Photostabilization of phenoxyacetic acid herbicides MCPA and mecoprop by hydroxypropyl-β-cyclodextrin

New strategies to reduce the environmental and economic costs of pesticides use are currently under study. Microencapsulation has been used as a versatile tool for the production of controlled release agricultural formulations. In this study, the photochemical degradation of the herbicides MCPA and mecoprop has been investigated in different aqueous media such as ultrapure and river water under simulated solar irradiation. To explore the possibility of introducing cyclodextrins in the herbicide formulations, the photodegradation study of the inclusion complexes of MCPA and mecoprop with (2-hydroxypropyl)-β-cyclodextrin (HP-β-CD) was also performed. The half-lives of MCPA and mecoprop inclusion complexes were increased approximately by a factor of three related to the free molecules. Additionally, it has been shown that the photodegradation of MCPA and mecoprop is influenced by their structural features. The additional methyl group existing in mecoprop molecular structure has a positive influence on the stabilization of the radical intermediate formed in the first stage of photodegradation of both herbicides. The results found indicated that MCPA and mecoprop form inclusion complexes with HP-β-CD showing higher photostability compared to free herbicides indicating that HP-β-CD may serve as ingredient in these herbicide formulations.

Microencapsulation of herbicide MCPA with native beta-cyclodextrin and its methyl and hydroxypropyl derivatives: An experimental and theoretical investigation

When a pesticide is released into the environment, most of it is lost before it reaches its target. An effective way to reduce environmental losses of pesticides is by using controlled release technology. Microencapsulation becomes a promising technique for the production of controlled release agricultural formulations. In this work, the microencapsulation of chlorophenoxy herbicide MCPA with native b-cyclodextrin and its methyl and hydroxypropyl derivatives was investigated. The phase solubility study showed that both native and b-CD derivatives increased the water solubility of the herbicide and inclusion complexes are formed in a stoichiometric ratio of 1:1. The stability constants describing the extent of formation of the complexes have been determined by phase solubility studies. 1H NMR experiments were also accomplished for the prepared solid systems and the data gathered confirm the formation of the inclusion complexes. 1H NMR data obtained for the MCPA/CDs complexes disclosed noticeable proton shift displacements for OCH2 group and H6 aromatic proton of MCPA provided clear evidence of inclusion complexation process, suggesting that the phenyl moiety of the herbicide was included in the hydrophobic cavity of CDs. Free energy molecular mechanics calculations confirm all these findings. The gathered results can be regarded as an essential step to the development of controlled release agricultural formulations containing herbicide MCPA.

Electrochemical sensor for simultaneous determination of herbicide MCPA and its metabolite 4-chloro-2-methylphenol. Application to photodegradation environmental monitoring

The development and application of a polyaniline/carbon nanotube (CNT) cyclodextrin matrix (PANI-β-CD/MWCNT)-based electrochemical sensor for the quantitative determination of the herbicide 4-chloro-2-methylphenoxyacetic acid (MCPA) and its main transformation product 4-chloro-2-methylphenol in natural waters are described. A simple cyclic voltammetry-based electrochemical methodology, in phosphate buffer solution at pH 6.0, was used to develop a method to determine both MCPA and 4-chloro-2-methylphenol, without any previous extraction or derivatization steps. A linear concentration range (10 to 50 μmol L−1) and detection limits of 1.1 and 1.9 μmol L−1, respectively, were achieved using optimized cyclic voltammetric parameters. The proposed method was successfully applied to the determination of MCPA and 4-chloro-2-methylphenol in natural water samples with satisfactory recoveries (94 to 107 %) and in good agreement with the results obtained by an established high-performance liquid chromatography technique, no significant differences being found between the methods. Interferences from ionic species and other herbicides used for broad-leaf weed control were shown to be small. The newly developed methodology was also successfully applied to MCPA photodegradation environmental studies.

