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The results of the study of toxicity level for the lamb's quarters (Chenopodium album L.) of well-known herbicides used in soybean crops as well as herbicides promising for soybean production in the Far East are presented. Lamb's quarters (Chenopodium album L.) is a highly noxious weed species that is widespread in the south of the region. Weediness monitoring conducted in 2006-2020 showed that this species was present in the Primorsky Territory on 63-100% of fields occupied by soybeans with an average growth density of 3-117 pcs/m2. In 2013-2020 in a series of experiments, conducted in the conditions of the vegetation house and on the experimental fields of the institute, the toxicity level for Common lamb's quarters was determined by 20 herbicides (based on 14 active substances) used in soybean crops. It was found that the control of this weed species is most effective when applying herbicides in the soil and treating plants with a height of up to 4-11 cm at the early stages of development - 1-3 pairs of leaves. Reliable suppression of lamb's quarters in experiments was provided by the soil (before sowing or before soybean seedlings) use of herbicides containing active substances metribuzin, imazetapir, trifluralin, pendimetalin and acetochlor. Among the active ingredients of the tested foliar herbicides, fomesafen, combinations of bentazone + acifluorfen and imazamox + chlorimuron-ethyl were the most toxic for lamb's quarters. It is recommended to use the following herbicidal preparations to control the spread of lamb's quarters: before sowing or sprouting of soybeans - Lazurit, Zenkor Ultra, Pivot, Proponit, Fabian; for processing vegetative plants - Flex, Galaksi Top, Concept, Classic Forte (Harmony Classic), Proponit.

Six genotypes of head cabbage (‘Flexima’, ‘Mutsuma’, ‘Septima’, ‘Zeus’, DC6, ‘Ancoma’) have been studied to find an effective method of obtaining polyploids using trifluralin. The effect of various trifluralin concentrations (0.5, 1.0, 1.5, and 2.0 mg·L−1) and treatment conditions (24 °C/24 h, 24 °C/48 h, 30 °C/24 h, and 30 °C/48 h) on organogenesis of organogenic callus, hypocotyl, and seeds was tested. Ploidy level was detected using a flow cytometer. As a result, low survival of hypocotyls and calli was obtained. The shoots induced from calli and hypocotyls failed to develop quality plantlets and most calli and hypocotyls remained recalcitrant for further differentiation. Treated seeds seemed to be promising objects basing on a considerable percentage of plant regeneration. With the increased concentration of trifluralin and increasing press of treatment conditions, the seed germination rate was significantly reduced. Shoot induction from germinated seeds varied among genotypes and treatments. The root induction was independent of treatments but also reduced considerably compared to the control. Under tested conditions, none of the polyploids resulted. Further studies are required to be successful in protocol development.

In this study, various concentrations of oryzalin (0, 5, 10, 50, and 100 mg l−1), trifluralin (0, 5, 10, 50, and 100 mg l−1), and colchicine (0, 250, 500, 1000, and 5000 mg l−1) each for 12, 24, or 48 h were tested on auto-tetraploid production in Zea mays L. inbred lines "S61" and "K1263/17". The highest rates of tetraploids were obtained in the inbred line "S61" following 50 mg l−1 oryzalin for 24 h (11.1%) and colchicine treatment at 1000 mg l−1 for 24 h (12.2%) and 48 h (15.8%). In the inbred line "K1263/17", tetraploids were detected with trifluralin at 10 mg l−1 (2.3%) and 50 mg l−1 (6.9%) both for 24 h. High rate of tetraploid production (19.5%) was also observed in the presence of 1000 mg l−1 colchicine for 24 h. Compared to their diploid counterparts, the regenerated tetraploids exhibited relatively lower survival rate, and only 33.3% of the inbred line "K1263/17"-derived tetraploids survived and developed. Moreover, all survived regenerants kept their altered ploidy level until the seventh-leaf stage, and then turned the ploidy level back to the diploid state prior to the anthesis. Except for plant height, basal stem diameter, and total chlorophyll content of the seventh leaves, the diploid and re-diploidized tetraploids were not significantly different in terms of other morphological features assessed. Although it was possible to induce polyploidy in Z. mays L., the auto-tetraploids re-diploidized their genome simultaneously before anthesis, offering a precious platform to study the mechanism of genome downsizing in greater detail in this species. Auto-tetraploids can be induced in Zea mays L. using colchicine and dinitroaniline herbicides. The generated tetraploids either die or downsize their genome prior to anthesis.

Amphibians are the most important vulnerable non-target vertebrate group that are affected by pesticides. Most previous studies have confirmed the destructive effects of pesticides. But, so far, no comprehensive studies have been carried out in Iran. Therefore, to estimate the mortality rate of frogs during the growing season in different cultivating systems, we examined the presence of pesticides in water and substrate as indicators of habitat quality and in the liver tissue of Marsh frog Pelophylax ridibundus (Pallas, 1771), enclosed in the prepared cages at five rice paddy fields in Mazandaran province, Iran. The measurement of pollution was done using mass gas chromatography method and statistical analyses by Minitab software. Furthermore, the probable movement pattern of free frogs was analyzed using capture-mark-recapture method. Thirteen pesticides were detected both in the habitat and in frogs’ liver tissue. Among them s-Mevinphos, Fenitrothion, Bromofos, and Trifluralin had the most frequent occurrence in liver tissue, and Diazinon with concentrations up to 517.8 μg/Kg had the highest concentration. Furthermore, there is a significant correlation (R2 > 0.96) between water quality and frogs’ contamination, whereas, no correlation was observed between substrate pollution and frogs’ contamination. Pesticide concentrations were higher in two stations but lower than lethal doses to frogs, so that no mortality was observed at any of the stations. However, some specimens had a considerable muscle atrophy. Despite no significant movement pattern was detected, we can expect that if this trend continues, in a long term, they will face a reduction in the survival rate.

Rapid and widespread evolution of multiple herbicide resistance in global weed species endowed by increased capacity to metabolize (degrade) herbicides (metabolic resistance) is a great threat to herbicide sustainability and global food production. Metabolic resistance in the economically damaging crop weed species Lolium rigidum is well known but a molecular understanding has been lacking. We purified a metabolic resistant (R) subset from a field evolved R L. rigidum population. The R, the herbicide susceptible (S) and derived F2 populations were used for candidate herbicide resistance gene discovery by RNA sequencing. A P450 gene CYP81A10v7 was identified with higher expression in R vs. S plants. Transgenic rice overexpressing this Lolium CYP81A10v7 gene became highly resistant to acetyl-coenzyme A carboxylase- and acetolactate synthase-inhibiting herbicides (diclofop-methyl, tralkoxydim, chlorsulfuron) and moderately resistant to hydroxyphenylpyruvate dioxygenase-inhibiting herbicide (mesotrione), photosystem II-inhibiting herbicides (atrazine and chlorotoluron) and the tubulin-inhibiting herbicide trifluralin. This wide cross-resistance profile to many dissimilar herbicides in CYP81A10v7 transgenic rice generally reflects what is evident in the R L. rigidum. This report clearly showed that a single P450 gene in a cross-pollinated weed species L. rigidum confers resistance to herbicides of at least five modes of action across seven herbicide chemistries.

