Japanese Foxtail Research Articles

Herbicidal Activity of Cinmethylin against Grass Weeds and Its Safety for Use with Different Wheat Varieties

Cinmethylin is a potential pre-emergence herbicide that could be used to control grass weeds in winter cereals. To determine the herbicidal activity of cinmethylin against common gramineous weeds in wheat fields in China and its level of safety on wheat, we conducted the following experiments: (i) assessing the efficacy of cinmethylin against 11 grass weeds and (ii) determining its safety against 19 wheat varieties. The results showed that cinmethylin had good herbicidal efficacy against annual bluegrass (Poa annua L.), shortawn foxtail (Alopecurus aequalis Sobol.), slender meadow foxtail (Alopecurus myosuroides Huds.), Japanese foxtail (Alopecurus japonicus Steud.), Italian ryegrass (Lolium multiflorum Lam.), British timothy (Phleum paniculatum Huds.), Asia Minor bluegrass (Polypogon fugax Nees ex Steud.), Helictotrichon tibeticum (Roshev.) Holub., and wild oat (Avena fatua L.), with a GR50 (the herbicide dose resulting in 50% growth inhibition) value of 4.50–99.21 g a.i. ha−1 in plant height and 1.43–70.34 g a.i. ha−1 in fresh weight. However, cinmethylin cannot control Japanese brome (Bromus japonicus L.) or Tausch’s goatgrass (Aegilops tauschii Coss.) at a dose of 200 g a.i. ha−1. Different wheat varieties varied in their phytotoxicity to cinmethylin. Overall, there is a phytotoxicity risk when using cinmethylin on wheat, mainly to wheat roots, with a reduction in root length of 40.81–64.09% at a dose of 400 g a.i. ha−1. These findings indicate that the pre-emergence herbicide cinmethylin provides good efficacy against most grass weeds and may possess potential for weed management in wheat fields. However, attention should be given to the application dosage and the sensitivity of wheat varieties when using cinmethylin in wheat fields.

Confirmation and chemical control of acetyl-CoA carboxylase- and acetolactate synthase-resistant Japanese foxtail in China

Japanese foxtail (Alopecurus japonicus Steud.) has developed resistance to multiple post-emergence herbicides in China. Hence, identification of alternative strategies for weed control using pre-emergence herbicides is urgently required. Therefore, we conducted greenhouse dose–response and field experiments over two wheat growing seasons at two sites in Xinyang City (CaiWan and LaoHuShan) to study resistance levels and effective chemical control strategies using the pre-emergence herbicides flufenacet and pyroxasulfone individually and tank-mixed. Greenhouse studies indicated that the CaiWan and LaoHuShan Japanese foxtail populations had a 5.1–35.3-fold resistance to acetyl-CoA carboxylase inhibitors and a 3.7–9.9-fold resistance to acetolactate synthase inhibitors, respectively. Field experiment data analyses revealed that pre-emergence herbicides applied before wheat sowing and immediately after wheat sowing provided >76% weed density and >82% weed biomass control efficiencies 30 and 90 days after sowing, respectively. They showed significantly higher results than herbicides applied after wheat was sown. Compared to the weedy check (control), flufenacet (300 and 450 g/ha), pyroxasulfone (180 and 240 g/ha), and their tank-mix (flufenacet at 300 + pyroxasulfone at 180 g/ha) significantly reduced weed density and biomass (P < 0.05). The tank-mix of flufenacet + pyroxasulfone consistently provided the highest weed control efficiency (85.3–94.5%). The tank-mix applied immediately after sowing resulted in the highest weed control efficiency (88.9–95.6%), the highest wheat yield (5470.3–5527.3 kg/ha), and no observed wheat injuries. However, multiple strategies, such as pre- and post-emergence herbicides, must be adopted to cope with high-density resistant weeds.

