Different Mutations Providing Target Site Resistance to ALS- and ACCase-Inhibiting Herbicides in Echinochloa spp. from Rice Fields

Echinochloa spp. is one of the most invasive weeds in rice fields worldwide. Acetolactate synthase (ALS) and acetyl-CoA carboxylase (ACCase) inhibiting herbicides are two of the most widely used rice herbicides. However, overuse has led to the resistance evolution of Echinochloa spp. to penoxsulam (ALS-inhibitor) and cyhalofop-methyl (ACCase-inhibitor). In this work, 137 different Echinochloa spp. populations were collected in different rice fields in Extremadura (western Spain) where lack of control was detected. Target-site based resistance (by sequencing ALS and ACCase gene) and characterization of Echinochloa species at the molecular level (based on PCR-RFLP analyses) were carried out in those populations. Most of the populations studied (111 of 137) belong to the E. oryzicola/E. oryzoides group. Three-point mutations were identified in ALS genes: Pro197Ser, Pro197Thr, and Ser653Asn, the first being the most frequent substitution in resistant plants. In the ACCase gene, the Ile1781Leu substitution was found. In both ALS and ACCase sequencing, evidence of heterozygosity was also observed. To assess whether cross-resistance patterns differed between mutations, two populations belonging to the E. oryzicola/E. oryzoides group had its most frequent mutations (Pro197Ser, population ech3-14 and Ile1781Leu, population ech114-10) chosen to be carried out in a dose-response assay. It was confirmed that Pro197Ser conferred resistance to triazolopyrimidine, imidazolinone, sulfonylurea, and pyrimidinyl benzoate families. On the other hand, the Ile1781Leu change gave resistance to aryloxyphenoxypropionate and cyclohexanedione families. Of the authorized herbicides in rice in Spain, more that 80% belong to these families. It is therefore important that farmers carry out an integrated control system that combines both chemical and non-chemical tools.

Download Full-text

Target-Site Resistance Mechanisms to Tribenuron-methyl and Cross-resistance Patterns to ALS-inhibiting Herbicides of Catchweed Bedstraw (Galium aparine) with Different ALS Mutations

Weed Science ◽

10.1017/wsc.2018.70 ◽

2018 ◽

Vol 67 (2) ◽

pp. 183-188 ◽

Cited By ~ 2

Author(s):

Wei Deng ◽

Yingjie Di ◽

Jingxuan Cai ◽

Yueyang Chen ◽

Shuzhong Yuan

Keyword(s):

Acetolactate Synthase ◽

Resistance Mechanisms ◽

Target Site ◽

Cross Resistance ◽

Susceptible Population ◽

Resistance Evolution ◽

Galium Aparine ◽

Resistance Patterns ◽

Target Site Resistance ◽

Tribenuron Methyl

AbstractCatchweed bedstraw (Galium aparine L.) is a problematic dicot weed that occurs in major winter wheat (Triticum aestivum L.) fields in China. Tribenuron-methyl has been widely used to control broadleaf weeds since 1988 in China. However, overuse has led to the resistance evolution of G. aparine to tribenuron-methyl. In this study, 20 G. aparine populations collected from Shandong and Henan provinces were used to determine tribenuron-methyl resistance and target-site resistance mechanisms. In dose–response experiments, 12 G. aparine populations showed different resistance levels (2.92 to 842.41-fold) to tribenuron-methyl compared with the susceptible population. Five different acetolactate synthase (ALS) mutations (Pro-197-Leu, Pro-197-Ser, Pro-197-His, Asp-376-Glu, and Trp-574-Leu) were detected in different resistant populations. Individuals heterozygous for Pro-197-Ser and Trp-574-Leu mutations were also observed in a resistant population (HN6). In addition, pHB4 (Pro-197-Ser), pHB7 (Pro-197-His), pHB8 (Pro-197-Leu), pHB5 (Asp-376-Glu), and pHB3 (Trp-574-Leu) subpopulations individually homozygous for specific ALS mutations were generated to evaluate the cross-resistance to ALS-inhibiting herbicides. The pHB4, pHB7, pHB8, pHB5, and pHB3 subpopulations all were resistant to sulfonylurea, pyrazosulfuron-ethyl, triazolopyrimidine, flumetsulam, sulfonylamino-carbonyl-triazolinone, flucarbazone-sodium, pyrimidinyl thiobenzoate, pyribenzoxim, and the imidazolinone imazethapyr. These results indicated the diversity of the resistance-conferring ALS mutations in G. aparine, and all these mutations resulted in broad cross-resistance to five kinds of ALS-inhibiting herbicides.

