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.

Interação entre herbicidas inibidores da accase e diferentes formulações de glyphosate no controle de capim-amargoso

O glyphosate é um herbicida não seletivo que inibe a produção de aminoácidos aromáticos essenciais à sobrevivência de plantas. Esse herbicida é disponibilizado no mercado em distintas formulações que podem apresentar diferenças no controle de espécies. A mistura de herbicidas é uma ferramenta importante no controle de plantas daninhas resistentes, sendo uma opção à mistura de inibidores da ACCase ao herbicida glyphosate. Este trabalho contou com a aplicação de diferentes misturas de quatro graminicidas e três formulações de glyphosate em dois estádios vegetativos de plantas de capim-amargoso resistente a esse herbicida. O controle químico foi avaliado por notas visuais de controle, aos 7, 14 e 28 dias após a aplicação, e pela medida de biomassa fresca das plantas, aos 28 dias depois da aplicação. As misturas foram avaliadas como antagonísticas, sinergísticas ou aditivas, segundo o modelo proposto por Colby. Dos tratamentos utilizados, conclui-se que todos são efetivos no manejo da resistência de Digitaria insularis, com exceção do uso de glyphosate isolado, porém o controle deve ser feito em estádios iniciais de desenvolvimento da planta. Em estádios mais desenvolvidos, o melhor controle se deu na associação de quizalofop às formulações sal de amônio e sal potássico de glyphosate; contudo, outras medidas de controle, além da associação de produtos, devem ser tomadas.

O desenvolvimento de culturas tolerantes ao herbicida diclorofenoxiacetato: revisão de literatura

O composto diclorofenoxiacetato (2,4-D) foi o primeiro herbicida orgânico, sistêmico, seletivo e de aplicação em pós-emergência desenvolvido no mundo. Juntamente com a revolução verde, ele contribuiu para elevar a produção dos cereais nas décadas posteriores a 1950. Esse produto é uma auxina sintética que pode ser utilizada como regulador do crescimento vegetal ou, ainda, como herbicida para o controle de espécies daninhas dicotiledôneas. Várias espécies infestantes dicotiledôneas que apresentam dificuldade de controle com outros herbicidas são suscetíveis ao 2,4-D. Contudo, a utilização desse herbicida fica restrita pela falta de seletividade em algumas culturas agrícolas. Nas últimas décadas, a descoberta de genes relacionados à tolerância ao 2,4-D em bactérias encontradas no solo e a sua transferência para culturas possibilitaram o desenvolvimento de linhagens tolerantes ao produto. Os objetivos desta revisão de literatura foram apresentar os genes e a atividade das enzimas responsáveis pela tolerância ao herbicida 2,4-D; ilustrar os mecanismos envolvidos na seletividade ao 2,4-D e a outros herbicidas; e equacionar algumas implicações para o manejo de plantas daninhas. O primeiro gene de tolerância ao 2,4-D descoberto foi o tfdA, encontrado no plasmídeo pJP4 da bactéria Cupriavidus necator. Este gene codifica a enzima 2,4-D/oxoglutarato dioxigenase, a qual realiza a conversão do 2,4-D em 2,4-diclorofenol e glioxilato. No final da década de 1980, foi realizada a primeira inserção do gene tfdA em plantas de Nicotiana tabacum, mediada por Agrobacterium tumefaciens. Isso conferiu tolerância de plantas de fumo ao 2,4-D. Resultados similares foram obtidos com inserções posteriores deste gene em plantas de Gossypium hirsutum, Brassica juncea e Vitis vinifera. Com a continuidade dos estudos de bactérias de solo, identificaram-se outros dois genes: o gene rdpA de Sphingobium herbicidivorans MH, que codifica a enzima ariloxialcanoato dioxigenase-1 (AAD-1); e o sdpA de Delftia acidovorans MC1, que codifica a enzima ariloxialcanoato dioxigenase-1(AAD-12). Essas duas enzimas são similares, mas têm cinética enzimática diferenciada e são capazes de degradar o 2,4-D e outros herbicidas. A enzima AAD-1 degrada o 2,4-D e, surpreendentemente, alguns herbicidas inibidores da acetil-CoA carboxilase (ACCase) do grupo dos ariloxifenoxipropionatos (FOPs). A enzima AAD-12 apresenta alta afinidade de ligação com os auxínicos 2,4-D, MCPA, triclopyr e fluroxypyr. Atualmente os genes que codificam estas enzimas estão sendo utilizados para o desenvolvimento de cultivares de soja, algodão e milho tolerantes ao 2,4-D e FOPs. Plantas de soja com o transgene sdpA se mostraram tolerantes ao 2,4-D. Plantas de milho contendo o gene rdpA também são tolerantes aos herbicidas FOPs. Trabalhos realizados com as espécies daninhas Conyza bonariensis, Conyza canadensis e Amaranthus palmeri resistentes ao herbicida glyphosate têm mostrado controle adequado com o 2,4-D. Portanto, os genes sdpA e rdpA são bons candidatos no desenvolvimento de culturas tolerantes ao 2,4-D e deverão ampliar as opções de controle de espécies daninhas de difícil manejo com outros herbicidas.

