Search Results (21)

Acetohydroxy acid synthase (AHAS) are key targets for herbicide resistance breeding in Brassica crops, yet the evolutionary origin and functional role of AHAS2 genes in Brassica napus (AACC) and B. carinata (BBCC) remain poorly understood. Here, we investigated the distribution, ancestry, and agronomic trait associations of AHAS2 across 227 accessions representing six Brassica species. Bra.AHAS2 was amplified in 21 of 42 B. rapa (AA) accessions, and Bol.AHAS2 in 10 of 15 B. oleracea (CC) accessions. In B. napus, BnaA.AHAS2 and BnaC.AHAS2 were amplified in 73/131 and 30/131 accessions, respectively, with 19 accessions showing amplification of both homologs. All seven B. carinata accessions amplified BcaC.AHAS2. No AHAS2 homologs were amplified in three B. nigra (BB) or 29 B. juncea (AABB) accessions. Phylogenetic and gene structure analyses revealed that BnaA.AHAS2 (in B. napus) originated from Bra.AHAS2 of B. rapa, whereas BnaC.AHAS2 (in B. napus) and BcaC.AHAS2 (in B. carinata) derived from Bol.AHAS2 of B. oleracea. Association analysis showed the amplification of BnaA.AHAS2 or BnaC.AHAS2 was not associated with tribenuron-methyl resistance. However, amplification of BnaA.AHAS2 was significantly associated with reduced plant height, branching height, silique number on the terminal raceme, seed yield per plant, and thousand-seed weight in B. napus. Furthermore, haplotypes of BnaA.AHAS2 (BnaA05g03070D) were significantly associated with eicosenoic acid content, oleic acid content, flowering time, and cadmium translocation. Collectively, these findings resolve the diploid progenitor origins of AHAS2 in Brassica allotetraploids and reveal previously unrecognized associations of AHAS2 with agronomic and stress-related traits, offering valuable insights for molecular breeding in oilseed Brassica crops. Full article

Biological remediation of agricultural soils contaminated with oil is complicated by the presence of residual amounts of chemical plant protection products, in particular, herbicides, which, like oil, negatively affect the soil microbiome and plants. In this work, we studied five strains of bacteria of the genera Pseudomonas and Acinetobacter, which exhibited a high degree of oil biodegradation (72–96%). All strains showed resistance to herbicides based on 2,4-D, imazethapyr and tribenuron-methyl, the ability to fix nitrogen, phosphate mobilization, and production of indole-3-acetic acid. The presence of pollutants affected the growth-stimulating properties of bacteria in different ways. The most promising strain P. citronellolis N2 was used alone and together with oat and lupine plants for soil remediation of oil, including herbicide-treated oil-contaminated soil. Combined contamination was more toxic to plants and soil microorganisms. Bacterization stimulated the formation of chlorophyll and suppressed the synthesis of abscisic acid and malonic dialdehyde in plant tissues. The combined use of bacteria and oat plants most effectively reduced the content of hydrocarbons in the soil (including in the presence of herbicides). The results obtained can be used to develop new methods for bioremediation of soils with polychemical pollution. Full article

Flixweed (Descurainia sophia) is a weed that seriously affects wheat fields in China. Over the past 20 years, it has evolved resistance to the herbicide tribenuron-methyl. In the present study, a resistant D. sophia population with a Pro-197-Thr mutation of acetolactate synthetase (ALS) was found to have a resistance index of 457.37 for tribenuron-methyl. Under the same growth conditions, the seeds of resistant (R) and susceptible (S) populations exhibited similar vitality but the germination rates of R seeds were higher than those of S seeds. This result demonstrated that seed dormancy periods were shorter in the R seeds. RNA-Seq transcriptome analysis was then used to choose candidate genes that could regulate seed dormancy pathways in the R population. A total of 504,976,046 clean reads were selected from nine RNA-Seq libraries and assembled into 79,729 unigenes. Among these, 33,476 unigenes were assigned to 51 GO subgroups, and 26,117 unigenes were assigned to 20 KEGG secondary metabolic pathways. Next, 2473 differentially expressed genes (DEGs) were divided into three groups, as follows: G-24 h (germinating seeds) vs. D (dormant seeds); G-48 h (germinated seeds) vs. D; and G-48 h vs. G-24 h. From these 2473 DEGs, 8 were selected as candidate dormancy unigenes for the R population if their expression levels continuously decreased during the seed germination progress and their functional annotations were related to plant seed dormancy. One candidate unigene was annotated as CYP707A2; two unigenes were annotated as the transcription factors TGA4 and TGA2; one unigene was annotated as the cystathionine beta-synthase gene; and four unigenes could not be annotated as any gene listed in the six public databases. However, qRT-PCR-validated results showed that, during the germination of R seeds, the expression of the three candidate unigenes first decreased and then increased, indicating that they may have other growth-regulating functions in R populations. In brief, the dormancy function of the eight candidate dormancy unigenes needs to be further studied. Full article

