[N,N??-bis-(mercaptoethyl)-1,5-diazacyclooctanato]NiII, Ni-1, is known to undergo metallation reactions with numerous metals. [N,N??-bis-(mercaptoethyl)-1,5-diazacycloheptanato]NiII, (bme-dach)Ni or Ni-1??, differs from Ni-1 by one less carbon in its diazacycle backbone ring producing subtle differences in N2S2Ni geometry. Metallation of Ni-1?? with PdCl2, Pd(NO3)2, and NiBr2 produced three structural forms: Ni2Pd basket, Ni4Pd2 C4-paddlewheel, and Ni3 slant chair. In attempts to provide a rationale for the heterogeneity in the active site of Acetyl coA Synthase, metal ion capture studies of Ni-1 in methanol found a qualitative ranking of metal ion preference: Zn2+ < Ni2+ < Cu+. Formation constants for metal ion capture of Ni-1?? in water were determined for Pb2+, Ni2+, Zn2+, Cu+, and Ag+. A quantitative estimate places copper some 15 orders of magnitude above nickel or zinc in binding affinity. Sulfur dioxide uptake by Ni-1?? is characterized by significant color change, improved adduct solubility, and reversible binding of two equivalents of SO2. These combined properties establish Ni-1?? as a suitable model for gas uptake at nickel thiolate sites and as a possibly useful chemical sensor for this poisonous gas. Comparisons of molecular structures,&#61472; &#957;(SO) stretching frequencies, and thermal gravimetric analyses are made to reported adducts including the diazacyclooctane derivative, Ni-1&#903;2SO2. Visual SO2 detection limits of Ni-1 and Ni-1?? are established at 25 ppm and 100 ppm, respectively. Structural studies of products resulting from reaction at the nucleophilic S-sites of (bme-dach)Ni and [(bme-dach)Zn]2 included acetyl chloride and sodium iodoacetate as electrophiles are shown. The acetyl group is a natural electrophile important to the citric acid cycle. Acetylation of (bme-dach)Ni produces a five coordinate, paramagnetic species. Iodoacetate is a cysteine modification agent known to inhibit enzymatic activity. The reaction of (bme-dach)Ni and sodium iodoacetate yields a blue, six coordinate nickel complex in a N2S2O2 donor environment. The bismercaptodiazacycloheptane ligand binds lead(II) forming an unprecedented structural form of N2S2M dimers, in which Pb2+ is largely bound to sulfur in a highly distorted trigonal geometry. Its unusual structure is described in comparison to other derivatives of the bme-daco ligand. The synthesis and structural characterization of square pyramidal (bme-dach)GaCl are also given and compared to the analogous (bme-daco)GaCl.

The A-cluster of acetyl-CoA synthase (ACS) catalyzes the formation of acetyl- CoA from CO, coenzyme-A, and a methyl group donated by a corrinoid iron-sulfur protein. Recent crystal structures have exhibited three different metals, Zn, Cu, and Ni, in the proximal site, which bridges a square-planar nickel site and a [Fe4S4] cubane. Contradicting reports supported both the nickel and copper containing forms as representing active enzyme. The results presented here indicate that copper is not necessary or sufficient for catalysis and that copper addition to ACS is deleterious. Several proposed mechanisms exist for the synthesis of acetyl-CoA, the two most prominent are the ‘paramagnetic’ and ‘diamagnetic’ mechanisms. The ‘diamagnetic’ mechanism proposes a two electron activation that precedes methylation to produce an EPR silent Ni2+-CH3 species. This then reacts with CO and coenzyme-A to form acetyl- CoA and regenerate the starting species. The ‘paramagnetic’ mechanism assumes a one electron activation prior to the methylation of the paramagnetic Ni1+-CO state to form an unstable Ni3+-acetyl species. This is immediately reduced by an electron shuttle. Results are presented here that no shuttle or external redox mediator is necessary for catalysis. This supports the ‘diamagnetic’ mechanism, specifically that a two-electron reductive activation is necessary and that the Ni1+-CO species is not an intermediate. The two-electron reductive activation required by the ‘diamagnetic’ mechanism results in an unknown electronic state. Two proposals have been made to describe this form of the A-cluster. The first hypothesis from Brunold et al involves a one-electron reduction of the [Fe4S4]2+ cube and a one-electron reduction of the Nip 2+. This should result in a spin-coupled state that is S = integer. The Ni0 hypothesis requires both electrons to localize on the Nip 2+ forming a zero-valent proximal nickel. Mössbauer spectroscopy has been used to probe the oxidation state and spin state of the [Fe4S4] cube in the reduced active form. No integer spin system is found and this is interpreted as supporting the Ni0 hypothesis. Additionally, spectra are presented that indicate the heterogeneous nature of the A-cluster is not caused by the occupancy of the proximal site.

