Next Article in Journal
Detecting Irrigation Events over Semi-Arid and Temperate Climatic Areas Using Sentinel-1 Data: Case of Several Summer Crops
Next Article in Special Issue
The Resistance Levels and Target-Site Based Resistance Mechanisms to Glyphosate in Eleusine indica from China
Previous Article in Journal
Varietal Variances of Grain Nitrogen Content and Its Relations to Nitrogen Accumulation and Yield of High-Quality Rice under Different Nitrogen Rates
Previous Article in Special Issue
Survey Reveals Frequency of Multiple Resistance to Tribenuron-Methyl, Bensulfuron-Methyl and Halosulfuron-Methyl in Cleavers (Galium aparine L.)
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Agronomy
Volume 12
Issue 11
10.3390/agronomy12112724
agronomy-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Metabolic Resistance to Acetyl-CoA Carboxylase-Inhibiting Herbicide Cyhalofop-Butyl in a Chinese Echinochloa crus-galli Population

by
Qian Yang
1,
Xia Yang
2,
Jinlei Zhu
1,
Tian Wei
1,
Min Lv
1,* and
Yongfeng Li
2,*
1
Institute of Agricultural Sciences for Lixiahe Region in Jiangsu, Yangzhou 225007, China
2
Institute of Crop Germplasm and Biotechnology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
*
Authors to whom correspondence should be addressed.
Agronomy 2022, 12(11), 2724; https://doi.org/10.3390/agronomy12112724
Submission received: 27 September 2022 / Revised: 22 October 2022 / Accepted: 1 November 2022 / Published: 2 November 2022
(This article belongs to the Special Issue Herbicides Toxicology and Weeds Herbicide-Resistant Mechanism)
Browse Figures
Versions Notes

Abstract

:
A population of Echinochloa crus-galli (L.) P. Beauv obtained from direct-seeding rice fields in Jiangxi Province, China, exhibited high resistance levels (13.5-fold) to the acetyl-CoA carboxylase (ACCase)-inhibiting herbicide cyhalofop-butyl. Compared with the susceptible (S) population, this resistant (R) population evolved a cross-resistance to aryloxyphenoxypropionates (APPs) herbicides metamifop (2.9-fold) and fenoxapro-p-ethyl (4.1-fold), cyclohexanediones (CHDs) herbicide clethodim (4.7-fold), phenyl pyrazoline (DEN) herbicide pinoxaden (6.4-fold), and evolved multiple-resistance to acetolactate synthase (ALS)-inhibiting herbicide penoxsulam (3.6-fold), and auxin mimic herbicides quinclorac (>34.7-fold) and florpyrauxifen-benzyl (2.4-fold). ACCase gene sequencing did not reveal the existence of any known mutation point conferring with herbicide resistance. In addition, three metabolic inhibitors—one glutathione—S-transferase (GST) inhibitor (NBD-Cl), and two cytochrome P450 inhibitors (malathion and PBO)—did not reverse the cyhalofop-butyl resistance. Furthermore, enhanced metabolic rates of more than 60% 24 h after treatment with the active compound cyhalofop acid was observed in R plants compared to S plants. Hence, enhanced metabolism activity endows a non-target-site resistance to cyhalofop-butyl in the R population of E. crus-galli. Future research will be required to determine what metabolizing enzyme genes are responsible for cyhalofop-butyl resistance in E. crus-galli.

1. Introduction

Rice (Oryza sativa L.) is one of the main grain crops in China. In recent years, direct-seeding rice has become one of the main rice cultivation methods in China due to its high mechanization and low cost [1]. Echinochloa crus-galli (L.) P. Beauv infected direct-seeding rice fields widely and severely in China. Studies have shown that E. crus-galli, with a density of 4 plants m−2, reduced the rice grain yield by 55.2%, and the rice taste quality declined significantly [2]. At present, chemical herbicides are still the main means to control E. crus-galli in rice fields in China, and acetyl-CoA carboxylase (ACCase)-inhibiting herbicides are one of most frequently used due to their high efficiency in controlling grass weeds.
ACCase-inhibiting herbicides include three different chemical classes: aryloxy phenoxy-propionates (APPs), cyclohexanediones (CHDs), and phenyl pyrazoline (DEN) [3]. Cyhalofop-butyl is the only herbicide of APPs with high safety to rice, and it has a good control effect on grass weeds, including E. crus-galli and Leptochloa chinensis in rice fields. Cyhalofop-butyl can be absorbed by the leaf and sheath of plants and translocated through the plant phloem. Afterwards, it can be rapidly degraded into the active compound cyhalofop acid [4]. Then, cyhalofop acid inhibits the activity of the ACCase enzyme, so that the fatty acid synthesis stops and cell growth and division cannot proceed normally, leading to the death of the weeds. ACCase-inhibiting herbicides are prone to herbicide resistance evolution [5]. Several grass weeds from rice fields, including L. chinensis, Diplachne fusca, and Eleusine indica, have been reported to be evolving a resistance to cyhalofop-butyl [6,7,8,9,10,11,12]. The cases of cyhalofop-butyl-resistant E. crus-galli have been increasing in China over recent years [13,14].
Weed resistance to ACCase-inhibiting herbicides is mainly caused by ACCase gene mutations and/or enhanced herbicide metabolism [15,16]. The Ile-1781-Leu and Asp-2078-Glu mutation in the ACCase gene of E. crus-galli have been demonstrated to confer target-site resistance (TSR) to ACCase inhibitors [13,17]. In addition, some studies found that ACCase mutations were not detected in herbicide-resistant E. crus-galli populations, and non-target-site resistance (NTSR) mechanisms may be involved in these populations [14,18,19]. NTSR mainly involves herbicide absorption, translocation, and metabolism [19]. In particular, the NTSR mechanism by enhanced metabolism is associated with plant detoxifying enzymes, such as cytochrome P450, glutathione-S-transferase (GST), and glucosyltransferases [20]. Compared to TSR mechanisms, very little genes were identified in NTSR mechanisms in Echinochloa spp. species [21,22,23,24]. In addition, potential NTSR in weeds can be due to multiple resistances to widespread herbicides, causing more difficulties to control resistant weeds.
In this study, we found an E. crus-galli population exhibiting high resistance level to cyhalofop-butyl. Therefore, we aimed to: (1) determine the cross- or multiple-resistance patterns to ACCase-inhibiting, ALS-inhibiting, and auxin mimic herbicides; (2) investigate the potential resistance mechanisms; (3) confirm the effect of three metabolic enzymes inhibitors—malathion, PBO, and NBD-Cl—on the resistance to cyhalofop-butyl.

