Detached leaf culture: viability to evaluate 2,4-D toxicity symptoms in cotton apex leaves

To study the viability of detached leaf culture technique, studies were carried out with detached leaves from cotton apex (true trilobed leaves). The prepared leaves were sprayed with 2,4-D amine and ester, at rates of 10, 30, 70, and 100% of the recommended doses. Detached leaves without herbicide spray were used as controls. Simultaneously, a greenhouse experiment was conducted with the same treatments as used for the detached leaves experiment. Toxicity was measured through a 0-to-5 grading according to the percentage of affected leaf area in the detached leaves experiment or examining the affected rate of whole plant as indicated in the greenhouse. Results showed that the ester form of the herbicide induced earlier and more severe toxicity symptoms in detached leaves and greenhouse grown plants. Positive and significant correlations (p < 0.001) were found between toxicity results obtained at 7 and 14 days after application in detached leaves and greenhouse plants (r = 0.97 and 0.92, respectively). Negative, significant correlations (p < 0.005) were found between the toxicity levels found at 7 and 14 days after application in detached leaves and dry matter of cotton plants grown in the greenhouse (r= - 0.92 and -0.92, respectively).     

Vapour of the free acid of the herbicide 2,4-D is toxic to tomato and lettuce plants

Tomato and lettuce plants were exposed to vapour of the free acid of [14C-phenyl] 2,4-D at concentrations in the range 1-600 pg litre(-1) for periods of 6, 24 or 72 h. The rate of uptake of radiolabel by tomato was about twice that by lettuce at the same vapour concentration. Uptake rates were linearly related to external vapour concentration. The relationship between uptake and vapour concentration of 2,4-D for the two species was similar to published values for the butyl and iso-octyl esters. The distribution of herbicide residue in the plant immediately after exposure indicated that the apical leaves of lettuce are particularly active in assimilating vapour, whereas for tomato, leaf position had no influence. Forty days after exposure, both species showed symptoms of toxicity and reduction in shoot dry weight typical of similar doses of 2,4-D esters. It is concluded that the vapour of 2,4-D represents a potential hazard to susceptible plants, and that further work is needed to determine the conditions likely to lead to the production of vapour of the free acids of phenoxyalkanoic herbicides following spraying.     

Detached Leaf Culture

Abstract

To study the viability of detached leaf culture technique, studies were carried out with detached leaves from cotton apex (true trilobed leaves). The prepared leaves were sprayed with 2,4-D amine and ester, at rates of 10, 30, 70, and 100% of the recommended doses. Detached leaves without herbicide spray were used as controls. Simultaneously, a greenhouse experiment was conducted with the same treatments as used for the detached leaves experiment. Toxicity was measured through a 0-to-5 grading according to the percentage of affected leaf area in the detached leaves experiment or examining the affected rate of whole plant as indicated in the greenhouse. Results showed that the ester form of the herbicide induced earlier and more severe toxicity symptoms in detached leaves and greenhouse grown plants. Positive and significant correlations (p < 0.001) were found between toxicity results obtained at 7 and 14 days after application in detached leaves and greenhouse plants (r = 0.97 and 0.92, respectively). Negative, significant correlations (p < 0.005) were found between the toxicity levels found at 7 and 14 days after application in detached leaves and dry matter of cotton plants grown in the greenhouse (r = ?0.92 and ?0.92, respectively).     

The impact of SO2 and SO2 + ascorbic acid treatments on growth and partitioning of dry matter in Trigonella foenum-graecum L

To study the impact of SO(2) and SO(2) + ascorbic acid on growth and partitioning of dry matter in Trigonella foenum-graecum L., two-week-old plants were exposed to SO(2) for 2 h daily over a 42 day period. One of the exposed sets was treated with ascorbic acid. Plants were grown in a wire house and unexposed plants were used as controls for comparison. The parameters measured, such as dry weights of leaf, stem and root per plant, were found to be lower in the exposed sets than in the controls. The reductions were greater in dry weights of stem and root as compared with weights of leaves, indicating that the partitioning of the dry matter was altered. Greater amounts of soluble sugars and starch in the leaves of exposed plants, compared with the stem, also revealed that translocation was hampered. Reductions were greater in fruiting than in flowering, suggesting that fruit abortion was high. Although ascorbic acid treatment could mitigate the effect of SO(2), the differences were not found to be statistically significant. Significant changes were seen in fruit yield, suggesting that the effect of ascorbic acid is cumulative. The impact of SO(2) and SO(2) + ascorbic acid on partitioning of dry matter to different 'sinks' is discussed.     

