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Benzobicyclon tolerance in rice (Oryza sativa L.) is dependent on the presence of a functional HIS1 gene, but the level of sensitivity might vary among different cultivars. Greenhouse, laboratory, and field experiments were conducted to further explore the role of HIS1 in cultivated rice tolerance and to exploit findings toward optimizing benzobicyclon activity on weedy rice (unwanted rice; Oryza sativa L.). In a heredity experiment, benzobicyclon tolerance was confirmed to be a semidominant trait conferred by HIS1 based on the intermediate response (ED50 values) of HIS1 heterozygous F1 plants. The spatial–temporal expression of HIS1 was next studied in tissue types (blades, sheaths, and whorls) across tolerant cultivars (‘Roy J’, ‘Diamond’, ‘LaKast’, ‘CLXL745’, and ‘XL753’) and growth stages (2- to 3- compared with 5- to 6-leaf). The relative expression of HIS1 was tissue specific and highest in whorls, followed by blades and then sheaths. Minimal differences in expression across cultivars and growth stages were observed. Furthermore, HIS1 was not largely upregulated at 6 h after benzobicyclon treatment. In the same experiment, cultivar tolerance to benzobicyclon at the label rate of 371 g ha−1 was found to be growth stage dependent. Plant growth was reduced by ∼35% when rice plants were at the 2- to 3- compared with 5- to 6-leaf growth stages. These results show that differences in benzobicyclon tolerance among HIS1 homozygous cultivars is likely not directly correlated with the expression of HIS1. In this research a model was proposed and supported by a field proof of concept study, indicating benzobicyclon efficacy on weedy rice is a function of HIS1 zygosity by growth stage at application. Prior research indicates HIS1 is the dominant allele in weedy rice accessions in Arkansas, and thus, based on our model, benzobicyclon should be applied to weedy rice with ≤2 leaves for suppression.
The 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.
Palmer amaranth is one of the most difficult-to-control weeds in row crop systems and has evolved resistance to several herbicide sites of action (SOAs). A late-season weed-escape survey had been conducted earlier to determine the distribution of protoporphyrinogen oxidase–inhibitor resistant Palmer Amaranth in Arkansas. The objective of this study was to evaluate the susceptibility of Arkansas Palmer amaranth accessions to commonly used herbicide SOAs. The SOAs evaluated were group 2 + 9, 3, 4, 5, 10, 14, 15, and 27, and the representative herbicide from each group was imazethapyr + glyphosate (79 + 860 g ha−1), trifluralin (1,120 g ha−1), dicamba (280 and 560 g ha−1), atrazine (560 g ha−1), glufosinate (594 g ha−1), fomesafen (395 g ha−1), S-metolachlor (1,064 g ha−1), and tembotrione (92 g ha−1), respectively. Palmer amaranth mortality varied among accessions across SOAs. Averaged across accessions, the mortality rates, by treatment in order from lowest to highest, were as follows: glyphosate + imazethapyr (16%), tembotrione (51%), dicamba at 280 g ha−1 (51%), fomesafen (76%), dicamba at 560 g ha−1 (82%), atrazine (85%), trifluralin (87%), S-metolachlor (96%), and glufosinate (99.5%). This study provides evidence that Palmer amaranth accessions with low susceptibility to glyphosate + imazethapyr, fomesafen, and tembotrione are widespread throughout Arkansas. Of the remaining SOAs, most Palmer amaranth accessions were sensitive; however, within each herbicide SOA, except glufosinate, control of some accessions was less than expected and resistance is suspected.
S-Metolachlor is commonly used by soybean and cotton growers, especially with POST treatments for overlapping residuals, to obtain season-long control of glyphosate- and acetolactate synthase (ALS)–resistant Palmer amaranth. In Crittenden County, AR, reports of Palmer amaranth escapes following S-metolachlor treatment were first noted at field sites near Crawfordsville and Marion in 2016. Field and greenhouse experiments were conducted to confirm S-metolachlor resistance and to test for cross-resistance to other very-long-chain fatty acid (VLCFA)–inhibiting herbicides in Palmer amaranth accessions from Crawfordsville and Marion. Palmer amaranth control in the field (soil <3% organic matter) 14 d after treatment (DAT) was ≥94% with a 1× rate of acetochlor (1,472 g ai ha–1; emulsifiable concentrate formulation) and dimethenamid-P (631 g ai ha–1). However, S-metolachlor at 1,064 g ai ha–1 provided only 76% control, which was not significantly different from the 1/2× and 1/4× rates of dimethenamid-P and acetochlor (66% to 85%). In the greenhouse, Palmer amaranth accessions from Marion and Crawfordsville were 9.8 and 8.3 times more resistant to S-metolachlor compared with two susceptible accessions based on LD50 values obtained from dose–response experiments. Two-thirds and 1.5 times S-metolachlor at 1,064 g ha–1 were the estimated rates required to obtain 90% mortality of the Crawfordsville and Marion accessions, respectively. Data collected from the field and greenhouse confirm that these accessions have evolved a low level of resistance to S-metolachlor. In an agar-based assay, the level of resistance in the Marion accession was significantly reduced in the presence of a glutathione S-transferase (GST) inhibitor, suggesting that GSTs are the probable resistance mechanism. With respect to other VLCFA-inhibiting herbicides, Marion and Crawfordsville accessions were not cross-resistant to acetochlor, dimethenamid-P, or pyroxasulfone. However, both accessions, based on LD50 values obtained from greenhouse dose–response experiments, exhibited reduced sensitivity (1.5- to 3.6-fold) to the tested VLCFA-inhibiting herbicides.
