Isolation and selection of plant growth-promoting bacteria associated with sugarcane

v. 46, n. 2, p. 149-158, apr./jun. 2016.

Improving the stability of an oxime based electrochemical microsensor for organo-phosphate vapor detection

A micro gas sensor has been developed by our group for the detection of organo-phosphate vapors using an aqueous oxime solution. The analyte diffuses from the high flow rate gas stream through a porous membrane to the low flow rate aqueous phase. It reacts with the oxime PBO (1-Phenyl-1,2,3,-butanetrione 2-oxime) to produce cyanide ions, which are then detected electrochemically from the change in solution potential. Previous work on this oxime based electrochemistry indicated that the optimal buffer pH for the aqueous solution was approximately 10. A basic environment is needed for the oxime anion to form and the detection reaction to take place. At this specific pH, the potential response of the sensor to an analyte (such as acetic anhydride) is maximized. However, sensor response slowly decreases as the aqueous oxime solution ages, by as much as 80% in first 24 hours. The decrease in sensor response is due to cyanide which is produced during the oxime degradation process, as evidenced by the cyanide selective electrode. Solid phase micro-extraction carried out on the oxime solution found several other possible degradation products, including acetic acid, N-hydroxy benzamide, benzoic acid, benzoyl cyanide, 1-Phenyl 1,3-butadione, 2-isonitrosoacetophenone and an imine derived from the oxime. It was concluded that degradation occurred through nucleophilic attack by a hydroxide or oxime anion to produce cyanide, as well as a nitrogen atom rearrangement similar to Beckmann rearrangement. The stability of the oxime in organic solvents is most likely due to the lack of water, and specifically hydroxide ions. The reaction between oxime and organo-phosphate to produce cyanide ions requires hydroxide ions, and therefore pure organic solvents are not compatible with the current micro-sensor electrochemistry. By combining a concentrated organic oxime solution with the basic aqueous buffer just prior to being used in the detection process, oxime degradation can be avoided while preserving the original electrochemical detection scheme. Based on beaker cell experiments with selective cyanide sensitive electrodes, ethanol was chosen as the best organic solvent due to its stabilizing effect on the oxime, minimal interference with the aqueous electrochemistry, and compatibility with the current microsensor material (PMMA). Further studies showed that ethanol had a small effect on micro-sensor performance by reducing the rate of cyanide production and decreasing the overall response time. To avoid incomplete mixing of the aqueous and organic solutions, they were pre-mixed externally at a 10:1 ratio, respectively. To adapt the microsensor design to allow for mixing to take place within the device, a small serpentine channel component was fabricated with the same dimensions and material as the original sensor. This allowed for seamless integration of the microsensor with the serpentine mixing channel. Mixing in the serpentine microchannel takes place via diffusion. Both detector potential response and diffusional mixing improve with increased liquid residence time, and thus decreased liquid flowrate. Micromixer performance was studies at a 10:1 aqueous buffer to organic solution flow rate ratio, for a total rate of 5.5 μL/min. It was found that the sensor response utilizing the integrated micromixer was nearly identical to the response when the solutions were premixed and fed at the same rate.

Síntese de sensores, funcionalização de nanopartículas e fibras óticas para reconhecimento de aniões