Eucalyptus urophylla is an important species of the genus Eucalyptus that is used across the world, especially in Brazil. It is used for pulp and paper production, and charcoal production for iron and steel industry, thus making the development of biotechnological approaches essential for the breeding programs of this species. The polyploidy induction is one such approach that may introduce phenotypic characteristics of interest to the market and industry. This is the first study that shows the production, evaluation, and development of tetraploids (4x = 44) from a clone of E. urophylla. For this, the shoot apical meristems were cultured for elongation using in vitro conditions for 15 with 5 or 10 µM colchicine, trifluralin, or oryzalin. Following this, every 30 days, the elongated plantlets were transferred to JADS medium for multiplication using tufts as explants. Thereafter, four tufts containing six to eight buds each were inoculated in flasks followed by six subcultures and were then transferred to the greenhouse for acclimatization. The experiment was performed in a completely randomized design in a factorial scheme (3x2+1: antimitotic substance x concentrations + control), with 12 replicates. Analysis of nuclear DNA content and chromosome counting by flow cytometry, photosynthetic pigment quantification, stomatal size and density, survival, and rooting were performed for 110 days after acclimatization. Data were submitted for analysis of variance followed by Scott & Knott’s test (P ≤ 0.05). As a result, we observed that the oryzalin and trifluralin treatments were efficient in the induction of tetraploid plants, while colchicine proved to be ineffective. Also, trifluralin treatment at 5 µM provided a higher polyploidy induction (18.2% tetraploids) in comparison to the others. Diploid plants displayed 1.41 pg of nuclear DNA content with 22 chromosomes, while tetraploid plants showed 2.86 pg with 44 chromosomes. In conclusion, the tetraploid plants, as compared to the diploids (control), were found to show higher chlorophyll content, larger but fewer stomata, and lower rooting and survival, which can be mitigated nonetheless through silvicultural techniques. This study shall bring new perspectives for the breeding programs of E. urophylla

Standard ecological risk assessment practices often rely on larval and juvenile fish toxicity data as representative of the amphibian aquatic phase. Empirical evidence suggests that endpoints measured in fish early life stage tests are often sufficient to protect larval amphibians. However, the process of amphibian metamorphosis relies on endocrine cues that affect development and morphological restructuring and are not represented by these test endpoints. The present study compares developmental endpoints for zebrafish (Danio rerio) and the African clawed frog (Xenopus laevis), 2 standard test species, exposed to the herbicide trifluralin throughout the larval period. Danio rerio were more sensitive and demonstrated a reduction in growth measurements with increasing trifluralin exposure. Size of X. laevis at metamorphosis was not correlated with exposure concentration; however, time to metamorphosis was delayed relative to trifluralin concentration. Gene expression patterns indicate discrepancies in response by D. rerio and X. laevis, and dose-dependent metabolic activity suggests that trifluralin exposure perturbed biological pathways differently within the 2 species. Although many metabolites were correlated with exposure concentration in D. rerio, nontargeted hepatic metabolomics identified a subset of metabolites that exhibited a nonmonotonic response to trifluralin exposure in X. laevis. Linking taxonomic distinctions in cellular-level response with ecologically relevant endpoints will refine assumptions used in interspecies extrapolation of standard test effects and improve assessment of sublethal impacts on amphibian populations. Environ Toxicol Chem 2020;39:1797-1812. Published 2020. This article is a US government work and is in the public domain in the USA.

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.

Processing-tomato (Lycopersicon esculentum L.) cultivation in California has transitioned to the use of transplants, subsurface drip irrigation (SDI), and shallow tillage from direct-seeded, flood-...

Praxelis is an annual to short-lived perennial herb in the Asteraceae family and an emerging problematic weed species in Florida nurseries. The objective of these experiments was to determine efficacy of selected PRE and POST herbicides for control of praxelis. An additional experiment was conducted to determine efficacy of the same PRE herbicides for control of bluemink, a weed similar in appearance to praxelis that is also common in Florida. The granular herbicides dimethenamid + pendimethalin, flumioxazin, oxyfluorfen + pendimethalin, oxyfluorfen + prodiamine, and trifluralin + isoxaben were evaluated, along with spray-applied formulations of dimethenamid, indaziflam, and prodiamine + isoxaben. Flumioxazin consistently provided over 90% control of praxelis at both sites. Indaziflam control was inconsistent between the two sites, providing 100% control in Apopka but only a 22% reduction in weed counts in Balm. Oxyfluorfen + pendimethalin, oxyfluorfen + prodiamine, and prodiamine + isoxaben also provided control (57% to 97% reduction in shoot weight), albeit generally to a lesser degree than was observed with flumioxazin. All PRE herbicides provided similar control of both praxelis and bluemink, with the exception of dimethenamid and dimethenamid + pendimethalin, which reduced bluemink shoot weight more than praxelis. Clopyralid, glyphosate, and triclopyr all provided effective POST control of mature and flowering praxelis through 12 wk after treatment (WAT) and resulted in greater praxelis control than glufosinate. Results suggest that many commonly used PRE and POST herbicides would provide control of praxelis, but use of PRE and POST herbicides as well as sequential applications may be needed for long-term management.

Organophosphate insecticides, which have the capacity to inhibit specific herbicide-degrading (cytochrome P450) enzymes, have been used to explore metabolic herbicide-resistance mechanisms in weeds. This study investigates the response of seven field-selected rigid ryegrass (Lolium rigidum Gaudin) populations to herbicides from three different sites of action in the presence or absence of the P450 inhibitor phorate. Phorate antagonized the thiocarbamate herbicides triallate and prosulfocarb (8-fold increase in LD50) in multiple resistant L. rigidum populations with resistance to three different site-of-action herbicides. In contrast, phorate synergized trifluralin and propyzamide in some populations, reducing the LD50 by 50%. Conversely, treatment with phorate had no significant effect on the LD50 for S-metolachlor or pyroxasulfone (inhibitors of very-long-chain fatty-acid synthesis). Phorate has diverse effects that are herbicide and population dependant in field-selected L. rigidum, suggesting P450 involvement in the metabolism of trifluralin and failure to activate thiocarbamate herbicides in these populations. This research highlights the need for implementation of diverse approaches other than herbicide alone as part of a long-term integrated strategy to reduce the likelihood of metabolism-based resistance to PPI herbicides in L. rigidum.

A total of six field experiments were conducted in southwestern Ontario over a 3-yr period (2016, 2017, 2018) to evaluate the efficacy of trifluralin and halosulfuron applied preplant incorporated (PPI) for weed management in white bean. Trifluralin, halosulfuron, and trifluralin + halosulfuron applied PPI caused as much as 2%, 6%, and 8% white bean injury, respectively. Weed interference delayed maturity and reduced white bean yield 56% compared with the weed-free control. Weed interference with trifluralin and halosulfuron applied alone reduced white bean seed yield as much as 35% and 29%, respectively; however, white bean seed yield with the trifluralin + halosulfuron tankmixes was similar to the weed-free control. Trifluralin, halosulfuron, and trifluralin + halosulfuron applied PPI provided 6%–12%, 75%–92%, and 71%–95% control of velvetleaf; 89%–95%, 93%–98%, and 96%–99% control of pigweed species; 5%–18%, 82%–96%, and 90%–97% control of common ragweed; 90%–97%, 81%–97%, and 95%–99% control of common lambsquarters; 23%–43%, 55%–88%, and 83%–96% control of flower-of-an-hour; 4%–25%, 94%–100%, and 95%–100% control of wild mustard; 96%–100%, 18%–45%, and 97%–100% control of barnyardgrass; and 92%–98%, 21%–40%, and 93%–98% control of green foxtail, respectively. Results indicated that low rates of trifluralin tank-mixed with halosulfuron has the potential to control problematic weeds and improve white bean yields in Ontario.

It is exceptionally difficult to have productive farming without the use of herbicides. It has been found that they create difficulties in the growth and biological nitrogen fixation of some legume crops after their application. Adverse impacts may result from a direct effect of the herbicide on rhizobial growth and/or an indirect effect on plant growth. In order to study the effect of some PRE and POST herbicides on nodulation, nitrogen fixation, and growth characteristics of three chickpea cultivars, a greenhouse trial was conducted in a completely randomized design with factorial arrangement and three replications. Pyridate, imazethapyr, and trifluralin were applied with the recommended rates of 1100, 100, and 720 g ai ha−1, respectively, and also non-treated control on three chickpea cultivars (ILS482, Hashem, and Kaka). The results showed that trifluralin and pyridate herbicides had the highest and the least negative impact on growth characteristics of three examined cultivars. Pyridate showed a positive effect on vegetative characteristics but nodulation and total nitrogen contents in the shoots of chickpea cultivars decreased significantly. Results also revealed that root and shoot growth, nodulation, and nitrogen fixation diminished significantly when imazethapyr and trifluralin were applied to Hashem and ILC482 cultivars. It seems that pyridate is an appropriate herbicide for broadleaf weed control in chickpea fields.