Download Full-text

Point Mutations as Main Resistance Mechanism Together With P450-Based Metabolism Confer Broad Resistance to Different ALS-Inhibiting Herbicides in Glebionis coronaria From Tunisia

Frontiers in Plant Science ◽

10.3389/fpls.2021.626702 ◽

2021 ◽

Vol 12 ◽

Author(s):

Zeineb Hada ◽

Yosra Menchari ◽

Antonia M. Rojano-Delgado ◽

Joel Torra ◽

Julio Menéndez ◽

...

Keyword(s):

Enzyme Activity ◽

Dose Response ◽

Resistance Mechanism ◽

Point Mutations ◽

Acetolactate Synthase ◽

Target Site ◽

Cross Resistance ◽

P450 Monooxygenase ◽

Target Site Resistance ◽

Sites Of Action

Resistance to acetolactate synthase (ALS) inhibiting herbicides has recently been reported in Glebionis coronaria from wheat fields in northern Tunisia, where the weed is widespread. However, potential resistance mechanisms conferring resistance in these populations are unknown. The aim of this research was to study target-site resistance (TSR) and non-target-site resistance (NTSR) mechanisms present in two putative resistant (R) populations. Dose–response experiments, ALS enzyme activity assays, ALS gene sequencing, absorption and translocation experiments with radiolabeled herbicides, and metabolism experiments were carried out for this purpose. Whole plant trials confirmed high resistance levels to tribenuron and cross-resistance to florasulam and imazamox. ALS enzyme activity further confirmed cross-resistance to these three herbicides and also to bispyribac, but not to flucarbazone. Sequence analysis revealed the presence of amino acid substitutions in positions 197, 376, and 574 of the target enzyme. Among the NTSR mechanisms investigated, absorption or translocation did not contribute to resistance, while evidences of the presence of enhanced metabolism were provided. A pretreatment with the cytochrome P450 monooxygenase (P450) inhibitor malathion partially synergized with imazamox in post-emergence but not with tribenuron in dose–response experiments. Additionally, an imazamox hydroxyl metabolite was detected in both R populations in metabolism experiments, which disappeared with the pretreatment with malathion. This study confirms the evolution of cross-resistance to ALS inhibiting herbicides in G. coronaria from Tunisia through TSR and NTSR mechanisms. The presence of enhanced metabolism involving P450 is threatening the chemical management of this weed in Tunisian wheat fields, since it might confer cross-resistance to other sites of action.

Download Full-text

Target-Site and Non-target-Site Resistance Mechanisms Confer Multiple and Cross- Resistance to ALS and ACCase Inhibiting Herbicides in Lolium rigidum From Spain

Frontiers in Plant Science ◽

10.3389/fpls.2021.625138 ◽

2021 ◽

Vol 12 ◽

Author(s):

Joel Torra ◽

José María Montull ◽

Andreu Taberner ◽

Nawaporn Onkokesung ◽

Neil Boonham ◽

...

