Optimization of microwave-assisted extration of phenolic compounds from tomao by full factorial design coupled with response surface methodology

Tomato (Lycopersicon esculentum Mill.), apart from being a functional food rich in carotenoids, vitamins and minerals, is also an important source of phenolic compounds [1 ,2]. As antioxidants, these functional molecules play an important role in the prevention of human pathologies and have many applications in nutraceutical, pharmaceutical and cosmeceutical industries. Therefore, the recovery of added-value phenolic compounds from natural sources, such as tomato surplus or industrial by-products, is highly desirable. Herein, the microwave-assisted extraction of the main phenolic acids and flavonoids from tomato was optimized. A S-Ieve! full factorial Box-Behnken design was implemented and response surface methodology used for analysis. The extraction time (0-20 min), temperature (60-180 "C), ethanol percentage (0-100%), solidlliquid ratio (5-45 g/L) and microwave power (0-400 W) were studied as independent variables. The phenolic profile of the studied tomato variety was initially characterized by HPLC-DAD-ESIIMS [2]. Then, the effect of the different extraction conditions, as defined by the used experimental design, on the target compounds was monitored by HPLC-DAD, using their UV spectra and retention time for identification and a series of calibrations based on external standards for quantification. The proposed model was successfully implemented and statistically validated. The microwave power had no effect on the extraction process. Comparing with the optimal extraction conditions for flavonoids, which demanded a short processing time (2 min), a low temperature (60 "C) and solidlliquid ratio (5 g/L), and pure ethanol, phenolic acids required a longer processing time ( 4.38 min), a higher temperature (145.6 •c) and solidlliquid ratio (45 g/L), and water as extraction solvent. Additionally, the studied tomato variety was highlighted as a source of added-value phenolic acids and flavonoids.

Extraction of betacyanins from Gomphrena globosa L. flowers: choosing na acid as adjuvant

Betalains are plant derived natural pigments that are presently gaining popularity for use as natural colorants in food industry. Although being betalains from red beetroot already used as food colorant (E- 162), these compounds are not as well studied as compared to other natural pigments such as anthocyanins, carotenoids or chlorophylls (I]. Since food additives are on the focus of public interest, it is becoming increasingly important to meet consumers' expectations for natural and healthy products. Hence, the search for new plant-derived colorants for the food industry is still necessary [2]. Betalains were originally called 'nitrogenous anthocyanins', which incorrectly implied structural similarities between the two pigment classes. There are two structurally different types of betalains: the yellow/orange betaxanthins which are the condensation products of betalamic acid and assorted amino compounds, and the red betacyanins which are formed by glycosylation and acylation of cyclo-DOPA [3]. Looking at the chemical structure of the pigment, the addition of an acid to the extraction solvent will increase the affinity of the pigment with the solvent. The aim of this study was to use Gomphrena globosa L. flowers, as an alternative plant source to obtain these pigments and to evaluate the best acid to be used within the extraction procedure. For that purpose three different acids (acetic, hydrochloric and phosphoric acids, all ofthem allowed by the food industry), adjusted at the same pH, were tested during a maceration extraction procedure. After the extraction a purification through C18 column was performed in order to obtain a more concentrate extract in betacyanins. The results were analysed by HPLC-PDA-MSIESI. The betacyanin profile allowed the identification of gomphrenin IIJIII and isogomphrenin IIIIII and the best results were achieved by performing the extraction procedure using hydrochloric acid (6.6 mg/g extract), while phosphoric acid only presented trace amounts of these compounds. When acetic acid was used, the pigment extracted was 6.8 times less (0.97 mg/g extract) when compared to HCI. In conclusion hydrochloric acid can be considered the most suitable acid to be applied in the extraction procedure of these pigments.

