Target-site Basis for Fomesafen Resistance in Redroot Pigweed (Amaranthus retroflexus) from China

Weed Science ◽  
2021 ◽  
pp. 1-34
Author(s):  
Long Du ◽  
Xiao Li ◽  
Xiaojing Jiang ◽  
Qian Ju ◽  
Wenlei Guo ◽  
...  

Abstract Redroot pigweed (Amaranthus retroflexus L.) is a dominant weed in soybean [Glycine max (L.) Merr.] fields in Heilongjiang Province, China. High selective pressure caused by the extensive application of the protoporphyrinogen oxidase (PPO)-inhibiting herbicide fomesafen has caused A. retroflexus to evolve resistance to this herbicide. Two susceptible and two resistant populations (S1, S2, R1 and R2) were selected in this study to illustrate the target-site resistance mechanism in resistant A. retroflexus. Whole-plant bioassays indicated that R1 and R2 had evolved high-level resistance to fomesafen, with resistance factors of 27.0-27.9. Sequence alignment of the PPO gene showed an Arg128Gly substitution in PPX2. The basal expression differences of PPX1 and PPX2 between the S1 and R1 plants were essentially nonsignificant, whereas the basal expression of PPX2 in R2 plants was slightly lower than that in S1 plants. Compared to the PPX1 gene, the PPX2 gene maintained higher expression in the resistant plants after treatment with fomesafen. ELISA showed a similar basal PPO content between the susceptible and resistant plants without treatment. After fomesafen treatment, the PPO content decreased sharply in the susceptible plants compared with that in the resistant plants. Furthermore, after 24 h of treatment, the resistant plants showed an increased PPO content, whereas the susceptible plants had died. PPO2 mutation resulted in high extractable PPO activity and low sensitivity to fomesafen along with changes in the PPO enzyme kinetics. Although the mutant PPO2 exhibited increased Km values in the resistant plants, the Vmax values in these plants were also increased. Changes in the properties of the PPO enzyme due to an Arg128Gly substitution in PPX2, including changes in enzyme sensitivity and enzyme kinetics, are the target-site mechanism of resistance in A. retroflexus.

Weed Science ◽  
2019 ◽  
pp. 1-8
Author(s):  
Hao Wang ◽  
Hengzhi Wang ◽  
Ning Zhao ◽  
Baolin Zhu ◽  
Penglei Sun ◽  
...  

Abstract A redroot pigweed (Amaranthus retroflexus L.) population (HN-02) collected from Nenjiang County, Heilongjiang Province, exhibited multiple resistance to fomesafen and nicosulfuron. The purposes of this study were to characterize the herbicide resistance status of an HN-02 population for both acetolactate synthase (ALS) and protoporphyrinogen oxidase (PPO) inhibitors and the response to other herbicides and to investigate the target site-based mechanism governing fomesafen and nicosulfuron resistance. Three mutations, Ala-205-Val and Trp-574-Leu mutations in the ALS gene and an Arg-128-Gly mutation in the PPX2 gene, were identified in individual resistant plants. An HN-02F1-1 subpopulation homozygous for the Ala-205-Val and Arg-128-Gly mutations was generated, and whole-plant experiments confirmed multiple resistance to PPO inhibitors (fomesafen, fluoroglycofen-ethyl, and acifluorfen) and ALS inhibitors (imidazolinones [IMI], sulfonylureas [SU], and triazolopyrimidines [TP]) in the HN-02F1-1 plants, which presented resistance index values ranging from 8.3 to 110; however, these plants were sensitive to flumioxazin, fluroxypyr-meptyl, and 2,4-D butylate. In vitro ALS enzyme activity assays revealed that, compared with ALS from susceptible plants, ALS from the HN-02F1-1 plants was 15-, 28- and 320-fold resistant to flumetsulam, nicosulfuron, and imazethapyr, respectively. This study confirms the first case of multiple resistance to PPO and ALS inhibitors in A. retroflexus and determines that the target-site resistance mechanism was produced by Ala-205-Val and Arg-128-Gly mutations in the ALS gene and PPX2 gene, respectively. In particular, the Ala-205-Val mutation was found to endow resistance to three classes of ALS inhibitors: TP, SU, and IMI.


