Multiple resistance of Conyza sumatrensis to Chlorimuron­ethyl and to Glyphosate

Weed resistance to herbicides has been a major issue in Brazil, mainly due to the inefficiency of the herbicides used in no-till areas and to the high cost of these herbicide treatments. Failures in controlling the weed Conyza have been reported in Western and Northern grain crop areas in Paraná (Brazil). This work aimed to evaluate the potential occurrence of C. sumatrensis biotypes resistant to the herbicides chlorimuron-ethyl and glyphosate. Experiments were carried out under greenhouse conditions with four biotypes (Cascavel-2, Toledo-4, Tupãssi-6, and Assis Chateaubriand-7) possibly resistant to, as well as a population considered susceptible to chlorimuron-ethyl and glyphosate. To obtain dose-response curves, eight herbicide doses of chlorimuron-ethyl (0, 2.5, 5, 10, 20, 40, 80 and 160 g ha-1) and glyphosate (0, 90, 180, 360, 720, 1,440, 2,880 and 5,760 g e.a. ha-1) were applied and weed control and shoot biomass evaluations were made. Results provided evidence that two biotypes (Cascavel-2 and Tupãssi-6) were resistant to glyphosate and four biotypes (Cascavel-2, Toledo-4, Tupãssi-6 and Assis Chateaubriand-7) were resistant to chlorimuron­ethyl. Multiple resistance to glyphosate and chlorimuron was confirmed for biotypes Cascavel­2 and Tupãssi 6. This is the first report on multiple resistance in Conyza sumatrensis, worldwide.

Determination of photosynthetic pigments in fleabane biotypes susceptible and resistant to the Herbicide Glyphosate

Chlorophylls and carotenoids are the main photosynthetic pigment in plants. In the weeds, the greatest amount of photosynthetic pigments can result in high competitiveness of the species. The aim of this study was to quantify the content of photosynthetic pigments in biotypes of fleabane (Conyza bonariensis) susceptible and resistant to glyphosate, by two different methods, as well as a correlation between chlorophyll content obtained by portable and classical methodology (extractable chlorophyll). An experiment was conducted in greenhouse and laboratory, 2 x 5 factorial scheme, where factor A was equivalent to biotypes of fleabane (resistant and susceptible to glyphosate) and factor B to developmental stages plants (rosette vegetative I, II and III and reproduction). At all stages of development, fleabane plants were evaluated with the portable determiner (chlorophyll content) and then the same leaves were subjected to classical methodology laboratory (extractable pigments). The resistant biotype of fleabane showed higher contents of chlorophyll a, b, and total carotenoids, inferring a greater competitive potential regarding the susceptible population to the herbicide. The portable determiner of chlorophyll showed high correlation with the classical method of determination of photosynthetic pigments, and can thus be used to accurately assess this, saving time and reagents.

Micorrização, nodulação e produção da soja roundup ready após a aplicação de diferentes formulações de glyphosate

Objetivou-se neste trabalho avaliar o efeito de diferentes formulações à base de glyphosate sobre a colonização micorrízica e nodulação no cultivar de soja RR TMG 125. Os experimentos foram desenvolvidos em condições de campo, em Latossolo Vermelho-Amarelo, durante os anos agrícolas 2010/2011 e 2011/2012. Os tratamentos constaram da aplicação da dose de 720 g e.a. ha-1 das seguintes formulações de glyphosate: Trop®, Roundup Original®, Roundup Ultra®, Roundup WG®, Roundup Transorb R® e Zapp Qi®, além de duas testemunhas (capinada e não capinada). Utilizou-se o delineamento em blocos casualizados com quatro repetições. A matéria seca da parte aérea, nodulação e produtividade de soja não foram influenciadas pelas diferentes formulações de glyphosate. Para a colonização micorrízica, observou-se efeito positivo da formulação Roundup Original®, em relação à testemunha capinada; no entanto, esse efeito não foi contínuo nos dois anos de experimento. A variável número de nódulos foi mais afetada pelas formulações de glyphosate testadas.