Palmer amaranth is one of the most problematic weeds in cropping systems of North America, especially in midsouthern United States, because of its competitive ability and propensity to evolve resistance to several herbicide sites of action. Previously, we confirmed and characterized the first case of nontarget site resistance (NTSR) to fomesafen in a Palmer amaranth accession from Randolph County, AR (RCA). The primary basis of the present study was to evaluate the cross- and multiple-resistance profile of the RCA accession. The fomesafen dose-response assay in the presence of malathion revealed a lower level of RCA resistance when compared with fomesafen alone. The resistance index of the RCA accession, based on 50% biomass reduction, ranged from 63-fold (fomesafen alone) to 22-fold (malathion plus fomesafen), when compared with a 2007 susceptible, and 476-fold and 167-fold, respectively, relative to a 1986 susceptible check. The RCA accession was resistant to other protoporphyrinogen oxidase (PPO) inhibitors (i.e., flumioxazin, acifluorfen, saflufenacil) as well as the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor tembotrione and acetolactate synthase (ALS) inhibitor pyrithiobac sodium. Sequencing of the ALS gene revealed no point mutations, indicating that a target-site mechanism is not involved in conferring ALS-inhibitor resistance in the RCA accession. Of the three PPO-inhibiting herbicides tested in combination with the malathion, saflufenacil resulted in the greatest biomass reduction (80%; P < 0.05) and lowest survival rate (23%; P < 0.05) relative to nontreated plants. The application of cytochrome P450 or glutathione S-transferase inhibitors with fomesafen did not lead to any adverse effects on soybean, suggesting a possible role for these compounds for management of NTSR under field conditions. These results shed light on the relative unpredictability of NTSR in conferring herbicide cross- and multiple resistance in Palmer amaranth.
Benzobicyclon is a new pro-herbicide being evaluated in the Midsouth United States as a post-flood weed control option in rice. Applications of benzobicyclon to flooded rice are necessary for efficacious herbicide activity, but why this is so remains unknown. Two greenhouse experiments were conducted to explore how herbicide placement (foliage only, flood water only, foliage and flood water simultaneously) and adjuvants (nonionic surfactant, crop oil concentrate, and methylated seed oil [MSO]) affect herbicide activity. The first experiment focused on importance of herbicide placement. Little to no herbicidal activity (<18% visual control) was observed on two- to four-leaf barnyardgrass, Amazon sprangletop, and benzobicyclon-susceptible weedy rice with benzobicyclon treatments applied to weed foliage only. In contrast, applications made only to the flood water accounted for >82% of the weed control and biomass reduction achieved when benzobicyclon was applied to flood water and foliage simultaneously. The second experiment concentrated on adjuvant type and benzobicyclon efficacy when applied to foliage and flood water simultaneously. At 28 days after treatment, benzobicyclon alone at 371 g ai ha−1 provided 29% and 67% control of three- to five-leaf barnyardgrass and Amazon sprangletop, respectively. The inclusion of any adjuvant significantly increased control, with MSO providing near-complete control of barnyardgrass and Amazon sprangletop. Furthermore, we used the physiochemical properties of benzobicyclon and benzobicyclon hydrolysate to derive theories to explain the complex activity of benzobicyclon observed in our study and in field trials. Benzobicyclon applications should contain an oil-based adjuvant and must be applied to flooded rice fields for optimal activity.
Pennsylvania smartweed [Persicaria pensylvanica (L.) M. Gómez] is a common weed of rice (Oryza sativa L.) in the midsouthern United States and has recently become a concern for farmers because of reduced tillage systems. Acetolactate synthase (ALS) inhibitors have been extensively used for controlling smartweeds in imidazolinone-resistant and conventional rice. In the present study, we confirmed resistance to commonly used ALS inhibitors in rice and characterized the underlying resistance mechanism in a P. pensylvanica biotype from southeast Missouri. A dose–response experiment was conducted in the greenhouse using bensulfuron-methyl, imazethapyr, and bispyribac-sodium to determine the resistance index (resistance/susceptibility [R/S]) based on GR50 estimates. The target-site ALS gene was amplified from R and S plants, and sequences were analyzed for mutations known to confer ALS-inhibitor resistance. The P. pensylvanica biotype in question was found to be resistant to bensulfuron-methyl (R/S=2,330), imazethapyr (R/S=12), and bispyribac-sodium (R/S=6). Sequencing of the ALS gene from R plants revealed two previously known mutations (Pro-197-Ser, Ala-122-Ser) conferring resistance to sulfonylureas and imidazolinones. This is the first report of ALS-inhibitor resistance in P. pensylvanica.