There are a limited number of soil-applied herbicides available for broad-spectrum weed control in dry bean production in Ontario, Canada. Four experiments were conducted from 2017 to 2019 in southwestern Ontario to compare the efficacy of six soil-applied grass herbicides [trifluralin (600 g ai ha-1), ethalfluralin (810 g ai ha-1), pendimethalin (1080 g ai ha-1), S-metolachlor (1050 g ai ha-1), dimethenamid-p (544 g ai ha-1) and EPTC (3400 g ai ha-1)] and halosulfuron (35 g ai ha-1) applied alone and in combination, applied preplant incorporated (PPI), on white bean tolerance and yield, and weed control efficacy. There was no white bean injury from the herbicide treatments evaluated. Grass herbicides (trifluralin, ethalfluralin, pendimethalin, S-metolachlor dimethenamid-P and EPTC) controlled velvetleaf 0% - 82%, pigweeds 87% - 99%, common ragweed 0% - 93%, common lambsquarters 81% - 99%, wild mustard 0% - 71%, barnyardgrass 98% - 100% and green foxtail 98% - 99%. Halosulfuron controlled velvetleaf 98%, pigweeds 94%, common ragweed 90% - 94%, common lambsquarters 97%, wild mustard 98% - 100%, barnyardgrass 19% - 24% and green foxtail 20% - 25%. Tankmixes of halosulfuron with soil-applied grass herbicides provided ≥93% control of the weed species evaluated. Reduction in density and biomass generally followed the same trend as visible control with herbicide treatments evaluated. Weed interference reduced white bean seed yield 70%. Seed yield was 53% - 66% of the weed-free control with trifluralin, ethalfluralin, pendimethalin, S-metolachlor and dimethenamid-P, 81% of the weed-free control with EPTC, 58% of the weed-free control with halosulfuron, and 87% - 95% of the weed-free control with halosulfuron tankmixes with the grass herbicides evaluated. Based on these results, halosulfuron in combination with trifluralin, ethalfluralin, pendimethalin, S-metolachlor, dimethenamid-p and EPTC, applied PPI at rates evaluated, can be used to effectively control common annual grass and broadleaf weeds in white beans.

There are a limited number of herbicides registered for weed management in white bean production in Ontario, Canada. Five field experiments were completed in Ontario from 2016 to 2018 to compare the efficacy of trifluralin and ethalfluralin applied alone and in combination with halosulfuron, applied preplant incorporated (PPI), for weed control efficacy and white bean tolerance and seed yield. At 2 and 4 WAE, there was no white bean injury from the herbicide treatments evaluated. Trifluralin applied PPI provided up to 32%, 99%, 13%, 99%, 27%, 99% and 99% control of velvetleaf, redroot pigweed, common ragweed, common lambsquarters, wild mustard, barnyardgrass and green foxtail, respectively. Trifluralin and ethalfluralin provide similar control of velvetleaf, redroot pigweed, barnyardgrass and green foxtail control, however, ethalfluralin is slightly more efficacious on common ragweed, common lambsquarters and wild mustard. Halosulfuron (35 g∙ai∙ha−1), applied PPI, provided as much as 76%, 98%, 96%, 96%, 100%, 19% and 23% control of velvetleaf, redroot pigweed, common ragweed, common lambsquarters, wild mustard, barnyardgrass and green foxtail, respectively. Trifluralin (600 or 1155 g∙ai∙ha−1) + halosulfuron (35 g∙ai∙ha−1), applied PPI, provided up to 88%, 100%, 98%, 100%, 100%, 99% and 98% control of velvetleaf, redroot pigweed, common ragweed, common lambsquarters, wild mustard, barnyardgrass and green foxtail, respectively. Ethalfluralin (810 or 1080 ai∙ha−1) + halosulfuron (35 g∙ai∙ha−1) provided similar control. Weed interference decreased white bean seed yield 44% - 45% with trifluralin, 30% - 41% with ethalfluralin and 34% with halosulfuron. However, decreased weed interference with trifluralin and ethalfluralin applied in combination with halosulfuron resulted white bean seed yield that was similar to the weed-free control. Trifluralin or ethalfluralin co-applied with halosulfuron can be safely used in white bean production for the control of common annual grass and broadleaf weeds in Ontario.

To investigate the effect of herbicide Trifluralin and the role of magnetized water on enzymatic activities and content of proline and growth substances including gibberellin (GA3) and cytokinin (6- benzyl amino purine), an experiment was conducted using squash (Cucurbita pepo L. var. Shiraz Hybrid F1) seedlings. Four levels of trifluralin (0, 5, 15 and 25 ppm) and irrigation by distilled and magnetized water were applied in a completely randomized design in 3 replicates at Urmia University in 2016. Different levels of trifluralin were added to pots 2 days after planting. Seedlings were treated by modified Hoagland solution with diurnal temperature 30:18 °C, relative humidity of 70-80 percent and light period of 16:8 hr, during a 5-week growing period in growth chamber. The effect of magnetized water and trifluralin on ornithine amino-transferase (OAT), proline dehydrogenase (PDH) and content of proline in both shoots and roots as well as GA3 and cytokinin was significant (p>0.01) according to ANOVA. The content of proline and OAT in roots and shoots as well as GA3 and cytokinin increased significantly, although the PDH of roots and shoots decreased under the influence of magnetized water. The content of proline and OAT increased and GA3, cytokinin and the activity of PDH declined by the increase of the levels of trifluralin. Overall, it can be concluded that the growth and biochemical indices of squash plants were improved by magnetized water under toxicity of herbicide trifluralin.

Seven algal species were used to conduct toxicity assays with 12 herbicides to determine differences in species sensitivity. A fluorescence microplate toxicity assay was used as an efficient and economical high-throughput assay. The obtained toxicity data were standardized based on the species sensitivity distribution concept. The most-sensitive individual species differed among herbicides: Desmodesmus subspicatus was most sensitive to chloronitrofen and pendimethalin; Achnanthidium minutissimum was most sensitive to chlorpropham; Nitzschia palea was most sensitive to diquat, glyphosate, and dichlobenil; Navicula pelliculosa was most sensitive to trifluralin; and Pseudanabaena foetida was most sensitive to glufosinate, asulam, and 2,4-D. Surprisingly, Raphidocelis (formerly Pseudokirchneriella) subcapitata, a standard green alga, was not the most sensitive to any of the herbicides. The results clearly showed that a single algal species cannot represent the algal assemblage in terms of sensitivity. Therefore, multispecies algal toxicity data sets are essential for assessing the ecological effect of herbicides.

Plumbago auriculata Lam. (Plumbaginaceae) is an ornamental/medicinal flowering shrub. However, it could be stop growing under 5 °C resulting in its poor germplasm resources. Increasing the tolerance of polyploids to environmental stress will enrich its germplasm resources. In this study, we thus employed the stem segments and buds exposed to pendimethalin and trifluralin in vitro to conduct an effective polyploid protocol, in which buds were cultured for 7 days in MS medium with 800 μM. In this study, we have developed a novel polyploid identification method based on accuracy and efficiency. Early screening was performed based on the morphological characteristics of this plants, and then polyploidy was determined by flow cytometry. The number of chromosomes was determined by the chromosomal accounting. At the anatomical level, in comparison with diploids, the root parenchyma, stem collenchyma and tetraploid spongy tissues were thickened, the guard cell size and vascular bundles number were increased, and the stomata were widened; but the stomatal density was decreased. After 24 h of cold stress, the maximal quantum yield of photosystem II (Fv/Fm) and maximum primary yield of photochemistry of PSII (Fv/Fo) of diploids decreased significantly. Conversely, the minimal value of chlorophyll a fluorescence (Fo), relative electrical conductivity and malondialdehyde increased markedly. Accordingly, the cold tolerance of the tetraploids presented better than these of the diploids. Taken together, our study has first developed an effective protocol to obtain and identify the tetraploid of P. auriculate plants. Furthermore, it was proved that this polyploid has presented improving cold tolerance and enriching phenotypic properties. These findings could be useful for improving cold-tolerance breeding and enriching genetic diversity of P. auriculate plants. We used stem segments and buds exposed to pendimethalin and trifluralin in vitro to conduct polyploids of Plumbago auriculata and compared characterizations including cold tolerance between diploids and polyploids.