Habilidade competitiva, alterações no metabolismo secundário e danos celulares de soja competindo com Conyza bonariensis resistente e suscetível a glyphosate

Biótipos de buva resistentes aos herbicidas podem apresentar menor habilidade competitiva, comparado com o biótipo suscetível, quando em convivência com a cultura da soja. Os objetivos deste trabalho foram avaliar a habilidade competitiva de soja com biótipos de buva, resistente e suscetível ao herbicida glyphosate, e identificar as alterações no metabolismo secundário e danos celulares em função da competição. Foram realizados três experimentos em casa de vegetação, em delineamento inteiramente casualizado com quatro repetições, sendo o primeiro conduzido em série aditiva e os demais em série de substituição, com população de 315 plantas m-2. As variáveis avaliadas foram estatura, área foliar, massa seca, fenóis totais, peroxidação lipídica e extravasamento eletrolítico. A análise da competitividade foi feita por aplicação de diagramas e interpretações dos índices de competitividade. Os biótipos de buva resistente e suscetível ao glyphosate apresentam maior habilidade competitiva que o cultivar de soja CD 226 RR, ao passo que os biótipos de buva possuem similar capacidade em competir com a cultura. A interferência interespecífica causa maiores danos à cultura da soja, enquanto a interespecífica é mais prejudicial aos biótipos de buva. A cultura sofre alterações no seu metabolismo secundário em razão da competição com ambos os biótipos de buva.

Glyphosate as a tool to produce shikimic acid in plants

Oseltamivir phosphate is a potent viral inhibitor produced from shikimic acid extracted from seeds of Ilicium verum, the most important natural source. With the site of action 5-enolpyruvylshikimate-3-phosphate synthase (EPSP), glyphosate is the only compound capable of inhibiting its activity with the consequent accumulation of shikimic acid in plants. Corn and soybean plants were sprayed with reduced rates of glyphosate (0.0 to 230.4 g a.i. ha¹) and shikimic acid content in the dry mass was determined by HPLC 3, 7 and 10 days after application. Results showed shikimic acid accumulation in dry mass with increases of up to 969% in corn and 33,000% on soybeans, with peak concentrations 3 days after treatment (DAT). Industrial feasibility for shikimic acid production, combined with favorable climatic conditions for growing corn and soybean in virtually all over Brazil, favor the use of reduced rates of glyphosate in shikimic acid biosynthesis, with potential for use as an inducer in exploration of alternative sources for production of oseltamivir phosphate with low environmental impact.

Glyphosate tolerant volunteer corn control at two development stages

The loss of grains during the harvest of glyphosate tolerant corn may generate volunteer plants, which can interfere in the conventional or glyphosate crop in succession. The current work aim to evaluate the control of the volunteer corn glyphosate tolerant under two weed stages. Aimed to evaluate the control of volunteer glyphosate tolerant corn in two stages of development. There were conducted two experiments with hybrid 2B688 HR (lepidoptera and glyphosate tolerant), the application were at V5 and V8 stage. The experiment was randomized block design with four replicates, using the treatments: haloxyfop at 25, 50 and 62 g ha-1 alone and associated with 2,4-D at 670 g ha-1 or fluroxypyr at 200 g ha-1. The standard was clethodim at 84 g ha-1 with 2,4-D and fluroxypyr at same rates. The applications of haloxyfop and clethodim both isolated or in a mixture with 2,4-D and fluroxypyr at V5 stage showed total control (100%) at 32 and 39 days after the application, except for haloxyfop + 2,4-D (25 + 670 g ha-1) mixture, which did not provided adequate control. At V8 stage, haloxyfop + 2,4-D (50 + 670 g ha-1) and haloxyfop + 2,4-D (62 + 670 g ha-1) mixtures took up to 6 and 10 days or longer to reach adequate to excellent control, when compared to haloxyfop isolated applications in the same doses, respectively. Either isolated clethodim or mixed with 2, 4-D and fluroxypyr did not show adequate control. The treatments showed efficient control on volunteer corn plants at V5 stage, except for haloxyfop + 2, 4-D (25 + 670 g ha-1) mixture. At V8 stage applications, haloxyfop either isolated or mixture with fluroxypyr demonstrated excellent control on every evaluated dose. The mixture with 2, 4-D can reduce haloxyfop efficiency at low doses. Clethodim alone or mixed with 2,4-D or furoxypyr did not provide acceptable level of control.