Effet des pesticides MCPA et imidaclopride sur la régulation des voies de signalisation du récepteur à la dioxine (AhR) et au récepteur aux androgènes (AR)

Avec l’ère industrielle sont venus les polluants environnementaux. Ils sont de plus en plus pointés du doigt pour une variété d’effets indésirables en particulier pour leur potentiel à affecter la santé humaine. Les pesticides font partie de ces polluants et leurs usages ne font que croître depuis une vingtaine d’années. Ces produits qui servent à améliorer la production agricole en éliminant les pestes qui ravagent les récoltes sont souvent peu étudiés à long terme avant d’être homologués. L’effet perturbateur au niveau cellulaire et les effets à long terme de ces pesticides sont peu connus. Pour ce projet de maîtrise, nous avons observé l’effet de deux pesticides, l’imidaclopride et l’acide 2-methyl-4-chlorophenoxyacetic (MCPA), sur les voies de signalisation du récepteur à la dioxine (AhR) et du récepteur aux androgènes (AR). L’imidaclopride est un insecticide de la famille des néonicotinoïdes, une classe de plus en plus utilisée. Ce pesticide est surtout connu pour être en lien avec le déclin des colonies d’abeilles depuis une décennie. Le MCPA est un des herbicides les plus utilisés au Québec, il est persistant et souvent retrouvé dans les eaux de la province. Nous avons traité des cellules du cancer du sein et des cellules du cancer de la prostate avec ces pesticides et nous avons vérifié si leur présence perturbait les deux voies de signalisation cellulaire à l’étude. Le récepteur AhR est un facteur de transcription activé par un ligand. Le TCDD, une dioxine, est le meilleur ligand exogène connu à ce jour de ce récepteur. Par contre, ses ligands naturels, des dérivés du tryptophane ou des facteurs de virulence de bactéries, l’activent de façon beaucoup moins forte. Lors de l’activation de la voie AhR, les gènes CYP1A1 et CYP1B1 sont transcrits et codent pour des enzymes du cytochrome P450 qui transforment les ligands en produits plus facilement éliminables. Dans un contexte où de l’œstradiol (E2) est présent dans les cellules, il y a une interaction croisée entre le récepteur à l’œstrogène (ER) et le récepteur AhR, qui fait en sorte que l’expression de CYP1A1 est réprimée. Cela se traduit en un ratio d’enzyme CYP1A1 à CYP1B1 différent qui pourrait augmenter la possibilité d’une accumulation de métabolites génotoxiques. En effet, CYP1B1 hydroxyle le ligand d’AhR mais aussi l’œstradiol en 4-hydroxyœstradiol (4-OHE), dont l’accumulation peut amener des mutations dans l’ADN alors que l’enzyme CYP1A1 l’hydroxyle en 2-hydroxyœstradiol (2-OHE), qui n’as aucun effet néfaste répertorié sur la cellule. Dans les cellules du cancer du sein, le MCPA appliqué en champs induisait fortement l’expression de CYP1B1 comparable à l’échantillon traité au témoin positif (TCDD), alors que CYP1A1 l’était que très légèrement par rapport au témoin non-traité. Au niveau protéique, CYP1A1 n’était qu’exprimée dans le témoin positif (TCDD) et ce, en quantité moindre lorsqu’il y avait