2. Materials and Methods

2.1. Plant Material and Growth Conditions

The resistant (R) population of E. crus-galli was obtained in 2019 from a direct-seeding rice field in Jiangxi Province, China (116.09° E, 28.99° N), which was severely infected by this weed. The susceptible (S) population was from an area where herbicides have not been used (116.15° E, 28.98° N). Seeds of each population were collected from each field with at least 20 plants pooled.
E. crus-galli seeds were germinated at 30 °C. Then, germinated seedlings were transplanted to a mixture of organic matter and vermiculite (1:1 v:v) loaded in 7 cm-diameter pots, and grown in a greenhouse at 30 ± 2 °C with a 14 h: 10 h day: night photoperiod, and 65–75% relative humidity. When grown to a 2-to 3-leaf stage, the seedlings of each pot were thinned to 8 plants.

2.2. Dose-Response to Cyhalofop-Butyl

E. crus-galli plants with 3 true leaves were treated with cyhalofop-butyl using a sprayer delivering a water volume of 450 L ha−1 at 300 KPa with a fan nozzle positioned 58 cm above the plants. The cyhalofop-butyl rates for the S population were 0, 1.30, 3.89, 11.67, 35, 105 g ai ha−1, and for the R population were 0, 3.89, 11.67, 35, 105, 315 g ai ha−1. At 15 days after treatment, the aboveground fresh weights of S and R plants were measured. Three replications were set for each herbicide rate, and each experiment was carried out twice.

2.3. Response to ACCase Inhibitors

E. crus-galli seedlings were grown and treated by herbicide solutions at 3-leaf stage under the conditions described above. Four herbicides that have the same mechanism of action as cyhalofop-butyl were used to determine the cross-resistance to ACCase inhibitors, namely metamifop and fenoxaprop-p-ethyl (APPs), clethodim (CHDs), and pinoxaden (DEN). The herbicides’ commercial information and application rates for S and R populations are listed in Table 1. The fresh shoot weights were recorded 15 days later, and the experiment was conducted twice with three replicates.

2.4. Response to Other Herbicides Used in Direct-Seeding Rice Fields

Four herbicides used for controlling grass weeds with post-emergence in direct-seeding rice fields were chosen to test the susceptibility of E. crus-galli. Commercial formulations of two ALS inhibitors, penoxsulam and bispyribac-sodium, and two auxin mimic herbicides, quinclorac and florpyrauxifen-benzyl, were applied at the 3-leaf stage with the dose gradients listed in Table 1. The fresh shoot weights were measured 15 days later. Each experiment was conducted twice with three replicates.

2.5. ACCase Genes Isolation and Sequencing

Genomic DNA was extracted from leaf tissues according to the instruction of the DNAsecure Plant Kit (Tiangen Biotech, Beijing, China). The ACCase gene fragments of E. crus-galli were amplified with primers, as reported by Yang et al. [14], which could cover all known mutation sites (Ile-1781, Trp-1999, Trp-2027, Ile-2041, Asp-2078, Cys-2088, and Gly-2096) responsible for resistance to ACCase inhibitors. The PCR mixture contained 25 μL of 2 × Taq PCR Mix, 1 μL template DNA, 1 μL of forward primer, 1 μL of reverse primer, and 22 μL ddH2O. The PCR profile was carried out as described by Yang et al. [14]. Afterwards, the amplified PCR products were purified, cloned into the pLB vector, and transformed into TOP 10 chemical competent cells (Tiangen Biotech, Beijing, China). In total, 5 S and 10 R plants were analyzed, and at least 10 transformed clones of each sample were selected and sequenced by Sangon Biotechnology (Shanghai, China) to obtain all of the six ACCase gene copies.

