Efeito de herbicidas sobre plantas de Brachiaria plantaginea submetidas a estresse hídrico

Este trabalho objetivou avaliar a eficiência de controle de herbicidas inibidores da ACCase aplicados em pós-emergência em plantas de Brachiaria plantaginea submetidas a diferentes teores de água no solo. O estudo foi conduzido em casa de vegetação. O delineamento experimental foi o inteiramente casualizado, com quatro repetições, utilizando-se o esquema fatorial 9 x 4, sendo a combinação de três manejos hídricos (-0,03, -0,07 e -1,5 MPa) com três herbicidas (fluazifop-p-butil, haloxyfop-methyl e sethoxydim + óleo mineral Assist) e quatro doses destes (100, 50, 25 e 0% da dose recomendada). A aplicação dos herbicidas foi realizada em dois estádios vegetativos das plantas: 4-6 folhas e 2-3 perfilhos. As avaliações visuais de fitointoxicação das plantas de B. plantaginea foram realizadas aos 3, 7 e 14 dias após a aplicação, sendo determinada a massa seca das plantas no final do experimento. A eficiência de controle foi menor em plantas mantidas em potencial de água no solo de -1,5 MPa (manejo hídrico de 8%), independentemente do herbicida utilizado, nos dois estádios de aplicação, com exceção do herbicida haloxyfop-methyl aplicado no estádio de 2-3 perfilhos. Não houve diferenças de controle entre as aplicações com 100 e 50% da dose recomendada dos herbicidas nas plantas no estádio de 4-6 folhas, independentemente do manejo hídrico.

Effect of water stress on herbicide efficiency applied to Urochloa decumbens

M.R. Rocha-Pereira, A.E. Klar, D. Martins, G.S. Ferreira de Souza, and J. Villalba. 2012. Effect of water stress on herbicide efficiency applied to Urochloa decumbens. Cien. Inv. Agr. 39(1): 211-220. This project aimed to measure the control efficiency of Acctil Coenzime A Carboxilase (ACCase)-inhibiting herbicides post-emergence applied to Urochloa decumbens (Stapf) R.D. Webster under different soil water contents. The experiment was conducted in a greenhouse at the Department of Plant Production, Faculty of Agronomic Sciences, UNESP, Botucatu, Silo Paulo. The experimental design was a completely randomized design with four replications, consisting of a 9 x 4 factorial, combined with three water management systems (-0.03, -0.07 and -1.5 MPa) and three herbicides (fluazifop-p-butyl, haloxyfop-methyl and sethoxydim + oil using four doses (100, 50, 25 and 0% of the recommended dose). Herbicide applications were conducted at two vegetative stages for all species: a 4-6 leaf stage and a 2-3 tiller stage. The physiological parameters evaluated were as follows: photosynthetic rate, stomatal conductance, transpiration, leaf temperature and plant dry matter. The visual assessments of phytotoxicity were performed 28 days after herbicide application. The control efficiency was lower in plants grown under soil water potential conditions of -1.5 MPa, regardless of the herbicide used during the two application stages; however, none reached 100% control. Fractionation of the recommended herbicide doses reduced effectiveness, with the exception of the 50%-dose application of sethoxydim and fluazifop-p-butyl herbicides, which were also effective in the 4-6 leaf plant control under normal water conditions.

Características anatômicas foliares e controle químico em pós-emergência de Brachiaria decumbens e Brachiaria plantaginea

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Seleção de um suporte sintético para imobilizar células do Botryospaheria rhodina e comparação da produção de lacase por células livres e imobilizadas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Potenciais hídricos no solo sobre a eficácia de herbicidas em plantas daninhas monocotiledôneas

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Respostas de populações autógamas de Avena fátua à seleção por baixas doses de diclofop-methyl e glyphosate

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Influence of soil water potential in the action of herbicides on goosegrass (Eleusine indica (L) Gaertn)