Due to the low availability of herbicide active ingredients, the chemical weed management in lettuce crop is a challenge. The aim of this study was to evaluate the sensitivity of lettuce to the pre-planting application of the herbicides flumioxazin and trifluralin alone or in tank-mixture in lettuce, Lucy Brown cultivar. Two experiments were conducted. In experiment A, the soil was tilled before the herbicide application and it was tilled again before seedling transplanting. In experiment B, the soil was tilled just once before the herbicide application. The experimental design was in randomized blocks in a factorial scheme (3x3) +1 (2 factors with 3 levels and an additional control) with 4 replicates. We applied flumioxazin (120 g ha-1), trifluralin (450 g ha-1) and flumioxazin + trifluralin (120 + 450 g ha-1) in three different seasons before transplanting (0, 7 and 14 days). Lettuce productivity after trifluralin application (48.4 t ha-1) was higher than after flumioxazin application (16.8 t ha-1) in experiment A. In experiment B, flumioxazin decreased the number of lettuce leaves (25.6) at 14 DAT; no differences in lettuce productivity were verified, though. The herbicide trifluralin caused lower levels of phytotoxicity and was the most selective between the treatments when applied in the three seasons in both experiments. RESUMO Devido à baixa disponibilidade de ingredientes ativos de herbicidas, o manejo químico de plantas daninhas na cultura da alface é um desafio. Objetivou-se avaliar a sensibilidade da alface à aplicação dos herbicidas flumioxazin e trifluralin, isolados ou em mistura em pré-plantio de mudas de alface cultivar Lucy Brown. Dois experimentos foram realizados. O experimento A recebeu uma operação de revolvimento no solo antes da aplicação dos herbicidas e outra antes do transplantio das mudas; o experimento B recebeu apenas um revolvimento do solo anterior à aplicação do herbicida. O delineamento experimental foi em blocos casualizados em esquema fatorial (3x3) + 1 (2 fatores com 3 níveis e uma testemunha adicional) com 4 repetições. Ambos experimentos receberam os tratamentos herbicidas flumioxazin (120 g ha-1), trifluralin (450 g ha-1) e a associação flumioxazin + trifluralin (120 + 450 g ha-1) aplicados em três épocas distintas antecedendo o transplantio (0, 7 e 14 dias). A produtividade de alface após aplicação de trifluralin (48,4 t ha-1) foi maior do que sob aplicação de flumioxazin (16,8 t ha-1) no experimento A. No experimento B flumioxazin aos 14 DAT reduziu o número de folhas de alface (25,6), todavia não houve diferenças de produtividade. O herbicida trifluralin causou menores níveis de fitointoxicação na alface e foi o mais seletivo dentre os tratamentos quando aplicado nas três épocas nos dois experimentos.

Amaranthus palmeri S. Watson (Palmer amaranth) is considered a problematic and troublesome weed species in many crops in the USA, partly because of its ability to evolve resistance to herbicides. In this study, we explored the mechanism of resistance in a trifluralin-resistant A. palmeri accession collected from Arkansas, USA. Dose-response assays using agar plates demonstrated an EC50 (effective concentration that reduces root length by 50%) of 1.02 µM trifluralin compared to 0.39 µM obtained in the susceptible accession. Thus, under these conditions, the resistant accession required 2.6 times more trifluralin to inhibit root length by 50%. Seeds in the presence or absence of the cytochrome P450-inhibitior malathion displayed a differential response with no significant influence on root length, suggesting that resistance is not P450-mediated. In addition, application of 4-chloro-7-nitrobenzofurazan (NBD-Cl), a glutathione S-transferase (GST) inhibitor, showed significant differences in root length, indicating that GSTs are most likely involved in the resistance mechanism. Sequencing of α- and β-tubulin genes revealed no single nucleotide polymorphisms (SNPs) previously described between accessions. In addition, relative gene copy number of α- and β-tubulin genes were estimated, however, both resistant and susceptible accessions displayed similar gene copy numbers. Overall, our results revealed that GST-mediated metabolism contributes to trifluralin resistance in this A. palmeri accession from Arkansas.

Chloris virgata is a problematic weed in mungbean crops due to its high seed production, resistance to glyphosate and high dispersal ability. Pot and field experiments were conducted in 2020 and 2021 to evaluate a range of preemergent (PRE) herbicides for C. virgata control in mungbean. In the field and pot studies, isoxaflutole 75 g ai ha−1 caused crop injury, and in the field experiment, it reduced mungbean yield by 61% compared with the best treatment (pyroxasulfone 100 g ai ha−1). In the field and pot experiments, dimethenamid-P 720 g ai ha−1, pyroxasulfone 100 g ai ha−1 and S-metolachlor 1400 g ai ha−1 provided &gt, 88% control of C. virgata (for reduced biomass) and in the field experiment, these herbicides resulted in improved yield by 230%, 270% and 170%, respectively, compared with nontreated control (250 kg ha−1). Similarly, pendimethalin 1000 g ai ha−1 and trifluralin 600 g ai ha−1 provided &gt, 89% control (biomass) of C. virgata, and in the field experiment, these resulted in improved yields of 230% and 160%, respectively, compared with the nontreated control. PRE herbicides such as diuron 750 g ai ha−1, linuron 1100 g ai ha−1, metribuzin 360 g ha−1, terbuthylazine 750 g ai ha−1, imazapic 48 g ai ha−1 and imazethapyr 70 g ha−1 although did not cause crop injury, however, these herbicides did not control C. virgata. Flumioxazin 90 g ai ha−1 caused reduced biomass of C. virgata by 80% compared with the nontreated control, and in the field experiment, it resulted in improved yield by 140% compared with the nontreated control. This study suggests the potential use of herbicides, such as dimethenamid-P, pyroxasulfone and S-metolachlor in addition to pendimethalin and trifluralin, for C. virgata control in mungbean. Further studies are needed to determine the efficacy of dimethenamid-P, S-metolachlor and pyroxasulfone for controlling other troublesome weeds in mungbean.

Amaç: Bu çalışma; organoklorlu (Endosulfan) ve pyrethroid (Cypermethrin) pestisitler ile herbisitlerin (2,4-D ve Trifluralin) Hershberger metodu ile peri-pubertal erkek sıçan modelinde serum LH ve FSH düzeyleri ile androjen duyarlı dokular üzerine androjenik ve anti-androjenik etkilerini araştırmak amacıyla planlandı.Gereç ve Yöntem: Çalışmada 56 adet peri-pubertal 42 günlük erkek Sprague Dawley sıçan kullanıldı.İlk grup sham kastrasyon olarak ayrıldı. Geriye kalan 49 adet sıçan kastre edildi. On günlük iyileşme süresinin sonunda sıçanlar yedi gruba daha ayrıldı: Kastrasyon grubu; Testosteron propiyonat (TP) grubu; TP+Flutamide grubu; TP+Endosulfan grubu; TP+2,4-D grubu; TP+Cypermethrin grubu; TP+Trifluralin grubu. Son uygulamadan 24 saat sonra sıçanlar dekapite edilerek kan örnekleri toplandı. Tüm hayvanların prostat bezi, bulbouretral bezleri, seminal vezikülleri ve musculus levator ani (mLA) diseke edilerek tartıldı. Serum LH, FSH ve testosteron düzeyleri ELISA yöntemi ile belirlendi.Bulgular: Kastrasyon, tüm androjen-duyarlı doku ağırlıklarında azalmaya neden olurken (p0.05). 2,4-D, Cypermethrin ve Trifluralin serum FSH düzeylerinde anlamlı artışa yol açarken; serum LH düzeylerindeki artış sadece Trifluralin grubunda istatistiksel olarak anlamlı idi. Sonuç: Çalışmadan elde edilen bulgular, Hershberger metodunda Endosulfan, Cypermethrin ve Trifluralin’in bulbouretral bez üzerinde anti-androjenik; 2,4-D’nin ise vesicula seminalis üzerinde androjenik etkilere sahip olduğunu göstermiştir. Bu çevresel kirleticilerin erkek üreme fonksiyonlarını olumsuz yönde etkileyebileceği öngörülebilir., Aim: This study was aimed to investigate the androgenic and anti-androgenic effects of organochlorid (Endosulfan) and pyrethroid (Cypermethrin) pesticides and herbicides (2,4-D and Trifluralin), on serum LH and FSH levels and androgen-sensitive tissues in peri-pubertal rat model with Hershberger assay. Material and Method: 56 peri-pubertal 42 days old Sprague Dawley rats were used in the study. The first group was separated as sham castration. The remaining 49 rats were castrated. After 10 days recovery period, rats were divided into further seven groups: Castration group; Testosterone Propionate (TP) group; TP+Flutamide group; TP+Endosulfan group; TP+2,4-D group; TP+Cypermethrin group; TP+Trifluralin group. 24 hours after the last application, rats were decapitated and blood samples were obtained. Prostate, bulbourethral gland, vesicula seminalis and musculus levator ani (mLA) were dissected and weighed. Serum LH, FSH and testosteron levels were determined with ELISA. Result: Castration caused significant reduction in all androgen-sensitive tissue weights n(p0.05). 2,4-D, Cypermethrin and Trifluralin significantly increased serum FSH levels; but the increase in serum LH levels was only significant in Trifluralin group. Conclusion: Data obtained from this study showed that 2,4-D has androjenic effects on vesicula seminalis whereas Endosulfan, Cypermethrin and Trifluralin have anti-androgenic effects on bulbourethral gland. It can be suggested that, these environmental pollutants could affect male reproductive functions negatively.