Electronic and steric parameters for square planar NiN2S2 complexes as bidentate, S-donor ligands have been established. According to the (CO) stretching frequencies and associated computed Cotton-Kraihanzel force constants of (NiN2S2)W(CO)4 adducts, a ranking of donor abilities and a comparison with classical bidentate ligands are as follows: Ni(ema)= > { [NiN2S2]0 } > bipy phen > Ph2PCH2CH2PPh2 > Ph2PCH2PPh2. In addition, we have demonstrated that the NiN2S2 ligands are hemilabile as evidenced from CO addition to (NiN2S2)W(CO)4, which is in equilibrium with the resulting (NiN2S2)W(CO)5 species (Keq = 2.8 M-1, G = -1.4 kJ/mole at 50C). Complete NiN2S2 ligand displacement by CO-cleavage of the remaining W-S bond to form W(CO)6 was not observed, indicating that the remaining W-S bond is considerably strengthened upon ring-opening. Several new cluster compounds based on the NiN2S2 ligands bound to CuI, RhI, PdII and W0 are reported. Structural analysis of (NiN2S2)MLn complexes show a unique structural feature defined by the dihedral angle formed by the intersection of NiN2S2/WS2C2 planes; placing the NiN2S2 ligand in closer proximity to one side of the reactive metal center. This unique orientational feature of the NiN2S2 ligands in the series of bimetallic compounds contrasts with classical diphosphine or diimine ligands. The "hinge angle" ranges in value from 136 as in the (Ni-1*)W(CO)4 to 101 in the (Ni-1)Pd(CH3)(Cl) complexes. The rigidity of the SR hinge of the nickeldithiolate ligands suggests that they might be suitable for stereochemical and regioselective substrate addition to catalytically active metals such as RhI and PdII.. The structural as well as functional similarities of the acetyl CoA synthase enzyme (ACS) and a palladium-metal based industrial type catalyst led to the preparation of a [(Ni-1)Pd(CH3)]+ bimetallic complex. This complex facilitates CO and ethylene copolymerization to produce polyketone similar to conventional (diphosphine)Pd(X)2 catalysts. However, the diphosphine ligands produce more efficient catalysts as the electron-rich character of the NiN2S2 ligand favors the resting state of the catalyst, [(Ni-1)Pd(C(O)CH3)(CO)]+, over the reactive form (Ni-1)Pd(C(O)CH3)(2-C2H4)]+. An exploratory investigation with the Ni-Pd heterobimetallic showed that this complex also facilitated the C-S coupling reaction to form a thioester similar to the ACS enzyme.

Lignin is a major chemical component of plants and the second most abundant natural polymer after cellulose. The concerns and interests of agriculture and industry have stimulated the study of genes governing lignin content in plants in an effort to adapt plants to human purposes. Arabidopsis thaliana provides a convenient model for the study of the genes governing lignin content because of its short growth cycle, small plant size, and small completely sequenced genome. In order to identify the genes controlling lignin content in Arabidopsis accessions using Quantitative Trait Locus (QTL) analysis, a rapid micro-scale method of lignin determination is required. The acetyl bromide method has been modified to enable the rapid micro-scale determination of lignin content in Arabidopsis. Modifications included the use of a micro-ball mill, adoption of a modified rapid method of extraction, use of an ice-bath to stabilize solutions and reduction in solution volumes. The modified method was shown to be accurate and precise with values in agreement with those determined by the conventional method. The extinction coefficient for Arabidopsis lignin, dissolved using acetyl bromide, was determined to be 23.35 g-iLcm-1. This value is independent of the Arabidopsis accession, environmental growth conditions and is insensitive to syringyl/guaiacyl ratio. The modified acetyl bromide method was shown to be well correlated with the 72% sulfuric acid method once the latter had been corrected for protein contamination and acid-soluble lignin content (R² = 0.988, P < 0.0001). As determined by the newly developed acetyl bromide method and confirmed by the sulfuric acid method, lignin content in Arabidopsis was found to be a divergent property. Lignin content in Arabidopsis was found to be weekly correlated with growth rate among Arabidopsis accessions (R² = 0.48, P = 0.011). Lignin content was also found to be correlated with plant height among Arabidopsis accessions (R² = 0.491, P < 0.0001).

No description available.

