Eletrolite leakage as a technique to diagnose euphorbia heterophylla biotypes resistant to ppo-inhibitors herbicides

The action of herbicides that affect the integrity of cell membranes and cause leakage, like PPO-inhibitors, can be detected by measuring the electric conductivity (EC) of a solution in which the plant tissue target is incubated in the presence of herbicide. The objectives of this work were to confirm PPO resistance in a new Euphorbia heterophylla (EPHHL) biotype, and to compare the electrolyte leakage from R and S to PPO-inhibitors biotypes, using two different methods of incubation in a solution containing herbicides. One experiment was carried in greenhouse and three in laboratory, with a completely randomized design. In the greenhouse experiment, four biotypes of EPHHL were sprayed with seven rates of fomesafen to confirm resistance in suspected biotypes. Leaf disks from R and S EPHHL biotypes in the second and the third experiments and entire leaves in the fourth experiment were incubated in a solution containing PPO-inhibitors to subsequently determine EC of solution. The study confirmed the resistance to PPO-inhibitors in two EPHHL biotypes. There were no significant differences between S and R biotypes in the experiments with the incubation of leaf disks, but incubation of entire leaves of EPHHL S biotype showed higher EC when in a solution with fomesafen, in comparison to the R biotype. The results of this work are an indirect evidence that resistance to PPO-inhibitors is related to lower absorption of herbicide by the shoots and also to some kind of mechanism to cope with oxidative stress.

Optimization and validation of the solid-liquid extraction technique for determination of picloram in soils by high performance liquid chromatography

The objective of this study was to optimize and validate the solid-liquid extraction (ESL) technique for determination of picloram residues in soil samples. At the optimization stage, the optimal conditions for extraction of soil samples were determined using univariate analysis. Ratio soil/solution extraction, type and time of agitation, ionic strength and pH of extraction solution were evaluated. Based on the optimized parameters, the following method of extraction and analysis of picloram was developed: weigh 2.00 g of soil dried and sieved through a sieve mesh of 2.0 mm pore, add 20.0 mL of KCl concentration of 0.5 mol L-1, shake the bottle in the vortex for 10 seconds to form suspension and adjust to pH 7.00, with alkaline KOH 0.1 mol L-1. Homogenate the system in a shaker system for 60 minutes and then let it stand for 10 minutes. The bottles are centrifuged for 10 minutes at 3,500 rpm. After the settlement of the soil particles and cleaning of the supernatant extract, an aliquot is withdrawn and analyzed by high performance liquid chromatography. The optimized method was validated by determining the selectivity, linearity, detection and quantification limits, precision and accuracy. The ESL methodology was efficient for analysis of residues of the pesticides studied, with percentages of recovery above 90%. The limits of detection and quantification were 20.0 and 66.0 mg kg-1 soil for the PVA, and 40.0 and 132.0 mg kg-1 soil for the VLA. The coefficients of variation (CV) were equal to 2.32 and 2.69 for PVA and TH soils, respectively. The methodology resulted in low organic solvent consumption and cleaner extracts, as well as no purification steps for chromatographic analysis were required. The parameters evaluated in the validation process indicated that the ESL methodology is efficient for the extraction of picloram residues in soils, with low limits of detection and quantification.

Morphological and physiological alterations induced by lactofen in soybean leaves are reduced with nitric oxide

Lactofen is a diphenylether herbicide recommended to control broad-leaved weeds in soybean (Glycine max) fields and its mechanism of action is the inhibition of protoporphyrinogen-IX oxidase (Protox), which acts in the chlorophyll biosynthesis. This inhibition results in an accumulation of protoporphyrin-IX, which leads to the production of reactive oxygen species (ROS) that cause oxidative stress. Consequently, spots, wrinkling and leaf burn may occur, resulting in a transitory crop growth interruption. However, nitric oxide (NO) acts as an antioxidant in direct ROS scavenging. Thus, the aim of this work was to verify, through phytometric and biochemical evaluations, the protective effect of NO in soybean plants treated with the herbicide lactofen. Soybean plants were pre-treated with different levels of sodium nitroprusside (SNP), a NO-donor substance, and then sprayed with 168 g a.i. ha-1 lactofen. Pre-treatment with SNP was beneficial because NO decreased the injury symptoms caused by lactofen in young leaflets and kept low the soluble sugar levels. Nevertheless, NO caused slower plant growth, which indicates that further studies are needed in order to elucidate the action mechanisms of NO in signaling the stress caused by lactofen in soybean crop.