Ispaghul (Plantago ovata Forsk.) as an important medicinal plant has obtained a remarkable reputation due to therapeutic applications of seed mucilage. To determine the effect of in vitro-induced polyploidy on various characteristics of P. ovata, the terminal bud of two true leaves seedlings were separately treated with colchicine [0.1, 0.3 and 0.5% (w/v) for 6, 12 and 24 h] and trifluralin [7.5, 15 and 22.5% (w/v) for 24, 48 and 72 h] solutions. The ploidy level of induced tetraploids was determined via chromosome counting of root tip cells, and then confirmed through flow-cytometric analysis. Comparison the morphological, physiological, anatomical features of intact diploids and induced tetraploids revealed that tetraploids had considerable more height, thicker leaf, larger spike and seed, larger pollen grain and more seeds per spike. Moreover, the amount of chlorophyll (a, b, and total) and carotenoids, as well as chloroplast number in guard cells was further in tetraploids than diploids. Unlike density, stomata size in tetraploids was bigger than that one in diploids. It was also observed that seeds of tetraploids had more mucilage than diploids. In summary, we firstly developed the P. ovata tetraploids and suggested 0.3% colchicine for 24 h and 22.5% trifluralin for 72 h as the optimum treatments for inducing tetraploidy in P. ovata.

Field studies were conducted from 2009 through 2011 at the Sustainable Agriculture Research and Extension Center near Lingle, Wyoming, to evaluate great northern bean response to PRE flumioxazin mixed with either trifluralin, pendimethalin, or ethalfluralin. Seven treatments were arranged in a randomized complete block with three or four replicates y–1. The soil texture of the study site was loam in 2009 and 2011, and sandy loam in 2010. Soil organic matter ranged from 1.4% to 1.8%. Treatments included flumioxazin plus trifluralin, flumioxazin plus pendimethalin, flumioxazin plus ethalfluralin, ethalfluralin plus EPTC, imazamox plus bentazon (POST), hand-weeded control, and nontreated control. Dry bean density 4 wk after planting differed among herbicide treatments (P < 0.001). Treatments that included flumioxazin reduced dry bean density 54% compared with treatments without flumioxazin. Dry bean yield was influenced by dry bean density; on average, yield in flumioxazin-containing herbicide treatments was 30% less than treatments not containing flumioxazin, even though weed control was generally greater in flumioxazin treatments.Nomenclature: bentazon; EPTC; ethalfluralin; flumioxazin; imazamox; pendimethalin; trifluralin; dry bean; Phaseolus vulgaris L.

Two field experiments were conducted during 2018 at Paskeville and Arthurton, South Australia, to identify effective herbicide options for the control of thiocarbamate-resistant rigid ryegrass in wheat. Dose–response experiments confirmed resistance in both field populations (T1 and A18) of rigid ryegrass to triallate, prosulfocarb, trifluralin, and pyroxasulfone. T1 and A18 were 17.9- and 20-fold more resistant to triallate than susceptible SLR4. The level of resistance detected in T1 to prosulfocarb (5.9-fold) and pyroxasulfone (4-fold) was lower compared to A18, which displayed 12.1- and 7.8-fold resistance to both herbicides, respectively. Despite resistance, the mixture of two different preplant-incorporated (PPI) site-of-action herbicides improved rigid ryegrass control and wheat yield compared to a single PPI herbicide only. Prosulfocarb + triallate and prosulfocarb + S-metolachlor + triallate did not reduce rigid ryegrass seed set when compared to prosulfocarb applied alone at the higher rate (2,400 g ai ha–¹). Pyroxasulfone + triallate PPI followed by glyphosate (1,880 g ai ha⁻¹) as a weed seed set control treatment reduced rigid ryegrass seed production by 93% and 95% at both sites, respectively. These herbicides also significantly improved grain yield of wheat at Paskeville (22%) and Arthurton (38%) compared to the untreated. Nomenclature: Prosulfocarb; triallate; pyroxasulfone; S-metolachlor; trifluralin; glyphosate; rigid ryegrass, Lolium rigidum Gaudin LOLRI; wheat; Triticum aestivum L. ‘Chief CL Plus'

Plantago psylliumis is under development as the source of mucilage, but has not been entirely domesticated. Since mucilage content is low and variable in the medicinal plant, it is imperative to apply breeding approaches to accelerate the domestication process and improve the mucilage content in the seeds. One approach to achieve these purposes is polyploidy induction, which can increase the size of plants and their phyto-medicines. To conduct this approach and evaluate the effect of in vitro-induced polyploidy on different traits of P. psylliumis, a gradient of colchicine concentrations, 0.1, 0.3 and 0.5% (w/v), was used to treat the terminal buds for 6, 12 and 24 h. The terminal buds were separately treated with different trifluralin concentrations, 7.5, 15 and 22.5% (w/v), for 12, 24 and 48 h. The optimal treatments of 0.5% colchicine for 24 h, and of 22.5% (w/v) trifluralin for 72 h resulted in an induction efficiency of 23% and 19%, respectively. Chromosome counting revealed successful chromosome duplication in tetraploids (2n = 4x = 24), in contrast with intact diploids (2n = 2x = 12). Duplication of DNA size in tetraploids was also proved by flow-cytometry analysis. The tetraploids size was larger than their intact diploids for spike, seed and pollen grain, leaf thickness, and plant height. The stomata of tetraploids were larger with lower density than diploids. The chloroplast number in guard cells, along with chlorophyll and carotenoid content all were higher in tetraploids. In final, our study suggests that tetraploidization could be useful to improving the seed mucilage content for commercial use. We firstly fulfilled the in vitro induced tetraploidy in P. psyllium by 0.5% colchicine for 24 h and 22.5% (w/v) trifluralin for 72 h, and improved the mucilage content in the seeds.

Trifluralin, a pre-emergent herbicide, is widely used in Brazil in the weed grass management in restoration areas. The objective was to evaluate the tolerance of Piptadenia gonoacantha to t...

Seeds of chia (Salvia hispanica L.) are known worldwide for the benefits in human nutrition. However, from the agricultural point of view, weed interference is one of the main limitations in this crop, and restricted basic information about the management with herbicides. The aim of this work was to evaluate the selectivity of herbicides applied in pre-emergence of chia crop (S. hispanica). It was carried out an experiment in the field at the Universidade Estadual do Centro-Oeste, Guarapuava - PR, using randomized complete block design, with ten treatments and four replications. The treatments applied in the pre-emergence of chia were constituted by the herbicides: sulfentrazone (100, 200, 400 and 800 g ha-1), oxyfluorfen (720 g ha-1), s-metolachlor (1920 g ha-1), flumioxazin (50 g ha-1), diclosulan (25 g ha-1), trifluralin (1800 g ha-1) and a control without application. Oxyfluorfen, s-metolachlor, flumioxazin and diclosulan were not selective for application in chia crop in pre-emergence, unlike trifluralin, which showed high selectivity. Sulfentrazone presented viability only at the dose of 100 g ha-1.

Field research was conducted in 2012 and 2013 in Georgia, New York, and North Carolina to evaluate the effect of trifluralin PPI on turnip root production. Treatments included trifluralin PPI at 0, 0.42, 0.56, 0.84, 1.12, 1.68, 2.24, and 3.36 kg ai ha−1. Aboveground injury to turnip varied by location and increased from 0% to 85% as trifluralin rate increased from 0.42 to 3.36 kg ha−1. Trifluralin at 0.42 to 0.84 kg ha−1caused ≤7% injury, except at Clayton, NC, and Freeville, NY, where injury ≤32%. Trifluralin at 0.42 to 0.84 kg ha−1reduced turnip root yield ≤11% at all locations, except Clinton, NC, where yield was reduced 29% and 43% by 0.56 and 0.84 kg ha−1, respectively. Turnip roots were not injured internally by trifluralin. Our research results suggest that up to 0.84 kg ha−1trifluralin PPI is safe to use in turnip roots.