présence d’œstradiol. CYP1B1 était fortement exprimée dans l’échantillon de TCDD, ce qui était attendu, mais aussi dans tous les échantillons traités au MCPA de NuFarm. Ces effets ne sont pas notés avec l’ingrédient actif du MCPA. La présence d’un ou plusieurs autres produits ajoutés dans le MCPA de la compagnie NuFarm en combinaison avec l’ingrédient actif pourrait activer la voie de signalisation d’AhR et causer ce débalancement dans l’expression des gènes CYP1A1 et CYP1B1. Nos résultats indiquent que plusieurs concentrations de l’ingrédient actif de l’imidaclopride ne perturbe pas les voies cellulaires d’AhR ni AR, alors que, le MCPA perturbe ces deux voies cellulaires. Par contre, c’est seulement celui produit par la compagnie NuFarm qui est utilisé en champs. Cette formulation appliquée en terrain agricole inclut l’ingrédient actif ainsi que les antigels, les surfactants et les adjuvants qui permettent au produit d’être plus efficace. L’ingrédient actif du MCPA seul n’affectait pas les deux voies. Le récepteur aux androgènes (AR) est aussi un facteur de transcription qui se lie à l’ADN afin de réguler l’expression des gènes et il est particulièrement important pour le développement et le maintien du phénotype masculin. Depuis une vingtaine d’années, des problèmes de baisse de libido et de fertilité s’accentuent dans notre société et semblent être reliés à la baisse de testostérone des hommes (Travison et al. 2007). Cette molécule est d’ailleurs un des deux ligands du récepteur AR, le deuxième étant la 5-dihydrotestostérone (DHT). Le facteur environnemental plutôt que le mode de vie semble être un facteur déterminant dans l’étude qui portait sur ce déclin. Les pesticides ont déjà été soupçonnés pour avoir un potentiel anti-androgénique, mais aucune étude ne fait un lien de causalité direct. Dans le projet de maitrise présenté dans ce document, l’expression des gènes marqueurs PSA (antigène spécifique de la prostate) et PCA3 (antigène du cancer de la prostate) a été quantifiée pour savoir si les pesticides ont un effet perturbateur sur la voie du récepteur AR. Dans les cellules du cancer de la prostate, l’expression de PSA et PCA3 était semblable au non-traité dans l’échantillon traité au MCPA (NuFarm), et ce, même après l’ajout de DHT, qui active l’expression de ces deux gènes. Cette fois-ci, l’ingrédient actif seul faisait en sorte que les deux gènes marqueurs étaient moins exprimés lors de l’ajout de la DHT, par rapport au témoin. Il semblerait que l’ingrédient actif est à la base de ce changement d’expression de nos gènes marqueurs. Donc, le MCPA pourrait avoir un effet anti-androgénique dans les cellules du cancer de la prostate. Donc, le MCPA est un pesticide qui affecte les voies de signalisation cellulaires AhR et AR. Il est particulier de noter que le pesticide appliqué en champ perturbe nettement plus les voies cellulaires. Il sera important de continuer à étudier les effets des pesticides sur l’homme au niveau cellulaire et de comprendre comment ils pourraient contribuer au développement du cancer.