Flowers of Gomphrena globosa L. as an alternatve source of betacyanins: optimization of the extraction using response surface methodology

Betacyanins are betalain pigments that display a red-violet colour which have been reported to be three times stronger than the red-violet dye produced by anthocyanins [1]. The applications of betacyanins cover a wide range of matrices, mainly as additives or ingredients in the food industry, cosmetics, pharmaceuticals and livestock feed. Although, being less commonly used than anthocyanins and carotenoids, betacyanins are stable between pH 3 to 7 and suitable for colouring in low acid matrices. In addition, betacyanins have been reported to display interesting medicinal character as powerful antioxidant and chemopreventive compounds either in vitro or in vivo models [2]. Betacyanins are obtained mainly from the red beet of Beta vulgaris plant (between I 0 to 20 mg per I 00 g pulp) but alternative primary sources are needed [3]. In addition, independently of the source used, the effect of the variables that affect the extraction of betacyanins have not been properly described and quantified. Therefore, the aim of this study was to identifY and optimize the conditions that maximize betacyanins extraction using the tepals of Gomphrena globosa L. flowers as an alternative source. Assisted by the statistical technique of response surface methodology, an experimental design was developed for testing the significant explanatory variables of the extraction (time, temperature, solid-liquid ratio and ethanolwater ratio). The identification was performed using high-performance liquid chromatography coupled with a photodiode array detector and mass spectrometry with electron spray ionization (HPLC-PDAMS/ ESI) and the response was measured by the quantification of these compounds using HPLC-PDA. Afterwards, a response surface analysis was performed to evaluate the results. The major betacyanin compounds identified were gomphrenin 11 and Ill and isogomphrenin IJ and Ill. The highest total betacyanins content was obtained by using the following conditions: 45 min of extraction. time, 35•c, 35 g/L of solid-liquid ratio and 25% of ethanol. These values would not be found without optimizing the conditions of the betacyanins extraction, which moreover showed contrary trends to what it has been described in the scientific bibliography. More specifically, concerning the time and temperature variables, an increase of both values (from the common ones used in the bibliography) showed a considerable improvement on the betacyanins extraction yield without displaying any type of degradation patterns.

Transcription of alternative oxidase (AOX) and plastid terminal oxidase (PTOX) during stress-regulated root tissue growth in Daucus carota L. - An approach to identify functional marker candidates for breeding on carrot yield stability

A presente tese explora a hipótese de utilização dos genes da oxidase alternativa (AOX) e da oxidase terminal da plastoquinona (PTOX) como genes-alvo para o desenvolvimento de marcadores funcionais (MF) para avaliar a performance do crescimento em cenoura, fator determinante da produtividade. Para avaliar se os referidos genes estão associados com o crescimento da cenoura procedeu—se ao seu isolamento e posterior análise dos seus perfis de transcrição em diversos sistemas biológicos. O sistema in vitro selecionado, denominado sistema de culturas primárias, permitiu avaliar alterações na quantidade de transcritos desses genes durante os processos de reprogramação celular e crescimento. Ao nível da planta foi também estudado o efeito do frio na expressão precoce dos genes AOX. Ambos os genes DcAOX1 e DcAOX2a revelaram uma resposta rápida e um padrão semelhante apos stresse (inoculação in vitro e resposta ao frio). Foi igualmente verificado um incremento na expressão do gene DcPTOX durante a fase inicial do processo de reprogramação celular. Estudos de expressão dos genes AOX durante o desenvolvimento da raiz da cenoura revelaram que o gene DcAOX2a será potencialmente o gene mais envolvido neste processo. De modo a avaliar a hipótese de envolvimento do gene DcPTOX no crescimento da raíz procederam—se a estudos de expressão ao nível do tecido meristemático. Todavia, para um mais completo entendimento da ligação entre DcPTOX e o crescimento secundário e/ou acumulação de carotenos, a expressão do gene DcPTOX foi também avaliada em raízes de cenoura durante o desenvolvimento, utilizando cultivares caracterizadas por distintos conteúdos de carotenos. Os resultados obtidos demonstraram a associação do gene DcPTOX a ambos os processos. O envolvimento da PTOX no crescimento adaptativo da raiz foi analisado com um ensaio que permitiu identificar, no tecido meristemático, uma resposta precoce do gene DcPTOX face a uma diminuição da temperatura. Adicionalmente, foi efetuada a seleção de genes de referência para uma analise precisa da expressão génica por RT-qPCR em diversos sistemas biológicos de cenoura, e a importância do seu estudo ao nível do sistema biológico foi realçada. Os resultados desta tese são encorajadores para prosseguir os estudos de utilização dos genes AOX e PTOX como MF no melhoramento da performance do crescimento adaptativo em cenoura, fator determinante para a produtividade; ABSTRACT: This thesis explores the hypothesis of using the alternative oxidase (AOX) and theplastid terminal oxidase (PTOX) as target genes for functional marker (FM) development for yield-determining growth performance in carrot. To understand if these genes are associated to growth, different AOX gene family members and the single PTOX gene were isolated, and their expression patterns evaluated in diverse carrot plant systems. An in-vitro primary culture system was selected to study AOX and PTOX transcript changes during cell reprogramming and growth performance. At plant level, a putative early response of AOX to chilling was also evaluated. In fact, both DcAOXl and DcAOXZa were early responsive and showed similar patterns under stress conditions (in vitro inoculation and chilling). A role for DcPTOX during earliest events of cell reprogramming was also suggested. Next, the expression profiles of AOX gene family members during carrot tap root development were investigated. DcAOXZa was identified as the most responsive gene to root development. In order to evaluate if DcPTOX is associated with carrot tap root growth performance, DcPTOX transcript levels were measured in the central root meristem. To further understand whether DcPTOX is associated with secondary growth and/or carotenoids accumulation, DcPTOX expression was also studied in deveIOping carrot tap roots in cultivars with different carotenoids contents. The results indicated that DcPTOX associates to both carotenoid biosynthesis and secondary growth during storage root development. To obtain further insights into the involvement of PTOX on adaptive growth, the early effects of temperature decrease were explored in the root meristem, where a short—term early response in DcPTOX was found, probably associated with adaptive growth. Furthermore, a selection of the most suitable reference genes for accurate RT—qPCR analysis in several carrot experimental systems was performed and discussed. The present research provides the necessary toolbox for continuing studies in carrot AOX and PTOX genes as promising resources for FM candidates in order to assist breeding on yield—determining adaptive growth performance.