Herbicide-resistant corn poppy (Papaver rhoeas L.) is one of the most important broadleaved weeds and the number of resistant cases is still growing. The aims of this study were to confirm the resistance of P. rhoeas from Tunisia to ALS inhibitors and auxin mimics and investigate the mechanisms of Target-Site Resistance (TSR) and Non-Target Site Resistance (NTSR) involved. Dose–response trials to determine cross-resistance patterns for ALS inhibitors and auxin mimics were conducted in a greenhouse. In this study, multiple resistance to tribenuron-methyl and dicamba but not to 2,4-D was found in P. rhoeas populations. Cross-resistance to imazamox was confirmed as well. Sequence analysis of the ALS gene detected target-site mutations in codon 197 of the ALS gene, namely, Pro197His, Pro197Thr, Pro197Leu, and Pro197Asn. In this study, the metabolism experiments with malathion (a cytochrome P450 inhibitor) showed that malathion reduced resistance to imazamox, indicating that P450 is involved in the resistance. TSR and NTSR mechanisms to ALS inhibitors likely coexist. The findings of this study revealed a significant synergistic interaction between malathion and dicamba in particular populations, suggesting that the resistance to auxin mimics can be conferred by enhanced metabolism. Full article

Shepherd’s-purse (Capsella bursa-pastoris), a globally distributed noxious weed species often found in wheat, has evolved resistance to ALS-inhibiting herbicides mainly due to single mutations in the ALS gene. In the present study, dose–response bioassays showed that a shepherd’s-purse population (R), collected from Xinghua, Jiangsu Province, China, had high level of resistance to the ALS-inhibiting herbicide, mesosulfuron-methyl (800-fold), and even much higher resistance levels to other reported ALS-inhibiting herbicides, tribenuron-methyl (1313-fold), bensulfuron-methyl (969-fold) and penoxsulam (613-fold). Sequencing of the open reading frame of the ALS gene revealed a double ALS gene mutation (Pro197-Ser plus Trp574-Leu) conferring the high resistance in the R plants. Docking analysis of the ALS protein and mesosulfuron-methyl predicts that the two amino acid substitutions in the R samples reduces the binding energy to the herbicide by decreasing the hydrogen bonds (H-bonds) and other interactions, thus endowing resistance to ALS-inhibiting herbicides. These results demonstrate that the double ALS mutation confers high resistance levels to ALS-inhibiting herbicides. To our knowledge, this is the first evidence of the double ALS mutation in shepherd’s-purse endowing ALS-inhibiting herbicide resistance. Full article

Tribenuron-methyl is used to control broad-leaved weeds and has a promising application prospect in millet fields. However, its negative impact on soil ecology cannot be ignored. Brassinosteroids have been widely reported to enhance plant resistance to stress, but information on brassinosteroids for the remediation of pesticide-contaminated soils is limited. Under field conditions, brassinosteroids were applied to explore their effects on the residues of tribenuron-methyl, soil enzyme activity, soil microbiol community, and millet yield. After applying brassinosteroids according to the dose of 150 mL hm⁻², the degradation rate of tribenuron-methyl accelerated. Brassinolide stimulated the activities of catalase and dehydrogenase, while the activities of sucrase and alkaline phosphatase were inhibited. The results of high-throughput sequencing showed that brassinosteroids inhibited the growth of Verrucomicrobia, Ascomycota, and Mortierellomycota and promoted the abundance of cyanobacteria. Additionally, brassinosteroids could also significantly increase the diversity index and change the community structure of soil bacteria and fungi. Further, the predicted function results indicated that brassinosteroids changed some metabolic-related ecological functions of the soil. We also found that brassinolide could increase millet yields by 2.4% and 13.6%. This study provides a theoretical basis for the safe use of tribenuron-methyl in millet fields and a new idea for the treatment of pesticide residues in soil. Full article