Keyword(s):

Glutathione Transferase ◽

Point Mutations ◽

Acetolactate Synthase ◽

Target Site ◽

Cross Resistance ◽

Lolium Rigidum ◽

P450 Monooxygenase ◽

Winter Cereals ◽

Target Site Resistance ◽

Sites Of Action

Lolium rigidum is one the worst herbicide resistant (HR) weeds worldwide due to its proneness to evolve multiple and cross resistance to several sites of action (SoA). In winter cereals crops in Spain, resistance to acetolactate synthase (ALS)- and acetyl-CoA carboxylase (ACCase)-inhibiting herbicides has become widespread, with farmers having to rely on pre-emergence herbicides over the last two decades to maintain weed control. Recently, lack of control with very long-chain fatty acid synthesis (VLCFAS)-inhibiting herbicides has been reported in HR populations that are difficult to manage by chemical means. In this study, three Spanish populations of L. rigidum from winter cereals were confirmed as being resistant to ALS- and ACCase-inhibiting herbicides, with broad-ranging resistance toward the different chemistries tested. In addition, reduced sensitivity to photosystem II-, VLCFAS-, and phytoene desaturase-inhibiting herbicides were confirmed across the three populations. Resistance to ACCase-inhibiting herbicides was associated with point mutations in positions Trp-2027 and Asp-2078 of the enzyme conferring target site resistance (TSR), while none were detected in the ALS enzyme. Additionally, HR populations contained enhanced amounts of an ortholog of the glutathione transferase phi (F) class 1 (GSTF1) protein, a functional biomarker of non-target-site resistance (NTSR), as confirmed by enzyme-linked immunosorbent assays. Further evidence of NTSR was obtained in dose-response experiments with prosulfocarb applied post-emergence, following pre-treatment with the cytochrome P450 monooxygenase inhibitor malathion, which partially reversed resistance. This study confirms the evolution of multiple and cross resistance to ALS- and ACCase inhibiting herbicides in L. rigidum from Spain by mechanisms consistent with the presence of both TSR and NTSR. Moreover, the results suggest that NTSR, probably by means of enhanced metabolism involving more than one detoxifying enzyme family, confers cross resistance to other SoA. The study further demonstrates the urgent need to monitor and prevent the further evolution of herbicide resistance in L. rigidum in Mediterranean areas.

Download Full-text

Molecular basis of resistance to bensulfuron-methyl and cross resistance patterns to ALS-inhibiting herbicides in Ludwigia prostrata Roxb

Weed Technology ◽

10.1017/wet.2021.47 ◽

2021 ◽

pp. 1-20

Author(s):

Wei Deng ◽

Mengting Yang ◽

Zhiwen Duan ◽

Cheng Peng ◽

Zhiming Xia ◽

...

Keyword(s):

Molecular Mechanism ◽

Dose Response ◽

Molecular Basis ◽

Resistance Mutation ◽

Gene Sequencing ◽

Acetolactate Synthase ◽

Rice Fields ◽

Cross Resistance ◽

Nucleotide Mutation ◽

Resistance Patterns

Abstract Ludwigia prostrata Roxb. is a problematic weed in rice fields in China, and acetolactate synthase (ALS)-inhibiting herbicides (e.g., bensulfuron-methyl) are widely used for the management of broadleaf weeds. Recently, a L. prostrata biotype (JS-R) that failed to be controlled with ALS-inhibiting herbicides was found in Jiangsu Province, China. This study aims to determine the level and molecular mechanism of resistance to bensulfuron-methyl in this JS-R biotype, and evaluate the cross-resistance spectrums to other ALS-inhibiting herbicides. The dose-response assays indicated that the JS-R L. prostrata biotype had evolved 21.2-fold resistance to bensulfuron-methyl compared to the susceptible biotype (JS-S). ALS gene sequencing revealed that a nucleotide mutation (CCA to TCA) at codon 197, resulting in a Pro-197-Ser mutation, was detected in the resistant plants. Moreover, the JS-R biotype contained the Pro-197-Ser resistance mutation showed cross-resistance to pyrazosulfuron-ethyl (12.0-fold), but was sensitive to penoxsulam, bispyribac-sodium and imazethapyr, which may serve as alternative herbicides to control the resistant L. prostrata biotype. This is the first confirmation of a L. prostrata biotype resistant to bensulfuron-methyl due to a Pro-197-Ser resistance mutation in the ALS gene.