Sorption and desorption of picloram in soils under pastures in Brazil

The objective of this work was to determine the coefficients of sorption and desorption of picloram in Ultisol (PVA) and Oxisol (LVA), displaying different physical and chemical characteristics. Samples of soil were collected at the 0 20 cm depth in degraded pasture areas in Viçosa-MG. Firstly, the equilibrium time between the herbicide in solution and the herbicide which was sorbed in the soil was determined by the Batch Equilibrium method. The time required was 24 hours. Sorption and desorption studies were carried out under controlled laboratory conditions; the sorption evaluation consisted in adding 10.0 mL of herbicide solutions at different concentrations to tubes containing 2.00 g of soil, with vertical rotary agitation being maintained during the pre-determined equilibrium time. After centrifugation, supernatant extract cleaning and filtration, herbicide concentration was determined by high performance liquid chromatography (HPLC) with UV detection at 254 nm. Desorption was evaluated using the samples in the tubes after the sorption tests. The Freundlich model was used for interpretation of the sorption process. Ultisol showed higher adsorption coefficient (Kf a) compared with Oxisol, which may be attributed to the lower pH of the soil and its higher organic matter content. Desorption process occurred in both soils; the LVA allowed greater release of the previously sorbed molecules.

Toxicidade de herbicidas utilizados na cultura da cana-de-açúcar à bactéria diazotrófica Azospirillum brasilense

Objetivou-se neste trabalho identificar herbicidas utilizados na cultura da cana-de-açúcar que não alteram o crescimento ou a capacidade de fixação biológica de nitrogênio (FBN) da bactéria diazotrófica Azospirillum brasi lense. Dezoito herbicidas - paraquat, ametryn, amicarbazone, diuron, metribuzin, [hexazinone + diuron], [hexazinone + clomazone], clomazone, isoxaflutole, sulfentrazone, oxyfluorfen, imazapic, imazapyr, [trifloxysulfuron-sodium + ametryn], S-metolachlor, glyphosate, MSMA e 2,4-D - foram testados em suas doses comerciais quanto ao impacto sobre o crescimento da bactéria em meio líquido DIGs. As variáveis capacidade de suporte de crescimento (carrying capacity) do meio de cultura, duração da fase lag e tempo de geração de A. brasilense foram calculadas a partir de dados de densidade ótica obtidos, em intervalos regulares, durante a incubação de culturas por 55 h. O impacto dos herbicidas na atividade da nitrogenase de A. brasilense foi avaliado em meio semissólido NFb, sem N, pela técnica da atividade de redução do acetileno (ARA). Os efeitos dos herbicidas sobre as variáveis de crescimento e ARA foram comparados ao controle pelo teste de Dunnett. Paraquat, oxyfluorfen, [trifloxysulfuron-sodium + ametryn] e glyphosate reduziram a capacidade do meio DIGs em suportar o crescimento de A. brasilense. Esse efeito foi associado ao aumento da duração da fase lag e do tempo de geração para [trifloxysulfuron-sodium + ametryn] e ao aumento no tempo de geração para glyphosate. MSMA, paraquat e amicarbazone reduzem a FBN in vitro de A. brasilense, porém essa redução é mais severa na presença do paraquat. Os demais herbicidas não alteram o crescimento e a FBN de A. brasilense.