Lignin is a major chemical component of plants and the second most abundant natural polymer after cellulose. The concerns and interests of agriculture and industry have stimulated the study of genes governing lignin content in plants in an effort to adapt plants to human purposes. Arabidopsis thaliana provides a convenient model for the study of the genes governing lignin content because of its short growth cycle, small plant size, and small completely sequenced genome. In order to identify the genes controlling lignin content in Arabidopsis accessions using Quantitative Trait Locus (QTL) analysis, a rapid micro-scale method of lignin determination is required. The acetyl bromide method has been modified to enable the rapid micro-scale determination of lignin content in Arabidopsis. Modifications included the use of a micro-ball mill, adoption of a modified rapid method of extraction, use of an ice-bath to stabilize solutions and reduction in solution volumes. The modified method was shown to be accurate and precise with values in agreement with those determined by the conventional method. The extinction coefficient for Arabidopsis lignin, dissolved using acetyl bromide, was determined to be 23.35 g-iLcm-1. This value is independent of the Arabidopsis accession, environmental growth conditions and is insensitive to syringyl/guaiacyl ratio. The modified acetyl bromide method was shown to be well correlated with the 72% sulfuric acid method once the latter had been corrected for protein contamination and acid-soluble lignin content (R² = 0.988, P < 0.0001). As determined by the newly developed acetyl bromide method and confirmed by the sulfuric acid method, lignin content in Arabidopsis was found to be a divergent property. Lignin content in Arabidopsis was found to be weekly correlated with growth rate among Arabidopsis accessions (R² = 0.48, P = 0.011). Lignin content was also found to be correlated with plant height among Arabidopsis accessions (R² = 0.491, P < 0.0001).

Thesis (Ph.D.) - Carleton University, 2005. / Includes bibliographical references. Also available in electronic format on the Internet.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1956. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaf 133).

Submitted by Fábio Henrique Krenchinski (fhkrenchinski@gmail.com) on 2018-09-02T21:16:29Z No. of bitstreams: 1 dissertação_normas_versão final.pdf: 4468078 bytes, checksum: 834889e17086c0871a6f34bdef5de98a (MD5) / Approved for entry into archive by Maria Lucia Martins Frederico null (mlucia@fca.unesp.br) on 2018-09-03T11:31:15Z (GMT) No. of bitstreams: 1 krenchinski_fh_me_botfca.pdf: 4468078 bytes, checksum: 834889e17086c0871a6f34bdef5de98a (MD5) / Made available in DSpace on 2018-09-03T11:31:15Z (GMT). No. of bitstreams: 1 krenchinski_fh_me_botfca.pdf: 4468078 bytes, checksum: 834889e17086c0871a6f34bdef5de98a (MD5) Previous issue date: 2018-07-31 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O gene fosfinotricina acetyltransferase (pat) produz a enzima PAT, que por n-acetilação é capaz de metabolizar o glufosinate, transformando-o em n-acetyl-L-glufosinate (NAG). Assim, plantas transgênicas contendo esse gene resistem às aplicações desse herbicida. No milho, esse gene foi inserido como marcador de seleção e mais estudos precisam ser realizados a fim de validar o uso dessa tecnologia. Nesse contexto, os objetivos do presente trabalho foram avaliar se a expressão do gene pat é proporcional ao nível de resistência de tecnologias de milho à aplicação de glufosinate e avaliar a seletividade de herbicidas em associação ao glufosinate em milho com o gene pat. Durante o primeiro trabalho, foram utilizados híbridos de milho com as tecnologias Herculex®; Agrisure TL®; Herculex Yieldgard®; Leptra®; Viptera 3®; Power Core® com o gene pat e VT PRO® sem o gene pat. Para isso, um experimento foi realizado para avaliar a expressão relativa do gene pat nos híbridos de milho, por meio de PCR em tempo real. Em outro estudo, foram aplicadas doses de glufosinate (0, 500, 1000, 2000 e 4000 g i.a ha-1) sobre os híbridos de milho, no qual foram avaliados os teores de glufosinate e NAG, assim como acúmulo de amônia, taxa de transporte de elétrons (ETR), injúria visual e acúmulo de biomassa. Em campo, foi realizada a aplicação de 500 g i.a ha-1 de glufosinate durante o estádio V4 do milho, e foi avaliado o rendimento de grãos. Um segundo estudo foi realizado a campo, adotando-se a tecnologia Power Core®, e os tratamentos foram: glufosinate; glyphosate; glufosinate + glyphosate; glufosinate + nicosulfuron; glufosinate + atrazine; glufosinate + tembotrione; glufosinate + mesotrione; glufosinate + carfentrazone ethyl; glufosinate + bentazon; glufosinate + 2,4-D; testemunha sem capina e testemunha capinada. As avaliações realizadas foram: injúria visual, ETR, quantificação de amônia, altura, rendimento de grãos, além de nota de controle visual de plantas daninhas. As tecnologias VT PRO®, Herculex®, Agrisure TL® e Viptera 3® apresentaram menor expressão do gene pat, e consequentemente menor teor de NAG, maiores teores de glufosinate e amônia, maior injúria visual, assim como maior redução em ETR e acúmulo de biomassa. O rendimento de grãos não foi afetado negativamente pela aplicação de 500 g i.a ha-1 em nenhum dos híbridos testados. Conclui-se que a expressão do gene pat é proporcional ao nível de resistência de cada tecnologia nos híbridos de milho. A adoção da aplicação de glufosinate em associação a outros herbicidas não proporcionou redução de produtividade no milho. / The phosphinothricin acetyltransferase (pat) gene produces the enzyme PAT, which by n-acetylation is able to metabolize the glufosinate, transforming it into n-acetyl-L-glufosinate (NAG). So transgenic plants containing this gene resist the applications of this herbicide. In corn, this gene was inserted as a selection marker and further studies are needed to be performed to validate this technology use. In this context, the objectives of the present work were to evaluate if the expression of the pat gene is proportional to the resistance level of maize technologies submitted to the application of glufosinate and also to evaluate the selectivity of some others herbicides in association with glufosinate in maize which presents the pat gene. For the first work, the technologies used were: Herculex®; Agrisure TL®; Herculex Yieldgard®; Leptra®; Viptera 3®; Power Core® with the pat gene and VT PRO® without the pat gene. For this, an experiment was carried out to evaluate the relative expression of the pat gene in the technologies by RT- PCR. In another study, glufosinate (0, 500, 1000, 2000 and 4000 g a.i ha-1) doses were applied to the technologies, in which the glufosinate and NGA contents, ammonia accumulation, electron transportation rate (ETR), visual injury and biomass accumulation were evaluated. In the field, the spraying of 500 g a.i ha-1 of glufosinate in the V4 stage of maize was carried out in the technologies and grain yield was measured. For the second study the Power Core® technology was adopted, and the experiment was carried out in the field. The treatments were: glufosinate; glyphosate; glufosinate + glyphosate; glufosinate + nicosulfuron; glufosinate + atrazine; glufosinate + tembotrione; glufosinate + mesotrione; glufosinate + carfentrazone ethyl; glufosinate + bentazon; glufosinate + 2,4-D; no-weeding control and weeding control. Evaluations of visual injury, ETR, ammonia quantification, plant height and yield were carried out, as well as weed control. The VT PRO®, Herculex®, Agrisure TL® and Viptera 3® technologies showed lower pat gene expression, and consequently lower NAG content, as well as higher glufosinate content, ammonia accumulation, visual injury, reduced ETR and lower biomass accumulation. The yield was not affected by the application of 500 g a.i ha-1. We conclude that the pat gene expression is proportional to the resistance level of each technology.The glufosinate use in combination with other herbicides did not provide a reduction in corn yield.