Chemical composition of cultivated seaweed ulva clathrata (Roth) C. Agardh

Samples of cultivated Ulva clathrata were collected from a medium scale system (MSS, 1.5 1.5 m tank), or from a large scale system (LSS, 0.8 ha earthen pond). MSS samples were dried directly while the LSS sample was washed in freshwater and pressed before drying. Crude protein content ranged 20–26%, essential amino acids accounting for 32–36% of crude protein. The main analysed monosaccharides were rhamnose (36–40%), uronic acids (27–29%), xylose (10–13%) and glucose (10–16%). Some notable variations between MSS and LSS samples were observed for total dietary fibre (26% vs 41%), saturated fatty acids (31% vs 51%), PUFAS (33% vs 13%), carotenoids (358 vs 169 mg kg1 dw) and for Ca (9 vs 19 g kg1 ), Fe (0.6 vs 4.2 g kg1 ), Cu (44 vs 14 mg kg1 ), Zn (93 vs 17 mg kg1 ) and As (2 vs 9 mg kg1 ). The chemical composition of U. clathrata indicates that it has a good potential for its use in human and animal food.

Bioactive compounds of citrus as health promoters

Citrus are a group of fruit species, quite heterogeneous in many aspects, including chemical composition of the fruit. Since ancient times, some citrus fruits were used to prevent and cure human diseases. In the recent decades, it has been demonstrated that fruits can actually help prevent and cure some diseases and above all, they are essential in a balanced diet. Citrus fruits, as one of the groups of fruit species, with greater importance in the world, have been studied for their effects on human health. Some species of citrus were referred as potential antioxidant based therapy for heart disease, cancer and inflammation. Fruit peels and seeds have also high antioxidant activity. The health benefits of citrus fruit have mainly been attributed to the high level of bioactive compounds, such as phenols (e.g., flavanone glycosides, hydroxycinnamic acids), carotenoids and vitamin C. These compounds are present in the fruit pulp and hence in the juice. But some bioactive compounds can be found in parts of the fruit which usually are not used for human food. The content of bioactive compounds depends on the species and cultivar, but also depends on the production system followed in the orchard. Citrus fruits, their derivatives and their by-products (peel, pulp and oil) are reach in different bioactive compounds and its maturity, postharvest and agroindustry processes influence their composition and concentration. The aim of this chapter was to review the main bioactive compounds of the different components of citrus and their relationship to health.