Herbicide-resistant weeds have been identified and recorded on every continent where croplands are available. Despite the diversity of weed communities, it is of interest how selection has led to the same consequences in distant regions. Brassica rapa is a widespread naturalized weed that is found throughout temperate North and South America, and it is a frequent weed among winter cereal crops in Argentina and in Mexico. Broadleaf weed control is based on glyphosate that is used prior to sowing and sulfonylureas or mimic auxin herbicides that are used once the weeds have already emerged. This study was aimed at determining whether a convergent phenotypic adaptation to multiple herbicides had occurred in B. rapa populations from Mexico and Argentina by comparing the herbicide sensitivity to inhibitors of the acetolactate synthase (ALS), 5-enolpyruvylshikimate-3-phosphate (EPSPS), and auxin mimics. Five B. rapa populations were analyzed from seeds collected in wheat fields in Argentina (Ar1 and Ar2) and barley fields in Mexico (Mx1, Mx2 and MxS). Mx1, Mx2, and Ar1 populations presented multiple resistance to ALS- and EPSPS-inhibitors and to auxin mimics (2,4-D, MCPA, and fluroxypyr), while the Ar2 population showed resistance only to ALS-inhibitors and glyphosate. Resistance factors ranged from 947 to 4069 for tribenuron-methyl, from 1.5 to 9.4 for 2,4-D, and from 2.7 to 42 for glyphosate. These were consistent with ALS activity, ethylene production, and shikimate accumulation analyses in response to tribenuron-methyl, 2,4-D, and glyphosate, respectively. These results fully support the evolution of the multiple- and cross-herbicide resistance to glyphosate, ALS-inhibitors, and auxinic herbicides in B. rapa populations from Mexico and Argentina. Full article

The cultivation of herbicide-resistant crops is an effective tool for weed management in agriculture. Weed control in flax (Linum usitatissimum L.) remains challenging due to the lack of available herbicide-resistant cultivars. In this study, a mutant resistant to acetolactate synthase (ALS)-inhibiting herbicides was obtained by ethyl methanesulphonate (EMS) mutagenesis using an elite cultivar, Longya10. Whole-plant dose–response assays revealed that, compared to Longya10, the mutant was 11.57-fold more resistant to tribenuron-methyl (TBM) and slightly resistant to imazethapyr (resistance index (mutant/Longya10) < 3). In vitro acetolactate synthase assays showed that the relative resistance of the mutant was 12.63 times more than that of Longya10. A biochemical analysis indicated that there was a Pro197Ser (relative to the Arabidopsis thaliana ALS sequence) substitution within the LuALS1, conferring high resistance to sulfonylurea herbicides in the mutant. Additionally, two cleaved amplified polymorphic sequence (CAPS) markers, BsaI-LuALS1 and EcoO109I-LuALS1, were developed based on the mutation site for marker assistant selection in breeding. Moreover, the mutant did not cause losses in natural field conditions. We find a mutant with ALS-inhibiting herbicide resistance chemically induced by EMS mutagenesis, providing a valuable germplasm for breeding herbicide-resistant flax varieties. Full article

Herbicide resistance in weeds, including corn poppy (Papaver rhoeas L.), is an increasing problem compromising global crop production. The aims of this study were to evaluate the susceptibility of P. rhoeas populations in Poland to acetolactate synthase (ALS) inhibitors and elucidate their mechanisms of resistance. Between 2017 and 2020, 157 seed samples were collected nationwide and a dose-response study with various ALS-inhibiting herbicides was performed in glasshouses. This revealed 14 resistant populations with R/S ranges of 2.3–1450.2, 9.5–398.5 and 2–2.5 for tribenuron, iodosulfuron and florasulam, respectively. Eight of them were cross-resistant to both tribenuron and iodosulfuron, three and one populations were singly resistant to tribenuron and iodosulfuron, respectively, and one population had reduced susceptibility to florasulam only. In one population, cross-resistance to tribenuron, iodosulfuron and florasulam was identified. The ED₅₀ of many populations susceptible to ALS inhibitors was close to half the recommended dose of the herbicides tested. In seven out of eight resistant P. rhoeas populations analysed, target-site resistance was identified. Six amino acid replacements were found (Ala197, Arg197, His197, Leu197, Ser197 and Thr197). In one population resistant to ALS inhibitors, no mutations in the ALS gene were detected. An efficient anti-resistance strategy is needed to reduce the development of herbicide resistance in P. rhoeas in Poland. Full article

The biological efficacy of herbicides MCPA+tribenuron-methyl (code name: MT-565 SG) and diflufenican+chlorotoluron (Legato Pro 425 SC) was estimated in eighteen field experiments on winter cereals in Poland and Germany to control broadleaf weeds. Postemergence application of tribenuron-methyl in combination with MCPA, applied at the 3-leaf stage to 3 tillers detectable in autumn in winter cereals, resulted in the majority of weed species occurring in autumn being effectively eliminated with MCPA+tribenuron-methyl applied at 1.0 kg∙ha^–1. It also provided an acceptable (82.4–94.1%) and comparable level of control to commonly occurring weeds Brassica napus, Capsella bursa-pastoris, Centaurea cyanus, Lamium purpureum, Tripleurospermum inodorum, Stellaria media, and Thlaspi arvense. A satisfactory level of control of 66.3 to 88.3% was confirmed for Veronica persica, Viola arvensis, and Galium aparine. According to these results, the formulation of tribenuron-methyl combined with MCPA can be recommended for application in winter cereals in the autumn as an alternative to commonly available herbicides. Full article