Weed Science ◽  
2020 ◽  
pp. 1-20
Author(s):  
Yi Cao ◽  
Shouhui Wei ◽  
Hongjuan Huang ◽  
Wenyu Li ◽  
Chaoxian Zhang ◽  
...  

Abstract Redroot pigweed (Amaranthus retroflexus L.) is a troublesome dicot weed species widely distributed across China. A population of A. retroflexus that survived the recommended label rate of thifensulfuron-methyl was collected from the main soybean [Glycine max (L.) Merr.] production area in China. Whole-plant dose-response assays indicated that the resistant (R) population was highly resistance (61.80-fold) to thifensulfuron-methyl compared to the S1 population. In vitro ALS activity experiments showed that the thifensulfuron-methyl I50 value for the R population was 40.17 times higher than that for the S1 population. A preliminary malathion treatment study indicated that the R population might have cytochrome P450-mediated metabolic resistance. The R population exhibited a high level of cross-resistance to representative ALS herbicides (imazethapyr, flumetsulam, and bispyribac-sodium) and multiple resistance to the commonly used PPO-inhibiting herbicides lactofen and fomesafen. Two common mutations, Trp-574-Leu in ALS and Arg-128-Gly in PPO2, were identified within the R population. This study identified possible enhanced metabolism of thifensulfuron-methyl co-existing with target site mutations in both ALS and PPO2 in a multiple-resistant A. retroflexus population.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ingvild Loubet ◽  
Laëtitia Caddoux ◽  
Séverine Fontaine ◽  
Séverine Michel ◽  
Fanny Pernin ◽  
...  

AbstractAmbrosia artemisiifolia L. (common ragweed) is a globally invasive, allergenic, troublesome arable weed. ALS-inhibiting herbicides are broadly used in Europe to control ragweed in agricultural fields. Recently, ineffective treatments were reported in France. Target site resistance (TSR), the only resistance mechanism described so far for ragweed, was sought using high-throughput genotyping-by-sequencing in 213 field populations randomly sampled based on ragweed presence. Additionally, non-target site resistance (NTSR) was sought and its prevalence compared with that of TSR in 43 additional field populations where ALS inhibitor failure was reported, using herbicide sensitivity bioassay coupled with ALS gene Sanger sequencing. Resistance was identified in 46 populations and multiple, independent resistance evolution demonstrated across France. We revealed an unsuspected diversity of ALS alleles underlying resistance (9 amino-acid substitutions involved in TSR detected across 24 populations). Remarkably, NTSR was ragweed major type of resistance to ALS inhibitors. NTSR was present in 70.5% of the resistant plants and 74.1% of the fields harbouring resistance. A variety of NTSR mechanisms endowing different resistance patterns evolved across populations. Our study provides novel data on ragweed resistance to herbicides, and emphasises that local resistance management is as important as mitigating gene flow from populations where resistance has arisen.


2020 ◽  
Vol 34 (3) ◽  
pp. 447-453
Author(s):  
Sushila Chaudhari ◽  
Vijay K. Varanasi ◽  
Sridevi Nakka ◽  
Prasanta C. Bhowmik ◽  
Curtis R. Thompson ◽  
...  

AbstractThe evolution of resistance to multiple herbicides in Palmer amaranth is a major challenge for its management. In this study, a Palmer amaranth population from Hutchinson, Kansas (HMR), was characterized for resistance to inhibitors of photosystem II (PSII) (e.g., atrazine), acetolactate synthase (ALS) (e.g., chlorsulfuron), and EPSP synthase (EPSPS) (e.g., glyphosate), and this resistance was investigated. About 100 HMR plants were treated with field-recommended doses (1×) of atrazine, chlorsulfuron, and glyphosate, separately along with Hutchinson multiple-herbicide (atrazine, chlorsulfuron, and glyphosate)–susceptible (HMS) Palmer amaranth as control. The mechanism of resistance to these herbicides was investigated by sequencing or amplifying the psbA, ALS, and EPSPS genes, the molecular targets of atrazine, chlorsulfuron, and glyphosate, respectively. Fifty-two percent of plants survived a 1× (2,240 g ai ha−1) atrazine application with no known psbA gene mutation, indicating the predominance of a non–target site resistance mechanism to this herbicide. Forty-two percent of plants survived a 1× (18 g ai ha−1) dose of chlorsulfuron with proline197serine, proline197threonine, proline197alanine, and proline197asparagine, or tryptophan574leucine mutations in the ALS gene. About 40% of the plants survived a 1× (840 g ae ha−1) dose of glyphosate with no known mutations in the EPSPS gene. Quantitative PCR results revealed increased EPSPS copy number (50 to 140) as the mechanism of glyphosate resistance in the survivors. The most important finding of this study was the evolution of resistance to at least two sites of action (SOAs) (~50% of plants) and to all three herbicides due to target site as well as non–target site mechanisms. The high incidence of individual plants with resistance to multiple SOAs poses a challenge for effective management of this weed.


2021 ◽  
Vol 12 ◽  
Author(s):  
Zeineb Hada ◽  
Yosra Menchari ◽  
Antonia M. Rojano-Delgado ◽  
Joel Torra ◽  
Julio Menéndez ◽  
...  

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.


2020 ◽  
pp. 1-7
Author(s):  
Jacob S. Montgomery ◽  
Darci A. Giacomini ◽  
Patrick J. Tranel

Abstract During the 2017 to 2019 growing seasons, samples of waterhemp and Palmer amaranth that had reportedly survived field-rate applications of protoporphyrinogen oxidase (PPO)–inhibiting herbicides were collected from the American Midwest and tested for target-site mutations known at the time to confer resistance. Target-site resistance was identified in nearly all (135 of 145) tested common waterhemp populations but in only 8 of 13 Palmer amaranth populations. Follow-up research on one population of Palmer amaranth (W-8), which tested negative for all such mutations, confirmed it was resistant to lactofen, with a magnitude of resistance comparable to that conferred by the ΔG210 PPO2 mutation. Gene sequences from both isoforms of PPO (PPO1 and PPO2) were compared between W-8 and known PPO inhibitor–sensitive sequence. A glycine-to-alanine substitution at the 399th amino acid position (G399A) of PPO2, recently identified to reduce target-site herbicide sensitivity, was observed in a subset of resistant W-8 plants. Because no missense mutation completely delimited resistant and sensitive sequences, we initially suspected the presence of a secondary, non-target-site resistance mechanism in this population. To isolate G399A, a segregating F2 population was produced and screened with a delimiting rate of lactofen. χ2 goodness-of-fit analysis of dead/alive ratings indicated single-locus inheritance of resistance in the F2 population, and molecular markers for the W-8 parental PPO2 coding region co-segregated tightly, but not perfectly, with resistance. More research is needed to fully characterize Palmer amaranth PPO inhibitor–resistance mechanisms, which appear to be more diverse than those found in common waterhemp.


2020 ◽  
Vol 29 (2) ◽  
pp. 79-96
Author(s):  
Sava Vrbničanin

Weed resistance to herbicides represents the acquired resistance of individuals to complete the life cycle and leave offspring in the conditions of extended exposure to the same herbicide, i.e. herbicides of the same mechanism of action to which they were sensitive at the beginning of the application. Based on the herbicide resistance mechanisms, all processes can be grouped as follows: target-site resistance, non-target-site resistance, cross-resistance and multiple-resistance. Currently, herbicide resistance has been reported in 514 cases (species x site of action) worldwide, in 262 weed species (152 dicotyledons, 110 monocotyledons). Many of those biotypes are resistant to als inhibitors, PS II inhibitors, EPSPS inhibitors and ACC-ase inhibitors. The higher degree of resistance to als inhibitors has been confirmed in the following weed species: Amaranthus retroflexus, Sorghum halepense, Ambrosia artemisiifolia and Helianthus annuus.


Weed Science ◽  
2017 ◽  
Vol 66 (2) ◽  
pp. 149-158 ◽  
Author(s):  
Vijay K. Varanasi ◽  
Chad Brabham ◽  
Jason K. Norsworthy ◽  
Haozhen Nie ◽  
Bryan G. Young ◽  
...  

Palmer amaranth is one of the most problematic weeds in the midsouthern United States, and the evolution of resistance to protoporphyrinogen oxidase (PPO) inhibitors in biotypes already resistant to glyphosate and acetolactate synthase (ALS) inhibitors is a major cause of concern to soybean and cotton growers in these states. A late-season weed-escape survey was conducted in the major row crop–producing counties (29 counties) to determine the severity of PPO-inhibitor resistance in Arkansas. A total of 227 Palmer amaranth accessions were sprayed with fomesafen at 395 g ha−1to identify putative resistant plants. A TaqMan qPCR assay was used to confirm the presence of the ΔG210 codon deletion or the R128G/M (homologous to R98 mutation in common ragweed) target-site resistance mechanisms in thePPX2gene. Out of the 227 accessions screened, 44 were completely controlled with fomesafen, and 16 had only one or two severely injured plants (≥98% mortality) when compared with the 1986 susceptible check (100% mortality). The remaining 167 accessions were genotypically screened, and 82 (49%) accessions were found to harbor the ΔG210 deletion in thePPX2gene. The R128G was observed in 47 (28%) out of the 167 accessions screened. The mutation R128M, on the other hand was rare, found in only three accessions. About 13% of the accessions were segregating for both the ΔG210 and R128G mutations. Sixteen percent of the tested accessions had mortality ratings <90% and did not test positive for the ΔG210 or the R128G/M resistance mechanisms, indicating that a novel target or non–target site resistance mechanism is likely. Overall, PPO inhibitor–resistant Palmer amaranth is widespread in Arkansas, and the ΔG210 resistance mechanism is especially dominant in the northeast corridor, while the R128G mutation is more prevalent in counties near Memphis, TN.


2012 ◽  
Vol 26 (2) ◽  
pp. 341-347 ◽  
Author(s):  
Michael J. Walsh ◽  
Karrie Stratford ◽  
Kent Stone ◽  
Stephen B. Powles

The synergistic interaction between mesotrione, a hydroxyphenylpyruvate dioxygenase (HPPD)-inhibiting herbicide, and atrazine, a photosystem II (PS II)-inhibiting herbicide, has been identified in the control of several weed species. A series of dose–response studies examined the synergistic effect of these herbicides on a susceptible (S) wild radish population. The potential for this interaction to overcome target-sitepsbA gene-based atrazine resistance in a resistant (R) wild radish population was also investigated. Control of S wild radish with atrazine was enhanced by up to 40% when low rates (1.0 to 1.5 g ha−1) of mesotrione were applied in combination. This synergistic response was demonstrated across a range of atrazine–mesotrione rate combinations on this S wild radish population. Further, the efficacy of 1.5 g ha−1mesotrione increased control of the R population by a further 60% when applied in combination with 400 g ha−1of atrazine. This result clearly demonstrated the synergistic interaction of these herbicides in overcoming the target-site resistance mechanism. The mechanism responsible for the observed synergistic interaction between mesotrione and atrazine remains unknown. However, it is speculated that an alternate atrazine binding site may be responsible. Regardless of the biochemical nature of this interaction, evidence from whole-plant bioassays clearly demonstrated that synergistic herbicide combinations improve herbicide efficiency, with lower application rates required to control weed populations. This, combined with the potential to overcomepsbA gene-based triazine resistance, and, thereby, regain the use of these herbicides, will result in more sustainable herbicide use.


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