Made available in DSpace on 2016-06-02T18:55:28Z (GMT). No. of bitstreams: 1 6148.pdf: 667547 bytes, checksum: f68fc760491d5919cffc86069dd54cf8 (MD5) Previous issue date: 2014-03-21 / Financiadora de Estudos e Projetos / The Roundup Ready (RR) soybean facilitated weed management by allowing the use of glyphosate in all stages of crop development. But the continuous use of this herbicide can select resistant weed biotypes and tolerant species, suggesting the need of developing new management practices. Thus, the aim of this proposal was to evaluate the effectiveness of different managements in soybean using residual and post-emergent herbicides in the control of weeds and the interference caused by straw and rainfall on the efficacy of the herbicides diclosulam and sulfentrazone. In the first experiment, in the field, was used the randomized block design with four repetitions and 14 treatments, involving the application of glyphosate (712 g ha-1) + 2,4-D (670 g ha-1) in the burndown application of weeds, 14 days before planting soybeans, with and without the presence of residual herbicides, diclosulam (29.4 g ha-1) and sulfentrazone (600 g ha-1), and treatments with glyphosate in post emergence at a single time in stages V2 (Second fully expanded trifoliate) or V4 (Fourth fully expanded trifoliate) at the dose (712 g ha-1) or in both stages V2 and V4 (356 g ha-1 in each one). In the second trial, in the greenhouse, was used a randomized block design with four repetitions in a factorial 2 x 3 x 5, being two herbicides diclosulam (29.4 g ha-1) and sulfentrazone (600 g ha-1), three levels of corn straw on the soil surface (0, 5 and 10 t ha-1) and five simulations of 20mm of rain (without rain, 24, 72, 120 and 240 hours after application of the herbicide), each experimental unit consisted of a vase and after rain simulation was sown in pots one biomarker, which was assessed for phytotoxicity. In the field experiment, it was noted the importance of applying a residual herbicide in the burndown, to reduce applications of glyphosate in post emergence, the treatment with diclosulam and a single application of glyphosate in V4 promoted the highest soybean yield of 3484,93 kg ha-1 against 719 kg ha-1 of the untreated treatment. In green house interaction was observed between the amount of straw (which can intercept the jet sprayed) and the time of occurrence of rain after application of herbicides and in this case diclosulam proved to be less dependent of water to reach its target. / O cultivo da soja transgênica Roundup Ready (RR) facilitou o manejo das plantas infestantes por permitir o uso do glifosato em todos os estádios de desenvolvimento da cultura. Porém, o uso contínuo desse herbicida pode selecionar biótipos resistentes ou espécies tolerantes de plantas infestantes sugerindo a necessidade do desenvolvimento de novas práticas de manejo. Dessa forma, o objetivo deste trabalhofoi avaliar a eficácia de diferentes manejos de plantas infestantes na cultura da soja, utilizando herbicidas residuais e pós-emergentes e verificar a interferência da palha e da precipitação sobre a eficácia dos herbicidas diclosulam e sulfentrazone. No primeiro experimento, em campo, utilizou-se o delineamento em blocos ao acaso com 4 repetições e 14 tratamentos, envolvendo a aplicação de glifosato (712 g ha-1) + 2,4-D (670 g ha-1) na dessecação das plantas infestantes, 14 dias antes da semeadura da soja (MonSoy 7211 RR) de forma isolada ou combinada com herbicidas residuais diclosulam (29,4 g ha-1) e sulfentrazone (600 g ha-1) e tratamentos com a aplicação de glifosato em pós emergência em uma única vez nos estádios de desenvolvimento da soja V2 (2° trifólio totalmente expandido) ou V4 (4° trifólio totalmente expandido) na dose de (712 g ha-1) ou parceladamente nos estádios de desenvolvimento V2 e V4 (356 g ha- 1 em cada). No segundo ensaio, em casa de vegetação, foi utilizado o delineamento em blocos ao acaso, com 4 repetições no esquema fatorial 2 x 3 x 5, sendo dois herbicidas diclosulam (29,4 g ha-1) e sulfentrazone (600 g ha-1), três níveis de palha de milho sobre o solo (0; 5 e 10 t ha-1) e cinco épocas de simulação de 20 mm de chuva, (sem chuva; 24; 72; 120 e 240 horas após a aplicação do herbicida) cada unidade experimental foi composta por um vaso e após a simulação de chuva foi semeado nos vasos o bioindicador Cucumis sativus, que foi avaliado com relação a fitotoxicidade. No experimento de campo, observou-se a importância de aplicar herbicida residual na dessecação, para diminuir as aplicações de glifosato em pós emergência. O tratamento com diclosulam e uma única aplicação de glifosato em V4 promoveu a maior produtividade da soja 3484,93 kg ha-1 contra 719 kg ha-1 da testemunha. Em casa de vegetação observou-se interação entre quantidade de palha (que pode interceptar o jato pulverizado) e o tempo de ocorrência da chuva após a aplicação dos herbicidas e neste caso o diclosulam mostrou ser menos dependente da água para atingir seu alvo.