Download Full-text

Investigating the resistance levels and mechanisms to penoxsulam and cyhalofop-butyl in barnyardgrass (Echinochloa crus-galli) from Ningxia province, China

Weed Science ◽

10.1017/wsc.2021.37 ◽

2021 ◽

pp. 1-25

Author(s):

Qian Yang ◽

Xia Yang ◽

Zichang Zhang ◽

Jieping Wang ◽

Weiguo Fu ◽

...

Keyword(s):

Herbicide Resistance ◽

Molecular Mechanisms ◽

Acetolactate Synthase ◽

Rice Fields ◽

Target Site ◽

Herbicide Resistant ◽

Susceptible Populations ◽

High Level ◽

Als Inhibitor ◽

Encoding Genes

Abstract Barnyardgrass (Echinochloa crus-galli) is a noxious grass weed which infests rice fields and causes huge crop yield losses. In this study, we collected twelve E. crus-galli populations from rice ﬁelds of Ningxia province in China and investigated the resistance levels to acetolactate synthase (ALS) inhibitor penoxsulam and acetyl-CoA carboxylase (ACCase) inhibitor cyhalofop-butyl. The results showed that eight populations exhibited resistance to penoxsulam and four populations evolved resistance to cyhalofop-butyl. Moreover, all of the four cyhalofop-butyl-resistant populations (NX3, NX4, NX6 and NX7) displayed multiple-herbicide-resistance (MHR) to both penoxsulam and cyhalofop-butyl. The alternative herbicides bispyribac-sodium, metamifop and fenoxaprop-P-ethyl cannot effectively control the MHR plants. To characterize the molecular mechanisms of resistance, we ampliﬁed and sequenced the target-site encoding genes in resistant and susceptible populations. Partial sequences of three ALS genes and six ACCase genes were examined. A Trp-574-Leu mutation was detected in EcALS1 and EcALS3 in two high-level (65.84- and 59.30-fold) penoxsulam-resistant populations NX2 and NX10, respectively. In addition, one copy (EcACC4) of ACCase genes encodes a truncated aberrant protein due to a frameshift mutation in E. crus-galli populations. None of amino acid substitutions that are known to confer herbicide resistance were detected in ALS and ACCase genes of MHR populations. Our study reveals the widespread of multiple-herbicide resistant E. crus-galli populations at Ningxia province of China that exhibit resistance to several ALS and ACCase inhibitors. Non-target-site based mechanisms are likely to be involved in E. crus-galli resistance to the herbicides, at least in four MHR populations.

Download Full-text

Target-Site Mutations and Expression of ALS Gene Copies Vary According to Echinochloa Species

Genes ◽

10.3390/genes12111841 ◽

2021 ◽

Vol 12 (11) ◽

pp. 1841

Author(s):

Silvia Panozzo ◽

Elisa Mascanzoni ◽

Laura Scarabel ◽

Andrea Milani ◽

Giliardi Dalazen ◽

...

Keyword(s):

Cropping Systems ◽

Point Mutations ◽

Common Species ◽

Target Site ◽

The Other ◽

Gene Copy ◽

Resistance Level ◽

Resistance Evolution ◽

Gene Copies ◽

Als Inhibitors

The sustainability of rice cropping systems is jeopardized by the large number and variety of populations of polyploid Echinochloa spp. resistant to ALS inhibitors. Better knowledge of the Echinochloa species present in Italian rice fields and the study of ALS genes involved in target-site resistance could significantly contribute to a better understanding of resistance evolution and management. Using a CAPS-rbcL molecular marker, two species, E. crus-galli (L.) P. Beauv. and E. oryzicola (Vasinger) Vasing., were identified as the most common species in rice in Italy. Mutations involved in ALS inhibitor resistance in the different species were identified and associated with the ALS homoeologs. The relative expression of the ALS gene copies was evaluated. Molecular characterization led to the identification of three ALS genes in E. crus-galli and two in E. oryzicola. The two species also carried different point mutations conferring resistance: Ala122Asn in E. crus-galli and Trp574Leu in E. oryzicola. Mutations were carried in the same gene copy (ALS1), which was significantly more expressed than the other copies (ALS2 and ALS3) in both species. These results explain the high resistance level of these populations and why mutations in the other ALS copies are not involved in herbicide resistance.