Activated Carbons Produced from Mixtures of Synthetic Polymers by Physical and Chemical Activation: Application on MCPA Removal

The production of AC was achieved using the most common industrial and consumer solid waste, namely PET, alone or blended with other synthetic polymer such PAN. The PET-PAN mixture (1:1 W/W %) was subjected to carbonization, with a pyrolysis yield off 31.9%, between that obtained with PET (16.9%) or PAN (42.6%) separately. By mixing PET, as a raw material, with PAN (different ratio), an improvement in the final yield of the AC production, for the same activation time, with CO2, was found.

Activated Carbons Produced from Mixtures of Synthetic Polymers by Physical and Chemical Activation: Application on MCPA Removal

The production of AC was achieved using the most common industrial and consumer solid waste, namely PET, alone or blended with other synthetic polymer such PAN. The PET-PAN mixture (1:1 W/W %) was subjected to carbonization, with a pyrolysis yield off 31.9%, between that obtained with PET (16.9%) or PAN (42.6%) separately. By mixing PET, as a raw material, with PAN (different ratio), an improvement in the final yield of the AC production, for the same activation time, with CO2, was found.

Obesidade e cancro da próstata: Avaliação do efeito dos adipócitos nas células RM1 de carcinoma da próstata

O cancro da próstata é o segundo cancro mais frequente e a sexta causa de morte mundial por cancro no sexo masculino. A obesidade tem sido associada ao aumento da incidência e mortalidade por cancro, com alguma controvérsia. As alterações nas expressões de adipocinas associadas à obesidade têm sido um dos diversos mecanismos propostos para explicar a associação entre a obesidade e o cancro da próstata, nomeadamente na promoção do desenvolvimento e progressão celular do tumor. O objetivo deste trabalho é avaliar o efeito dos fatores produzidos pelos pré-adipócitos e os adipócitos na proliferação, migração e invasão das células de carcinoma da próstata independentes dos androgénios. As células RM1 foram cultivadas na presença de diferentes concentrações de insulina e leptina, bem como em meio condicionado (MC) de pré-adipócitos e adipócitos e co-cultivadas em sistema de transwells, com as mesmas células. A proliferação celular das RM1 foi avaliada recorrendo a contagem celular em camara de Neubauer e em citometro de fluxo, e aos ensaios metabólicos alamar blue e XTT. Efetuou-se um ensaio de migração por dano nas células RM1 na presença dos meios condicionados. A invasão das células foi avaliada recorrendo a um sistema de transwells, com membrana de matrigel, quando cultivadas com pré-adipócitos e adipócitos. A insulina aumentou significativamente a proliferação celular, ao contrário da leptina que não teve efeito. O meio condicionado dos pré-adipócitos aumentou ligeiramente a proliferação, enquanto meio condicionado dos adipócitos de 1 e 2 dias aumentou significativamente a proliferação das células RM1 (p<0.01), quando avaliada por XTT. Na câmara de Neubauer não se verificaram diferenças significativas na proliferação celular. Relativamente à migração celular, observou-se um aumento significativo da migração das células RM1 cultivadas com meio condicionado de adipócitos (MCA) e pré-adipócitos (MCPA) em comparação com o controlo (p<0.01). Observou-se um aumento significativo da invasão de células RM1 cultivadas com adipócitos e pré-adipócitos (p <0.05). Os adipócitos aumentaram significativamente a proliferação das células RM1 em co-cultura (p<0.01). Em conclusão, as células RM1 parecem ser influenciadas por fatores secretados pelos adipócitos, capazes de aumentar a sua capacidade de proliferar, invadir e migrar.

Effects of Chlorophenoxy Herbicides and Their Main Transformation Products on DNA Damage and Acetylcholinesterase Activity

Persistent pesticide transformation products (TPs) are increasingly being detected among different environmental compartments, including groundwater and surface water. However, there is no sufficient experimental data on their toxicological potential to assess the risk associated with TPs, even if their occurrence is known. In this study, the interaction of chlorophenoxy herbicides (MCPA, mecoprop, 2,4-D and dichlorprop) and their main transformation products with calf thymus DNA by UV-visible absorption spectroscopy has been assessed. Additionally, the toxicity of the chlorophenoxy herbicides and TPs was also assessed evaluating the inhibition of acetylcholinesterase activity. On the basis of the results found, it seems that AChE is not the main target of chlorophenoxy herbicides and their TPs. However, the results found showed that the transformation products displayed a higher inhibitory activity when compared with the parent herbicides. The results obtained in the DNA interaction studies showed, in general, a slight effect on the stability of the double helix. However, the data found for 4-chloro-2-methyl-6-nitrophenol suggest that this transformation product can interact with DNA through a noncovalent mode.

Mutation analysis of the different tfd genes for degradation of chloroaromatic compounds in Ralstonia eutropha JMP134.