Acetohydroxyacid synthase is a multisubunit enzyme that catalyses the first committed step in the biosynthesis of the branched chain amino acids viz., valine, leucine and isoleucine. In order to understand the structural basis for the observed allosteric feedback inhibition in AHAS, the regulatory subunit of AHAS isozymes I from E. coli was cloned, expressed, purified and the conditions were optimized for solution NMR spectroscopy. IlvN was found to exist as a dimer both in the presence and absence of the feedback inhibitor. Using high-resolution multidimensional, multinuclear NMR experiments, the structure of the dimeric valine-bound 22 kDa IlvN was determined. The ensemble of twenty low energy structures shows a backbone root mean square deviation of 0.73 ± 0.13 Å and a root mean square deviation of 1.16 ± 0.13 Å for all heavy atoms. Furthermore, greater than 98% of the backbone φ, ψ dihedral angles occupy the allowed and additionally allowed regions of the Ramachandran map. Each protomer exhibits a βαββαβα topology that is a characteristic feature of the ACT domain fold that is observed in regulatory domains of metabolic enzymes. In the free form, IlvN exists as a mixture of conformational states that are in intermediate exchange on the NMR timescale. Important structural properties of the unliganded state were probed by H-D exchange studies by NMR, alkylation studies by mass spectrometry and other biophysical methods. It was observed that the dynamic unliganded IlvN underwent a coil-to-helix transition upon binding the effector molecule and this inherent conformational flexibility was important for activation and valine-binding. A mechanism for allosteric regulation in the AHAS holoenzyme was proposed. Study of the structural and conformational properties of IlvN enabled a better understanding of the mechanism of regulation of branched chain amino acid biosynthesis. Solution structural studies of 32 kDa Protease-VPg (PVPg) from Sesbania mosaic virus (SeMV) Polyprotein processing is a commonly found mechanism in animal and plant viruses, by which more than one functional protein is produced from the same polypeptide chain. In Sesbania Mosaic Virus (SeMV), two polyproteins are expressed that are catalytically cleaved by a serine protease. The VPg protein that is expressed as a part of the polyprotein is an intrinsically disordered protein (by recombinant expression) that binds to various partners to perform several vital functions. The viral protease (Pro), though possessing the necessary catalytic residues and the substrate binding pocket is unable to catalyse the cleavage reactions without the VPg domain fused at the C-terminus. In order to determine the structural basis for the aforementioned activation of protease by VPg I undertook the structural studies of the 32 kDa PVPg domains of SeMV by solution NMR spectroscopy. NMR studies on this protein were a challenge due to the large size and spectral overlap. Using a combination of methods such as deuteration, TROSY-enhanced NMR experiments and selective ‘reverse-labelling’, the sequence specific assignments were completed for ~80% of the backbone and 13C nuclei. NMR studies on mutants such as the C-terminal deletion mutant, I/L/V to A mutants in VPg domain were conducted in order to identify the residues important for aliphatic-aromatic interactions observed in PVPg. Attempts were made to obtain NOE restraints between Pro and VPg domains through ILV labelled samples; however these proved unsuccessful. It was observed that ‘natively unfolded’ VPg possessed both secondary and tertiary structure in PVPg. However, 30 residues at the C-terminus were found to be flexible. Even though atomic-resolution structure could not be determined, the region of interaction between the domains was determined by comparing NMR spectra of Pro and PVPg. The conditions for reconstitution of the Protease-VPg complex by recombinantly expressed Pro and VPg proteins were standardised. These studies lay the foundation for future structural investigations into the Protease-VPg complex.