Dissipation of 2,4-D in soils of the Humid Pampa region, Argentina: a microcosm study

Phenoxy herbicides like 2,4-dichlorophenoxyacetic acid (2,4-D) are widely used in agricultural practices. Although its half life in soil is 7-14d, the herbicide itself and its first metabolite 2,4-dichlorophenol (2,4-DCP) could remain in the soil for longer periods, as a consequence of its intensive use. Microcosms assays were conducted to study the influence of indigenous microflora and plants (alfalfa) on the dissipation of 2,4-D from soils of the Humid Pampa region, Argentina, with previous history of phenoxy herbicides application. Results showed that 2,4-D was rapidly degraded, and the permanence of 2,4-DCP in soil depended on the presence of plants and soil microorganisms. Regarding soil microbial community, the presence of 2,4-D degrading bacteria was detected even in basal conditions in this soil, possibly due to the adaptation of the microflora to the herbicide. There was an increment of two orders of magnitude in herbicide degraders after 15d from 2,4-D addition, both in planted and unplanted microcosms. Total heterotrophic bacteria numbers were about 1x10(8) CFUg(-1) dry soil and no significant differences were found between different treatments. Overall, the information provided by this work indicates that the soil under study has an important intrinsic degradation capacity, given by a microbial community adapted to the presence of phenoxy herbicides.     

The effect of propanil co-application on 2,4-D sorption by soil

The herbicide 2,4-D is often applied as a tank mixture in combination with other herbicide products. However, current information on 2,4-D sorption by soil is largely based on batch-equilibrium experiments without considering the competition of other herbicides for sorption sites by soil. This study quantified the effect of the herbicide propanil on the sorption of 2,4-D in soil. Results indicated that propanil competed with 2,4-D for sorption sites, particularly in soils with an organic carbon content greater than 3.6%. The decrease in 2,4-D sorption by soil, as a result of propanil competition, was most notably for herbicide concentrations that are typical of recommended field rates. We conclude that herbicide co-applications on agricultural fields have the potential to increase the mobility of herbicides in soil.     

Transient effect of the herbicide flazasulfuron on carbohydrate physiology in Vitis vinifera L

In the Champagne vineyard, most of the areas treated in early 2000 with the newly approved herbicide flazasulfuron had vines with altered growth and yellow leaves throughout the growing season. In order to clarify the physiological perturbations caused on the non-target grapevine and their potential consequences, C nutrition of grape plants grown in vineyards treated or not with flazasulfuron in 2000 was characterized during the following season. Vines from treated areas exhibited yellow leaves and an alteration of photosynthetic activity, characterized by declines in leaf gas exchanges (by 85%) and photosynthetic pigment concentrations (by 88%), and a marked disorganization of the leaf plastids. The herbicide also caused a decrease in leaf starch and soluble carbohydrate levels (-74% and -90%, respectively). Surprisingly, some vines re-greened after bloom, then exhibiting similar carbohydrate physiology to those grown in a non-treated area. Thus, recovery of CO(2) fixation rates, plastid ultra-structure, pigment concentrations and carbohydrate levels was found in re-greening leaves. Unlike the informations available in the literature, our results showed that flazasulfuron may be phytotoxic for grapevine. However, this toxicity was overcome the following year, indicating that vines have the potential to recover from this herbicide stress after one season.     