Download Full-text

Target site–based penoxsulam resistance in barnyardgrass (Echinochloa crus-galli) from China

Weed Science ◽

10.1017/wsc.2019.5 ◽

2019 ◽

Vol 67 (3) ◽

pp. 281-287 ◽

Cited By ~ 1

Author(s):

Jiapeng Fang ◽

Tingting Liu ◽

Yuhua Zhang ◽

Jun Li ◽

Liyao Dong

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Acetolactate Synthase ◽

Target Site ◽

Cross Resistance ◽

Predicted Amino Acid Sequence ◽

Sequence Analyses ◽

Als Inhibitors ◽

Higher Sensitivity

AbstractBarnyardgrass [Echinochloa crus-galli (L.) P. Beauv.] is acknowledged to be the most troublesome weed in rice fields in Anhui and Jiangsu provinces of China. It cannot be effectively controlled using certain acetolactate synthase (ALS)-inhibiting herbicides, including penoxsulam. Echinochloa crus-galli samples with suspected resistance to penoxsulam were collected to identify the target site–based mechanism underlying this resistance. Populations AXXZ-2 and JNRG-2 showed 33- and 7.3-fold resistance to penoxsulam, respectively, compared with the susceptible JLGY-3 population. Cross-resistance to other ALS inhibitors was reported in AXXZ-2 but not in JNRG-2, and occasionally showed higher sensitivity than JLGY-3. In vitro ALS activity assays revealed that penoxsulam concentrations required to inhibit 50% of ALS activity were 11 and 5.2 times greater in AXXZ-2 and JNRG-2, respectively, than in JLGY-3. DNA and predicted amino acid sequence analyses of ALS revealed Ala-205-Val and Ala-122-Gly substitutions in AXXZ-2 and JNRG-2, respectively. Our results indicate that these substitutions in ALS are at least partially responsible for resistance to penoxsulam.

Download Full-text

Resistance to Bensulfuron-Methyl in Water Plantain (Alisma plantago-aquatica) Populations from Chilean Paddy Fields

Weed Technology ◽

10.1614/wt-08-078.1 ◽

2008 ◽

Vol 22 (4) ◽

pp. 602-608 ◽

Cited By ~ 2

Author(s):

Rodrigo Figueroa ◽

Marlene Gebauer ◽

Albert Fischer ◽

Marcelo Kogan

Keyword(s):

Weed Management ◽

Wide Spectrum ◽

Acetolactate Synthase ◽

Rice Fields ◽

Integrated Weed Management ◽

Cross Resistance ◽

Content Type ◽

Als Inhibitors

Bensulfuron-methyl (BSM) has been one of the most widely used herbicides in Chilean rice fields because it controls a wide spectrum of weeds and does not require field drainage for application. However, failures of BSM to control water plantain in rice fields have been noted since 2002. We assessed BSM effects on suspected resistant (CU1 and CU2) and susceptible (AN1) water plantain accessions collected in Chilean rice fields during 2004 and 2005. BSM rates resulting in 50% growth reduction (GR50) of CU2 and CU1 plants were 12- and 33-fold higher than for AN1 plants, respectively. Acetolactate synthase (ALS) activity assays in vitro suggested resistance in CU1 and CU2 was due to an ALS enzyme with reduced BSM sensitivity compared to the AN1 biotype. Resistance indices (RI), or ratios of the resistant to susceptibleI50values (BSM rate to inhibit ALS-enzyme activity by 50%), were 266 (CU2/AN1) and > 38,462 (CU1/AN1). This agreed with in vivo ALS activity assays whereRIwere 224 (CU2/AN1) and > 8,533 (CU1/AN1). Resistance levels detected in whole-plant or in vivo ALS activity assays were orders of magnitude lower than those detected in in vitro ALS activity studies suggesting nontarget site mechanisms may have mitigated BSM toxicity. However, a consistent ranking of BSM sensitivity levels (AN1 > CU2 > CU1) throughout all three types of assays suggests resistance is primarily endowed by low target site sensitivity. We conclude that susceptible and resistant water plantain biotypes coexist in Chilean paddies, and the use of integrated weed management involving herbicides with a different mode of action would be imperative to prevent further evolution of resistance to BSM and possibly cross-resistance to other ALS inhibitors. In vitro ALS-enzyme assays provided the best discrimination of resistance levels between biotypes.