Ralstonia eutropha JMP134 possesses two sets of similar genes for degradation of chloroaromatic compounds, tfdCDEFB (in short: tfdI cluster) and tfdDII CII EII FII BII (tfdII cluster). The significance of two sets of tfd genes for the organism has long been elusive. Here, each of the tfd genes in the two clusters on the original plasmid pJP4 was replaced by double recombination with a gene fragment in which a kanamycin resistance gene was inserted into the respective tfd gene's reading frame. The insertion mutants were all tested for growth on 2,4-dichlorophenoxyacetic acid (2,4-D), 2-methyl-4-chlorophenoxyacetic acid (MCPA), and 3-chlorobenzoate (3-CBA). None of the tfdDII CII EII FII BII genes appeared to be essential for growth on 2,4-D or on 3-CBA. Mutations in tfdC, tfdD and tfdF also did not abolish but only retarded growth on 2,4-D, indicating that they were redundant to some extent as well. Of all tfd genes tested, only tfdE and tfdB were absolutely essential, and interruption of those two reading frames abolished growth on 2,4-D, 3-CBA ( tfdE only), and MCPA completely. Interestingly, strains with insertion mutations in the tfdI cluster and those in tfdDII, tfdCII, tfdEII and tfdBII were severely effected in their growth on MCPA, compared to the wild-type. This indicated that not only the tfdI cluster but also the tfdII cluster has an essential function for R. eutropha during growth on MCPA. In contrast, insertion mutation of tfdDII resulted in better growth of R. eutropha JMP134 on 3-CBA, which is most likely due to the prevention of toxic metabolite production in the absence of TfdDII activity.

LOSS OF EXTRACTION CAPACITY OF MEHLICH-1 AND MONOCALCIUM PHOSPHATE AS A VARIABLE OF REMAINING P AND ITS RELATIONSHIP TO CRITICAL LEVELS OF SOIL PHOSPHORUS AND SULFUR

The Mehlich-1 (M-1) extractant and Monocalcium Phosphate in acetic acid (MCPa) have mechanisms for extraction of available P and S in acidity and in ligand exchange, whether of the sulfate of the extractant by the phosphate of the soil, or of the phosphate of the extractant by the sulfate of the soil. In clayey soils, with greater P adsorption capacity, or lower remaining P (Rem-P) value, which corresponds to soils with greater Phosphate Buffer Capacity (PBC), more buffered for acidity, the initially low pH of the extractants increases over their time of contact with the soil in the direction of the pH of the soil; and the sulfate of the M-1 or the phosphate of the MCPa is adsorbed by adsorption sites occupied by these anions or not. This situation makes the extractant lose its extraction capacity, a phenomenon known as loss of extraction capacity or consumption of the extractant, the object of this study. Twenty soil samples were chosen so as to cover the range of Rem-P (0 to 60 mg L-1). Rem-P was used as a measure of the PBC. The P and S contents available from the soil samples through M-1 and MCPa, and the contents of other nutrients and of organic matter were determined. For determination of loss of extraction capacity, after the rest period, the pH and the P and S contents were measured in both the extracts-soils. Although significant, the loss of extraction capacity of the acidity of the M-1 and MCPa extractants with reduction in the Rem-P value did not have a very expressive effect. A “linear plateau” model was observed for the M-1 for discontinuous loss of extraction capacity of the P content in accordance with reduction in the concentration of the Rem-P or increase in the PBC, suggesting that a discontinuous model should also be adopted for interpretation of available P of soils with different Rem-P values. In contrast, a continuous linear response was observed between the P variables in the extract-soil and Rem-P for the MCPa extractor, which shows increasing loss of extraction capacity of this extractor with an increase in the PBC of the soil, indicating the validity of the linear relationship between the available S of the soil and the PBC, estimated by Rem-P, as currently adopted.