O trabalho descrito nesta dissertação envolve a síntese e caracterização de novos macrociclos tetrapirrólicos e afins com potencial aplicação como quimiossensores de aniões, tanto em solução como quando suportados em diferentes materiais. As porfirinas e ftalocianinas ocupam um lugar de destaque nesta dissertação, pelo que no primeiro capítulo, é feita uma revisão bibliográfica acerca das suas metodologias de síntese bem como das suas principais características e aplicações, nomeadamente como quimiossensores de aniões. No segundo capítulo é discutida a síntese e caracterização dos compostos porfirínicos e ftalocianinicos com grupos amina ou poliamina, posteriormente utilizados como hospedeiros de aniões. Descrevem-se, pormenorizadamente, os métodos de síntese, purificação e caracterização estrutural dos diversos compostos sintetizados. No terceiro capítulo realizaram-se os estudos de complexação com aniões em solução e determinaram-se as respetivas constantes de afinidade. Os compostos sintetizados apresentam capacidade de interagir com diferentes aniões. As porfirinas testadas apresentam elevadas constantes de afinidade para o anião di-hidrogenofosfato, mesmo em soluções aquosas quando testadas com cristais piezoelétricos. No caso das ftalocianinas verificou-se que estas interagem com vários aniões e apresentam propriedades cromogénicas, podendo mesmo distinguir aniões cianeto em soluções contendo água. No quarto capítulo estudou-se a imobilização dos quimiossensores, que demonstraram maior eficácia nos estudos de reconhecimento em solução, em diferentes materiais. Primeiro foi estudada a imobilização dos quimiossensores em nanopartículas de sílica (com e sem núcleo magnético) e testada a sua capacidade como sensor de aniões em solução. Numa segunda parte foi estudada a imobilização em fibras óticas. Estas, além das suas excecionais propriedades físico-químicas, têm a vantagem de poderem ser integradas em diferentes estruturas e/ou equipamentos de análise. Na ultima parte desta dissertação encontra-se a descrição da síntese e caracterização de novos conjugados porfirina-C60-OligoDNA com potencial aplicação em transferência eletrónica. Foram sintetizados e caracterizados novos compostos porfirina-OligoDNA e C60-OligoDNA. Esta parte do trabalho foi realizada no “Institute of Advanced Energy” na Universidade de Quioto, Japão.

Involvement of Mitochondrial Activity and OXPHOS in ATP Synthesis During the Motility Phase of Spermatozoa in the Pacific Oyster, Crassostrea gigas

In the Pacific oyster, spermatozoa are characterized by a remarkably long movement phase (i.e., over 24 h) sustained by a capacity to maintain intracellular ATP level. To gain information on oxidative phosphorylation (OXPHOS) functionality during the motility phase of Pacific oyster spermatozoa, we studied 1) changes in spermatozoal mitochondrial activity, that is, mitochondrial membrane potential (MMP), and intracellular ATP content in relation to motion parameters and 2) the involvement of OXPHOS for spermatozoal movement using carbonyl cyanide m-chlorophenyl hydrazone (CCCP). The percentage of motile spermatozoa decreased over a 24 h movement period. MMP increased steadily during the first 9 h of the movement phase and was subsequently maintained at a constant level. Conversely, spermatozoal ATP content decreased steadily during the first 9 h postactivation and was maintained at this level during the following hours of the movement phase. When OXPHOS was decoupled by CCCP, the movement of spermatozoa was maintained 2 h and totally stopped after 4 h of incubation, whereas spermatozoa were still motile in the control after 4 h. Our results suggest that the ATP sustaining flagellar movement of spermatozoa may partially originate from glycolysis or from mobilization of stored ATP or from potential phosphagens during the first 2 h of movement as deduced by the decoupling by CCCP of OXPHOS. However, OXPHOS is required to sustain the long motility phase of Pacific oyster spermatozoa. In addition, spermatozoa may hydrolyze intracellular ATP content during the early part of the movement phase, stimulating mitochondrial activity. This stimulation seems to be involved in sustaining a high ATP level until the end of the motility phase.

Tratamiento químico y biológico de efluentes mineros cianurados a escala laboratorio

El cianuro es el compuesto químico empleado por excelencia para la lixiviación de oro en la industria minera. Sin embargo, es altamente tóxico para los organismos que se desarrollan alrededor de las industrias mineras, y para el medio ambiente. Con el fin de reducir los niveles de cianuro libre en efluentes provenientes de la minería, el trabajo se enfocó en determinar las condiciones óptimas para la degradación de cianuro empleando compuestos químicos y un consorcio microbiano. Los ensayos químicos y biológicos se realizaron por separado, utilizando muestras de efluentes provenientes de la minería a diferentes concentraciones de cianuro (280 y 10 mg/l CN-). Para la degradación química se utilizó tres oxidantes diferentes: hipoclorito de sodio, peróxido de hidrógeno y ácido de caro en diferentes concentraciones, pH (10-11) y tiempos de degradación (4,71 y 20,75 h). Para los ensayos de biodegradación se empleó un consorcio microbiano en matraces que contenían el efluente cianurado y medio líquido a pH (11), agitación (200 rpm) y temperatura (20±5°C). Se midió la concentración de cianuro libre, pH y la concentración de biomasa. Los resultados del tratamiento químico mostraron que el mejor compuesto oxidante fue el peróxido de hidrógeno (8:1 gH2O2/gCN-) a pH (10), obteniendo un 92,7% remoción de cianuro libre en 45 minutos (280 mg/l CN-) y un 91,0% de remoción en 25 minutos (10 mg/l CN-). Mientras que en la degradación biológica en matraces la remoción fue del 73,7% (280 mg/l CN-) en 384 h y de 78,6% (10 mg/l CN-) en 240 h.