Silene conoidea L. is an annual troublesome broadleaf weed in winter wheat fields in China. In recent years, field applications of tribenuron-methyl have been ineffective in controlling S. conoidea in Hebei Province, China. The aim of this study was to determine the molecular basis of tribenuron-methyl resistance in S. conoidea. Whole-plant response assays revealed that the resistant population (R) exhibited a higher level of resistance (382.3-fold) to tribenuron-methyl. The R population also showed high cross-resistance to other acetolactate synthase (ALS) inhibitors, including imazethapyr, bispyribac-sodium and florasulam. However, the R population could be controlled by the field-recommended rates of bentazone, MCPA, fluroxypyr, carfentrazone-ethyl and bromoxynil. In vitro ALS activity assays indicated that the tribenuron-methyl I₅₀ value for the R population was 18.5 times higher than those for the susceptible population (S). ALS gene sequencing revealed an amino acid mutation, Trp-574-Leu, in the R population. Pretreatment with the P450 inhibitor malathion indicated that the R population might have cytochrome P450-mediated metabolic resistance. These results suggest that the Trp-574-Leu mutation and P450-mediated enhanced metabolism coexist in S. conoidea to generate tribenuron-methyl resistance. This is the first time that target-site and non-target-site resistance to tribenuron-methyl has been reported in S. conoidea. Full article

Competitive ability of cereals against segetal weeds depends among other things, on soil properties and the weather. Concerning cornflower (Centaurea cyanus L.), this issue is poorly recognized, as there are no reports on the impact of environmental conditions on the competitiveness of wheat against susceptible and resistant biotypes. Our study aimed to evaluate the effects of site and weather conditions on the competitive effects between winter wheat (WW) and two cornflower biotypes, either florasulam and tribenuron-methyl resistant (R) or sensitive (S). The experiment was conducted in a replacement series model at six sites across Poland in three growing seasons. The competitive relations were determined on the basis of two indices, i.e., the relative biomass and the number of seeds produced by the tested plants. The relative yield of wheat and weed were plotted on graphs and fitted to one of five competition models. In addition, a competitive ratio (CR) was calculated on the basis of fresh plant biomass and the number of seeds. Correlation coefficients were determined between the length of the plant, yield, biomass, the number of seeds per plant, hydrothermal coefficient K, and soil texture. Biometric parameters of wheat for its competition with two cornflower biotypes were analyzed using canonical variate analysis (CVA). The number of days to WW emergence and the day-difference between WW and cornflower (B) emergence were also calculated. The environmental characteristics of the sites, i.e., hydrothermal coefficient K and soil texture, were used as categorizing variables. Drought generally favored the greater competitive ability of WW against B for both biomass accumulation and seed production. During the first season of the research (relatively dry), only in one case out of 12 cases cornflower was more competitive than wheat. In the second year of the experiment (dry season), the competition of WW against B for resources was lower. It depended more on the site than on the cornflower biotype or the proportion of plants in the mixture. Under high or optimal rainfalls (the third year of the experiment), the competitiveness of WW toward B was significantly lower than in years with rainfall deficiency. In addition, the ability of wheat competition against the weed may have been influenced by the earlier emergence of wheat than cornflower. Even though it was sown together with wheat, cornflower emerged 0–12 days later than the tested cereal. It was also noticed that wheat was more competitive on light soils against the herbicide-susceptible (S) biotype. In contrast, greater WW competitiveness was observed against herbicide-resistant (R) cornflower on heavy soils. In conclusion, winter wheat competitiveness against herbicide-resistant or herbicide-sensitive cornflower biotypes is significantly dependent on weather and soil conditions. It is therefore reasonable to study this phenomenon in more detail. It would also be interesting to learn more about the underground competition on varying soil types under different water availability. Full article