Ultrafast transient absorption spectroscopy: principles and applications

The study of photophysical and photochemical processes crosses the interest of many fields of research in physics, chemistry and biology. In particular, the photophysical and photochemical reactions, after light absorption by a photosynthetic pigment-protein complex, are among the fastest events in biology, taking place on timescales ranging from tens of femtoseconds to a few nanoseconds. Among the experimental approaches developed for this purpose, the advent of ultrafast transient absorption spectroscopy has become a powerful and widely used technique.[1,2] Focusing on the process of photosynthesis, it relies upon the efficient absorption and conversion of the radiant energy from the Sun. Chlorophylls and carotenoids are the main players in the process. Photosynthetic pigments are typically arranged in a highly organized fashion to constitute antennas and reaction centers, supramolecular devices where light harvesting and charge separation take place. The very early steps in the photosynthetic process take place after the absorption of a photon by an antenna system, which harvests light and eventually delivers it to the reaction center. In order to compete with internal conversion, intersystem crossing, and fluorescence, which inevitably lead to energy loss, the energy and electron transfer processes that fix the excited-state energy in photosynthesis must be extremely fast. In order to investigate these events, ultrafast techniques down to a sub-100 fs resolution must be used. In this way, energy migration within the system as well as the formation of new chemical species such as charge-separated states can be tracked in real time. This can be achieved by making use of ultrafast transient absorption spectroscopy. The basic principles of this notable technique, instrumentation, and some recent applications to photosynthetic systems[3] will be described. Acknowledgements M. Moreno Oliva thanks the MINECO for a “Juan de la Cierva-Incorporación” research contract. References [1] U. Megerle, I. Pugliesi, C. Schriever, C.F. Sailer and E. Riedle, Appl. Phys. B, 96, 215 – 231 (2009). [2] R. Berera, R. van Grondelle and J.T.M. Kennis, Photosynth. Res., 101, 105 – 118 (2009). [3] T. Nikkonen, M. Moreno Oliva, A. Kahnt, M. Muuronen, J. Helaja and D.M. Guldi, Chem. Eur. J., 21, 590 – 600 (2015).

Variação no conteúdo protéico e pigmentar em variantes cromáticas de Gracilaria domingensis nas populações naturais de Rio do Fogo-RN, BRASIL

The seaweed Gracilaria domingensis is a common species in the coast of Rio Grande do Norte. This species lives in the intertidal zone, where colour strains (red, green and brown) co-occur during the whole year. Seaweeds that live in this region are exposed to daily changes and to the rhythm of the tide. During the low tide they are exposed to dissection, hiper-or hipo-osmotic shock, high temperatures and high irradiance. The aim of this study was to analyze whether the pigment and protein content of the colour strains of G. domingensis is affected by some environmental parameters in a temporal scale. The seaweeds were collected during 10 months in the seashore of Rio do Fogo (RN). The total soluble proteins and the phycobiliprotein were extracted in phosphate buffer and the carotenoids were analyzed by a standardized method through HPLC-UV. The pigments analysis showed that phycoerithrin is the most abundant pigment in the three strains. This pigment was strongly correlated with nitrogen and the photosynthetically active radiation. Chlorophyll presented higher concentrations than carotenoids during the whole, but the ratio carotenoid/chlorophyll-a was modified by incident radiation. The most abundant carotenoid was ß-carotene and zeaxanthin, which had higher concentrations in the higher radiation months. The concentration increase of zeaxanthin in this period indicated a photoprotective response of the seaweed. The three strains presented a pigment profile that indicates different radiation tolerance profile. Our results pointed that the green strain is better adapted to high irradiance levels than the red and brown strains

Serum levels of lycopene, beta-carotene, and retinol and their correlation with sperm DNA damage in normospermic and infertile men