Glyphosate drift affects arbuscular mycorrhizal association in coffee

Mycorrhizal association promotes better survival and nutrition of colonized seedling on field, and consequently, increasing of productivity. However, the weed management can interfere on this association, due to incorrect use of glyphosate. This work has assessed the effects of glyphosate drift on the growth and nutrition of arabica coffee plants (Catuaí Vermelho - IAC 99) colonized with arbuscular mycorrhizal fungi (AMF). The experiment was conducted in 2 x 5 factorial scheme, and included inoculated and non-inoculated plants, and five glyphosate subdoses (0.0, 57.6, 115.2, 230.4, and 460.8 g ha-1 of glyphosate), in randomized blocks with five replication. The inoculation was carried during the greenhouse phase of seedlings production with a mixture of Rhizophagus clarus and Gigaspora margarita, and after to transplanting, when the plants had seven pairs of leaves, glyphosate subdoses were applied. The product caused intoxication in up to 60% of non-inoculated and 45% on inoculated plants, when the highest dose of 460.8 g a.e. ha-1 was applied. A negative effect was noted on the growth and phosphorus content of coffee plants, this effect increased depending on glyphosate subdose, but regardless of inoculation. Glyphosate drift reduces the growth and nutrition of plants colonized by species of AMF and native fungi, negatively affecting root colonization of plants treated.

How glyphosate may affect transgenic soybean in different soil and phosphorus levels

The technology that employs genetic modifications brought a significant increase in the utilization of glyphosate. Transgenic soybean has been suffering injury, even though it possesses a resistance mechanism to glyphosate. Currently, there are only a few studies on the dynamics of glyphosate in transgenic soybean planted in soils with different textures interacting with phosphorus concentrations. This study focused on assessing the effects of glyphosate in transgenic soybean plants on different types of soil and at different phosphorus levels. The experimental design was completely randomized, in factorial design: 2 x 6 x 3, that being 2 soil types, 6 doses of glyphosate and 3 levels of phosphorus, and four replications. Plants were cultivated for thirty days in pots with two types of soil, one being clayey (Red-Yellow Latosol) and the other sandy (Quartzarenic Neosol). They received one, two, and three times the maintenance dose of fertilization of phosphorus, corresponding to: 170, 250 and 330 kg of P2O5 ha-1 to QN, and 380, 460 and 540 kg P2O5 ha-1 to RYL, respectively. Glyphosate was applied at six different doses: 0, 1,200, 2,400, 12,000, 60,000 and 120,000 g ha-1 of active ingredient. Plant height, a and b chlorophyll, and shoot were lower for the plants that received lower doses of glyphosate, regardless of the type of soil. Greater availability of phosphorus and lower amount of glyphosate used in Quartzarenic Neosol soil provided for less phytointoxication symptoms in transgenic soybean.

Behavior of RR soybeans subjected to different formulations and rates of glyphosate in the reproductive period

Recent studies indicate that glyphosate applied in post-emergence in RR soybean can eventually cause phytotoxic effects. However, there are many questions that need to be clarified in the scientific and technical contexts, involving the issue of RR soybeans regarding the use of glyphosate. This study has assessed the impact of the application of different doses and formulations of glyphosate in the reproductive period of RR soybean (R1 stage). For that purpose, an experiment in the field was conducted in two harvests (2011/12 and 2012/13), in which a 2 x 5 factorial design was used (formulations versus doses) totaling 10 treatments. In these two experiments the variables related to agronomic performance were: phytotoxicity (7, 14, 21 and 28 days after application), plant height, number of pods per plant, yield and weight of 100 grains (end of soy cycle). The results obtained allowed characterizing phytotoxicity and damages to the height and yield in RR soybean, with increasing rates of glyphosate applied in the reproductive period.

Chemical control of different Digitaria insularis populations and management of a glyphosate-resistant population

This study aimed to control different populations of Digitaria insularisby glyphosate herbicide, isolated and mixed, besides the combination of methods (chemical and mechanical) to manage resistant adult plants. Three experiments were conducted, one in pots which were maintained under non-controlled conditions and two under field conditions. In the experiment in pots, twelve populations of D. insularis were sprayed with isolated glyphosate (1.44 and 2.16 kg a.e. ha-1) and mixed (1.44 and 2.16 kg a.e. ha-1) with quizalofop-p tefuryl (0.12 kg i.a. ha-1). The treatment of 1.44 kg a.e. ha-1 of glyphosate plus 0.12 kg a.i. ha-1 of quizalofop was sufficient for adequate control (>95%) of all populations. Population 11 (area of grain production in Itumbiara, GO) was considered sensitive to glyphosate. Others populations were moderately sensitive or tolerant to the herbicide. In the field, the plants of D. insularis of one of the experiments were mowed and, in the other, there were not. Eight treatments with herbicides [isolated glyphosate (1.44 and 2.16 kg a.e. ha-1) and mixed (1.44 and 2.16 kg a.e. ha-1) with quizalofop-p-tefuryl at 0.12 kg a.i. ha-1), clethodim at 0.108 kg a.i. ha-1) or nicosulfuron at 0.06 kg a.i. ha-1)] were assessed, in combination with or without sequential application of the standard treatment, sprayed 15 days after the first application. The combination of the mechanic control with the application of glyphosate (2.16 and 1.44 kg a.e. ha-1) plus quizalofop-p-tefuryl (0.12 kg a.i. ha-1) or clethodim (0.108 kg a.i. ha-1), associated to the sequential application, was the most effective strategy for the management of adult plants of resistant D. insularis.