Currently, the use of herbicides is essential in a practical and common in agricultural areas, but efficiency of these herbicides can be compromised when applied on plants that thrive in water deficit conditions, due to low uptake and translocation of the product. Therefore, the aim of this study was to compare the efficiency of control ACCase inhibiting herbicides applied post-emergence in plants of Eleusine indica under different soil water contents. The experiment was conducted in a greenhouse and the experimental design was completely randomized design with four replications, consisting of a 9x4 factorial, with the combination of three soil water potentials (-0.03, -0.07 and -1.5 MPa) three herbicides (fluazifop-p -butyl, haloxyfop-methyl and sethoxydim + oil) and four doses (0, 25, 50, and 100 % of the recommended dose). Herbicide application was made in plants in vegetative stage 2-3 tillers. The soil water potential was initiated in the development stage of two leaves, and the water was supplemented until the soil reaches the potential of -0.01 MPa, when it came to minimum pre-determined for each water management. The physiological parameters evaluated were: photosynthetic rate, stomatal conductance, transpiration leaf temperature and plant dry mass. The visual assessments of phytotoxicity were performed at 7 and 14 days after application. The herbicides behaved in different ways according to the used water management. In severe water stress conditions (soil moisture at 8%) only fluazifop-p-butyl herbicide achieved satisfactory control (> 90%) in E. indica plants.

Manejo de azevém (Lolium multiflorum Lam.) resistente ao glyphosate com o uso de diferentes herbicidas

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Controle químico em pós-emergência de espécies de Brachiaria em três estádios vegetativos

The aim of this study was to correlate the chemical control efficacy in post-emergence of Brachiaria decumbens Stapf. (signal grass) and Brachiaria plantaginea Hitchc. (alexandergrass) through ACCase-inhibitor enzyme herbicide application in function of the stage of development of the plant in order to contribute to the chemical management of these weed species. B. decumbens and B. plantaginea were sown in plastic pots filled with soil and kept in a greenhouse. For chemical control evaluation, the following herbicides were tested (g ha-1): fluazifop-p-butyl at 150, haloxyfop-methyl at 50, and sethoxydim at 230. The herbicides were applied in all three plant development phases: stage 1 (plants presenting 4-6 leaves at 15 days after emergence), stage 2 (plants presenting 3-4 tillers at 23 days after emergence), and stage 3 (adult plants in the beginning of flowering at 48 days after emergence). Evaluations were done at 4, 7, 10, 14, 17, 21, 24, and 28 days after herbicide application. The developmental stage of the two Brachiaria species was instrumental in the efficiency of the control provided by different herbicides. Sethoxydim was the most efficient herbicide in controlling Brachiaria species in the three evaluated development stages.

Wheat cytosolic acetyl-CoA carboxylase complements an ACC1 null mutation in yeast

Spores harboring an ACC1 deletion derived from a diploid Saccharomyces cerevisiae strain, in which one copy of the entire ACC1 gene is replaced with a LEU2 cassette, fail to grow. A chimeric gene consisting of the yeast GAL10 promoter, yeast ACC1 leader, wheat cytosolic acetyl-CoA carboxylase (ACCase) cDNA, and yeast ACC1 3′ tail was used to complement a yeast ACC1 mutation. The complementation demonstrates that active wheat ACCase can be produced in yeast. At low concentrations of galactose, the activity of the “wheat gene” driven by the GAL10 promoter is low and ACCase becomes limiting for growth, a condition expected to enhance transgenic yeast sensitivity to wheat ACCase-specific inhibitors. An aryloxyphenoxypropionate and two cyclohexanediones do not inhibit growth of haploid yeast strains containing the yeast ACC1 gene, but one cyclohexanedione inhibits growth of the gene-replacement strains at concentrations below 0.2 mM. In vitro, the activity of wheat cytosolic ACCase produced by the gene-replacement yeast strain is inhibited by haloxyfop and cethoxydim at concentrations above 0.02 mM. The activity of yeast ACCase is less affected. The wheat plastid ACCase in wheat germ extract is inhibited by all three herbicides at concentrations below 0.02 mM. Yeast gene-replacement strains will provide a convenient system for the study of plant ACCases.