Background: Oxidative stress in reproductive system leads to sperm DNA damage and sperm membrane lipid peroxidation and may play an important role in the pathogenesis of male infertility, especially in idiopathic cases. Antioxidants such as carotenoids function against free radical damages. Objective: The aim of this study was to determine the levels of lycopene, beta-carotene and retinol in serum and their relationship with sperm DNA damage and lipid peroxidation in infertile and normospermic males. Materials and Methods: Sixty two infertile men and 71 normospermic men participated in this study. Blood and semen samples were collected from all subjects. Sperm DNA damage was measured using TUNEL method. Carotenoids, retinol, and malonedildehyde in serum were also determined. Results: DNA fragmentation was higher in infertile group comparing to control group. Serum levels of lycopene, beta-carotene and, vitamin A in infertile men were significantly lower than normospermic men (p< 0.001, =0.005, and =0.003 respectively). While serum MDA was not significantly different between two groups, MDA in seminal plasma of infertile men was significantly higher than control group (p< 0.001). Conclusion: We concluded that lycopene, beta-carotene, and retinol can reduce sperm DNA fragmentation and lipid peroxidation through their antioxidant effect. Therefore the DNA fragmentation assay and determination of antioxidants factors such as lycopene, beta-carotene and retinol, along with sperm analysis can be useful in diagnosis and treatment of men with idiopathic infertility.

Effect of drying on the physical, chemical and sensorial properties of kiwi

Kiwi fruit is a highly nutritional fruit due to the high level of vitamin C and its strong antioxidant capacity due to a wide number of phytonutrients including carotenoids, lutein, phenolics, flavonoids and chlorophyll [1]. Drying consists of a complex process in which simultaneous heat and mass transfer occur. Several alterations occur during the drying of foods at many levels (physical, chemical, nutritional or sensorial) which are influenced by a number of factors, including processing conditions [2]. Temperature is particularly important because of the effects it produces at the chemical and also at the physical level, particularly colour and texture [3]. In the present work were evaluated the changes in sliced kiwi when exposed to air drying at different temperatures (50, 60, 70, 80 ºC), namely in terms of some chemical properties like ascorbic acid or phenolic compounds, physical characteristics like colour and texture and also at the sensorial level. All experiments followed standard established procedures and several replicates were done to assess each property. The results obtained indicated that moisture was reduced with drying by 74 to 87%, depending on the temperature. Also ascorbic acid decreased with drying, being 7% for 50 ºC and increasing up to 28% for the highest temperature (80 ºC). The phenolic compounds and antioxidant activity were also very much affected by the drying temperature. The water activity of the dried samples varied from 0.658 to 0.753, being compatible with a good preservation. Regarding colour, the total colour difference between the dried samples and the fresh sample was found to vary in the range 9.45 – 17.17. The textural parameters were also much affected by drying, namely hardness which decreased by 45 to 72 %, and all other parameters increased: cohesiveness (approximately doubled), springiness (increased 2 to 3 times) and chewiness which increased up to 2.5 times that off the fresh sample. Adhesiveness, which was observed for the fresh samples (-4.02 N.s) disappeared in all the dried samples. The sensorial analysis made to the dried samples allowed establishing the sensorial profiles as shown in Figure 1.

Tomato Lycopene: Functional Proprieties and Health

The growing concerns for physical wellbeing and health have been reflected in the way we choose food in our table. Nowadays, we are all more informed consumers and choose healthier foods. On the other hand, stroke, cancer and atherosclerosis may be somehow minimized by the intake of some bioactive compounds present in food, the so-called nutraceuticals and functional foods. The aim of this work was to make a revision of the published studies about the effects of some bioactive compounds, namely lycopene in human health, in the prevention of diseases, thus playing the role of a functional food. Free radical in human body can induce cell damage and consequently can be responsible for the development of some cancers and chronic diseases. Lycopene is one of the most powerful antioxidants known, being the predominant carotenoid in tomato. The respective chemistry, bioavailability, and its functional role in the prevention of several diseases will be object of this work. On the other hand, the inclusion of lycopene in some foods can also be made by biotechnology and represents a way to recover the wastes in the tomato industry with nutritional positive effects in health.

A first insight on the biodegradation of limestone - the case of the World Heritage Convent of Christ

The present study is a multidisciplinary approach applied to architectural stone materials of the Convent of Christ in Tomar (Portugal) in order to understand and mitigate the active decay processes. The structure and appearance of the stonework from the Convent of Christ are strongly affected by stains, biofilms and structural degradation. To investigate these phenomena, a multianalytical approach comprising X-ray microdiffraction, scanning electron microscopy, microRaman and microinfrared spectroscopy was applied to the examination of altered outdoor stone areas being detected calcium oxalates, carotenoids and microbial proliferation. The presence of these alteration products seems to be correlated with the microbial activity of bacteria, microalgae, cyanobacteria and filamentous fungi. This work showed that the application of complementary methodologies is an efficient strategy to characterise the stone decay, and constitute a starting point for successful conservation intervention plans that are urgent to ensure the preservation and safeguard of this emblematic monument.