2.6. Cyhalofop-Butyl Metabolism in S and R Plants of E. crus-galli

When the plants had grown to the 3- to 4-leaf stage, they were foliar treated with cyhalofop-butyl at 1/3 of the field recommended application rate (35 g ai ha−1); 97% cyhalofop-butyl technical (Jiangsu Fenghua Chemical Industrial Co., Ltd., Yancheng, China) was dissolved in 1 mL acetone, diluted to the application rate with 0.2% Tween-80 aqueous solution, and sprayed under the conditions described above. Aboveground tissue samples of S and R plants were harvested at 2, 24, and 48 h after treatment (HAT), and frozen using liquid nitrogen. Five plants were pooled as a replicate, and each harvest time point was conducted with four replications.
Cyhalofop-butyl and its active metabolite cyhalofop acid were extracted from S and R samples according to the methods of Deng et al. [25]. The final supernatant obtained was analyzed by UPLC-MS/MS (Agilent Technologies 1200 Triple Quad 6460, Santa Clara, CA, USA) after being filtered through a 0.22 μm membrane. The extraction was separated with an Agilent C18 column (5 μm, 2.1 × 150 mm). A 0.01% formamide aqueous solution-acetonitrile mixture (v:v 20:80) was used as the mobile phase. The fragment ions of cyhalofop-butyl and cyhalofop acid are 256.2/120.2 m/z and 228.1/208.0 m/z, respectively.

2.7. Metabolic Inhibitors Effect on Resistance to Cyhalofop-Butyl

To preliminary evaluate the involvement of a metabolic enzyme in the metabolic resistance mechanism, plants at the 2- to 3-leaf stage from S and R populations were each pre-sprayed with malathion (J&K Scientific, Beijing, China), piperonylbutoxide (PBO, Shanghai Aladdin Biochemical Technology, Shanghai, China), and 4-chloro-7-nitrobenzofurazan (NBD-Cl, Shanghai Aladdin Biochemical Technology, Shanghai, China). The pre-experiment applications of malathion, PBO, and NBD-Cl at rates of 1000, 2000, and 300 g ai ha−1, respectively, displayed no visual effects on the growth of E. crus-galli. Malathion was prepared in deionized water. PBO and NBD-Cl were dissolved in 1 mL acetone, and diluted to the application rates with 0.2% Tween-80 aqueous solution. Cyhalofop-butyl was sprayed with the same doses as indicated by the dose-response curve. Malathion, PBO and NBD-Cl were sprayed 2 h, 4 h, and 48 h, before cyhalofop-butyl application [25,26]. At 21 days, the fresh shoot weights were measured, and the GR50 of S and R population to cyhalofop-butyl in the presence and absence of metabolic inhibitors was determined.

2.8. Statistical Analysis

The dose-response data of two repeated experiments were pooled as they showed no significance with the t-test (p < 0.05) (SPSS 20.0, IBM, Chicago, IL, USA). The herbicide dose causing 50% growth reduction (GR50) of plants was calculated using Equation (1) and SigmaPlot software (version 12.0, Systat Software, San Jose, CA, USA) [27].
y = C + (D − C)/[1 + (x/GR50)b]
In this equation, C is the lower limit, D is the upper limit, x is the herbicide rate, y is the fresh above-ground weight relative to non-treated control (%) response to x, and b is the slope around GR50. The GR50 ratio of the R population divided by the S population was calculated as the resistance index (RI). Data on cyhalofop-butyl metabolism were analyzed using SPSS 20.0 software and the analysis of variance (ANOVA).

3. Results

3.1. Resistance level to Cyhalofop-Butyl

The resistance level to cyhalofop-butyl of the R population was confirmed by the dose-response experiment (Figure 1). The S population was completely killed by 1/3 of the field recommended application dose (35 g ai ha−1), and the GR50 was calculated to be 5.6 g ai ha−1. The R population was still all alive at the highest rate (315 g ai ha−1) and exhibited > 20% growth relative to the untreated control at that rate. The GR50 value of the R population was 75.8 g ai ha−1 and the resistance index (RI) was 13.5.

3.2. Resistance Pattern to Different Herbicides

The R population exhibited cross-resistance to three classes of ACCase inhibitors (Table 2). All the four ACCase inhibitors tested could kill the S plants completely under the field recommended rate. Compared to the S population, the R population was moderately resistant to fenoxapro-p-ethyl (APPs, RI = 4.1), clethodim (CHDs, RI = 4.7) and pinoxaden (DEN, RI = 6.4), and lowly resistant to metamifop (APPs, RI = 2.9) (Table 2).
The susceptibility of herbicides with different modes of action was also determined with the whole plant experiments. The ALS inhibitor bispyribac-sodium completely controlled both the S and R plants at the field rate (45 g ai ha−1). On the contrary, the other three herbicides exhibited poor control effects on the R plants. The R population exhibited high-level resistance to auxin mimic herbicide quinclorac (RI > 34.7), moderate-level resistance to ALS inhibitor penoxsulam (RI = 8.9), and low-level resistance to auxin mimic herbicide florpyrauxifen-benzyl (RI = 2.4) (Table 2).

3.3. ACCase Gene Sequencing

Partial sequences of the carboxyltransferase domain of ACCase genes were amplified and examined. As E. crus-galli is hexaploid, all six transcripts were amplified. Sequence alignment results showed that ACCase 1 to ACCase 6 have more than 99% similarity in the nucleotide sequence. However, none of the mutation sites in ACCase genes conferring resistance were detected in the R population.

3.4. Enhanced Cyhalofop-Butyl Metabolism in R Plants

According to the results shown in Table 3, the transformation rate of cyhalofop-butyl into cyhalofop acid was similar in the two biotypes, because there was no significant difference in its content between S and R plants. The cyhalofop acid in S was 2.7- and 3-fold richer than that in R plants at 24 and 48 HAT, respectively, which means that the E. crus-galli plants of the R population could detoxify cyhalofop acid faster than that of S population.