Link between light and fatty acid synthesis: Thioredoxin-linked reductive activation of plastidic acetyl-CoA carboxylase

Fatty acid synthesis in chloroplasts is regulated by light. The synthesis of malonyl-CoA, which is catalyzed by acetyl-CoA carboxylase (ACCase) and is the first committed step, is modulated by light/dark. Plants have ACCase in plastids and the cytosol. To determine the possible involvement of a redox cascade in light/dark modulation of ACCase, the effect of DTT, a known reductant of S-S bonds, was examined in vitro for the partially purified ACCase from pea plant. Only the plastidic ACCase was activated by DTT. This enzyme was activated in vitro more efficiently by reduced thioredoxin, which is a transducer of redox potential during illumination, than by DTT alone. Chloroplast thioredoxin-f activated the enzyme more efficiently than thioredoxin-m. The ACCase also was activated by thioredoxin reduced enzymatically with NADPH and NADP-thioredoxin reductase. These findings suggest that the reduction of ACCase is needed for activation of the enzyme, and a redox potential generated by photosynthesis is involved in its activation through thioredoxin as for enzymes of the reductive pentose phosphate cycle. The catalytic activity of ACCase was maximum at pH 8 and 2–5 mM Mg2+, indicating that light-produced changes in stromal pH and Mg2+ concentration modulate ACCase activity. These results suggest that light directly modulates a regulatory site of plastidic prokaryotic form of ACCase via a signal transduction pathway of a redox cascade and indirectly modulates its catalytic activity via stromal pH and Mg2+ concentration. A redox cascade is likely to link between light and fatty acid synthesis, resulting in coordination of fatty acid synthesis with photosynthesis.

Herbicide sensitivity determinant of wheat plastid acetyl-CoA carboxylase is located in a 400-amino acid fragment of the carboxyltransferase domain

A series of chimeral genes, consisting of the yeast GAL10 promoter, yeast ACC1 leader, wheat acetyl-CoA carboxylase (ACCase; EC 6.4.1.2) cDNA, and yeast ACC1 3′-tail, was used to complement a yeast ACC1 mutation. These genes encode a full-length plastid enzyme, with and without the putative chloroplast transit peptide, as well as five chimeric cytosolic/plastid proteins. Four of the genes, all containing at least half of the wheat cytosolic ACCase coding region at the 5′-end, complement the yeast mutation. Aryloxyphenoxypropionate and cyclohexanedione herbicides, at concentrations below 10 μM, inhibit the growth of haploid yeast strains that express two of the chimeric ACCases. This inhibition resembles the inhibition of wheat plastid ACCase observed in vitro and in vivo. The differential response to herbicides localizes the sensitivity determinant to the third quarter of the multidomain plastid ACCase. Sequence comparisons of different multidomain and multisubunit ACCases suggest that this region includes part of the carboxyltransferase domain, and therefore that the carboxyltransferase activity of ACCase (second half-reaction) is the target of the inhibitors. The highly sensitive yeast gene-replacement strains described here provide a convenient system to study herbicide interaction with the enzyme and a powerful screening system for new inhibitors.

Plastid-localized acetyl-CoA carboxylase of bread wheat is encoded by a single gene on each of the three ancestral chromosome sets

5′-End fragments of two genes encoding plastid-localized acetyl-CoA carboxylase (ACCase; EC 6.4.1.2) of wheat (Triticum aestivum) were cloned and sequenced. The sequences of the two genes, Acc-1,1 and Acc-1,2, are 89% identical. Their exon sequences are 98% identical. The amino acid sequence of the biotin carboxylase domain encoded by Acc-1,1 and Acc-1,2 is 93% identical with the maize plastid ACCase but only 80–84% identical with the cytosolic ACCases from other plants and from wheat. Four overlapping fragments of cDNA covering the entire coding region were cloned by PCR and sequenced. The wheat plastid ACCase ORF contains 2,311 amino acids with a predicted molecular mass of 255 kDa. A putative transit peptide is present at the N terminus. Comparison of the genomic and cDNA sequences revealed introns at conserved sites found in the genes of other plant multifunctional ACCases, including two introns absent from the wheat cytosolic ACCase genes. Transcription start sites of the plastid ACCase genes were estimated from the longest cDNA clones obtained by 5′-RACE (rapid amplification of cDNA ends). The untranslated leader sequence encoded by the Acc-1 genes is at least 130–170 nucleotides long and is interrupted by an intron. Southern analysis indicates the presence of only one copy of the gene in each ancestral chromosome set. The gene maps near the telomere on the short arm of chromosomes 2A, 2B, and 2D. Identification of three different cDNAs, two corresponding to genes Acc-1,1 and Acc-1,2, indicates that all three genes are transcriptionally active.