Degradación bacteriana de cianuro y compuestos nitrogenados tóxicos

La ruta de asimilación de cianuro en P. pseudoalcaligenes CECT5344 transcurre a través de un nitrilo formado por la reacción química del cianuro con el oxalacetato, siendo este último acumulado como consecuencia de la acción conjunta de una malato:quinona oxidoreductasa (MQO) y la oxidasa terminal resistente a cianuro (CioAB) (Luque-Almagro et al., 2011b). Los nitrilos pueden ser convertidos en amonio por la acción de una nitrilasa o un sistema nitrilo hidratasa/amidasa. Con el objetivo de elucidar la ruta de asimilación de cianuro en P. pseudoalcalígenes CECT5344, se ha analizado el proteoma de este microorganismo en condiciones cianotróficas frente a nitrato como fuente de nitrógeno como control. En este estudio se identificaron proteínas relacionadas con la ruta de asimilación de cianuro en la estirpe CECT5344, que aparecían inducidas por cianuro, como NitB y NitG, cuyos genes se encuentran localizados en la agrupación génica nit1C. Además de NitB y NitG, de función desconocida, la agrupación génica nit1C codifica un regulador transcripcional del tipo Fis dependiente de σ54 (NitA), una nitrilasa (NitC), una proteína que pertenece a la superfamilia S-adenosilmetionina (NitD), un miembro de la superfamilia N-aciltransferasa (NitE), un polipéptido de la familia AIRS/GARS (NitF) y una oxidorreductasa dependiente de NADH (NitH). Un análisis transcripcional mediante RT-PCR determinó que los genes nitBCDEFGH se cotranscriben, mientras que el gen regulador nitA se transcribe de forma divergente. Además, resultados obtenidos por RT-PCR confirman que la expresión de los genes nitBCDEFGH está inducida por cianuro y reprimida por amonio. La relación entre el cianuro y el grupo de genes nit1C queda patente por el fenotipo de los mutantes deficientes nitA, nitB y nitC, incapaces de usar complejos cianuro-metálicos o 2-hidroxinitrilos como única fuente de nitrógeno. Todos estos datos indican que la nitrilasa NitC, junto con la proteína NitB, utilizan de forma específica determinados nitrilos alifáticos como sustrato, entre los que se encuentran el formado durante la asimilación de cianuro (Estepa et al., 2012). Además, entre las proteínas inducidas por cianuro se identificaron una dihidropicolinato sintasa (DapA), una fosfoserina transaminasa (SerC) y una proteína de función desconocida (Orf1), las tres codificadas por genes del operón cio, una cianasa (CynS), la proteína S6 de la subunidad ribosomal 30S (RpsF), una superóxido dismutasa (SodB), la ferritina (Dps), una oxidorreductasa (Fpr) y un factor de elongación P (EF-P). Una vez identificadas, estas proteínas se han analizado funcionalmente y se han localizado en el genoma de P. pseudoalcaligenes CECT5344 los genes correspondientes, así como los genes adyacentes. La inducción de estas proteínas en condiciones cianotróficas sugiere que el metabolismo del cianuro incluye, además de la resistencia y asimilación de este tóxico, otros procesos biológicos relacionados con el metabolismo del cianato y de algunos aminoácidos, el estrés oxidativo y la homeostasis de hierro, entre otros. Por otra parte, el conocimiento en profundidad y la interpretación de la secuencia génica de P. pseudoalcaligenes CECT5344, así como el análisis comparativo frente a organismos no cianotrofos ha permitido entender algunos de los mecanismos implicados en la resistencia y asimilación de cianuro, lo que permitiría conducir a la posterior mejora del proceso de biodegradación de cianuro. Además, el estudio del genoma de la estirpe CECT5344 permitirá explorar la capacidad de este organismo para ser utilizado en procesos de biorremediación de residuos cianurados en los que se encuentran metales y otros tóxicos (Luque-Almagro et al., 2013; Wibberg et al., 2014). En este trabajo se muestran y discuten los resultados de la secuenciación del genoma de P. pseudoalcaligenes, así como el estudio del análisis filogenético y evolutivo de la cepa, estableciéndose de esta manera relaciones con otras especies en base a los genomas secuenciados de las mismas, entre las que destaca P. mendocina ymp relacionada con P. pseudoalcaligenes CECT5344. El estudio de las características del genoma de P. pseudoalcaligenes CECT5344 ha sido completado con un análisis comparativo frente a los genomas de otras especies de Pseudomonas, encontrándose así semejanzas y diferencias en cuanto a la distribución génica funcional. Por último, se muestra un análisis del genoma de P. pseudoalcaligenes CECT5344 en relación con los genes implicados probablemente en los procesos de asimilación de cianuro y residuos cianurados, tales como los codificantes de nitrilasas y aquellos implicados en la resistencia a cianuro como los constituyentes del operón cio que codifican la oxidasa terminal insensible a cianuro. Finalmente, se discute la presencia de genes implicados posiblemente en otros procesos con una alto potencial biotecnológico, tales como la producción de bioplásticos y la biodegradación de diversos contaminantes.