3.5. Metabolic Inhibitors Eeffect on Cyhalofop-Butyl Resistance

To investigate the involvement of metabolic enzymes in the metabolic resistance, two P450 inhibitors (malathion and PBO) and a GST inhibitor (NBD-Cl) were applied prior to cyhalofop-butyl treatment. For the S population, there were no significant differences on plant growth between inhibitors pre-treatment and cyhalofop-butyl treatment alone, with the GR50 ranging from 2.7 to 4.3 g ai ha−1 (Figure 2). For the R population, the GR50 values to cyhalofop-butyl in the presence of malathion, PBO, and NBD-Cl were 37.5, 44.1, and 32.7 g ai ha−1, respectively, and showed no significant difference to that in the absence of inhibitors (33.2 g ai ha−1) (Figure 2). Hence, the three metabolic enzyme inhibitors did not weaken the resistance of the R E. crus-galli population to cyhalofop-butyl.

4. Discussion

This study demonstrated that a population of E. crus-galli, which was collected from direct-seeding rice fields in Jiangxi Province, China, induced a high resistance level to ACCase-inhibiting herbicide cyhalofop-butyl, with a RI value of 13.5. Dose-response experiments confirmed that the R population at the whole-plant level was resistant to three groups of ACCase inhibitors—metamifop and fenoxapro-p-ethyl (APPs), clethodim (CHDs), and pinoxaden (DEN). Furthermore, the R population also displayed multiple-resistances to herbicides with other modes of action used in rice fields, especially more than 34.7-fold resistance to the auxin mimic herbicide quinclorac and 8.9-fold resistance to the ALS-inhibiting herbicide penoxsulam, making it very challenging to control the population of E. crus-galli.
No known mutation sites responsible for herbicide resistance were detected in any of the six ACCase transcripts, which indicates the possible participation of NTSR mechanisms. In Echinochloa spp., there are only two reports of ACCase mutations, namely, Ile-1781-Leu and Asp-2078-Glu substitution [13,17,28]. In contrast, more resistant cases are involved in NTSR mechanisms [18,19,29]. A similar result was reported in a Lolium rigidum population from Spain, which showed multiple- and cross-resistance to ALS and ACCase inhibiting herbicides, and without mutations in the ACCase gene [30]. Our results also support this phenomenon. This may be associated with the dilution effect of multiple ACCase genes. Yu et al. [31] reported that Avena fatua carrying one mutant allele in the three ACCase copies only exhibited low-level resistance to ACCase-inhibiting herbicides. Therefore, NTSR may be more likely to evolve in weed species with multiple copies of target-site genes.
As a typical APP herbicide, cyhalofop-butyl is rapidly converted into its active metabolite, cyhalofop acid, after being absorbed by plants [4]. In our study, similar contents of cyhalofop-butyl were observed in S and R plants, indicating the same absorption and transformation rate of cyhalofop-butyl in the two populations. However, the content of cyhalofop acid in R plants was significantly reduced by 63% and 67% at 24 and 48 HAT, respectively, compared to the S plants, suggesting enhanced metabolism was the prime reason conferring resistance to cyhalofop-butyl. Our results are in accordance with the researches of other ACCase inhibitor resistant weeds [32,33,34,35,36,37]. Similar results were also observed in resistance to the auxin mimic herbicide florpyrauxifen-benzyl in this species, in which significant herbicide absorption and metabolism both contribute to the inability to control E. crus-galli [38].
Nevertheless, none of the tested metabolic enzyme inhibitors significantly affected resistance to cyhalofop-butyl in the R population. Similar results were reported in Echinochloa colona resistance to fenoxaprop-p-ethyl, which was not reversed by any of three metabolic inhibitors [26]. This is in contrast to some types of grass weeds, such as Bromus rigidus, L. rigidum, and Polypogon fugax, in which resistance could be greatly reduced by malathion or PBO treatment [39,40,41,42]. In L. chinensis, where a resistance mechanism with an enhanced herbicide metabolism has been identified, none of malathion, PBO, or NBD-Cl inhibited cyhalofop-butyl resistance [25]. This suggests that these three metabolic inhibitors may not inhibit the activity of all enzymes involved in weed resistance. Although one cytochrome P450 enzyme, CYP81A68, has been identified in E. crus-galli to endow metabolic resistance to ACCase-inhibiting herbicides [24], there are still many enzymes responsible for NTSR need to be verified. Therefore, additional work, such as RNA-sequence analyses and gene expression assays, will be required to determine what metabolizing enzyme genes are responsible for cyhalofop-butyl resistance in E. crus-galli.
In conclusion, the E. crus-galli population collected from direct-seeding rice fields in China exhibited a high resistance level to cyhalofop-butyl, and the enhanced metabolism conferred an NTSR in this population. It should be noted that the R population had also evolved cross-resistance to three classes of ACCase inhibitors and multiple-resistance to ALS inhibitors and auxin mimic herbicides, which highlights the difficulties in selecting alternative post-emergence herbicides to control the resistant weeds. The emergence of multiple-resistance E. crus-galli populations were reported in several regions of China in recent years, including Anhui, Jiangsu, Hunan, and Ningxia Province [13,14,24,43], and should remind Chinese growers of the evolving multiple herbicide resistance risk in E. crus-galli populations to most available herbicides used in rice fields of China. Therefore, non-chemical methods, such as reducing the seed bank and using bioherbicides, and integrated management strategies should be adopted to reduce the pressure of herbicide selection and dependence, particularly with regard to dealing with this species that is highly prone to herbicide resistance.