A Multisubunit Acetyl Coenzyme A Carboxylase from Soybean1

A multisubunit form of acetyl coenzyme A (CoA) carboxylase (ACCase) from soybean (Glycine max) was characterized. The enzyme catalyzes the formation of malonyl CoA from acetyl CoA, a rate-limiting step in fatty acid biosynthesis. The four known components that constitute plastid ACCase are biotin carboxylase (BC), biotin carboxyl carrier protein (BCCP), and the α- and β-subunits of carboxyltransferase (α- and β-CT). At least three different cDNAs were isolated from germinating soybean seeds that encode BC, two that encode BCCP, and four that encode α-CT. Whereas BC, BCCP, and α-CT are products of nuclear genes, the DNA that encodes soybean β-CT is located in chloroplasts. Translation products from cDNAs for BC, BCCP, and α-CT were imported into isolated pea (Pisum sativum) chloroplasts and became integrated into ACCase. Edman microsequence analysis of the subunits after import permitted the identification of the amino-terminal sequence of the mature protein after removal of the transit sequences. Antibodies specific for each of the chloroplast ACCase subunits were generated against products from the cDNAs expressed in bacteria. The antibodies permitted components of ACCase to be followed during fractionation of the chloroplast stroma. Even in the presence of 0.5 m KCl, a complex that contained BC plus BCCP emerged from Sephacryl 400 with an apparent molecular mass greater than about 800 kD. A second complex, which contained α- and β-CT, was also recovered from the column, and it had an apparent molecular mass of greater than about 600 kD. By mixing the two complexes together at appropriate ratios, ACCase enzymatic activity was restored. Even higher ACCase activities were recovered by mixing complexes from pea and soybean. The results demonstrate that the active form of ACCase can be reassembled and that it could form a high-molecular-mass complex.

An isoleucine/leucine residue in the carboxyltransferase domain of acetyl-CoA carboxylase is critical for interaction with aryloxyphenoxypropionate and cyclohexanedione inhibitors

cDNA fragments encoding the carboxyltransferase domain of the multidomain plastid acetyl-CoA carboxylase (ACCase) from herbicide-resistant maize and from herbicide-sensitive and herbicide-resistant Lolium rigidum were cloned and sequenced. A Leu residue was found in ACCases from herbicide-resistant plants at a position occupied by Ile in all ACCases from sensitive grasses studied so far. Leu is present at the equivalent position in herbicide-resistant ACCases from other eukaryotes. Chimeric ACCases containing a 1000-aa fragment of two ACCase isozymes found in a herbicide-resistant maize were expressed in a yeast ACC1 null mutant to test herbicide sensitivity of the enzyme in vivo and in vitro. One of the enzymes was resistant/tolerant, and one was sensitive to haloxyfop and sethoxydim, rendering the gene-replacement yeast strains resistant and sensitive to these compounds, respectively. The sensitive enzyme has an Ile residue, and the resistant one has a Leu residue at the putative herbicide-binding site. Additionally, a single Ile to Leu replacement at an equivalent position changes the wheat plastid ACCase from sensitive to resistant. The effect of the opposite substitution, Leu to Ile, makes Toxoplasma gondii apicoplast ACCase resistant to haloxyfop and clodinafop. In this case, inhibition of the carboxyltransferase activity of ACCase (second half-reaction) of a large fragment of the Toxoplasma enzyme expressed in Escherichia coli was tested. The critical amino acid residue is located close to a highly conserved motif of the carboxyltransferase domain, which is probably a part of the enzyme active site, providing the basis for the activity of fop and dim herbicides.