Shikimate accumulation, glyphosate absorption and translocation in horseweed biotypes

In Brazil, few research works on mechanisms of weed resistance to glyphosate have been conducted so far. Therefore, this research aimed to study analytical procedures determining the relation between the concentration of plant shikimate after glyphosate application and the plant resistance to this herbicide; and evaluate the glyphosate absorption and translocation into two resistant ® and susceptible (S) horseweed biotypes to glyphosate. Horseweed plants with nine true leaves received glyphosate (720 g a.e. ha-1), and 2, 3, 4, 7 and 10 days after application (DAA) the concentration of shikimic acid was measured by HPLC. In another experiment, plants were treated with radiolabeled glyphosate (14C) (1.456 MBq mmol-1 specific activity) and radioactivity was measured 4, 8, 24, 48 and 72 hours after treatment (HAT) by liquid scintillation spectrometry. The shikimate concentration in plants increased 16,351.14 and 7,892.25 mg kg-1 of dry weight, for R and S plants respectively, at seven DAA. Therefore, the procedure for quantification of shikimic acid was suitable for R and S plants differentiation to glyphosate, indicating that the R population is actually resistant to glyphosate. On average, 98% of glyphosate applied was absorbed by the studied biotypes, at 72 HAT. Around 68% of the absorbed radioactivity remained on the biotypes leaves treated, the S biotype showing the highest translocation. Therefore, the R biotype resistance mechanism studied is associated to the differential translocation.

Glyphosate Effectiveness in the Control of Macrophytes Under a Greenhouse Condition

The aim of this study was to assess the effectiveness of glyphosate, as Rodeo(r) formulation, to control Eichhornia crassipes, Pistia stratiotes, Salvinia molesta, Salvinia herzogii and Urochloa subquadripara, under greenhouse conditions. The doses assessed were (480, 960, 1440, 1920, 2400, 2880, 3360 and 3840 g ha-1 of glyphosate) with 0.5% of the Aterbane(r) BR surfactant and a control, with no herbicide application. All experiments were conducted in a completely randomized experimental design with ten replications. Applications were carried out by precision backpack sprayer at a CO2 constant pressure of 25 psi and spray solution consumption of 200 L ha-1. Product effectiveness was determined by assessing the growth inhibitions and phytotoxicity signals during the periods of 3, 7, 15, 21, 30 and 45 days after application (DAA). At 45 DAA, the highest glyphosate dose (3840 g ha-1) displayed 100% effectiveness for all four macrophyte species. Under greenhouse conditions, the glyphosate in formulation Rodeo(r) with 0.5% of Aterbarne(r) BR surfactant showed excellent effectiveness, inhibiting the growth of the floating aquatic macrophytes. This procedure can be included on the tillage plans for these species.

Extraction and Simultaneous Determination of Glyphosate, AMPA and Compounds of the Shikimic Acid Pathway in Plants