Author Contributions

Conceptualization, Y.L. and M.L.; methodology, Q.Y.; software, X.Y.; validation, Q.Y., X.Y. and T.W.; formal analysis, Q.Y.; investigation, Q.Y.; resources, X.Y.; data curation, Q.Y.; writing—original draft preparation, Q.Y.; writing—review and editing, X.Y.; visualization, J.Z.; supervision, J.Z.; project administration, Q.Y.; funding acquisition, Y.L. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the National Natural Science Foundation, grant number 31901904 and 32072435.

Data Availability Statement

The data for this study are available upon request to the corresponding author.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Dong, L.Y.; Gao, Y.; Fang, J.P.; Chen, G.Q. Research progress on the herbicide-resistance of weeds in rice fields in China. Plant Prot. 2018, 44, 69–76. [Google Scholar]
  2. Zhang, Z.; Gu, T.; Zhao, B.; Yang, X.; Peng, Q.; Li, Y.; Bai, L. Effects of common Echinochloa varieties on grain yield and grain quality of rice. Field Crops Res. 2017, 203, 163–172. [Google Scholar] [CrossRef]
  3. Sasaki, Y.; Konishi, T.; Nagano, Y. The compartmentation of acetyl-coenzyme A carboxylase in plants. Plant Physiol. 1995, 108, 445–449. [Google Scholar] [CrossRef] [PubMed]
  4. Ottis, B.V.; Mattice, J.D.; Talbert, R.E. Determination of antagonism between cyhalofop-butyl and other rice (Oryza sativa) herbicides in barnyardgrass (Echinochloa crus-galli). J. Agric. Food Chem. 2005, 53, 4064–4068. [Google Scholar] [CrossRef] [PubMed]
  5. Peterson, M.A.; Collavo, A.; Ovejero, R.; Shivrain, V.; Walsh, M.J. The challenge of herbicide resistance around the world: A current summary. Pest Manag. Sci. 2018, 74, 2246–2259. [Google Scholar] [CrossRef]
  6. Yu, J.; Gao, H.; Pan, L.; Yao, Z.; Dong, L. Mechanism of resistance to cyhalofop-butyl in Chinese sprangletop (Leptochloa chinensis (L.) Nees). Pestic. Biochem. Physiol. 2017, 143, 306–311. [Google Scholar] [CrossRef]
  7. Deng, W.; Cai, J.; Zhang, J.; Chen, Y.; Chen, Y.; Di, Y.; Yuan, S. Molecular basis of resistance to ACCase-inhibiting herbicide cyhalofop-butyl in Chinese sprangletop (Leptochloa chinensis (L.) Nees) from China. Pestic. Biochem. Physiol. 2019, 158, 143–148. [Google Scholar] [CrossRef]
  8. Yuan, S.; Di, Y.; Chen, Y.; Chen, Y.; Cai, J.; Deng, W. Target-site resistance to cyhalofop-butyl in bearded sprangletop (Diplachne fusca) from China. Weed Sci. 2019, 67, 534–538. [Google Scholar] [CrossRef]
  9. Deng, W.; Yang, Q.; Chen, Y.; Yang, M.; Xia, Z.; Zhu, J.; Chen, Y.; Cai, J.; Yuan, S. Cyhalofop-butyl and glyphosate multiple-herbicide resistance evolved in an Eleusine indica population collected in Chinese direct-seeding rice. J. Agric Food Chem. 2020, 68, 2623–2630. [Google Scholar] [CrossRef]
  10. Peng, Y.; Pan, L.; Liu, D.; Cheng, X.; Ma, G.; Li, S.; Liu, X.; Wang, L.; Bai, L. Confirmation and characterization of cyhalofop-butyl–resistant Chinese sprangletop (Leptochloa chinensis) populations from China. Weed Sci. 2020, 68, 253–259. [Google Scholar] [CrossRef]
  11. Chen, K.; Peng, Y.; Zhang, L.; Wang, L.; Mao, D.; Zhao, Z.; Bai, L.; Wang, L. Whole transcriptome analysis resulted in the identification of Chinese sprangletop (Leptochloa chinensis) genes involved in cyhalofop-butyl tolerance. BMC Genom. 2021, 22, 1–14. [Google Scholar] [CrossRef] [PubMed]
  12. Jiang, M.; Wang, Y.; Li, W.; Li, Q.; Zhang, J.; Liao, M.; Zhao, N.; Cao, H. Investigating resistance levels to cyhalofop-butyl and mechanisms involved in Chinese sprangletop (Leptochloa chinensis L.) from Anhui Province, China. Pestic. Biochem. Physiol. 2022, 186, 105165. [Google Scholar] [CrossRef] [PubMed]
  13. Fang, J.; He, Z.; Liu, T.; Li, J.; Dong, L. A novel mutation Asp-2078-Glu in ACCase confers resistance to ACCase herbicides in barnyardgrass (Echinochloa crus-galli). Pestic. Biochem. Physiol. 2020, 168, 104634. [Google Scholar] [CrossRef] [PubMed]
  14. Yang, Q.; Yang, X.; Zhang, Z.; Wang, J.; Fu, W.; Li, Y. Investigating the resistance levels and mechanisms to penoxsulam and cyhalofop-butyl in barnyardgrass (Echinochloa crus-galli) from Ningxia Province, China. Weed Sci. 2021, 69, 422–429. [Google Scholar] [CrossRef]