Lignin is a major chemical component of plants and the second most abundant natural polymer after cellulose. The concerns and interests of agriculture and industry have stimulated the study of genes governing lignin content in plants in an effort to adapt plants to human purposes. Arabidopsis thaliana provides a convenient model for the study of the genes governing lignin content because of its short growth cycle, small plant size, and small completely sequenced genome. In order to identify the genes controlling lignin content in Arabidopsis accessions using Quantitative Trait Locus (QTL) analysis, a rapid micro-scale method of lignin determination is required. The acetyl bromide method has been modified to enable the rapid micro-scale determination of lignin content in Arabidopsis. Modifications included the use of a micro-ball mill, adoption of a modified rapid method of extraction, use of an ice-bath to stabilize solutions and reduction in solution volumes. The modified method was shown to be accurate and precise with values in agreement with those determined by the conventional method. The extinction coefficient for Arabidopsis lignin, dissolved using acetyl bromide, was determined to be 23.35 g-iLcm-1. This value is independent of the Arabidopsis accession, environmental growth conditions and is insensitive to syringyl/guaiacyl ratio. The modified acetyl bromide method was shown to be well correlated with the 72% sulfuric acid method once the latter had been corrected for protein contamination and acid-soluble lignin content (R² = 0.988, P < 0.0001). As determined by the newly developed acetyl bromide method and confirmed by the sulfuric acid method, lignin content in Arabidopsis was found to be a divergent property. Lignin content in Arabidopsis was found to be weekly correlated with growth rate among Arabidopsis accessions (R² = 0.48, P = 0.011). Lignin content was also found to be correlated with plant height among Arabidopsis accessions (R² = 0.491, P < 0.0001). / Forestry, Faculty of / Graduate