2,4-dichlorophenoxyacetic acid (2,4-D) micropollutant herbicide removing from water using granular and powdered activated carbons: a comparison applied for water treatment and health safety

Abstract

Activated carbons are well-known porous materials as an effective adsorbent used for the removal of emerging contaminants, such as herbicides, which are increasingly present in water bodies. Most water treatment plants, specially in Brazil, are unable to completely remove such contaminants by the conventional process and advanced treatment using activated carbons is required. The aim of this paper was to verify the influence of the activated carbons granulometry and specific surface area on the 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide removal efficiency using distilled-deionized water and filtered water collected from a conventional Water Treatment Plant. Commercial activated carbons samples used in this work were obtained from two different manufacturers. Activated carbons were analyzed by the specific surface area, pore size and volume distribution, nuclear magnetic resonance, infrared and x-ray spectroscopy, moisture, volatile matter and ash contents. Batch adsorption isotherms experiments were used and performed by Langmuir and Freundlich models. Granular and powdered activated carbons removed over 99% of 2,4-D in distilled water and near to 99% using filtered water. The activated carbons evaluated in this work presented high performance and played a key role in water treatment by removing 2,4-D herbicide, ensuring the protection of human health and the ecosystem.     

Effects of 2,4-dichlorophenoxyacetic acid formulation on medulla spinalis of Poecilia reticulata: A histopathological study

This study investigated the possible effects of a commonly used foliar herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) formulation on medulla spinalis of lebistes. Fish were exposed to 2,4-D (15, 30, 45 mg L⁻¹), behavioral changes were monitored. Fish were fixed, histopathological examination was carried out on sections taken from the upper parts of the fish body. Histopathology showed increase in neuronal loss, swelling indicating formation of intracellular edema, vacuolization noticed as the formation of vacuoles within or adjacent to cells, deformation in the Nissl granules, pyknosis and gliosis in medulla spinalis. Behavioral changes were decreased general activity, grouping, shortness in breath, sudden rotations and jumping, loss of equilibrium and colour. In conclusion, this commercial formulation of 2,4-D is considerably neurotoxic to lebistes. Fish constitute the last link in the chain of the feeding cycle in aquatic eco-system, number of studies investigating acute and chronic neurotoxicity of various herbicides in fish should be increased.     

Effects of nozzle types and 2,4-D formulations on spray deposition

The objective of this study was to evaluate the effects of nozzle types and 2,4-D formulations on spray deposition on different targets. Two field experiments were carried out in a completely randomized design, and treatments were arranged in a factorial scheme. Species in experiment 1 were Sumatran fleabane (Conyza sumatrensis) and Brazil pusley (Richardia brasiliensis) and in experiment 2 were soybeans (Glycine max) and Benghal dayflower (Commelina benghalensis). For both experiments, the first factor corresponded to spray nozzles with different settings (AD 110.015 – 61 and 105 L ha^?1; AD 015-D – 75 and 146 L ha^?1; XR 110.0202 – 200 L ha^?1; and ADIA-D 110.02 – 208 L ha^?1) and the second factor consisted of two formulations of 2,4-D (amine and choline). The formulation of 2,4-D choline has contained Colex-D? Technology. Similar or higher spray deposition was observed on the leaves and artificial targets when using 2,4-D choline as compared to the 2,4-D amine formulation, and these differences in deposition were more evident for nozzles applying lower spray volumes. Deposition was more affected by nozzle type when amine formulation was used, compared to choline formulation.     

Dissipation and translocation of saisenxin in tobacco and soil under conventional field and controlled laboratory conditions

Abstract

The aim of this investigation was to investigate the fate and translocation characteristics of saisenxin (SSX), a novel organic zinc fungicide, in the environment and tobacco plants under conventional field and laboratory conditions. A rapid and sensitive analytical technique based on high-performance liquid chromatography was used for determination of SSX, in soil samples and tobacco leaf, stem and root samples. The method had satisfactiry linearity (R^2?=?0.9999) and the limits of detection and of quantitation of the target compound were 0.06 and 0.20?mg kg^?1, respectively. The average recoveries were in the range of 89.74–94.24% in soil, leaf, stem and root samples, with relative standard deviations of <8%. For conventional field trials, the half-life (t_1/2) of SSX was 5.9–6.5 days in soil and 4.8–5.3 days in tobacco leaves; the corresponding values under controlled laboratory conditions were extended to 7.1 and 7.6?days. The translocation factor (TF) values were in the range of 0–2.25 and 0–0.25 for foliage and root irrigation treatments, respectively. The TFs of SSX in tobacco indicated that tobacco had a high ability to transfer SSX upward.     