Effect of Japanese foxtail (Alopecurus japonicus) at varying densities on the physiological characteristics of wheat

To evaluate the effect of Japanese foxtail (Alopecurus japonicus Steud.) at different densities on the photosynthetic physiology of wheat, “Yangmai 16” wheat was cultivated in a field using two sowing rates (150 kg ha−1, conventional seeding rate; 225 kg ha−1, high seeding rate) and with different densities of A. japonicus for two years. A. japonicus affected the physiological characteristics of wheat, such as its leaf photosynthetic rate, fluorescence parameters, Hill reaction activity, Rubisco activity, yield and yield components, to varying degrees. However, the physiological responses of wheat at different seeding rates varied; the effect of A. japonicus on wheat was more severe at the conventional seeding rate than at the high seeding rate. A. japonicus affected wheat leaf morphology and inhibited photosynthetic processes, leading to decreased PSII electron transport, energy metabolism and photosynthate accumulation, ultimately reducing wheat yield. An appropriate increase in the wheat seeding rate could improve specific leaf weight, maintain Rubisco activity, reduce photoinhibition, and alleviate the stress caused by A. japonicus.

Cross-resistance to acetolactate synthase (ALS) inhibitors associated with different mutations in Japanese foxtail (Alopecurus japonicus)

AbstractJapanese foxtail (Alopecurus japonicus Steud.) is an invasive grass weed that severely threatens the production of wheat (Triticum aestivum L.) and canola (Brassica napus L.) crops in eastern Asia. Mesosulfuron-methyl is a highly efficient acetolactate synthase (ALS)-inhibiting herbicide widely used for control of this species in China. However, in recent years, some A. japonicus populations have evolved resistance to mesosulfuron-methyl by different amino acid substitutions (AASs) within the ALS gene. In the current study, 11 populations of A. japonicus were collected from Anhui Province, China, where the wheat fields were severely infested with this weed. Based on single-dose screening, eight of these populations evolved resistance to mesosulfuron-methyl, and gene sequencing revealed three AASs located in codon 197 or 574 of the ALS gene in the different resistant populations. Subsequently, three typical populations, AH-1, AH-4, and AH-10 with Trp-574-Leu, Pro-197-Thr, and Pro-197-Ser mutations, respectively, in ALS genes were selected to characterize their cross-resistance patterns to ALS inhibitors. Compared with the susceptible population AH-S, AH-1 showed broad-spectrum cross-resistance to sulfonylureas (SUs), imidazolinones (IMIs), triazolopyrimidines (TPs), and sulfonyl-aminocarbonyl-triazolinones (SCTs); whereas AH-4 and AH-10 were resistant to SUs, TPs, and SCTs but sensitive to IMIs. Moreover, all three resistant populations were sensitive to both photosystem II inhibitor isoproturon and 4-hydroxyphenylpyruvate dioxygenase inhibitor QYM201 (1-(2-chloro-3-(3-cyclopropyl-5-hydroxy-1-methyl-1H-pyrazole-4-carbonyl)-6-(trifluoromethyl)phenyl)piperidin-2-one). Based on the current state of knowledge, this study is the first report of A. japonicus evolving cross-resistance to ALS-inhibiting herbicides due to a Pro-197-Ser mutation in the ALS gene.

Japanese Foxtail (Alopecurus japonicus) Management in Wheat in China: Seed Germination, Seedling Emergence, and Response to Herbicide Treatments

AbstractLittle information is published related to seed germination and seedling emergence of Japanese foxtail, a troublesome annual grass weed widely distributed in winter wheat fields in China. Three Japanese foxtail populations were studied under laboratory and greenhouse conditions, to determine the effects of different environmental factors on seed germination or seedling emergence. Chemical control is absolutely necessary in integrated management, and efficacy of POST herbicides against different growth stages of Japanese foxtail was evaluated. Germination rate was 90% or more when temperature ranged from 5 to 25 C, with germination onset shortened as temperature increased. Light was not required for germination to occur. For pH values ranging from 5 to10 there was no effect on seed germination. Japanese foxtail seed germination was sensitive to osmotic stress and completely inhibited at an osmotic potential of -1.1 MPa. The ‘1513’ population of Japanese foxtail demonstrated tolerance to soil salinity, with 98% germination at 80 mM NaCl compared with 25 and 40% germination for populations ‘1532’ and ‘1544’, respectively. High amounts of crop residue (10 t ha−1) suppressed Japanese foxtail emergence 38 to 55%. Germination of seeds placed at 160 C for 5 min was completely inhibited for dry seeds, with a similar effect at 130 C for pre-soaked seeds. Seed burial in the soil from 0 to 4 cm had no effect on seedling emergence, but burial at 7 cm completely inhibited seedling emergence. POST herbicides mesosulfuron-methyl (13.5 g ai ha−1), clodinafop-propargyl (67.5 g ai ha−1), pyroxsulam (13.5 g ai ha−1), pinoxaden (67.5 g ai ha−1) and isoproturon (1125 g ai ha−1) reduced plant dry weight 80% or more when applied at three- to seven-leaf stage, but control declined with application at later growth stages. The information from this study helps to develop an integrated approach to Japanese foxtail management.