Efeitos de herbicidas nos teores de macronutrientes e nas características tecnológicas da cana-de-açúcar (Saccharum spp.). I - Misturas de herbicidas em pós-emergência

A presente pesquisa foi conduzida em um Latossol Roxo, com 3,9% de matéria orgânica, na Usina São Carlos, município de Jaboticabal - SP com o objetivo de verificar o comportamento das principais misturas de herbicidas aplicadas em pós-emergência e suas possíveis interferências nos teores de macronutrientes, e no desenvolvimento da variedade CB 41-14. Os tratamentos utilizados com as respectivas doses do ingrediente ativo em kg/ha foram: ametryne + 2,4 - D (formulação comercial) a 1,47 - 2,03, diuron + 2,4 -D a 1,60 + 2,16; ametryne + 2,4-D a 1,60 + 2,16; alachlor + 2,4-D a 1,09 + 1,80; asulam + ioxynil - 2,4-D a 2,80 + 1,75; MCPA + 2,4-D a 0,83 + 0,83; oxadiazon + 2,4 -D a 0,50 + 2,16; ametryne + simazine + 2,4-D a 1,25 + 1,25 + 2,16; ametryne + secbumetone + 2,4-D a 2,00 + 2,00 + 2,16; diuron + hexazinone + surfatante a 0,80 + 0,45 + 0,5% e MCPA + 2,3,6-TBA + pendimethalin + surfatante a 1,50 + 0,48 + 0,66 + 0,5%. O controle das plantas daninhas foi avaliado através de contagens por espécie botânica e por avaliações visuais . Sobre a cultura os efeitos foram constatados pela contagem da brotação inicial, avaliações visuais dos efeitos fitotóxicos, análise dos teores de macronutrientes aos 5 e 8 meses, medidas do comprimento dos colmos, análise dos teores de fibra, pol, brix, % de cana e pureza, assim como valores de pol/ha, e peso de colmos por hectare. Não houve interferência das diferentes misturas de herbicidas na brotação inicial, no comprimento médio dos colmos, e nos teores de fibra, pol, brix e pureza por ocasião da colheita. Em relação às principais plantas daninhas presentes na área, que eram capim-colchão (Digitaria sanguinalis (L). Scop.) capim-coloniâo (Panicum maximum Jacq), beldroega (Portulacca oleracea L.) e caruru (Amaranthus spp), as melhores porcentagens de controle foram obtidas com as misturas de ametryne + 2,4-D, MCPA + 2,4-D, oxadiazon + 2,4-D, ametryne + secbumetone, diuron + hexazinone e MCPA + 2,3,6-TBA + pendimethalin. Aos cinco meses o teor de P foi diminuido pela mistura de ametryne + secbumetone + 2,4-D, e o teor de Ca decresceu pelas misturas de diuron + 2,4-D, ametryne + 2,4-D, alachlor + 2,4-D, asulam + ioxynil + 2,4-D, MCPA + 2,4-D, oxadiazon + 2,4-D, ametryne + simazine + 2,4-D e ametryne + secbumetone + 2,4-D. Já aos oito meses não houve diferença estatística para qualquer dos nutrientes estudados. Os teores de pol/ha e produção de colmos/ha não foram significativamente afetados por qualquer dos tratamentos quando comparados com a testemunha com capina. Todavia, todos os tratamentos apresentaram produção inferior a esta, sugerindo certa precaução com as aplicações pós-emergentes em canade-açúcar.

Optimizing activity of herbicides at reduced rate on Emex spinosa campd. with adjuvants

In pot experiments, two adjuvants were evaluated for their efficacy in enhancing activity of five herbicides applied at reduced rates (75% of the recommended rates) on Emex spinosa at the cotyledon-leaf and at the two- to four- leaf stage. Herbicides (at recommended rates) including fluroxypyr+MCPA at 450 g a.i. ha-1, carfentrazone-ethyl at 20 g a.i. ha-1, bromoxynil+MCPA at 450 g a.i. ha-1, thifensulfuron-methyl at 75 g a.i. ha-1 and tribenuronmethyl at 75 g a.i. ha-1 alone and tank mixed at reduced rates with adjuvants, namely, alkyl ether sulphate sodium salt at 625 mL ha-1 or fatty alcohol ethoxylate at 375 mL ha-1. Addition of the adjuvants to reduced rates of fluroxypyr+MCPA and carfentrazone-ethyl, increased their efficacy with 100% mortality and biomass reduction of E. spinosa at the cotyledon- leaf stage and at the two- to four- leaf stage which was similar to their recommended rates without the adjuvants except for carfentrazone-ethyl at a reduced rate without adjuvants at the two- to four- leaf stage. Bromoxynil+MCPA at reduced rates with alkyl ether sulphate sodium salt also gave 100% control of E. spinosa over weedy check at the two- to four- leaf stage. Both the adjuvants generally increased the efficacy of tribenuron-methyl at reduced rates when sprayed at both leaf stages. These findings suggest that the use of adjuvants may increase the efficacy of the above mentioned herbicides against E. spinosa and it may be incorporated in an integrated weed management program.