Mobility and efficacy of 2,4-D herbicide from slow-release delivery systems based on organo-zeolite and organo-bentonite complexes

This research aimed to develop slow-release formulations (SRFs) of 2,4-dichlorophenoxyacetic acid (2,4-D) using zeolite and bentonite minerals modified with cetyltrimethylammonium (CTMA) surfactant. Adsorption–desorption, greenhouse bioassay and column experiments were carried out to assess the potential of the SRFs to control weeds while reducing the herbicide leaching losses to deep layers of soil. The results showed that only 6.5 mmol 2,4-D kg^?1 was retained by Na-bent, and the herbicide was not adsorbed by Na-zeol at all. The surface modification with CTMA surfactant, however, improved the 2,4-D adsorption capacity of the zeolite and bentonite up to 207.5 and 415.8 mmol kg^?1, respectively. The synthesized organo-minerals slowly released the retained 2,4-D discharging 22 to 64% of the adsorbed 2,4-D to the solution phase within 7 days. The SRFs significantly (P = 0.05) reduced the herbicide mobility within the soil columns keeping a great portion of the herbicide active ingredient in the upper 5 cm soil layer. The SRFs were significantly (P = 0.05) as effective as the free technical herbicide in weed control without harming the ryegrass as the main plant. Therefore, the synthesized SRFs could be considered as useful tools for weed control in sustainable agriculture.     

Toxic effects of the herbicide 2,4-dichlorophenoxyacetic acid on lymphoid organs of the rat

We investigated the toxicity of the widely used herbicide dimethylammonium salt of 2,4-dichlorophenoxyacetic acid (DMA-2,4-D) on the lymphoid system of rats after a single dose oral administration using histological, cytochemical and molecular methods. DMA-2,4-D destroyed in a dose- and time-dependent manner the vascular integrity of the thymus and caused cell depletion in the white pulp of the spleen and in the cortex of the thymus, which was at least partly due to programmed cell death. DMA-2,4-D appeared to have hemolytic activity and caused intravascular hemolysis of erythrocytes. An increased hemosiderin content in macrophages of the spleen indicated phagocytosis of lysed erythrocytes. We conclude that acute DMA-2,4-D herbicide poisoning results in severe damage of the lymphatic organs thymus and spleen.     

Absorption and translocation of sulfometuron-methyl in sugarcane (Saccharum officinarum L.) at different growth stages

ABSTRACT

In Brazil, weed management in sugarcane fields is mainly done with the use of selective herbicide formulations. For many years, diuron+hexazinone was one of the main herbicide mixture formulations used in sugarcane. Later, sulfometuron-methyl was included in the same mixture, which was marketed as a new herbicide formulation for residual in-season weed control in sugarcane. The mixture diuron+hexazinone+sulfometuron-methyl has been widely used in commercial sugarcane fields in Brazil. However, recent field observations have shown that sugarcane plants at different growth stages varied in their phytotoxicity levels after treatment with diuron+hexazinone+sulfometuron-methyl. Greenhouse and laboratory studies were conducted to determine ¹⁴Csulfometuron-methyl absorption and translocation, as well as ¹⁴C distribution in sugarcane at two growth stages, 2 to 3 leaves and 5 to 6 leaves. ¹⁴Csulfometuron-methyl absorption by sugarcane did not differ between the two growth stages. Different patterns of ¹⁴C accumulation were observed, which may explain variations in sulfometuron-methyl phytotoxic responses observed in the field.     