Identification of Reference Genes for Studying Herbicide Resistance Mechanisms in Japanese Foxtail (Alopecurus japonicus)

Herbicide-resistant weeds pose a considerable threat to agriculture, but their resistance mechanisms are poorly understood. Differential gene expression analysis of a weed subjected to herbicide treatment is a key step toward more mechanistic studies. Such an analysis, often involving quantitative real-time PCR (qPCR), requires suitable reference genes as internal controls. In this study, we identified optimal reference genes in the noxious weed, Japanese foxtail. This weed has evolved resistance to acetyl-coenzyme A carboxylase (ACCase) inhibitors. We analyzed the stability of eight commonly used candidate reference genes (glyceraldehyde-3-phosphate dehydrogenase [GAPDH]; ubiquitin [UBQ]; capsine phosphatase [CAP]; beta-tubulin [TUB]; eukaryotic initiation factor 4a [EIF4A]; elongation factor-1 alpha [EF1]; 18S ribosomal RNA [18S]; 25S ribosomal RNA [25S]) from root, stem, and leaf tissue of plants that were either resistant or sensitive to ACCase inhibitors, with or without herbicide stress, using qPCR. The results were further ranked and analyzed using geNorm, NormFinder, and BestKeeper software. These analyses identified EF1 and UBQ in roots, EF1, TUB, CAP, and 18S in stems, and EF1, GAPDH, and 18S in leaves as suitable references for qPCR normalization. We have identified a set of reference genes that can be used to study herbicide resistance mechanisms in Japanese foxtail.

Cross-resistance Patterns to Acetyl-CoA Carboxylase Inhibitors Associated with Different Mutations in Japanese Foxtail (Alopecurus japonicus)

Japanese foxtail is a grass weed in eastern China. This weed is controlled by fenoxaprop-P-ethyl, one of the most common acetyl-CoA carboxylase (ACCase)-inhibiting herbicides. Some Japanese foxtail populations have developed resistance to fenoxaprop-P-ethyl, owing to target-site mutations (amino acid substitutions) located within the carboxyl transferase domain of ACCase. In the present study, three mutations were detected in three fenoxaprop-P-ethyl–resistant Japanese foxtail populations: Ile-1781-Leu in JCJT-2, Ile-2041-Asn in JZJR-1, and Asp-2078-Gly in JCWJ-3. Two copies ofACCase(Acc1-1andAcc1-2) were identified, but mutations were detected only inAcc1-1. The derived cleaved amplified polymorphic sequence (dCAPS) method detected these mutations successfully in Japanese foxtail. The mutation frequencies in JCJT-2, JZJR-1, and JCWJ-3 were approximately 98%, 92%, and 87%, respectively. Different cross-resistance patterns to ACCase inhibitors were found in the three resistant populations. JCJT-2 (Ile-1781-Leu) and JZJR-1 (Ile-2041-Asn) showed cross-resistance to haloxyfop-R-methyl, clodinafop-propargyl, and pinoxaden, but were susceptible to clethodim. JCWJ-3 (Asp-2078-Gly) showed cross-resistance to all tested ACCase-inhibiting herbicides.