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.

Desenvolupament de metodologia analítica per al seguiment d'herbicides fenoxiacètics i cafeïna en el medi ambient

El control d'herbicides i altres anàlits orgànics presents en el medi ambient constitueix una pràctica habitual en els laboratoris des de l'establiment de legislacions que limiten la seva concentració. Per aquesta raó, cal el desenvolupament de noves metodologies analítiques per al seguiment de compostos orgànics en el medi. Molt sovint aquests anàlits es troben a nivells traça en aigües i sòls, conjuntament amb un alt contingut de substàncies húmiques i fúlviques. Així, un dels reptes existents és el tractament de la mostra (extracció, concentració i "clean-up" d'aquests anàlits per a una bona quantificació). Aquests processos han de venir complementats per tècniques cromatogràfiques que permetin la mesura final dels anàlits. La investigació que es presenta en aquesta tesi es centra en el desenvolupament d'un mètode per a la determinació de 2,4-D i MCPA i els seus metabòlits fenòlics i d'un altre per a la determinació de cafeïna. El primer dels procediments desenvolupats s'ha aplicat al seguiment dels herbicides i els metabòlits fenòlics en sòls d'un camp de golf, mentre que el segon s'ha emprat per a la determinació de cafeïna en aigües naturals i, posteriorment, en aigües residuals.

The use of hyphenated spectrometric techniques for the environmental forensic assessment of non-traditional pollutants and degradates in the greater Florida Everglades

A comprehensive investigation of sensitive ecosystems in South Florida with the main goal of determining the identity, spatial distribution, and sources of both organic biocides and trace elements in different environmental compartments is reported. This study presents the development and validation of a fractionation and isolation method of twelve polar acidic herbicides commonly applied in the vicinity of the study areas, including e.g. 2,4-D, MCPA, dichlorprop, mecroprop, picloram in surface water. Solid phase extraction (SPE) was used to isolate the analytes from abiotic matrices containing large amounts of dissolved organic material. Atmospheric-pressure ionization (API) with electrospray ionization in negative mode (ESP-) in a Quadrupole Ion Trap mass spectrometer was used to perform the characterization of the herbicides of interest. ^ The application of Laser Ablation-ICP-MS methodology in the analysis of soils and sediments is reported in this study. The analytical performance of the method was evaluated on certified standards and real soil and sediment samples. Residential soils were analyzed to evaluate feasibility of using the powerful technique as a routine and rapid method to monitor potential contaminated sites. Forty eight sediments were also collected from semi pristine areas in South Florida to conduct screening of baseline levels of bioavailable elements in support of risk evaluation. The LA-ICP-MS data were used to perform a statistical evaluation of the elemental composition as a tool for environmental forensics. ^ A LA-ICP-MS protocol was also developed and optimized for the elemental analysis of a wide range of elements in polymeric filters containing atmospheric dust. A quantitative strategy based on internal and external standards allowed for a rapid determination of airborne trace elements in filters containing both contemporary African dust and local dust emissions. These distributions were used to qualitative and quantitative assess differences of composition and to establish provenance and fluxes to protected regional ecosystems such as coral reefs and national parks. ^