhibitors (G/9)

United States (North Carolina)

2015

Golf courses

oxadiazon

PPO inhibitors (E/14)

United States (Virginia)

2015

Golf courses

oxadiazon

PPO inhibitors (E/14)

Literature about Similar Cases

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Takano, H., R. Mendes, L. Scoz, R. Ovejero, J. Constantin, T. Gaines, P. Westra, F. Dayan, and R. Oliveira. 2019. Proline 106 EPSPS mutation imparting glyphosate resistance in goosegrass (Eleusine indica) emerges in South America. Weed Science 67 : 48 - 56.

Glyphosate-resistant (GR) goosegrass [Eleusine indica (L.) Gaertn.] was recently identified in Brazil, but its resistance mechanism was unknown. This study elucidated the resistance mechanism in this species and developed a molecular marker for rapid detection of this target-site resistance trait. The resistance factor for the resistant biotype was 4.4-fold compared with the glyphosate-susceptible (GS) in greenhouse dose–response experiments. This was accompanied by a similar (4-fold) difference in the levels of in vitro and in planta shikimate accumulation in these biotypes. However, there was no difference in uptake, translocation, or metabolism of glyphosate between the GS and GR biotypes. Moreover, both biotypes showed similar values for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) copy number and transcription. Sequencing of a 330-bp fragment of the EPSPS gene identified a single-nucleotide polymorphism that led to a Pro-106-Ser amino acid substitution in the enzyme from the GR biotype. This mutation imparted a 3.8-fold increase in the amount of glyphosate required to inhibit 50% of EPSPS activity, confirming the role of this amino acid substitution in resistance to glyphosate. A quantitative PCR–based genotyping assay was developed for the rapid detection of resistant plants containing this Pro-106-Ser mutation..

Gherekhloo, J., Fernández-Moreno, P.T., Alcántara-de la Cruz, R., Sánchez-González, E., Cruz-Hipolito, H.E., Domínguez-Valenzuela, J.A. and De Prado, R.. 2017. Pro-106-Ser mutation and EPSPS overexpression acting together simultaneously in glyphosate-resistant goosegrass (Eleusine indica). Scientific Reports 7 : 6702 - .

Glyphosate has been used for more than 15 years for weed management in citrus groves in the Gulf of Mexico, at up to 3–4 applications per year. Goosegrass (Eleusine indica (L.) Gaertn.) control has sometimes failed. In this research, the mechanisms governing three goosegrass biotypes (Ein-Or from an orange grove, and Ein-Pl1 and Ein-Pl2 from Persian lime groves) with suspected resistance to glyphosate were characterized and compared to a susceptible biotype (Ein-S). Dose-response and shikimate accumulation assays confirmed resistance of the resistant (R) biotypes. There were no differences in glyphosate absorption, but the R biotypes retained up to 62–78% of the herbicide in the treated leaf at 96 h after treatment (HAT), in comparison to the Ein-S biotype (36%). The 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity in the Ein-Or and Ein-S biotypes was over 100-fold lower than the Ein-Pl1 and Ein-Pl2 ones. The latter showed a high EPSPS-basal activity, a mutation at Pro-106-Ser position in the EPSPS gene, and EPSPS overexpression. The EPSPS basal and EPSPS overexpression were positively correlated. The R goosegrass biotypes displayed poor glyphosate translocation. Furthermore, this grassweed showed, for the first time, two mechanisms at the target-site level (Pro-106-Ser mutation + EPSPS overexpression) acting together simultaneously against glyphosate..

Yu, Q., A. Jalaludin, H. Han, M. Chen, R.D. Sammons, and S. Powles. 2015. Evolution of a Double Amino Acid Substitution in the 5-Enolpyruvylshikimate-3-Phosphate Synthase in Eleusine indica Conferring High-Level Glyphosate Resistance. Plant Physiology 167 : 1440 - 1447.