Mechanisms of Resistance to Pyroxsulam and ACCase Inhibitors in Japanese Foxtail (Alopecurus japonicus)

Japanese foxtail is a predominant tetraploid grass weed in wheat and oilseed rape fields in eastern China. In China, pyroxsulam is mainly used to manage annual grass weeds, especially those resistant to acetyl coenzyme A carboxylase (ACCase)-inhibiting herbicides. Using dose–response studies, a pyroxsulam-resistant population, ACTC-1, was identified with a resistance index value of 58. Additionally, ACTC-1 was cross-resistant to sulfonylureas, imidazolinones, triazolopyrimidines, pyrimidinyl-benzoates, and sulfonylaminocarbonyl-triazolinones and multiresistant to ACCase and photosystem II inhibitors. Sequence analysis revealed four gene fragments encoding acetolactate synthase (ALS) from ACTC-1, and three from JNXW-1, a pyroxsulam-sensitive population. An Asp-376-Glu substitution was found in ALS1;2 and an Ile-2041-Asn in Acc1;1, which may be responsible for its resistance to pyroxsulam and ACCase inhibitors, respectively. In vitro assays of ALS activity revealed that in ACTC-1, the sensitivity of ALS to pyroxsulam was lower, and the basal ALS activity was twofold higher than that of sensitive population JNXW-1. Additionally, the combined application of pyroxsulam with malathion or piperonyl butoxide increased the sensitivity of ACTC-1 to pyroxsulam, although it could not completely overcome the resistance. It was inferred that both target-site-based resistance and nontarget-site-based resistance may be involved in the resistance to pyroxsulam.

Germination Requirements Differ between Fenoxaprop-P-ethyl Resistant and Susceptible Japanese Foxtail (Alopecurus japonicus) Biotypes

Herbicide-resistant (R biotype) and -sensitive (S biotype) individuals were identified from the same population, and seed was increased for each biotype for three generations. We conducted laboratory experiments to determine the effects of temperature, light, salt stress, osmotic stress, pH, and burial depth on the germination and emergence of resistant and sensitive biotypes of Japanese foxtail. The results revealed that there was no difference in the final germination rate between the two biotypes under different temperature conditions, but time to obtain 50% germination or emergence (tE50) and mean germination time of the R biotype were higher than that of the S biotype at 10 C and 15/10 C 12-h day/night regime. In dark conditions, the final germination rate of the S biotype was higher and lower than that of the R biotype at 10 and 25 C, respectively. The overall germination rate of the R biotype was lower than that in the S biotype, and extended germination time was required in extreme conditions, such as 250 mM NaCl and −0.4 MPa osmotic potential. The change in environmental pH had no effect on the germination of the two biotypes. Emergence of the R biotype was lower than the S biotype when seed was buried at least 8 cm deep in an organic matter substrate. This study demonstrated the pleiotropic effects of a resistance allele on seed germination and emergence under different environmental conditions. Deep tillage could be used to reduce the growth and spread of resistant Japanese foxtail individuals.

Cross-Resistance of Japanese Foxtail (Alopecurus japonicus) to ACCase Inhibitors in China

The increasing use of ACCase-inhibiting herbicides has resulted in evolved resistance in key grass weeds infesting cereal cropping systems worldwide. Japanese foxtail is one of the most important grass weed species in wheat in China. Most populations have evolved resistance to fenoxaprop-p-ethyl, which is one of the most common ACCase-inhibiting herbicides in wheat. The seeds of two Japanese foxtail populations were collected from wheat fields where farmers complained that control could not be effectively obtained with fenoxaprop-p-ethyl. Seeds from one susceptible population were collected from an area along a roadside where ACCase inhibitors had not been used to be used for validating cross-resistance and elucidating the mechanism of resistance. The experimental results showed that the two populations, Aloja-JS10-R1 and Aloja-JS10-R2, expressed high resistance to fenoxaprop-p-ethyl, with resistance indexes (RIs) of 29.2 and 27.9. These populations also expressed high cross-resistance to clodinafop-propargyl with RIs of 12.8 and 14.7, and moderate cross-resistance to clethodim and pinoxaden with RIs ranging from 2.6 to 11.4. Comparison of the ACCase carboxyl-transferase (CT) domain sequences of the susceptible and resistant populations with blackgrass revealed that tryptophan at position 2027 of the ACCase gene was substituted by cysteine in population Aloja-JS10-R1, and isoleucine at position 1781 of the ACCase gene was substituted by leucine in populations Aloja-JS10-R2. The study confirmed Japanese foxtail resistance to the ACCase inhibitor fenoxafop-p-ethyl, cross-resistance to other ACCase inhibitors, and the resistance mechanism being conferred by specific ACCase point mutations at amino acid position 1781 and 2027.