Metabolic consequences of knocking out UGT85B1, the gene encoding the glucosyltransferase required for synthesis of dhurrin in Sorghum bicolor (L. Moench)

Many important food crops produce cyanogenic glucosides as natural defense compounds to protect against herbivory or pathogen attack. It has also been suggested that these nitrogen-based secondary metabolites act as storage reserves of nitrogen. In sorghum, three key genes, CYP79A1, CYP71E1 and UGT85B1, encode two Cytochrome P450s and a glycosyltransferase, respectively, the enzymes essential for synthesis of the cyanogenic glucoside dhurrin. Here, we report the use of targeted induced local lesions in genomes (TILLING) to identify a line with a mutation resulting in a premature stop codon in the N-terminal region of UGT85B1. Plants homozygous for this mutation do not produce dhurrin and are designated tcd2 (totally cyanide deficient 2) mutants. They have reduced vigor, being dwarfed, with poor root development and low fertility. Analysis using liquid chromatography-mass spectrometry (LC-MS) shows that tcd2 mutants accumulate numerous dhurrin pathway-derived metabolites, some of which are similar to those observed in transgenic Arabidopsis expressing the CYP79A1 and CYP71E1 genes. Our results demonstrate that UGT85B1 is essential for formation of dhurrin in sorghum with no co-expressed endogenous UDP-glucosyltransferases able to replace it. The tcd2 mutant suffers from self-intoxication because sorghum does not have a feedback mechanism to inhibit the initial steps of dhurrin biosynthesis when the glucosyltransferase activity required to complete the synthesis of dhurrin is lacking. The LC-MS analyses also revealed the presence of metabolites in the tcd2 mutant which have been suggested to be derived from dhurrin via endogenous pathways for nitrogen recovery, thus indicating which enzymes may be involved in such pathways.

Ab Initio Quantum Chemical Studies on Neutral-Radical Reactions of Ethynyl (C2H) and Cyano (CN) with Unsaturated Hydrocarbons