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.

Natural variation in expression of genes associated with carotenoid biosynthesis and accumulation in cassava (Manihot esculenta Crantz) storage root.

ABSTRACT: BACKGROUND: Cassava (Manihot esculenta Crantz) storage root provides a staple food source for millions of people worldwide. Increasing the carotenoid content in storage root of cassava could provide improved nutritional and health benefits. Because carotenoid accumulation has been associated with storage root color, this study characterized carotenoid profiles, and abundance of key transcripts associated with carotenoid biosynthesis, from 23 landraces of cassava storage root ranging in color from white-to-yellow-to-pink. This study provides important information to plant breeding programs aimed at improving cassava storage root nutritional quality. RESULTS: Among the 23 landraces, five carotenoid types were detected in storage root with white color, while carotenoid types ranged from 1 to 21 in storage root with pink and yellow color. The majority of storage root in these landraces ranged in color from pale-to-intense yellow. In this color group, total ß-carotene, containing all-E-, 9-Z-, and 13-Z-ß-carotene isomers, was the major carotenoid type detected, varying from 26.13 to 76.72 %. Although no ?-carotene was observed, variable amounts of a ?-ring derived xanthophyll, lutein, was detected; with greater accumulation of ?-ring xanthophylls than of ß-ring xanthophyll. Lycopene was detected in a landrace (Cas51) with pink color storage root, but it was not detected in storage root with yellow color. Based on microarray and qRT-PCR analyses, abundance of transcripts coding for enzymes involved in carotenoid biosynthesis were consistent with carotenoid composition determined by contrasting HPLC-Diode Array profiles from storage root of landraces IAC12, Cas64, and Cas51. Abundance of transcripts encoding for proteins regulating plastid division were also consistent with the observed differences in total ß-carotene accumulation. CONCLUSIONS: Among the 23 cassava landraces with varying storage root color and diverse carotenoid types and profiles, landrace Cas51 (pink color storage root) had low LYCb transcript abundance, whereas landrace Cas64 (intense yellow storage root) had decreased HYb transcript abundance. These results may explain the increased amounts of lycopene and total ß-carotene observed in landraces Cas51 and Cas64, respectively. Overall, total carotenoid content in cassava storage root of color class representatives were associated with spatial patterns of secondary growth, color, and abundance of transcripts linked to plastid division. Finally, a partial carotenoid biosynthesis pathway is proposed.

Effect of post-emergent herbicides application on carotenoid concentration in green corn.

Considering the influence of herbicides in the metabolism of the carotenoids in corn, the objective of the present study was to evaluate the effect of herbicides and genotype on carotenoids concentration. The green corn hybrids BRS 1030 and P30F53 were subjected to a post-emergent herbicides application at 20 and 30 days after emergence. Carotenoids were extracted from corn grains and analyzed to quantify ?- and ?-carotene, lutein, zeaxanthin, ?-cryptoxanthin, total carotenoids (TC), and total of vitamin A carotenoids precursors (proVA). The application of foramsulfuron + iodosulfuron-methyl-sodium (40 + 2.6 g ha-1), nicosulfuron (20 g ha-1), mesotrione (120 g ha-1) and tembotrione (80 g ha-1 and 100 g ha-1) promoted higher concentration of carotenoids in fresh green corn. Lutein, zeaxanthin, ?-cryptoxanthin, ?-carotene, ?-carotene, proVA carotenoids, and TC concentration increased after foramsulfuron + iodosulfuron-methyl-sodium in late application (V5 to V6), nicosulfuron in both applications, mesotrione applied post-initial (V3 to V4), tembotrione (100 g ha-1) in both applications and tembotrione (80 g ha-1) in late post-application, at least for one hybrid. The content of carotenoids in the green corn kernels differed between ?BRS 1030? and ?P30F53?. Our results suggest a possibility of significant increase of carotenoids in green corn kernels through the handling of corn production with post-emergent herbicides.