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.

Manejo de plantas daninhas eudicotiledôneas na cultura da soja Roundup Ready

As espécies eudicotiledôneas sempre foram um desafio no manejo de plantas daninhas na cultura da soja. Essa dificuldade foi aumentada pelo surgimento de plantas de difícil controle com os herbicidas atualmente existentes no mercado. Diante disso, o objetivo deste trabalho foi o de buscar alternativas de manejo que diminuam a pressão de seleção devido ao uso contínuo de glifosato na soja Roundup Ready®. O trabalho foi implantado no delineamento em blocos casualizados, em esquema de parcelas sub-subdivididas, com quatro repetições. Foram avaliadas cinco coberturas de inverno (parcelas), seis manejos químicos em pré-semeadura da soja (subparcelas) e três manejos químicos em pós-emergência da soja (sub-subparcelas). Nenhum dos tratamentos avaliados mostrou fitotoxicidade visual às plantas de soja. No entanto, as aplicações sequenciais em pós-emergência diminuíram a altura da soja, mas sem impactar negativamente a sua produtividade. Nos tratamentos com cobertura de aveia, trigo ou azevém, qualquer combinação de aplicações em pré e pós-semeadura propiciou controle de buva acima de 90%. De maneira geral, os tratamentos que propiciaram os maiores controles para leiteiro foram as combinações de glifosato + diclosulam e glifosato + sulfentrazone em pré-semeadura. Quando a soja foi semeada em área de pousio de inverno, a produtividade foi em média 8% inferior do que quando semeada em áreas com cobertura de aveia e trigo. De modo geral, os tratamentos que resultaram em maiores produtividades foram aqueles em que se utilizou glifosato + diclosulam ou glifosato + sulfentrazone na pré-semeadura da soja.

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.

Misturas de agrotóxicos em tanque nas propriedades agrícolas do Brasil

As infestações de plantas daninhas, pragas e doenças, muitas vezes, ocorrem simultaneamente em uma mesma área agrícola. Nesse caso, a alternativa de controle mais econômica e, por isso, mais comumente adotada pelos produtores é a pulverização de mistura de agrotóxicos em tanque. Nesta pesquisa, a fim de subsidiar as discussões técnicas, são descritas e quantificadas as práticas de campo adotadas em relação às misturas em tanque de agrotóxicos em propriedades agrícolas do Brasil. Para isso, foi elaborado um questionário e enviado aos profissionais que lidam cotidianamente com tratamentos fitossanitários em diferentes regiões do País. Constatou-se que 97% dos entrevistados utilizam misturas em tanque, 95% das vezes variando de dois a cinco produtos. Na maioria das vezes, utilizam a dose cheia, isto é, as maiores doses constantes nos rótulos dos agrotóxicos. As aplicações de glyphosate em soja 86% das vezes são realizadas simultaneamente com inseticidas, fungicidas e outros herbicidas. A maioria dos participantes (72%) afirmou desconhecer ou considera insuficientes as informações sobre misturas, e 99% deles demonstraram interesse em recebê-las. Considerando que a mistura em tanque de agrotóxicos é prática comum entre os agricultores, mas pode apresentar vantagens e desvantagens, conclui-se que o tema deve ser discutido com urgência entre os órgãos governamentais competentes e os demais segmentos das cadeias produtivas.

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.