To develop an easy and reliable method for detecting pesticides and their residues in the Mekong Delta, a GC-MS analytical method was developed and validated according to European guidelines (SANTE/11945/2015) for the determination of residues of three pesticides (quinalphos, trifluralin and dichlorvos) in water. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.002 and 0.007 μg/L, respectively, for quinalphos and trifluralin, and 0.016 and 0.053 μg/L, respectively, for dichlorvos and quinalphos. The repeatability, the within-laboratory reproducibility as well as the trueness met the European criteria. The recovery rate ranged between 72% (for dichlorvos and quinalphos) and 82% (for trifluralin). The developed method was then applied for the analysis of 33 water samples, collected in April 2013, at the beginning of the rainy season in the Mekong Delta in Vietnam. Thirteen samples were from rice field, 10 were collected from cat fish ponds and from red tilapia cages. Results showed that only 9% of total water samples analysed contained residues of pesticides, but only in water from rice fish systems. From the 13 samples taken in these systems, quinalphos was detected in three samples. The other two pesticides were not detected. A comparison between analytical results obtained from GC-MS and an alternative method, that is GC-ECD indicated that GC-ECD is less sensitive than GC-MS, with LOQ ranging from 0.37 to 1.18 (depending on the pesticide). However, for samples with concentrations above these LOQ, no significant difference was observed between the results obtained from the two analytical methodologies.

Overreliance on herbicides for weed control is conducive to the evolution of herbicide resistance. Annual ryegrass (Lolium rigidum Gaud.) is a species that is prone to evolve resistance to a wide range of herbicide modes of action. Rapid detection of herbicide-resistant weed populations in the field can aid farmers to optimize the use of herbicides for their control. The feasibility of a portable agar-based test to rapidly and reliably detect annual ryegrass resistance to key herbicides such as clethodim, glyphosate, pyroxasulfone and trifluralin on-farm was investigated. The three research phases of this study show that: a) easy-to-interpret results are obtained with non-dormant seed from well-characterised susceptible and resistant populations, and resistance is detected as effectively as with traditional dose-response pot-based resistance assays. However, the test may not be suitable for portable use on-farm because of b) the low stability of some herbicides such as trifluralin and clethodim in agar and c) the tendency of seed dormancy in freshly-harvested seeds to confound the results. The agar-based test is best used as a research tool as a complement to confirm results obtained in traditional pot-based resistance screenings. Comprehensive agar test and / or whole-plant resistance tests by herbicide application at the recommended label rate (whole plants grown in pots) are the current benchmark for proactive in- and off-season resistance testing and should be promoted more widely to allow early detection of resistance, optimization of herbicide technology use and deploy appropriate weed management interventions.

BACKGROUND Lolium rigidum is the weed of greatest economic impact in Australia due to its formidable capacity to evolve herbicide resistance. In this study, 579 field-sampled L. rigidum populations were tested for resistance to 21 herbicides applied at the recommended rate. Nine herbicide treatments were binary mixtures. RESULTS A total of 15 876 individual resistance tests were conducted by screening two million seeds at the recommended label rate. The overall frequency of resistant populations was 31%, 14%, 71%, 6% and 0% in response to the post-emergence herbicide treatments clethodim, clethodim + butroxydim, imazamox + imazapyr, glyphosate and paraquat, respectively. The resistance frequency to stand-alone pre-emergence wheat-selective herbicides ranged from 10% to 34%. Conversely, the levels of resistance to pre-emergence mixtures or stand-alone propyzamide were significantly lower, ranging from 6% to 0%. In winter, the responses to glyphosate, paraquat, cinmethylin, prosulfocarb, pyroxasulfone and trifluralin were reassessed, with 7%, 0%, 0%, 21%, 21% and 28% as the respective resistance frequencies. South Australia and Victoria are identified as epicenters for L. rigidum population resistance to pyroxasulfone, whereas populations in New South Wales have the greatest resistance to glyphosate and in Western Australia to clethodim. CONCLUSIONS For the first time, resistance levels to stand-alone herbicides and binary mixtures are geographically ranked across the Australian continent by benchmark statistical analysis of resistance frequencies and distribution. The extension of these results will raise awareness of rapidly emerging patterns of herbicide resistance, encouraging the adoption of cost-effective modes of action and integration of diverse strategies for weed resistance management.

Dinitroanilines are microtubule inhibitors, targeting tubulin proteins in plants and protists. Dinitroaniline herbicides, such as trifluralin, pendimethalin and oryzalin, have been used as pre-emergence herbicides for weed control for decades. With widespread resistance to post-emergence herbicides in weeds, the use of pre-emergence herbicides such as dinitroanilines has increased, in part, due to relatively slow evolution of resistance in weeds to these herbicides. Target-site resistance (TSR) to dinitroaniline herbicides due to point mutations in α-tubulin genes has been confirmed in a few weedy plant species (e.g.,Eleusine indica,Setaria viridis, and recently inLolium rigidum). Of particular interest is the resistance mutation Arg-243-Met identified from dinitroaniline-resistantL. rigidumthat causes helical growth when plants are homozygous for the mutation. The recessive nature of the TSR, plus possible fitness cost for some resistance mutations, likely slows resistance evolution. Furthermore, non-target-site resistance (NTSR) to dinitroanilines has been rarely reported and only confirmed inLolium rigidumdue to enhanced herbicide metabolism (metabolic resistance). A cytochrome P450 gene (CYP81A10) has been recently identified inL. rigidumthat confers resistance to trifluralin. Moreover, TSR and NTSR have been shown to co-exist in the same weedy species, population, and plant. The implication of knowledge and information on TSR and NTSR in management of dinitroaniline resistance is discussed.

Terminalia actinophylla (Mart), commonly known as "chapada", is used in folk medicine for the treatment of diarrhea, diabetes and anti-inflammatory. However, studies on its toxicity are still incipient. The present work was carried out with the objective of evaluating the phytochemical profile and the cytogenotoxic effect of the ethanolic extract of the T. actinophylla bark using the Allium cepa L. seed. Seeds of A. cepa were subjected to different extract concentrations (1.25, 2.5, 5 and 10 mg/mL), a negative control (NC - distilled water) as well as trifluralin (PC - positive control), during 24 h of exposure. The phytochemical profile of the extract was obtained to identify the main secondary metabolites. The cytotoxicity (mitotic index) and the genotoxicity (chromosomal alterations) were analyzed by means of the collection of 5,000 meristematic cells. The statistical analysis was carried out using the Kruskal-Wallis test (p

In vitro regeneration systems provide a powerful tool for manipulating ploidy to facilitate breeding and development of new crops. Polyploid induction can expand breeding opportunities, assist with the development of seedless triploid cultivars, enhance ornamental characteristics and environmental tolerances, increase biomass and restore fertility in wide hybrids. In vitro ploidy manipulation is commonly induced using antimitotic agents such as colchicine, oryzalin and trifluralin, while many other antimitotic agents have been relatively unexplored. Successful induction requires a synergistic pairing of efficient penetration of the antimitotic agent and may be dependent the length of exposure and concentrations of antimitotic agents, tissue types, and interactions with basal media and plant growth regulators. In vitro conditions vary among taxa and individual genera, species, and cultivars, often requiring unique treatments to maximize polyploid induction. In some taxa, the induction of polyploidy influences in vitro growth, development, and root formation. Here we provide an overview of mitotic inhibitors and their application for in vitro ploidy manipulation for plant breeding and crop improvement.