Glyphosate is the most important and widely used herbicide in world agriculture. Intensive glyphosate selection has resulted in the widespread evolution of glyphosate-resistant weed populations, threatening the sustainability of this valuable once-in-a-century agrochemical. Field-evolved glyphosate resistance due to known resistance mechanisms is generally low to modest. Here, working with a highly glyphosate-resistant Eleusine indica population, we identified a double amino acid substitution (T102I + P106S [TIPS]) in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene in glyphosate-resistant individuals. This TIPSmutation recreates the biotechnology-engineered commercial first generation glyphosate-tolerant EPSPS in corn (Zea mays) and now in other crops. In E. indica, the naturally evolved TIPS mutants are highly (more than 180-fold) resistant to glyphosate compared with the wild type and more resistant (more than 32-fold) than the previously known P106S mutants. The E. indica TIPS EPSPS showed very high-level (2,647-fold) in vitro resistance to glyphosate relative to the wild type and is more resistant (600-fold) than the P106S variant. The evolution of the TIPSmutation in crop fields under glyphosate selection is likely a sequential event, with the P106S mutation being selected first and fixed, followed by the T102I mutation to create the highly resistant TIPS EPSPS. The sequential evolution of the TIPS mutation endowing high-level glyphosate resistance is an important mechanism by which plants adapt to intense herbicide selection and a dramatic example of evolution in action..

Chen, J., H. Huang, C. Zhang, S. Wei, Z. Huang, J. Chen, and X. Wang. 2015. Mutations and amplification of EPSPS gene confer resistance to glyphosate in goosegrass (Eleusine indica). Planta 242 : 859 - 868.

Glyphosate has been used widely in the south of China. The high selection pressure from glyphosate use has led to the evolution of resistance to glyphosate in weeds. We investigated the molecular mechanisms of three recently discovered glyphosate-resistant Eleusine indicapopulations (R1, R2 and R3). The results showed that R1 and R2 had double Thr102Ile and Pro106Ser mutation and a single mutation of Pro106Leu in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene, respectively. Escherichia coli containing the mutated EPSPS genes was tolerant to glyphosate. EPSPS activity in R1 and R2 plants was higher than in the sensitive plants. There was no amino acid substitution in EPSPS gene in R3. However, expression of EPSPS in R3 plants was higher than in glyphosate-susceptible (S) population (13.8-fold) after glyphosate treatment. EPSPS enzyme activity in both R3 and S plants was inhibited by glyphosate, while shikimate accumulation in R3 was significantly lower than for the S population. Further analysis revealed that the genome of R3 contained 28.3-fold more copies of the EPSPS gene than that of susceptible population. EPSPS expression was positively correlated with copy number of EPSPS. In conclusion, mutation of the EPSPS gene and increased EPSPS expression are part of the molecular mechanisms of resistance to glyphosate in Eleusine indica..

Cha ThyeSan ; Anne-Marie, K. ; Chuah TseSeng. 2014. Identification and characterization of RAPD-SCAR markers linked to glyphosate-susceptible and -resistant biotypes of Eleusine indica (L.) Gaertn. Molecular Biology Reports 41 : 823 - 831.

Eleusine indica is one of the most common weed species found in agricultural land worldwide. Although herbicide-glyphosate provides good control of the weed, its frequent uses has led to abundant reported cases of resistance. Hence, the development of genetic markers for quick detection of glyphosate-resistance in E. indica population is imperative for the control and management of the weed. In this study, a total of 14 specific random amplified polymorphic DNA (RAPD) markers were identified and two of the markers, namely S4R727 and S26R₆976 were further sequence characterized. Sequence alignment revealed that marker S4R727 showing a 12-bp nucleotides deletion in resistant biotypes, while marker S26R6976 contained a 167-bp nucleotides insertion in the resistant biotypes. Based on these sequence differences, three pairs of new sequence characterized amplified region (SCAR) primers were developed. The specificity of these primer pairs were further validated with genomic DNA extracted from ten individual plants of one glyphosate-susceptible and five glyphosate-resistant (R2, R4, R6, R8 and R11) populations. The resulting RAPD-SCAR markers provided the basis for assessing genetic diversity between glyphosate-susceptible and -resistant E. indica biotypes, as well for the identification of genetic locus link to glyphosate-resistance event in the species..

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