This study has aimed to develop a method for simultaneous extraction and determination by liquid chromatography and mass spectrometry (LC-MS/MS) of glyphosate, aminomethylphosphonic acid (AMPA), shikimic acid, quinic acid, phenylalanine, tyrosine and tryptophan. For the joint analysis of these compounds the best conditions of ionization in mass spectrometry and for chromatographic separation of the compounds were selected. Calibration curves and linearity ranges were also determined for each compound. Different extraction systems of the compounds were tested from plant tissues collected from sugarcane (Saccharum officinarum) and eucalyptus (Eucalyptus urophylla platiphylla) plants two days after the glyphosate application at the dose of 720 g a.e. ha-1. The plant material was dried in a forced air circulation drying oven and in a lyophilizer, and subsequently the extractions with acidified water (pH 2.5), acetonitrile-water (50:50) [v/v] and methanol-water (50:50) [v/v] were tested. To verify the recovery of the compounds in the plant matrix with acidified water as an extracting solution, the samples were fortified with a solution containing the mixture of the different analytical standards present so that this one presented the same levels of 50 and 100 μg L-1 of each compound. All experiments were conducted with three replicates. The analytical method developed was efficient for compounds quantifications. The extraction from the samples dried in an oven and using acidified water allowed better extraction levels for all compounds. The recovery levels of the compounds in the fortified samples with known amounts of each compound for both plants samples were rather satisfactory.

Influência de Adjuvantes e Pontas de Pulverização na Deriva de Aplicação do Glyphosate

Além do impacto ambiental, a deriva de glyphosate pode causar danos sobre culturas sensíveis a esse herbicida. O objetivo deste estudo foi avaliar a influência de adjuvantes e pontas de pulverização na deriva de glyphosate aplicado via terrestre. O estudo foi conduzido em campo, arranjado em esquema fatorial (3 x 9), sendo o primeiro fator constituído por três pontas de pulverização: uma de jato plano (Teejet XR 110.015) e duas de jato plano com indução de ar (Teejet AIXR 110.015 e Teejet TTI 110.015). O segundo fator foi composto por sete adjuvantes: Break Thru(r) (0,1% v.v.), Grap' Oil(r) (0,5% v.v.), Grap Super Gun(r) (0,03% v.v.), Iharol(r) (1% v.v.), NP-10(r) (0,075 L ha-1), Dash HC(r) (0,5% v.v.) e Emultec R(r) (0,05 L ha-1). Os adjuvantes foram adicionados à calda de aplicação juntamente com o herbicida Roundup Transorb R(r) na dose de 2,16 kg e.a. ha-1. Além desses, foram incluídos dois tratamentos testemunha: um sem aplicação e outro com aplicação de glyphosate sem adjuvante. As variáveis avaliadas foram a distância percorrida pelas gotas para fora do alvo, a fitotoxicidade causada pela deriva, o diâmetro mediano volumétrico, a densidade de gotas e o volume recuperado pela utilização de papéis hidrossensíveis. As pontas de pulverização TTI 110.015 e AIXR 110.015 proporcionaram menor deriva que a ponta XR 110.015. As pontas TTI e AIXR apresentaram menor densidade de gotas e maior diâmetro mediano volumétrico, quando comparadas à ponta XR. A utilização dos adjuvantes NP-10(r) e Emultec R(r) com a ponta de pulverização XR 110.015 aumentou a deriva quando comparada com a aplicação de glyphosate sem adjuvante, porém não diferiram dos demais adjuvantes. O adjuvante Emultec R(r) aumentou o DMV das gotas, quando geradas pela ponta XR 110.015. A utilização dos adjuvantes Grap' Oil(r), Grap Super Gun(r), Iharol(r) e Dash HC(r) diminuiu a densidade de gotas derivadas, quando geradas pela ponta XR 110.015.

MANAGEMENT OF MAIZE STOVER WITH UNDERDOSES RATES OF GLYPHOSATE APPLICATIONS IN NO-TILLAGE

ABSTRACTGlyphosate has significant effects on the growth and development of plants when in underdoses. This work was developed to verify the effect of the application of glyphosate in underdoses in lignin synthesis and consequently decomposition of maize stover. Two experiments were conducted; the first one in a greenhouse for underdoses adjustments and the second one in the production area. The experimental design of the first trial was completely randomized with four replications. The treatments consisted in the application of the underdoses: 0, 25, 50 and 100 g ha-1 of glyphosate. In the production area, the experimental design was a randomized block with four replications, in underdoses: 0, 12.5, 25 and 50 g ha-1. The results were submitted to analysis of variance and regression. The underdoses of 25 g a.e. ha-1in a greenhouse promoted 36% increase in productivity of stover, in addition to increasing the lignin content in 16%, with no change in the unwanted growth of maize plants. In the production area, the concentration of 12.5 g a.e. ha-1 of glyphosate reduced the lignin content and the other underdoses have not changed this feature in maize plants. None of the underdoses affected the height and biomass produced by the maize plants. The highest underdose tested promoted acceleration in the decomposition of maize stover.