Download Full-text

Multiple Pro197ALS Substitutions Endow Resistance to ALS Inhibitors within and among Mayweed Chamomile Populations

Weed Science ◽

10.1614/ws-d-10-00146.1 ◽

2011 ◽

Vol 59 (3) ◽

pp. 431-437 ◽

Cited By ~ 18

Author(s):

Suphannika Intanon ◽

Alejandro Perez-Jones ◽

Andrew G. Hulting ◽

Carol A. Mallory-Smith

Keyword(s):

Pacific Northwest ◽

Genotypic Variation ◽

Point Mutations ◽

Acetolactate Synthase ◽

Cross Resistance ◽

Collection Site ◽

Whole Plant ◽

The Pacific ◽

Als Inhibitors

Mayweed chamomile seeds were collected from six different fields across the Pacific Northwest. All populations (each collection site was considered a population) were suspected to have some level of acetolactate synthase (ALS) resistance. Greenhouse and laboratory studies were conducted to determine if these populations were resistant to three different classes of ALS inhibitors: sulfonylureas (SU), imidazolinones (IMI), and triazolopyrimidines (TP). A whole-plant dose–response andin vitroALS activity studies confirmed cross-resistance to thifensulfuron + tribenuron/chlorsulfuron (SU), imazethapyr (IMI), and cloransulam (TP); however, resistance varied by herbicide class and population. TwoALSisoforms of theALSgene (ALS1andALS2) were identified in mayweed chamomile; however, only mutations inALS1were responsible for resistance. No mutations were found inALS2. Sequence analysis of the partialALSgene identified four point mutations at position 197 (Pro197to Leu, Gln, Thr, or Ser) in the resistant populations. This study demonstrates genotypic variation associated with cross-resistance to ALS inhibitors within and between populations.

Download Full-text

The occurrence of herbicide-resistant Avena fatua (wild oats) populations to ACCase-inhibiting herbicides in Ireland

Irish Journal of Agricultural and Food Research ◽

10.15212/ijafr-2020-0127 ◽

2021 ◽

Author(s):

R. Byrne ◽

A.V. Vijaya Bhaskar ◽

J. Spink ◽

R. Freckleton ◽

P. Neve ◽

...

Keyword(s):

Herbicide Resistance ◽

Weed Management ◽

Resistance Mechanism ◽

Point Mutations ◽

Acetolactate Synthase ◽

Avena Fatua ◽

Integrated Weed Management ◽

Cross Resistance ◽

Wild Oats ◽

Als Inhibitor

Following growers’ reports of herbicide control problems, populations of 30 wild oats, Avena fatua, were collected from the south-east main arable counties of Ireland in 2016 and investigated for the occurrence and potential for herbicide resistance to acetyl-CoA carboxylase (ACCase) inhibitors pinoxaden, propaquizafop and cycloxydim, as well as acetolactate synthase (ALS) inhibitor mesosulfuron + iodosulfuron. Plant survival ≥20% was considered as the discriminating threshold between resistant and susceptible populations, when plants were treated with full recommended field rates of ACCase/ALS inhibitors. Glasshouse sensitivity screens revealed 2 out of 30 populations were cross-resistant to all three ACCase inhibitors. While three populations were cross-resistant to both pinoxaden and propaquizafop, and additionally, two populations were resistant to propaquizafop only. Different degree of resistance and cross-resistance between resistant populations suggest the involvement of either different point mutations or more than one resistance mechanism. Nevertheless, all populations including the seven ACCase-resistant populations were equally susceptible to ALS inhibitor. An integrated weed management (cultural/non-chemical control tactics and judicious use of herbicides) approach is strongly recommended to minimize the risk of herbicide resistance evolution.

Download Full-text