Tribenuron-methyl-resistant (TmR) cleavers (Galium aparine L.) have been reported around the major winter wheat farming region in China. From 2017 to 2020, cleavers seeds were collected from wheat production fields across Jiangsu Province to evaluate the frequency and distribution of tribenuron-methyl-, bensulfuron-methyl- and halosulfuron-methyl-resistant cleavers, and to assess the frequency of multiple resistance. Here we report resistance frequency as percent resistance within a population, and resistance distribution as the percentage and locations of populations classified as resistant to a discriminating herbicide dose. From 2017 to 2020, cleavers populations were screened with tribenuron-methyl, bensulfuron-methyl and halosulfuron-methyl. The percentages of tribenuron-methyl-resistant cleavers populations from 2017 to 2020 were 53.33%, 51.52%, 52.38% and 47.17%, respectively; and the percentages of cleavers populations with low tribenuron-methyl resistance were 23.33%, 26.67%, 30.00% and 36.67%, respectively. The percentages of bensulfuron-methyl-resistant cleavers populations from 2017 to 2020 were 36.67%, 39.39%, 35.71% and 33.96%, respectively; and the percentages of cleavers populations had low tribenuron-methyl resistance were 30.00%, 40.00%, 53.33% and 23.33%, respectively. The percentages of halosulfuron-methyl-resistant cleavers populations from 2017 to 2020 were 26.67%, 27.27%, 50.00% and 41.51%, respectively; and the percentages of cleavers population with low tribenuron-methyl resistance were 50.00%, 53.33%, 33.33% and 40.00%, respectively. Finally, 26.67%, 22.22%, 19.05% and 20.75% of cleavers populations had resistance to 2-methyl-4-chlorophenoxyacetic acid sodium (MCPA-Na) from 2017 to 2020, respectively; however, all populations were sensitive to fluroxypyr and carfentrazone-ethyl. This confirmation of multiple resistance in cleavers populations emphasizes the importance of diversity in herbicide sites of action as critical to extending the usefulness of remaining effective herbicides such as MCPA-Na, fluroxypyr and carfentrazone-ethyl for the management of this weed. Full article

Wild mustard (Sinapis arvensis L.) and milk thistle (Silybum marianum (L.) Gaertn.) are two competitive broad-leaved weeds commonly found in cereals in Europe, while several weed species have developed resistance to the main herbicides that are applied on these crops. Thus, the implementation of integrated weed management (IWM) programs is of great importance. Field experiments were conducted based on a split-plot design with two factors (sowing date and herbicides). Our results showed that the density of wild mustard and milk thistle was higher in the early sowing compared to the late sowing, while the total weed density was up to 75% higher in early sowing. Moreover, the herbicides florasulam + 2.4-D and bromoxynil + 2.4-D exhibited high efficacy (>98%) against milk thistle and wild mustard, while tribenuron-methyl and florasulam + clopyralid provided greater efficacy in the late sowing compared to the early sowing. Among the four herbicides, the lowest dry biomass and grain yield of wheat were observed in tribenuron-methyl and florasulam + clopyralid, while in the weed-infested treatment, the highest values of both parameters were recorded in late sowing. Finally, the results showed that the sowing date is a cultural weed control method that should be implemented in IWM programs, since it can affect both weed density and herbicide efficacy. Full article

Descurainia sophia L. (flixweeds) is a noxious broad-leaf weed infesting winter wheat fields in China that has evolved high resistance to tribenuron-methyl. In this work, a brand new gene CYP77B34 was cloned from tribenuron-methyl-resistant (TR) D. sophia and transferred into Arabidopsis thaliana, and the sensitivities of Arabidopsis with or without the CYP77B34 transgene to herbicides with a different mode of actions (MoAs) were tested. Compared to Arabidopsis expressing pCAMBIA1302-GFP (empty plasmid), Arabidopsis transferring pCAMBIA1302-CYP77B34 (recombinant plasmid) became resistant to acetolactate synthase (ALS)-inhibiting herbicide tribenuron-methyl, protoporphyrinogen oxidase (PPO)-inhibiting herbicides carfentrazone-ethyl and oxyfluorfen. Cytochrome P450 inhibitor malathion could reverse the resistance to tribenuron-methyl, carfentrazone-ethyl and oxyfluorfen in transgenic Arabidopsis plants. In addition, the metabolic rates of tribenuron-methyl in Arabidopsis expressing CYP77B34 were significantly higher than those in Arabidopsis expressing pCAMBIA1302-GFP. Other than that, the transgenic plants showed some tolerance to very-long-chain fatty acid synthesis (VLCFAs)-inhibiting herbicide pretilachlor and photosystem (PS) II-inhibiting herbicide bromoxynil. Subcellular localization revealed that the CYP77B34 protein was located in the endoplasmic reticulum (ER). These results clearly indicated that CYP77B34 mediated D. sophia resistance to tribenuron-methyl and may have been involved in D. sophia cross-resistance to carfentrazone-ethyl, oxyfluorfen, pretilachlor and bromoxynil. Full article