Calendula officinalis L. is known as an ornamental plant as well as a source of biochemical compounds used in cosmetics and industry. C. officinalis has a complex karyotype. Published chromosome numbers differ between 2n = 4x = 28 or 32. We have estimated genome sizes in nine commercial cultivars and evaluated the ploidy level by karyotyping and fluorescent in situ hybridization (FISH) using 5S and 45S rDNA loci. The detection of chromosome sets of two rather than four homologues would suggest that C. officinalis has an allotetraploid background. In addition, four signals for 45S but only two for 5S were found by using FISH. Artificial chromosome doubling is a common technique in plant breeding, as polyploidization results in several consequences for plant growth and development. Especially the suggested allotetraploid background in C. officinalis is interesting when examining the effect of chromosome doubling on the plant phenotype. Here we describe chromosome doubling of three allotetraploid cultivars of C. officinalis, 'Nova,' 'WUR 1553-7' and 'Orange Beauty'. Three antimitotic agents - colchicine, oryzalin and trifluralin - were used in different concentrations to find the combination of the best agent and the best dosage to obtain octaploids. For all three cultivars a few octaploids were obtained. A concentration of 200 and 400 ppm of colchicine was most efficient for chromosome doubling in 'Nova' and 'Orange Beauty,' respectively. For 'WUR 1553-7' the treatment with 20 ppm oryzalin was also effective. Cell numbers and first observations of the phenotype in the chromosome doubled plants show thicker leaves and bigger cells, as commonly observed after ploidy doubling. Due to the low number of chromosome doubled plants obtained more elaborate phenotyping will be performed on following generations cultivated under field conditions.

Challenges for wheat doubled haploid (DH) production using anther culture include genotype variability in green plant regeneration and spontaneous chromosome doubling. The frequency of chromosome doubling in our program can vary from 14% to 80%. Caffeine or trifluralin was applied at the start of the induction phase to improve early genome doubling. Caffeine treatment at 0.5 mM for 24 h significantly improved green plant production in two of the six spring wheat crosses but had no effect on the other crosses. The improvements were observed in Trojan/Havoc and Lancer/LPB14-0392, where green plant numbers increased by 14% and 27% to 161 and 42 green plants per 30 anthers, respectively. Caffeine had no significant effect on chromosome doubling, despite a higher frequency of doubling in several caffeine treatments in the first experiment (67&ndash, 68%) compared to the control (56%). In contrast, trifluralin significantly improved doubling following a 48 h treatment, from 38% in the control to 51% and 53% in the 1 &micro, M and 3 &micro, M trifluralin treatments, respectively. However, trifluralin had a significant negative effect on green plant regeneration, declining from 31.8 green plants per 20 anthers (control) to 9&ndash, 25 green plants per 20 anthers in the trifluralin treatments. Further work is required to identify a treatment regime with caffeine and/or anti-mitotic herbicides that consistently increases chromosome doubling in wheat without reducing green plant regeneration.

BACKGROUND Resistance to the dinitroaniline herbicide trifluralin in Lolium rigidum (annual ryegrass) often is mediated by the enhanced capacity to metabolize the herbicide to less toxic polar conjugates and/or by functionally recessive target-site mutations in α-tubulin. RESULTS In two L. rigidum populations possessing enhanced trifluralin metabolism, resistance was largely reversed by recurrent selection with the thiocarbamate herbicide prosulfocarb (i.e. plant survival was two- to >20-fold lower). Their ability to metabolize trifluralin was significantly decreased (by ≈2.3-fold) following recurrent prosulfocarb selection, to levels comparable to those observed in susceptible plants or when trifluralin metabolism was inhibited by treatment with the insecticide phorate. CONCLUSIONS This study provides evidence that trait(s) enabling efficient trifluralin metabolism in L. rigidum are purged from the population under prosulfocarb recurrent selection. The level of trifluralin metabolism in vitro and its inhibition caused by phorate action on trifluralin-metabolizing enzyme(s) is equivalent to the effect produced by prosulfocarb selection. The hypothetical link between the two phenomena is that the putative monooxygenase(s) conferring trifluralin metabolic resistance also mediate the activation of prosulfocarb to its toxic sulfoxide. Thus, we speculate that survival to prosulfocarb via a lack of metabolic herbicide activation, and survival to trifluralin conferred by enhanced herbicide metabolism, are mutually exclusive. These findings not only open up a new research direction in terms of the interaction between different herbicide resistance mechanisms in L. rigidum, but also offer strategies for immediate management of the population dynamics of metabolism-based resistance in the field. © 2020 Society of Chemical Industry.

Herbicides and insecticides are widely used in modern agriculture. It has been reported in various studies that application of insecticides can increase tolerance of herbivorous insects to insecticides. However, limited information exists on susceptibility to insecticides when insects are exposed to herbicides. This study was conducted to investigate the potential impact of the herbicides trifluralin and 2-methyl-4-chlorophenoxyacetic acid sodium salt (MCPA-Na) on the susceptibility of the nocturnal moth Spodoptera litura to the insecticides λ-cyhalothrin, phoxim and bifenthrin. We found that larvae exposed to trifluralin or MCPA-Na became significantly less susceptible to both insecticides than non-exposed control larvae. Herbicide-treated larvae did not show altered growth under the used test conditions. However, heads of herbicide-treated larvae showed increased expression of the acetylcholinesterase genes SlAce1 and SlAce2. Moreover, the fat body and midgut of herbicide-treated larvae displayed elevated expression of detoxification genes (the carboxylesterase gene SlCarE; the glutathione S-transferase genes SlGSTe2 and SlGSTe3; the cytochrome P450 monooxygenase genes CYP6B48, CYP9A40 and CYP321B1). The CYP6B48 gene exhibited highest inducibility. In conclusion, the data of this study suggest that exposure of S. litura larvae to herbicides may stimulate detoxification mechanisms that compromise the efficacy of insecticides.

In order to identify combinations of effective preemergent herbicides for weeds in sugarcane, an experiment was conducted at the Agricultural School of Concepcion; Concepcion, Paraguay. The experimental model was subdivided plots, with two factors: A1 without herbicide for broadleaf weeds, A2 diuron (dose 3 t ha-1), A3 atrazine (dose 4 kg ha-1), A4 sulfentrazone (dose 1.2 t ha-1) and B1 without herbicide for weeds, B2 trifluralin (dose 3 l ha-1), B3 acetochlor (dose 1 l ha-1), forming twelve treatments with three repetitions. They were applied in preemergence, where the evaluations were from 20 to 41 days after application (DDA), for: degree of control, residual effect and control of predominant weeds. The combination of diuron and acetochlor, produced high weed control (more 90% up to 41 days DDA), being superior to the other treatments. Sida spp was better controlled with diuron; whereas Mollugo verticillata was effectively controlled with most combinations; Ipomoea nil was more sensitive to sulfentrazone, while; Cenchrus echinatus was effectively controlled, with the combination of diuron plus acetochlor. It is concluded that with the combination of diuron and acetochlor, a large variety of weeds was controlled and the highest residual effect was recorded. In addition, atrazine combined with graminicides (trifluralin or acetochlor) or diuron with acetochlor, provided total control of weeds. It is recommended: for Sida spp. diuron with acetochlor or atrazine. For Mollugo verticillata, sulfentrazone alone or combined with graminicides. Ipomoea nil, sulfentrazone. Cenchrus echinatus, diuron and acetochlor combined. Key words: Saccharum officinarum, weeds, preemergence herbicides.

An effective chromosome doubling protocol was established in essential garden crop of cucumber (Cucumis sativus L.) Cv. Hi Power. The different concentrations of colchicine (0, 250, 500, 750, and 1500 mg/L), oryzalin (0, 5, 15, 25, 50, 75, and 150 mg/L) and trifluralin (0, 5, 15, 25, 50, 75, and 150 mg/L) were applied on parthenogenesis-induced haploid nodal and shoot tip explants of cucumber for 18 and 38 h in three independent factorial experiments. Increasing concentrations of applied antimitotic agents led to the significant reduction in the survival rate of both shoot tip and nodal explants, especially in longer exposure duration. Three ploidy levels including haploid, mixoploid, and doubled haploid were regenerated form both explant types treated with colchicine, oryzalin, and trifluralin. Flow cytometry analysis proved successful chromosome doubling of haploid plants. Based on the results obtained, the highest number of regenerated doubled haploid plants (92.31%) and fruit set (86.21%) were related to immersion of nodal explants in 50 mg/L oryzalin for 18 h. The highest doubled haploid regeneration for colchicine and trifluralin antimitotic agents were 58.33 and 83.33%, respectively. The leaf size of doubled haploid plants was larger than their correspond haploids. The optimized chromosome doubling protocol would be applicable for doubled haploid production in garden crops of Cucurbitaceae family, which is recalcitrant to the spontaneous doubling, and also for in vitro polyploidy induction studies.