Determination of ploidy level and isolation of genes encoding acetyl-CoA carboxylase in Japanese Foxtail (Alopecurus japonicus).

Ploidy level is important in biodiversity studies and in developing strategies for isolating important plant genes. Many herbicide-resistant weed species are polyploids, but our understanding of these polyploid weeds is limited. Japanese foxtail, a noxious agricultural grass weed, has evolved herbicide resistance. However, most studies on this weed have ignored the fact that there are multiple copies of target genes. This may complicate the study of resistance mechanisms. Japanese foxtail was found to be a tetraploid by flow cytometer and chromosome counting, two commonly used methods in the determination of ploidy levels. We found that there are two copies of the gene encoding plastidic acetyl-CoA carboxylase (ACCase) in Japanese foxtail and all the homologous genes are expressed. Additionally, no difference in ploidy levels or ACCase gene copy numbers was observed between an ACCase-inhibiting herbicide-resistant and a herbicide-sensitive population in this study.

Mutations at codon position 1999 of acetyl‐CoA carboxylase confer resistance to ACCase‐inhibiting herbicides in Japanese foxtail (Alopecurus japonicus)

The intensive and global application of ACCase-inhibiting herbicides has resulted in the evolution of resistance in a growing number of grass weeds. Among the mutations implicated in conferring resistance, limited knowledge is available regarding mutations at codon position 1999. In addition, multiple copies of genes encoding plastidic ACCase have been ignored in previous studies of resistance in Alopecurus japonicus. Dose-response tests indicated that the population JLGY-4 had evolved high-level resistance to fenoxaprop-P-ethyl. The carboxyltransferase domain of the ACCase gene in A. japonicus was sequenced and compared. Two loci encoding plastidic ACCase were isolated from both the resistant and sensitive populations. Simultaneously, two resistance-endowing mutations at codon position 1999 of ACCase were determined (W1999C and W1999L). Moreover, a molecular study was conducted to determine the mechanism of resistance to some ACCase-inhibiting herbicides. The W1999C mutation conferred resistance to fenoxaprop and moderate resistance to pinoxaden. The W1999L mutation conferred resistance to fenoxaprop. This study revealed that A. japonicus had multiple copies of genes encoding plastidic ACCase, and each gene was able to carry its own mutation. It also established the clear importance of the W1999C and W1999L mutations in conferring resistance to ACCase-inhibiting herbicides.

Mechanism of resistance to fenoxaprop in Japanese foxtail (Alopecurus japonicus) from China

Japanese foxtail is one of the most common and troublesome weeds infesting cereal and oilseed rape fields in China. Repeated use during the last three decades of the ACCase-inhibiting herbicide fenoxaprop-P-ethyl to control this weed has resulted in the occurrence of resistance. Dose–response tests established that a population (AHFD-1) from eastern China had evolved high-level resistance to fenoxaprop-P-ethyl. Based on the resistance index, this resistant population of A. japonicus is 60.31-fold resistant to fenoxaprop-P-ethyl. Subsequently, only a tryptophan to cysteine substitution was identified to confer resistance to fenoxaprop-P-ethyl in this resistant population. ACCase activity tests further confirmed this substitution was linked to resistance. This is the first report of the occurrence of Trp-2027-Cys substitution of ACCase in A. japonicus. From whole-plant pot dose–response tests, we confirmed that this population conferred resistance to other APP herbicides, including clodinafop-propargyl, fluazifop-P-butyl, quizalofop-P-ethyl, haloxyfop-R-methyl, cyhalofop-butyl, metamifop, DEN herbicide pinoxaden, but not to CHD herbicides clethodim, sethoxydim. There was also no resistance observed to ALS-inhibiting herbicides sulfosulfuron, mesosulfuron-methyl, flucarbazone-sodium, pyroxsulam, Triazine herbicide prometryne and glyphosate. However, this resistant population was likely to confer slightly (or no) resistant to Urea herbicides chlortoluron and isoproturon.