  15. Powles, S.B.; Yu, Q. Evolution in action: Plants resistant to herbicides. Ann. Rev. Plant. Physiol. 2010, 61, 317–347. [Google Scholar] [CrossRef] [Green Version]
  16. Kaundun, S.S. Resistance to acetyl-CoA carboxylase-inhibiting herbicides. Pest Manag. Sci. 2014, 70, 1405–1417. [Google Scholar] [CrossRef]
  17. Huan, Z.B.; Jin, T.; Zhang, S.Y.; Wang, J.X. Cloning and sequence analysis of plastid acetyl-CoA carboxylase cDNA from two Echinochloa crusgalli biotypes. J. Pestic. Sci. 2011, 36, 461–466. [Google Scholar] [CrossRef] [Green Version]
  18. Iwakami, S.; Hashimoto, M.; Matsushima, K.; Watanabe, H.; Hamamura, K.; Uchino, A. Multiple-herbicide resistance in Echinochloa crus-galli var. formosensis, an allohexaploid weed species, in dry-seeded rice. Pestic. Biochem. Physiol. 2015, 119, 1–8. [Google Scholar] [CrossRef]
  19. Hwang, J.I.; Norsworthy, J.K.; González-Torralva, F.; Piveta, L.B.; Priess, G.L.; Barber, L.T.; Butts, T.R. Absorption, translocation, and metabolism of florpyrauxifen-benzyl and cyhalofop-butyl in cyhalofop-butyl-resistant barnyardgrass [Echinochloa crus-galli (L.) P. Beauv.]. Pestic. Biochem. Physiol. 2020, 180, 104999. [Google Scholar] [CrossRef]
  20. Ghanizadeh, H.; Harrington, K.C. Non-target Site Mechanisms of Resistance to Herbicides. Crit. Rev. Plant Sci. 2017, 36, 24–34. [Google Scholar] [CrossRef]
  21. Iwakami, S.; Endo, M.; Saika, H.; Okuno, J.; Nakamura, N.; Yokoyama, M.; Watanabe, H.; Toki, S.; Uchino, A.; Inamura, T. Cytochrome P450 CYP81A12 and CYP81A21 are associated with resistance to two acetolactate synthase inhibitors in Echinochloa phyllopogon. Plant Physiol. 2014, 165, 618–629. [Google Scholar] [CrossRef] [PubMed]
  22. Iwakami, S.; Kamidate, Y.; Yamaguchi, T.; Ishizaka, M.; Endo, M.; Suda, H.; Nagai, K.; Sunohara, Y.; Toki, S.; Uchino, A.; et al. CYP81A P450s are involved in concomitant cross-resistance to acetolactate synthase and acetyl-CoA carboxylase herbicides in Echinochloa phyllopogon. New Phytol. 2019, 221, 2112–2122. [Google Scholar] [CrossRef] [PubMed]
  23. Dimaano, N.G.; Yamaguchi, T.; Fukunishi, K.; Tominaga, T.; Iwakami, S. Functional characterization of cytochrome P450 CYP81A subfamily to disclose the pattern of cross-resistance in Echinochloa phyllopogon. Plant Mol. Biol. 2020, 102, 403–416. [Google Scholar] [CrossRef]
  24. Pan, L.; Guo, Q.; Wang, J.; Shi, L.; Yang, X.; Zhou, Y.; Yu, Q.; Bai, L. CYP81A68 confers metabolic resistance to ALS and ACCase-inhibiting herbicides and its epigenetic regulation in Echinochloa Crus-Gall. J. Hazard. Mater. 2022, 428, 128225. [Google Scholar] [CrossRef] [PubMed]
  25. Deng, W.; Yang, M.; Li, Y.; Xia, Z.; Chen, Y.; Yuan, S.; Yang, Q. Enhanced metabolism confers a high level of cyhalofop-butyl resistance in a Chinese sprangletop (Leptochloa chinensis (L.) Nees) population. Pest Manag. Sci. 2021, 77, 2576–2583. [Google Scholar] [CrossRef]
  26. Wright, A.A.; Nandula, V.K.; Grier, L.; Showmaker, K.C.; Bond, J.A.; Peterson, D.G.; Ray, J.D.; Shaw, D.R. Characterization of fenoxaprop-p-ethyl-resistant junglerice (Echinochloa colona) from Mississippi. Weed Sci. 2016, 64, 588–595. [Google Scholar] [CrossRef]
  27. Seefeldt, S.S.; Jensen, J.E.; Fuerst, E.P. Log-logistic analysis of herbicide dose-response relationships. Weed Technol. 1995, 9, 218–227. [Google Scholar] [CrossRef]
  28. Amaro-Blanco, I.; Romano, Y.; Palmerin, J.A.; Gordo, R.; Palma-Bautista, C.; Prado, R.D.; Osuna, M.D. Different mutations providing target site resistance to ALS-and ACCase-inhibiting herbicides in Echinochloa spp. from rice fields. Agriculture 2021, 11, 382. [Google Scholar] [CrossRef]
  29. Hwang, J.; Norsworthy, J.K.; González-Torralva, F.; Piveta, L.B.; Barber, L.T.; Butts, T.R. Cross-resistance of barnyardgrass [Echinochloa crus-galli (L.) P. beauv.] to aryloxyphenoxypropionate herbicides. Pestic. Biochem. Physiol. 2022, 184, 105089. [Google Scholar] [CrossRef]