Leaching of Picloram in ultisol under different rainfall volumes

The present study aimed to evaluate the leaching potential of Picloram in Ultisol columns under different rainfall amounts. For such, 30 treatments were evaluated (one soil associated with three levels of rainfall and ten depths).The experiments were arranged in a split-plot design, in a completely randomized design, with four replications. PVC columns of 10 cm in diameter and 50 cm in length were filled with these soils, moistened, and placed upright for 48 hours to drain the excess water. The herbicide was applied and rainfall simulations were carried out at specified intensities, according to the treatments, to force Picloram leaching. After 72 hours, all the columns were arranged in a horizontal position and opened lengthwise. Then, soil sampling was carried out every 5 cm of depth for subsequent herbicide extraction and quantification and analysis by high performance liquid chromatography. The remaining soil samples were placed in plastic pots, and, at the respective depths, the indicator species Cucumis sativus was sown. Twenty-one days after the emergence (DAE) of the indicator plants, evaluations were conducted to verify the symptoms of toxicity caused by Picloram in the plants. It was concluded that Picloram leaching is directly dependent on the volume of rain applied. The herbicide reached the deepest regions in the soil with the highest intensity of rain. The results obtained by bioassay were in agreement with those found by liquid chromatography.

Potential of macrophytes for removing atrazine from aqueous solution

The potential of three macrophytes, Azolla caroliniana, Salvinia minima, and Lemna gibba was assessed in this study to select plants for use in environmental remediation contaminated with atrazine. Experiments were carried out in a greenhouse over six days in pots containing Hoagland 0.25 strength nutritive solution at the following atrazine concentrations: 0; 0.01; 0.1; 1.0; 10.0 mg L-1. Decrease in biomass accumulation was observed in the three macrophytes, as well as toxic effects evidenced by the symptomatology developed by the plants which caused their deaths. The chlorosis and necrosis allowed to observe in the plants the high sensitivity of the three species to the herbicide. Plants presented low potential for removal of atrazine in solution when exposed to low concentrations of the herbicide. However, at the 10.0 mg L-1 atrazine concentration, L. gibba and A. caroliniana showed potential to remove the herbicide from the solution (0.016 and 0.018 mg atrazine per fresh mass gram, respectively). This fact likely resulted from the processes of atrazine adsorption by the dead material. The percentage of atrazine removed from the solution by the plants decreased when the plants were exposed to high concentrations of the pollutant. Azolla caroliniana, S. minima, and L. gibba were not effective in removing the herbicide from solution. The use of these species to remedy aquatic environments was shown to be limited.

Effects of early compost application on no-till organic soybean

Weed control has always been an important issue in agriculture. With the advent of no-till systems, soil erosion was reduced but herbicide use was increased. Organic no-till systems try to adjust reduced erosion to the no use of herbicides. Nevertheless, this adjustment is limited by the cost of mechanical weed control. This cost may be reduced by improved cultural weed control with cover crops mulches. In this paper we report a study on the application of compost manure on an oats winter cover crop, preceding soybean, instead of on the soybean summer crop. Treatments comprised a control without compost manure, and compost manure doses of 4 and 8 Mg ha-1 applied either on oats in winter or soybean in summer, organized in a randomized block design, with five replications. In summer, plots were split into weed-controlled or not controlled subplots. The timing of application and the manure doses did not affect the oats biomass or the soybean performance. However, in summer, without water stress, the application of manure at 8 Mg ha-1 directly on soybean has reduced weed biomass in this crop.

Efficacy of herbicides applied to Digitaria horizontalis plants under different water conditions

This project aimed to relate the control efficiency of ACCase inhibiting herbicides applied post-emergence to Digitaria horizontalis plants under different soil water contents. The experiments were conducted in a greenhouse, with the application of three different herbicides (fluazifop-p-butyl, haloxyfop-methyl, and sethoxydim + mineral oil Assist). The experimental design used for each herbicide was completely randomized, with four replications, consisting of a 3 x 4 factorial, with the combination of water management strategies (-0.03, -0.07 and -1.5 MPa) and four doses of these products (100%, 50%, 25%, and 0% of the recommended dose). Herbicide application was made at two vegetative stages, 4-6 leaves and 2-3 tillers. The visual phytotoxicity evaluations were performed at 14 days after application and the plant dry weight at the end of the study was evaluated. The control efficiency was not affected by water management strategies when applied to the recommended dose of the herbicides in early stages of plant development (4-6 leaf stage). In late applications (2-3 tiller stage) the plants held under drought stress showed less phytotoxicity.