Molecular basis of resistance to mesosulfuron-methyl in Japanese foxtail, &lt;i&gt;Alopecurus japonicus&lt;/i&gt;

Molecular basis of resistance to mesosulfuron-methyl in Japanese foxtail, &lt;i&gt;Alopecurus japonicus&lt;/i&gt;

Japanese Foxtail (Alopecurus japonicus) Resistance to Fenoxaprop and Pinoxaden in China

Japanese foxtail is one of the most common and competitive annual grass weeds of wheat in China. Whole-plant dose-response experiments were conducted with fenoxaprop and pinoxaden to confirm and characterize resistant and susceptible Japanese foxtail populations and to elucidate the basis of resistance to these herbicides. The resistant Japanese foxtail population was 49-fold resistant to fenoxaprop and 16-fold (cross) resistant to pinoxaden relative to the susceptible population, which was susceptible to both fenoxaprop and pinoxaden herbicides. Molecular analysis of resistance confirmed that the Ile1781to Leu mutation in the resistant population conferred resistance to both fenoxaprop and pinoxaden. This is the first report of cross resistance of Japanese foxtail to pinoxaden in the world and of a target site mutation that corresponded to resistance to both fenoxaprop and pinoxaden in Japanese foxtail. Prior selection pressure from fenoxaprop could result in evolution of resistance to fenoxaprop and cross resistance to pinoxaden in Japanese foxtail population.

Molecular bases for resistance to acetyl‐coenzyme A carboxylase inhibitor in Japanese foxtail (Alopecurus japonicus)

Haloxyfop-R-methyl is a widely used herbicide to control Poaceae weeds. Alopecurus japonicus, a widespread annual grass, can no longer be controlled by haloxyfop-R-methyl after continuous use of this herbicide for several years. Dose-response experiments have established that the Js-R biotype of A. japonicas has evolved resistance to aryloxyphenoxypropionates (APPs). Target-site enzyme sensitivity experiments have established that the haloxyfop (free acid) rate causing 50% inhibition of acetyl-CoA carboxylase (ACCase) activity (I(50)) for the resistant (Js-R) biotype is 11 times higher than that for the susceptible (Js-S) biotype. In many cases, resistance to ACCase-inhibiting herbicides is due to a resistant ACCase enzyme. Full-length DNA and mRNA sequences of the plastidic ACCase gene were amplified. Eight single-nucleotide differences were detected in this region. Four of the nucleotide changes were silent mutations. However, the other four nucleotide mutations caused four amino acid substitutions, replacing Arg-1734 with Gly, Met-1738 with Leu, Thr-1739 with Ser and Ile-2041 with Asn in the R biotype respectively; the substitution at position 2041 had been reported, while the other three had not. The ACCase in the Js-R biotype was less susceptible to haloxyfop-R-methyl than that in the Js-S biotype. Moreover, the amino acid substitution of Ile-2041 with Asn might confer resistance to haloxyfop-R-methyl in A. japonicas.

Identification of Japanese Foxtail (Alopecurus Japonicus) Resistant to Haloxyfop Using Three Different Assay Techniques

The objective of this study was to investigate the resistance level of Japanese foxtail to haloxyfop, an acetyl coenzyme A carboxylase (ACCase; EC 6.4.1.2)–inhibiting herbicide. Eleven biotypes were collected from oilseed rape fields in different areas in Jiangsu and Anhui provinces where haloxyfop had been continuously applied for various periods. Biotypes were assessed by two different methods, a seed bioassay and whole-plant assay, to identify the most resistant and susceptible biotypes for further studies on the activity of the target enzyme ACCase. A good correlation was obtained between the two different bioassay methods. The Jurong and Chuzhou biotypes were the most resistant and susceptible biotypes, respectively, whereas the other nine biotypes showed variable and relatively low degrees of haloxyfop resistance. Furthermore, target-site enzyme sensitivity results confirmed that the Jurong biotype was resistant to haloxyfop with a concentration of herbicide causing 50% inhibition of ACCase activity (IC50) of 9.19 µM, whereas the IC50 of the susceptible biotype (Chuzhou) was 0.76 µM, giving a resistance index of 12.