An Ab Initio/RRKM study of the reaction mechanism and product branching ratios of neutral-radical ethynyl (C2H) and cyano (CN) radical species with unsaturated hydrocarbons is performed. The reactions studied apply to cold conditions such as planetary atmospheres including Titan, the Interstellar Medium (ISM), icy bodies and molecular clouds. The reactions of C2H and CN additions to gaseous unsaturated hydrocarbons are an active area of study. NASA’s Cassini/Huygens mission found a high concentration of C2H and CN from photolysis of ethyne (C2H2) and hydrogen cyanide (HCN), respectively, in the organic haze layers of the atmosphere of Titan. The reactions involved in the atmospheric chemistry of Titan lead to a vast array of larger, more complex intermediates and products and may also serve as a chemical model of Earth’s primordial atmospheric conditions. The C2H and CN additions are rapid and exothermic, and often occur barrierlessly to various carbon sites of unsaturated hydrocarbons. The reaction mechanism is proposed on the basis of the resulting potential energy surface (PES) that includes all the possible intermediates and transition states that can occur, and all the products that lie on the surface. The B3LYP/6-311g(d,p) level of theory is employed to determine optimized electronic structures, moments of inertia, vibrational frequencies, and zero-point energy. They are followed by single point higher-level CCSD(T)/cc-vtz calculations, including extrapolations to complete basis sets (CBS) of the reactants and products. A microcanonical RRKM study predicts single-collision (zero-pressure limit) rate constants of all reaction paths on the potential energy surface, which is then used to compute the branching ratios of the products that result. These theoretical calculations are conducted either jointly or in parallel to experimental work to elucidate the chemical composition of Titan’s atmosphere, the ISM, and cold celestial bodies.

Mechanisms of Chloroperoxidase-catalyzed Enantioselective Reactions as Probed by Site-directed Mutagenesis and Isotopic Labeling

Chloroperoxidase (CPO) is a heme-containing glycoprotein secreted by the marine fungus Caldariomyces fumago. Chloroperoxidase contains one ferriprotoporphyrin IX prosthetic group per molecule and catalyzes a variety of reactions, such as halogenation, peroxidation and epoxidation. The versatile catalytic activities of CPO coupled with the increasing demands for chiral synthesis have attracted an escalating interest in understanding the mechanistic and structural properties of this enzyme. In order to better understand the mechanisms of CPO-catalyzed enantioselective reactions and to fine-tune the catalytic properties of chloroperoxidase, asparagine 74 (N74) located in the narrow substrate access channel of CPO was replaced by a bulky, nonpolar valine and a polar glutamine using site-directed mutagenesis. The CPO N74 mutants displayed significantly enhanced activity toward nonpolar substrates compared to wild-type CPO as a result of changes in space and polarity of the heme distal environment. More interestingly, N74 mutants showed dramatically decreased chlorination and catalase activity but significantly enhanced epoxidation activity as a consequence of improved kinetic perfection introduced by the mutation as reflected by the favorable changes in kcat and kcat/KM of these reactions. It is also noted that the N74V mutant is capable of decomposing cyanide, the most notorious poison for many hemoproteins, as judged by the unique binding behavior of N74V with potassium cyanide. Histidine 105 (H105) was replaced by a nonpolar amino acid alanine using site-directed mutagenesis. The CPO H105 mutant (H105A) displayed dramatically decreased chlorination and catalase activity possibly because of the decreased polarity in the heme distal environment and loss of the hydrogen bonds between histidine 105 and glutamic acid 183. However, significantly increased enantioselectivity was observed for the epoxidation of bulky styrene derivatives. Furthermore, my study provides strong evidence for the proposed histidine/cysteine ligand switch in chloroperoxidase, providing experimental support for the structure of the 420-nm absorption maximum for a number of carbon monoxide complexes of heme-thiolate proteins. For the NMR study, [dCPO(heme)] was produced using 90% deuterated growth medium with excess heme precursors and [dCPO(Phe)] was grown in the same highly deuterated medium that had been supplemented with excess natural phenylalanine. To make complete heme proton assignments, NMR spectroscopy has been performed for high-resolution structural characterization of [dCPO(heme)] and [dCPO(Phe)] to achieve unambiguous and complete heme proton assignments, which also allows important amino acids close to the heme active center to be determined.

Engineering design of an integrated sustainable process system for cassava biobased materials