Uptake and distribution of Zn, Cu, Cd, and Pb in an aquatic plant Potamogeton natans

A better understanding of metal uptake and translocation by aquatic plants can be used to enhance the performance of constructed wetland systems for stormwater treatment. Specifically, this study examines whether the uptake of Zn, Cu, Cd, and Pb by Potamogeton natans is via the leaves, stems, or roots, and whether there is translocation from organs of uptake to other plant parts. Competition between the metals at uptake and at the level of the cell wall-bound part of the metals accumulated in stem and leaf tissue was also examined. The results show that Zn, Cu, Cd, and Pb were taken up by the leaves, stems, and roots, with the highest accumulation found in the roots. At the elevated metal concentrations common in stormwater the uptake of Cu, but not of Zn, Cd, or Pb, by the roots was somewhat limited at uptake due to competition with other metals. Between 24% and 59% of the metal content was bound to the cell walls of the plant. Except in the case of Pb, the cell wall-bound fraction was generally smaller in stems than in leaves. No translocation of the metals to other parts of the plant was found, except for Cd which was translocated from leaf to stem and vice versa. Dispersion of metals from sediment to water through P. natans is therefore unlikely.     

Evaluating the degradation of the herbicides picloram and 2,4-D in a compartmentalized reactive biobarrier with internal liquid recirculation

Tordon is a widely used herbicide formulation of 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-amino-3,5,6-trichloropicolinic acid (picloram), and it is considered a toxic herbicide. The purposes of this work were to assess the feasibility of a microbial consortium inoculated in a lab-scale compartmentalized biobarrier, to remove these herbicides, and isolate, identify, and evaluate their predominant microbial constituents. Volumetric loading rates of herbicides ranging from 31.2 to 143.9 g m^?3 day^?1, for 2,4-D, and 12.8 to 59.3 g m^?3 day^?1 for picloram were probed; however, the top operational limit of the biobarrier, detected by a decay in the removal efficiency, was not reached. At the highest loading rates probed, high average removal efficiencies of 2,4-D, 99.56?±?0.44; picloram, 94.58?±?2.62; and chemical oxygen demand (COD), 89.42?±?3.68, were obtained. It was found that the lab-scale biofilm reactor efficiently removed both herbicides at dilution rates ranging from 0.92 to 4.23 day^?1, corresponding to hydraulic retention times from 1.087 to 0.236 days. On the other hand, few microbial strains able to degrade picloram are reported in the literature. In this work, three of the nine bacterial strains isolated cometabolically degrade picloram. They were identified as Hydrocarboniphaga sp., Tsukamurella sp., and Cupriavidus sp.     

Fungal bioconversion of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenol (2,4-DCP)

Ninety strains of fungi from the collection of our mycology laboratory were tested in Galzy and Slonimski (GS) synthetic liquid medium for their ability to degrade the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and its by-product, 2,4-dichlorophenol (2,4-DCP) at 100 mg l(-1), each. Evolution of the amounts of each chemical in the culture media was monitored by HPLC. After 5 days of cultivation, the best results were obtained with Aspergillus penicilloides and Mortierella isabellina for 2,4-D and with Chrysosporium pannorum and Mucor genevensis for 2,4-DCP. The data collected seemed to prove, on one hand, that the strains responses varied with the taxonomic groups and the chemicals tested, and, on the other hand, that 2,4-D was less accessible to fungal degradation than 2,4-DCP. In each case, kinetics studies with the two most efficient strains revealed that there was a lag phase of 1 day before the onset of 2,4-D degradation, whereas there was none during 2,4-DCP degradation. Moreover, 2,4-DCP was detected transiently during 2,4-D degradation. Finally, M. isabellina improved its degradation potential in Tartaric Acid (TA) medium relative to GS and Malt Extract (ME) media.     