Glyphosate-resistant (GR) Italian ryegrass is one of the most troublesome weeds in Mississippi row crop production. Fall-applied residual herbicide applications are recommended for control of GR Italian ryegrass. However, carryover of residual herbicides applied in fields for rice production can have a negative impact on rice performance. Field studies were conducted in Stoneville, MS, to determine the effects of fall-applied residual herbicides on rice growth and yield. Herbicide treatments included suggested use rates (1×) of clomazone at 840 g ai ha–1, pyroxasulfone 170 g ai ha–1,S-metolachlor 1,420 g ai ha–1, and trifluralin 1,680 g ai ha–1, and two times (2×) the suggested use rates in the fall before rice seeding. Pooled across application rate, pyroxasulfone,S-metolachlor, and trifluralin injured rice to an extent 28% to 36% greater than clomazone 14 d after emergence (DAE). Rice seedling density and height 14 DAE and rice maturity were negatively affected by all fall-applied herbicides except clomazone. Applications at 2× rates reduced rough rice yields in plots treated with pyroxasulfone,S-metolachlor, and trifluralin compared with clomazone. Pyroxasulfone applied at the 2× rate reduced rough rice yield 22% compared with the 1× rate. Rough rice yield was 90% or greater of the nontreated control in plots treated with either rate ofS-metolachlor, and these were comparable with rough rice yields from plots treated with both rates of trifluralin and the 1× rate of pyroxasulfone. Early-season injury and reductions in seedling density and height 14 DAE, would preclude even 1× applications of pyroxasulfone,S-metolachlor, and trifluralin from being viable options for residual herbicide treatments targeting GR Italian ryegrass in the fall before rice seeding. Of the herbicides evaluated, only clomazone should be utilized as a fall-applied residual herbicide treatment targeting GR Italian ryegrass before seeding rice.

Growth chamber experiments were conducted to evaluate the response of sesame to PRE and POST applications of soil residual herbicides. PRE applications of acetochlor andS-metolachlor at 1.26 and 1.43 kg ai·ha−1showed little or no sesame injury (0 to 1%) 4 wks after herbicide treatments (WAT). POST treatments of acetochlor and trifluralin made 3 wks after planting (WAP) resulted in greater sesame injury (40%) compared to applications at bloom (18%). Field studies were conducted in Texas and Oklahoma during the 2014 and 2015 growing seasons to determine sesame response to clethodim, diuron, fluometuron, ethalfluralin, quizalofop-P, pendimethalin, pyroxasulfone, trifluralin, and trifloxysulfuron-sodium applied 2, 3, or 4 weeks after planting (WAP). Late-season sesame injury with the dinitroaniline herbicides consisted of a proliferation of primary branching at the upper nodes of the sesame plant (in the shape/form of a broom). Ethalfluralin and trifluralin caused more “brooming” effect than pendimethalin. Some yield reductions were noted with the dinitroaniline herbicides. Trifloxysulfuron-sodium caused the greatest injury (up to 97%) and resulted in yield reductions from the untreated check. Early-season diuron injury (leaf chlorosis and necrosis) decreased as application timing was delayed, and late-season injury was virtually nonexistent with only slight chlorosis (

Resistance to the pre-emergence herbicide trifluralin is increasing in Australian annual ryegrass (Lolium rigidum) populations. Three L. rigidum populations (R1, R2, and R3) collected from Australian grain fields were identified with trifluralin resistance. Both target-site and nontarget-site resistance mechanisms were investigated. No target-site α-tubulin mutations were detected in populations R1 and R3, while an Arg-243-Lys mutation was found in R2. Compared with the three trifluralin-susceptible populations, enhanced [14C]-trifluralin metabolism, quantified by measuring the amount of [14C] label partitioning into the polar phase of a hexane:methanol system, was identified in all the three resistant populations. This is the first report of metabolic resistance to trifluralin. Coevolution of target-site and nontarget-site resistance to trifluralin is occurring, and metabolic resistance is not rare in L. rigidum populations in Australia. A method was established for trifluralin metabolic resistance detec...

Large amounts of trifluralin are applied each year for weed control; however, its effects on soil microbial communities and functions are unknown. Two agricultural soils, one silty loam and one silty clay were spiked with TFL (0, 0.84, 8.4, and 84 mg kg–1) and studied the effects using a laboratory microcosm approach. The half-lives were 44.19–61.83 d in all cases. Bacterial abundance increased 1.12–5.56 times by TFL, but the diversity decreased. From the next-generation sequencing results, TFL altered the bacterial community structure, which initially diverged from the control community structure, then recovered, and then diverged again. Linear discriminant analysis effect size indicated that Sphingomonas and Xanthomonadaceae were the predominant species on day 7 and 15 in TFL treatments. N2-fixing bacteria were initially increased, then decreased, and then recovered, and it was positively correlated with NH4+-N content. Compared with the control, ammonia-oxidizing bacteria were decreased by 25.51–92.63%, ammonia-oxidizing archaea were decreased by 17.12–85.21% (except day 7), and the NO3−-N concentration was also inhibited. In contrast to bacteria, fungal abundance was inhibited without any observable effects on fungal diversity or community structure. These results suggest that TFL impacts soil bacterial community and alters functional microorganisms involved in soil N processing.

A population of rigid ryegrass (Lolium rigidumGaudin) from a field on the Eyre Peninsula, South Australia, was suspected of resistance to thiocarbamate herbicides. Dose–response studies were conducted on this population (EP162) and two susceptible populations (SLR4 and VLR1). The resistant population exhibited cross-resistance to triallate, prosulfocarb, EPTC, and thiobencarb with higher LD50to triallate (14.9-fold), prosulfocarb (9.4-fold), EPTC (9.7-fold), and thiobencarb (13.6-fold) compared with the susceptible populations SLR4 and VLR1. The resistant population also displayed resistance to trifluralin, pyroxasulfone, and propyzamide. The LD50of the resistant population was higher for trifluralin (13.8-fold), pyroxasulfone (8.1-fold), and propyzamide (2.7-fold) compared with the susceptible populations. This study documents the first case of field-evolved resistance to thiocarbamate herbicides inL. rigidum.

Annual crops and crotalaria intercropping has been used to control nematodes. The evaluation of the susceptibility of crotalaria species to herbicides is an important step to make this technology viable. The aim of this experiment was to determine the tolerance of Crotalaria juncea, C. ochroleuca, C. spectabilis and C. breviflora to pre and post-emergence herbicides. Post-emergence (bentazon, ametryn, amicarbazone, 2,4-D amine and mesotrione) and pre-emergence herbicides (atrazine + mesotrione, atrazine + simazine, atrazine, isoxaflutole, pendimenthalin, s-metolachlor and trifluralin) were evaluated in doses of 0.5D; 0.75D; 1D and 1,25D; in which D was the commercial dose of each product. The results demonstrated that the species of crotalaria differ on the susceptibility to pre and post emergence herbicides. Among the post-emergence herbicides, bentazon showed higher selectivity. However, reduced doses of some post-emergence herbicides can make the intercropping with some species of crotalaria viable. The most selective pre-emergence herbicides were pendimenthalin, s-metolachlor and trifluralin. In general, C. spectabilis was the most susceptible to the evaluated herbicides.

The purpose of this study was to clarify the kinetic bioaccumulation potential of herbicides in the earthworm, Pheretima spp., the most common earthworms throughout Asia, and Eisenia spp., litter-feeding earthworms included in the test species recommended by the Organization for Economic Co-operation and Development. The kinetic bioaccumulation factors of trifluralin and pendimethalin were estimated from an uptake test for 10 or 12 days and from an elimination test for 10 days. The time required to reach a steady state following herbicide exposure was 7 days for both herbicides in Eisenia spp. and 1 day in Pheretima spp. The uptake rate constant (g-soil/g-worm/day) and elimination rate constant (per day) for trifluralin were 2.1 and 0.23 in Eisenia spp. and 0.42 and 0.45 in Pheretima spp., respectively, and those for pendimethalin were 1.5 and 0.26 in Eisenia spp. and 0.27 and 1.0 in Pheretima spp., respectively. Kinetic bioaccumulation factors of both herbicides were relatively close to bioaccumulation factors in steady state and were higher in Eisenia spp. (8.9 for trifluralin and 5.7 for pendimethalin) than in Pheretima spp. (0.95 and 0.26). These results demonstrated that the herbicide bioaccumulation risk is lower for Pheretima spp. than for Eisenia spp. because of the lower uptake rate and higher elimination rate in Pheretima spp.