Cassava contributes significantly to biobased material development. Conventional approaches for its bio-derivative-production and application cause significant wastes, tailored material development challenges, with negative environmental impact and application limitations. Transforming cassava into sustainable value-added resources requires redesigning new approaches. Harnessing unexplored material source, and downstream process innovations can mitigate challenges. The ultimate goal proposed an integrated sustainable process system for cassava biomaterial development and potential application. An improved simultaneous release recovery cyanogenesis (SRRC) methodology, incorporating intact bitter cassava, was developed and standardized. Films were formulated, characterised, their mass transport behaviour, simulating real-distribution-chain conditions quantified, and optimised for desirable properties. Integrated process design system, for sustainable waste-elimination and biomaterial development, was developed. Films and bioderivatives for desired MAP, fast-delivery nutraceutical excipients and antifungal active coating applications were demonstrated. SRRC-processed intact bitter cassava produced significantly higher yield safe bio-derivatives than peeled, guaranteeing 16% waste-elimination. Process standardization transformed entire root into higher yield and clarified colour bio-derivatives and efficient material balance at optimal global desirability. Solvent mass through temperature-humidity-stressed films induced structural changes, and influenced water vapour and oxygen permeability. Sevenunit integrated-process design led to cost-effectiveness, energy-efficient and green cassava processing and biomaterials with zero-environment footprints. Desirable optimised bio-derivatives and films demonstrated application in desirable in-package O2/CO2, mouldgrowth inhibition, faster tablet excipient nutraceutical dissolutions and releases, and thymolencapsulated smooth antifungal coatings. Novel material resources, non-root peeling, zero-waste-elimination, and desirable standardised methodology present promising process integration tools for sustainable cassava biobased system development. Emerging design outcomes have potential applications to mitigate cyanide challenges and provide bio-derivative development pathways. Process system leads to zero-waste, with potential to reshape current style one-way processes into circular designs modelled on nature's effective approaches. Indigenous cassava components as natural material reinforcements, and SRRC processing approach has initiated a process with potential wider deployment in broad product research development. This research contributes to scientific knowledge in material science and engineering process design.

Caracteres morfo-agronômicos e bioquímicos de clones elite de mandioca de mesa com raízes de polpas amarelada e rosada.