Tolerância de cana-de-açúcar a herbicidas avaliada pela diferença dos tratamentos

O objetivo deste trabalho foi avaliar a tolerância dos cultivares RB855453, RB845257, SP90(-3)414, SP90(-1)638, SP89(-1)115, SP81(-3)250, IAC91-2218 e IAC91-5155 de cana-de-açúcar em pós-emergência inicial quanto à aplicação dos herbicidas sulfentrazone + diuron + hexazinone, metsulfuron-methyl + sulfentrazone, diuron + hexazinone + clomazone, metribuzin + diuron + hexazinone, diuron + hexazinone + MSMA e ametryn + trifloxysulfuronsodium + diuron + hexazinone, utilizando-se da diferença com a testemunha pareada, considerando a minimização da área experimental. O experimento foi conduzido assumindo que as testemunhas pareadas ao respectivo tratamento facilitam a avaliação da tolerância dos cultivares aos tratamentos herbicidas. A instalação do experimento foi em blocos casualizados, utilizando-se o esquema fatorial 8 (cultivares) x 6 (herbicidas). As parcelas foram constituídas por seis linhas de cana-de-açúcar, sendo uma linha central destinada ao tratamento herbicida (TH) e outra à testemunha pareada (TP); as demais linhas foram usadas como bordaduras. Para verificar se as diferenças médias (TP-TH) não diferem de zero, utilizouse a estatística t= (TP-TH)/((QMRes/n)1/2 ou, de forma equivalente, a diferença mínima significativa com zero, dms0(TP-TH)=|t/( (QMRes/n)1/2)|, em que QMRes é o quadrado médio da análise de variância (com 47 graus de liberdade e p<0,05, t=2) e n representa o número de repetições da respectiva média: 2 para comparações de herbicidas em um cultivar e 2 x 6 = 12 para comparações da média geral de cultivares. Pela metodologia proposta, os cultivares foram tolerantes às associações de herbicidas sem prejuízo final da produtividade e da qualidade da matéria-prima, embora ocorram diferenças iniciais entre cultivares quanto a sintomas de intoxicação, teor de clorofila e altura das plantas.

Weed mapping using a machine vision system

Weed mapping is a useful tool for site-specific herbicide applications. The objectives of this study were (1) to determine the percentage of land area covered by weeds in no-till and conventionally tilled fields of common bean using digital image processing and geostatistics, and (2) to compare two types of cameras. Two digital cameras (color and infrared) and a differential GPS were affixed to a center pivot structure for image acquisition. Sample field images were acquired in a regular grid pattern, and the images were processed to estimate the percentage of weed cover. After calculating the georeferenced weed percentage values, maps were constructed using geostatistical techniques. Based on the results, color images are recommended for mapping the percentage of weed cover in no-till systems, while infrared images are recommended for weed mapping in conventional tillage systems.

Ecotoxicidade de herbicidas para a macrófita aquática (Azolla caroliniana)

Os objetivos deste estudo foram avaliar Azolla caroliniana como planta-teste em estudos ecotoxicológicos e estimar a CL50;7d dos herbicidas 2,4-D, glyphosate, clomazone e oxyfluorfen. As plantas foram aclimatadas em sala de bioensaio. Para isso, foram selecionadas cinco plantas em 50 mL de meio de cultivo Hoagland. Após esse período, foram adicionados 50 mL de Hoagland mais o herbicida, completando o volume para 100 mL. A concentração letal de 50% (CL50;7d) para A. caroliniana exposta ao herbicida 2,4-D foi de 708,35 mg L-1; ao glyphosate (formulação Scout®), de 23,66 mg L-1; ao glyphosate (formulação Trop®), de 38,91 mg L-1; ao clomazone, de 129,63 mg L-1; e ao oxyfluorfen, de 80,50 mg L-1. Os herbicidas glyphosate (Scout® e Trop®) e oxyflourfen foram classificados como moderadamente tóxicos a A. caroliniana, e o clomazone e o 2,4-D, como praticamente não tóxicos. Conclui-se que A. caroliniana pode ser utilizada como planta bioindicadora de herbicidas à base de glyphosate e oxyfluorfen.