Palmer amaranth (Amaranthus palmeri S. Watson), a dioecious summer annual species, is one of the most troublesome weeds in U.S. cropping systems. The evolution of resistance to protoporphyrinogen oxidase inhibitors in A. palmeri biotypes is a major cause of concern to soybean [Glycine max (L.) Merr.] and cotton (Gossypium hirsutum L.) growers in the midsouthern United States. The objective of this study was to confirm and characterize the non–target site mechanism in a fomesafen-resistant accession from Randolph County, AR (RCA). A dose–response assay was conducted to assess the level of fomesafen resistance, and based on the GR50 values, the RCA accession was 18-fold more resistant to fomesafen than a susceptible (S) biotype. A TaqMan allelic discrimination assay and sequencing of the target-site genes PPX2 and PPX1 revealed no known or novel target-site mutations. An SYBR Green assay indicated no difference in PPX2 gene expression between the RCA and S biotypes. To test whether fomesafen resistance is metabolic in nature, the RCA and the S biotypes were treated with different cytochrome P450 (amitrole, piperonyl butoxide [PBO], malathion) and glutathione S-transferase (GST) (4-chloro-7-nitrobenzofurazan [NBD-Cl]) inhibitors, either alone or in combination with fomesafen. Malathion followed by (fb) fomesafen in RCA showed the greatest reduction in survival (67%) and biomass (86%) compared with fomesafen alone (45% and 66%, respectively) at 2 wk after treatment. Interestingly, NBD-Cl fb fomesafen also resulted in low survival (35%) compared with the fomesafen-only treatment (55%). Applications of malathion or NBD-Cl preceding fomesafen treatment resulted in reversal of fomesafen resistance, indicating the existence of cytochrome P450– and GST-based non–target site mechanisms in the RCA accession. This study confirms the first case of non–target site resistance to fomesafen in A. palmeri.
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−1 to 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 the PPX2 gene. 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 the PPX2 gene. 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.
Palmer amaranth is the most problematic weed in agronomic crop production fields in the United States. A Palmer amaranth biotype was not controlled with sequential applications of glyphosate in glyphosate-resistant (GR) soybean production field in south-central Nebraska. The seeds of the putative GR Palmer amaranth biotype were collected in the fall of 2015. The objectives of this study were to (1) confirm GR Palmer amaranth and determine the level of resistance in a whole-plant dose-response bioassay, (2) determine the copy number of 5-enolpyruvylshikimate-3-phosphate (EPSPS) gene, the molecular target of glyphosate, and (3) evaluate the response of GR Palmer amaranth biotype to POST corn and soybean herbicides with different modes-of-action. Based on the effective dose required to control 90% of plants (ED90), the putative GR Palmer amaranth biotype was 37- to 40-fold resistant to glyphosate depending on the glyphosate-susceptible (GS) used as a baseline population. EPSPS gene amplification was present in the GR Palmer amaranth biotype with up to 32 to 105 EPSPS copies compared to the known GS biotypes. Response of GR Palmer amaranth to POST corn and soybean herbicides suggest reduced sensitivity to atrazine, hydroxyphenylpyruvate dioxygenase (HPPD)- (mesotrione, tembotrione, and topramezone), acetolactate synthase (ALS)- (halosulfuron-methyl), and protoporphyrinogen oxidase (PPO)- (carfentrazone and lactofen) inhibitors. GR Palmer amaranth was effectively controlled (>90%) with glufosinate applied at 593 g ai ha−1 with ≥95% reduction in biomass. More research is needed to determine whether this biotype exhibits multiple resistant to other group of herbicides and evaluate herbicide programs for effective management in corn and soybean.
Henbit is a facultative broadleaf winter annual in the Lamiaceae family.
Acetolactate synthase (ALS) inhibitors are primarily used to control a broad
spectrum of weeds, including henbit. During 2012 to 2013, field applications
of ALS-inhibiting herbicides were ineffective in controlling a henbit
population from Marion County, KS (MCK). To confirm field-evolved resistance
to ALS inhibitors, response of MCK henbit and a known susceptible henbit
population from Kansas (DPS) to varying doses of three different ALS
inhibitors were examined: chlorsulfuron, imazamox, and propoxycarbazone.
Results of the dose–response experiments suggest that the MCK population is
highly resistant to chlorsulfuron (resistance index [R/S] > 1,000) and
propoxycarbazone (R/S = 331) but is susceptible to imazamox. A full-length
ALS gene sequence obtained using the 5′- and 3′- rapid
amplification of complementary DNA ends approach revealed a
Pro197 to Arg point mutation (a common mutation that confers
resistance to sulfonylurea herbicides, e.g., chlorsulfuron) in the MCK
henbit. No other known resistance-conferring mutations were found in the
study. Evolved resistance to major classes of ALS inhibitors in the MCK
henbit will reduce herbicide options for its control. To our knowledge, this
is the first case of evolution of herbicide resistance in henbit.
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