A mandioca (Manihot esculenta Crantz) é considerada uma espécie relevante como fonte alimentícia para a população mundial, principalmente para os países subdesenvolvidos e emergentes. A mandioca é fornecedora de energia a partir do amido acumulado em suas raízes de reserva, mas é também importante destacar a presença dos carotenóides com atividade antioxidante. Nesse contexto, o presente trabalho teve como objetivo caracterizar, por meio de descritores morfológicos, agronômicos e bioquímicos, clones elite de mandioca de mesa de polpa aparelhada e rosada do programa de melhoramento genético de mandioca da Embrapa Cerrados. Foram caracterizados durante duas safras, 13 clones de mandioca de mesa com polpa amarelada e 8 clones com polpa rosada, em comparação com a variedade testemunha IAC 576-70 (BGMC 753). Para avaliar as características morfológicas foram obtidos 40 descritores qualitativos para cada clone. Tanto nos clones de polpas amarelada quanto naqueles de raízes de polpas rosada, houve diferenças morfológicas, demostrando que nenhum clone apresentou 100% de similaridade. O fator ano/safra não influenciou a expressão fenotípica dos caracteres aferidos. Com base no coeficiente cofenético, verificou-se elevado ajuste entre a representação gráfica via dendrograma de r = 0,80 nas raízes de polpa amarelada e r = 0,92 na rosada e a matriz de dissimilaridade genética. Entre os caracteres aferidos, os que apresentaram maior entropia nas raízes amarelada foram, a coloração da epiderme externa, forma do lóbulo central da folha e cor do córtex da raiz, ao passo que na rosada foi à cor do disco, forma do lóbulo central e cor do pecíolo. Foi realizada também a caracterização com base na altura da planta, altura da primeira ramificação, peso da parte aérea sem a cepa, produtividade em raízes, índices de amido nas raízes determinados por meio do método da balança hidrostática, tempo para a cocção e teor de ácido cianídrico nas raízes. Com base nos caracteres avaliados, os clones que se destacaram com polpa amarelada e rosada respectivamente, no caractere altura da primeira ramificação (273/08 e 259/08) e (390/08, 345/08 e a testemunha IAC 576-70), altura da planta (90/08, 272/08, 273/08, 497/08, 259/08 e 450/08) e (390/08, 345/08 e 378/08), peso da parte aérea sem a cepa (94/08 e 272/08) e (390/08, 406/08, 390/08, 378/08 e 341/08), porcentagem de amido nas raízes (26/08, 272/08, 259/08 e 450/08) e (378/08, 413/08, 390/08 e a testemunha IAC 576-70), produtividade de raízes (215/08) e (testemunha IAC 576-70, 341/08, 406/08, 390/08 e 387/08). Com relação ao tempo de cocção na safra 2011/2012, todos os clones necessitaram de tempo inferior a 30 minutos. Em relação ao teor de carotenóides totais nas raízes os clones de amarelada que se destacaram foram 91/08, 94/08, 215/08, 246/08, 272/08 e 497/08, e, naqueles de raízes rosada, os clones 406/08 e 341/08. Em relação ao teor de proteínas nas raízes amarelada, os clones 26/08, 90/08 e 91/08, foram os melhores enquanto nas raízes rosada se destacaram os clones 406/08 e a testemunha IAC 576-70. Os teores de HCN nas raízes de reserva de mandioca foram inferiores a 100 mg kg-1 em todos os clones avaliados. Diferenças significativas entre clones de mandioca de polpas amarelada e rosada foram verificadas para todas as características agronômicas, morfológicas e bioquímicas avaliadas. Os clones tiveram bom desempenho nas avaliações para o cultivo comercial na região do Cerrado e, alguns destes, têm potencial para utilização no melhoramento visando o incremento de carotenóides. ABSTRACT: Cassava (Manihot esculenta Crantz) is considered a relevant species as a food source for the world's population, particularly for developing and emerging countries. The cassava is a provider of energy from starch accumulated in their reserve roots, but it is also important to highlight the presence of carotenoids with antioxidant activity. In this context, this study aimed to characterize, using morphological, agronomic and biochemical, descriptors elite clones from sweet cassava of yellowish and pinkish pulps from the cassava breeding program at Embrapa Cerrados. They were characterized for two crops, 13 edible cassava clones with yellowish pulp and 8 clones with pinkish pulp, compared with the control variety IAC 576-70 (BGMC 753). To evaluate the morphological characteristics were obtained 40 qualitative descriptors for each clone. Both clones the yellowish pulp as those the roots the pinkish pulp, there was morphological differences among clones, showing that no clone showed 100% similarity. The year / crop factor did not influence the phenotypic expression of measured characters. Based on cofenetic coefficient, was found high fit between the graphical representation via dendrogram of r = 0.80 in the roots of yellowish pulp and r = 0.92 in the pinkish of genetic dissimilarity matrix. Among the measured characters, those with the highest entropy in the yellowish roots were, the color of the outer epidermis, the central lobe shape of the leaf and root cortex color, whereas the pinkish was the color to disc, central lobe shape and petiole color. We also performed the characterization based on plant height, the first branch point, and shoot weight without strain, productivity in roots, and index of starch in the roots determines by the method of hydrostatic balance, time for cooking and acid cyanide content in the roots. Based on the evaluated characters, clones stood out with pulps yellowish and pinkish respectively, characters height of the first branch (273/08 and 259/08) and (390/08, 345/08 and the witness IAC 576-70), plant height (90 / 08, 272/08, 273/08, 497/08, 259/08 and 450/08) and (390/08, 345/08 and 378/08), shoot weight without strain (94/08 and 272/08) and (390/08, 406/08, 390/08, 378/08 and 341/08), percentage of starch in the roots (26/08, 272/08, 259/08 and 450/08) and (378/08, 413/08, 390/08 and the witness IAC 576-70), roots of productivity (215/08) and (witnesses IAC 576-70, 341/08, 406/08, 390/08 and 387/08). Regarding the cooking time in the 2011/2012 harvest, all clones showed time less than 30 minutes. Regarding the total carotenoid content in the pulps clones of yellowish roots that stood out were 91/08, 94/08, 215/08, 246/08, 272/08 and 497/08, and, those the clones with pulp pinkish 406/08 and 341/08. Regarding the protein content in yellowish roots the clones 26/08, 90/08 and 91/08, was the best while the pinkish roots highlight clones 406/08 and witness IAC 576-70. The levels of HCN in reserve roots of cassava were less than 100 mg kg-1em all evaluated clones. Significant differences between yellowish and pinkish of pulps cassava clones were checked for all agronomic, morphological and biochemical characteristics evaluated. The clones had well in the ratings for commercial cultivation in the Cerrado region and some of these, clones has potential for use in breeding aimed at increase of carotenoids.