  30. Torra, J.; Montull, J.M.; Taberner, A.; Onkokesung, N.; Boonham, N.; Edwards, R. Target-site and non-target-site resistance mechanisms confer multiple and cross-resistance to ALS and ACCase inhibiting herbicides in Lolium rigidum from spain. Front. Plant Sci. 2021, 12, 625138. [Google Scholar] [CrossRef]
  31. Yu, Q.; Ahmad-Hamdani, M.S.; Han, H.; Christoffers, M.J.; Powles, S.B. Herbicide resistance-endowing ACCase gene mutations in hexaploidy wild oat (Avena fatua): Insights into resistance evolution in a hexaploidy species. Heredity 2013, 110, 220–231. [Google Scholar] [CrossRef] [PubMed]
  32. Han, H.; Yu, Q.; Owen, M.; Cawttray, G.R.; Powles, S.B. widespread occurrence of both metabolic and target-site herbicide resistance mechanisms in Lolium rigidum populations. Pest Manag. Sci. 2016, 72, 255–263. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  33. Chen, G.; Xu, H.; Zhang, H.; Bai, C.; Dong, L. Fenoxaprop-p-ethyl resistance conferred by cytochrome P450s and target site mutation in Alopecurus japonicas. Pest Manag. Sci. 2018, 74, 1694–1703. [Google Scholar] [CrossRef] [PubMed]
  34. Bai, S.; Zhao, Y.; Zhou, Y.; Wang, M.; Li, Y.; Luo, X.; Li, L. Identification and expression of main genes involved in non-target site resistance mechanisms to fenoxaprop-p-ethyl in Beckmannia syzigachne. Pest Manag. Sci. 2020, 76, 2619–2626. [Google Scholar] [CrossRef] [PubMed]
  35. Yanniccari, M.; Gigón, R.; Larsen, A. Cytochrome P450 herbicide metabolism as the main mechanism of cross-resistance to ACCase- and ALS-inhibitors in Lolium spp. populations from Argentina: A molecular approach in characterization and detection. Front. Plant Sci. 2020, 11, 600301. [Google Scholar] [CrossRef] [PubMed]
  36. Vazquez-Garcia, J.G.; Torra, J.; Palma-Bautista, C.; Cruz, R.A.; Prado, R.D. Point mutations and cytochrome P450 can contribute to resistance to ACCase-inhibiting herbicides in three Phalaris species. Plants 2021, 10, 1703. [Google Scholar] [CrossRef]
  37. Li, Z.; Liu, H.; Wang, J.; Chen, W.; Bai, L.; Pan, L. Enhanced metabolism evolved high-level resistance to fenoxaprop-p-ethyl in Alopecurus japonicas. Agronomy 2022, 12, 2172. [Google Scholar] [CrossRef]
  38. Hwang, J.; Norsworthy, J.K.; González-Torralva, F.; Priess, G.L.; Barber, L.T.; Butts, T.R. Non-target-site resistance mechanism of barnyardgrass [Echinochloa crus-galli (L.) P. Beauv.] to florpyrauxifen-benzyl. Pest Manag. Sci. 2022, 78, 287–295. [Google Scholar] [CrossRef]
  39. Owen, M.J.; Goggin, D.E.; Powles, S.B. Non-target-site-based resistance to ALS-inhibiting herbicides in six Bromus rigidus populations from Western Australian cropping fields. Pest Manag. Sci. 2012, 68, 1077–1082. [Google Scholar] [CrossRef]
  40. Yu, Q.; Han, H.; Cawthray, G.R.; Wang, S.F. Powles, S.B. Enhanced rates of herbicide metabolism in low herbicide-dose selected resistant Lolium rigidum. Plant Cell Environ. 2013, 36, 818–827. [Google Scholar] [CrossRef]
  41. Yu, Q.; Powles, S.B. Metabolism-based herbicide resistance and cross-resistance in crop weeds: A threat to herbicide sustainability and global crop production. Plant Physiol. 2014, 166, 1106–1118. [Google Scholar] [CrossRef] [PubMed]
  42. Zhao, N.; Yang, J.; Jiang, M.; Liao, M.; Cao, H. Identification of essential genes involved in metabolism-based resistance mechanism to fenoxaprop-p-ethyl in Polypogon fugax. Pest Manag. Sci. 2022, 78, 1164–1175. [Google Scholar] [CrossRef] [PubMed]
  43. Yang, X.; Zhang, Z.; Tao Gu, T.; Dong, M.; Peng, Q.; Bai, L.; Li, Y. Quantitative proteomics reveals ecological fitness cost ofmulti-herbicide resistant barnyardgrass (Echinochloa crus-galli L.). J. Proteom. 2017, 150, 160–169. [Google Scholar] [CrossRef] [PubMed]
Agronomy 12 02724 g001 550
Figure 1. Cyhalofopbutyl dose-response curves of S and R E. crus-galli populations.
Figure 1. Cyhalofopbutyl dose-response curves of S and R E. crus-galli populations.
Agronomy 12 02724 g001
Agronomy 12 02724 g002 550
Figure 2. Metabolic enzyme inhibitor effect on cyhalofop-butyl resistance in the S and R populations of E. crus-galli.
Figure 2. Metabolic enzyme inhibitor effect on cyhalofop-butyl resistance in the S and R populations of E. crus-galli.
Agronomy 12 02724 g002
Table