Chemical composition and ecophysiological responses of Empetrum nigrum to aboveground element application

Empetrum nigrum L. (crowberry) is one of the plants surviving near the Cu-Ni smelters in Finland and Russia. According to field observations, the fine roots of E. nigrum are situated below 40 cm depth and the root biomass is reduced in the polluted sites. This could cause a reduced root uptake of macronutrients and trace elements in the field and, therefore, the possible element uptake by aboveground parts of E. nigrum was studied in a greenhouse. Six different treatment solutions containing various heavy metal and macronutrient concentrations were applied to the stems and leaves of E. nigrum and the chemical composition and ecophysiological parameters were measured. Heavy metal concentrations in the leaves and stem bark, and Cu concentrations in the stems, increased with increasing metal concentrations in the spraying solutions. The bark and leaves had higher heavy metal concentrations than the stems of comparable age classes. The macronutrient and Mn concentrations in E. nigrum did not change significantly with increasing element concentrations in the spraying solution. Neither the stem water potential nor the leaf chlorophyll concentrations showed any clear response to element applications. Therefore, the element uptake by aboveground parts of E. nigrum was not confirmed by this study. However, there was a tendency to a decrease in CO2 exchange rate and increase in foliar abscisic acid content in plants treated with the highest element concentrations.     

2,4-Dichlorophenoxy Acetic Acid Mineralization in Amended Soil

The application of municipal biosolid or liquid hog manure to agricultural soils under laboratory conditions at 20°C influenced the fate of the herbicide 2,4-D [2,4-(dichlorophenoxy)acetic acid] in soil. When 2,4-D was added to soil at agronomic rates immediately after the addition of manure or biosolids to a coarse-textured soil, the percentage of 2,4-D mineralized at 100 days was about 47% for both treatments, compared to only 31% for control soils without amendments. The enhanced 2,4-D mineralization as a result of amendment addition was due to an increased heterotrophic microbial activity, with the greatest increases in soil respiration occurring for soils amended with biosolids. When additions of 2,4-D were delayed for one, two, or four weeks after the amendments were applied, the additions of amendments generally reduced 2,4-D mineralization in soil, particularly for manure, indicating that the effect of amendments on enhancing soil microbial activities diminished over time. In contrast, the mineralization of 2,4-D in control soils was less dependent on when 2,4-D was applied in relation to pre-incubations of soil for zero, one, two, or four weeks. The effect of manure on decreasing 2,4-D mineralization in specific soils was as large as the effect of soil texture on differences in 2,4-D mineralization across soils. Because manure was not found to impact 2,4-D sorption by soil, it is possible that 2,4-D mineralization decreased because 2,4-D transformation products were strongly sorbed onto organic carbon constituents in manure-amended soils and were therefore less accessible to microorganisms. Alternatively, microorganisms were less likely to metabolize the herbicide because they preferentially consumed the type of organic carbon in manure that is a weak sorbent for 2,4-D.     

2,4-Dichlorophenoxy acetic acid mineralization in amended soil

The application of municipal biosolid or liquid hog manure to agricultural soils under laboratory conditions at 20 degrees C influenced the fate of the herbicide 2,4-D [2,4-(dichlorophenoxy)acetic acid] in soil. When 2,4-D was added to soil at agronomic rates immediately after the addition of manure or biosolids to a coarse-textured soil, the percentage of 2,4-D mineralized at 100 days was about 47% for both treatments, compared to only 31% for control soils without amendments. The enhanced 2,4-D mineralization as a result of amendment addition was due to an increased heterotrophic microbial activity, with the greatest increases in soil respiration occurring for soils amended with biosolids. When additions of 2,4-D were delayed for one, two, or four weeks after the amendments were applied, the additions of amendments generally reduced 2,4-D mineralization in soil, particularly for manure, indicating that the effect of amendments on enhancing soil microbial activities diminished over time. In contrast, the mineralization of 2,4-D in control soils was less dependent on when 2,4-D was applied in relation to pre-incubations of soil for zero, one, two, or four weeks. The effect of manure on decreasing 2,4-D mineralization in specific soils was as large as the effect of soil texture on differences in 2,4-D mineralization across soils. Because manure was not found to impact 2,4-D sorption by soil, it is possible that 2,4-D mineralization decreased because 2,4-D transformation products were strongly sorbed onto organic carbon constituents in manure-amended soils and were therefore less accessible to microorganisms. Alternatively, microorganisms were less likely to metabolize the herbicide because they preferentially consumed the type of organic carbon in manure that is a weak sorbent for 2,4-D.