Alterações nas características fisiológicas de cultivares de cana-de-açúcar submetida à aplicação de herbicidas

Avaliou-se neste trabalho a influência de herbicidas nas características fisiológicas de três cultivares de cana-de-açúcar. O experimento foi conduzido no delineamento em blocos ao acaso, com três repetições, no esquema fatorial 3 x 8. O fator A foi composto por três cultivares de cana-de-açúcar (RB867515, RB855156 e SP80-1816), e o B, pelos herbicidas (tembotrione, MSMA, diuron + hexazinone, sulfentrazone, trifloxysulfuron-sodium, tebuthiuron, clomazone) e uma testemunha sem uso de herbicidas. A taxa transpiratória das plantas do cultivar RB867515 foi afetada negativamente quando foram aplicados os herbicidas sulfentrazone, tebuthiuron e clomazone em comparação à testemunha. Com relação à eficiência do uso da água, não se observaram diferenças entre os cultivares e a aplicação de herbicidas. A condutância estomática dos cultivares RB867515 e SP80-1816 não foi alterada pelos herbicidas aplicados. Apenas o cultivar RB867515 apresentou taxa fotossintética menor quando se aplicou o sulfentrazone. Os herbicidas testados afetam de forma diferenciada as características fisiológicas nos três cultivares avaliados. O cultivar RB867515 foi o que apresentou menor variação na taxa fotossintética na presença dos produtos testados, em relação à testemunha.

Tolerância de espécies de Mucuna a herbicidas utilizados na cultura da cana-de-açúcar

Os fluxos de emergência de Mucuna em canaviais, mesmo após a aplicação dos herbicidas para o manejo de plantas daninhas, permitiu elaborar a hipótese de que essas plantas são tolerantes aos herbicidas comumente utilizados na cultura. Para comprovar a hipótese, objetivou-se estudar a tolerância de Mucuna aterrima, Mucuna cinerea e Mucuna deeringiana a herbicidas de diferentes mecanismos de ação aplicados em pré e pós-emergência. O delineamento experimental utilizado foi o de blocos casualizados com nove tratamentos, em cinco repetições, dispostos em esquema fatorial 3x3, mais testemunhas. Na pré-emergência, o primeiro fator foi constituído pelos herbicidas sulfentrazone (800 g ha-1), imazapic (245 g ha-1) e amicarbazone (1.400 g ha-1), e o segundo, pelas três espécies de Mucuna, além de uma testemunha para cada espécie estudada. Na pós-emergência, alteraram-se os herbicidas para clomazone (1.100 g ha-1), ametryn+trifloxysulfuron-sodium (1.463 + 37 g ha-1) e 2,4-D (1.209 g ha-1). No manejo químico em pré-emergência, verificou-se que as espécies foram sensíveis ao herbicida amicarbazone, seguido de sulfentrazone, e tolerantes ao imazapic. Na pós-emergência, todas as espécies foram sensíveis ao ametryn+trifloxysulfuron-sodium e 2,4-D, mas tolerantes ao clomazone.

Resistance to ACCase inhibitors in Eleusine indica from Brazil involves a target site mutation

Eleusine indica (goosegrass) is a diploid grass weed which has developed resistance to ACCase inhibitors during the last ten years due to the intensive and frequent use of sethoxydim to control grass weeds in soybean crops in Brazil. Plant dose-response assays confirmed the resistant behaviour of one biotype obtaining high resistance factor values: 143 (fenoxaprop), 126 (haloxyfop), 84 (sethoxydim) to 58 (fluazifop). ACCase in vitro assays indicated a target site resistance as the main cause of reduced susceptibility to ACCase inhibitors. PCR-generated fragments of the ACCase CT domain of the resistant and sensitive reference biotype were sequenced and compared. A point mutation was detected within the triplet of aspartate at the amino acid position 2078 (referred to EMBL accession no. AJ310767) and resulted in the triplet of glycine. These results constitute the first report on a target site mutation for a Brazilian herbicide resistant grass weed.