Table 1. Herbicides’ information and treatment doses used for dose-response experiments of S and R populations.
Table 1. Herbicides’ information and treatment doses used for dose-response experiments of S and R populations.
TypeHerbicidesFormulation 1 and CompanyTreatment Doses
(g ai ha−1)		Field Recommended Dose
(g ai ha−1)
ACCase inhibitorsCyhalofop-butyl100 g L−1 EC, Dow AgroSciencesS: 0, 1.30 3.89, 11.67, 35, 105
R: 0, 3.89, 11.67, 35, 105, 315		105
Metamifop10% EC, FMCS: 0, 1.48, 4.4, 13.33, 40, 120
R: 0, 4.44, 13.33, 40, 120, 360		120
Fenoxaprop-p-ethyl69 g L−1 EW, Bayer CropScienceS: 0, 0.26, 0.77, 2.3, 6.9, 20.7
R: 0, 0.77, 2.3, 6.9, 20.7, 62.1		62.1
Clethodim120 g L−1 EC, Qingdao Modern AgrochemicalS: 0, 0.3, 0.89, 2.67, 8, 24
R: 0, 0.89, 2.67, 8, 24, 72		72
Pinoxaden5% EC, SyngentaS: 0, 0.56, 1.67, 5, 15, 45
R: 0, 1.67, 5, 15, 45, 135		45
ALS inhibitorsPenoxsulam25 g L−1 OD, Dow AgroSciencesS: 0, 0.37, 1.11, 3.33, 10, 30
R: 0, 1.11, 3.33, 10, 30, 90		30
Bispyribac-sodium10% SC, Jiangsu Hormone Research InstituteS: 0, 0.56, 1.67, 5, 15, 45
R: 0, 1.67, 5, 15, 45, 135		45
Auxin mimic herbicidesQuinclorac75% WG, Jiangsu Hormone Research InstituteS: 0, 5.56, 16.67, 50, 150, 450
R: 0, 50, 150, 450, 1350, 4050		450
Florpyrauxifen-benzyl3% EC, Dow AgroSciencesS: 0, 0.23, 0.67, 2, 6, 18
R: 0, 0.67, 2, 6, 18, 54		18
1 EC, emulsifiable concentrate; EW, emulsion in water; OD, oil dispersion; SC, suspension concentrate; WG, water dispersible granule.
Table
Table 2. GR50 and resistance index (RI) to ACCase-inhibiting herbicides and other herbicides used for direct-seeding rice of E. crus-galli populations.
Table 2. GR50 and resistance index (RI) to ACCase-inhibiting herbicides and other herbicides used for direct-seeding rice of E. crus-galli populations.
HerbicidesSRRI 2
GR50 ± SE 1pGR50 ± SEp
Cyhalofop-butyl5.6 ± 0.6<0.0175.8 ± 8.7<0.0113.5
Metamifop8.1 ± 0.5<0.0123.7 ± 1.3<0.012.9
Fenoxaprop-p-ethyl1.2 ± 0.1<0.014.9 ± 0.9<0.014.1
Clethodim2.1 ± 0.2<0.019.8 ± 0.8<0.014.7
Pinoxaden3.7 ± 0.5<0.0123.7 ± 3.2<0.016.4
Penoxsulam3.2 ± 1.10.0228.6 ± 2.19<0.018.9
Bispyribac-sodium11.64 ± 2.670.0214.18 ± 0.94<0.011.2
Quinclorac116.63 ± 17.170.01>4050.000.03>34.7
Florpyrauxifen-benzyl3.4 ± 0.2<0.0110.1 ± 0.6<0.013.0
1 GR50, herbicide dose causing 50% growth reduction. 2 RI = GR50 resistant biotype (R)/GR50 susceptible biotype (S).
Table
Table 3. Content of cyhalofop-butyl and cyhalofop acid in E. crus-galli plants from S and R populations after cyhalofop-butyl treatment.
Table 3. Content of cyhalofop-butyl and cyhalofop acid in E. crus-galli plants from S and R populations after cyhalofop-butyl treatment.
Time
(h)		Cyhalofop-Butyl (μg kg−1)Cyhalofop Acid (μg kg−1)
SRSR
20.85 ± 0.100.94 ± 0.120.71 ± 0.080.67 ± 0.07
240.42 ± 0.050.48 ± 0.060.57 ± 0.090.21 ± 0.05 *
480.027 ± 0.0080.024 ± 0.0040.24 ± 0.070.08 ± 0.02 **
*, ** significant difference at p < 0.05, 0.01, respectively.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Yang, Q.; Yang, X.; Zhu, J.; Wei, T.; Lv, M.; Li, Y. Metabolic Resistance to Acetyl-CoA Carboxylase-Inhibiting Herbicide Cyhalofop-Butyl in a Chinese Echinochloa crus-galli Population. Agronomy 2022, 12, 2724. https://doi.org/10.3390/agronomy12112724

AMA Style

Yang Q, Yang X, Zhu J, Wei T, Lv M, Li Y. Metabolic Resistance to Acetyl-CoA Carboxylase-Inhibiting Herbicide Cyhalofop-Butyl in a Chinese Echinochloa crus-galli Population. Agronomy. 2022; 12(11):2724. https://doi.org/10.3390/agronomy12112724

Chicago/Turabian Style

Yang, Qian, Xia Yang, Jinlei Zhu, Tian Wei, Min Lv, and Yongfeng Li. 2022. "Metabolic Resistance to Acetyl-CoA Carboxylase-Inhibiting Herbicide Cyhalofop-Butyl in a Chinese Echinochloa crus-galli Population" Agronomy 12, no. 11: 2724. https://doi.org/10.3390/agronomy12112724

APA Style

Yang, Q., Yang, X., Zhu, J., Wei, T., Lv, M., & Li, Y. (2022). Metabolic Resistance to Acetyl-CoA Carboxylase-Inhibiting Herbicide Cyhalofop-Butyl in a Chinese Echinochloa crus-galli Population. Agronomy, 12(11), 2724. https://doi.org/10.3390/agronomy12112724

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop