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Plant tissue culture allows the maintenance of species of interest under controlled conditions. There have been few studies related to the morphological changes that take place during callus formation and redifferentiation processes. The aim of this work was to analyze the morphological changes during dedifferentiation and redifferentiation processes in leaf explants of marigold (Tagetes erecta L.) using transmission electron microscopy, digital image analysis and concepts of fractal dimension. Leaf explants were sown in MS medium supplemented with 2.0 mg L−1 of benzyladenine and 2.0 mg L−1 of 2,4-dichlorophenoxyacetic acid to induce dedifferentiation (callus formation). The redifferentiation process (bud formation) was obtained with explants sown in MS medium supplemented with 3.0 mg L−1 indoleacetic acid and 0.5 mg L−1 benzyladenine. During the first 8 days, the dedifferentiation process showed gradual changes in terms of the area and perimeter of the explants. The fractal dimension of the perimeter corresponding to the projected image of the calli changed from smooth to rough at day 20. These features indicate variations in tissue morphometry as a result of the callus formation process and further formation of cellular aggregates. Structural observations during the dedifferentiation process showed changes in shape and cellular organization as callus formation became evident. Round-shaped cells were observed in the callus, with few organelles and a low degree of differentiation. Indirect organogenesis was achieved over a period of 10 days; in addition, plantlets developed from the obtained buds in 20 days in growth regulator-free MS medium.

Hibiscus sabdariffa has gained an important position in the food industry because of its high anthocyanins content, related to the persistent red calyx of its flowers as the major component. Moreover, it has been used as a food colorant and active ingredient to develop food with some health benefits. The effect of addition of microencapsulated anthocyanins from Hibiscus sabdariffa in a fish feed was evaluated using Carassius auratus as a study model. Anthocyanins were microencapsulated with maltodextrins (10 DE) and microcapsules structure was analyzed by scanning electron microscopy. Microcapsules were added to fish diet (D) at 150, 300, and 450 mg anthocyanin/kg of diet. Fantail goldfish were feeding for 8 weeks and the feed intake (FI), feed conversion ratio (FCR), growth rate (GR), specific growth rate (SGR), survival, and weight gain (WG) were evaluated. Skin chromatophores from the head, dorsal, and caudal fin of each treatment were observed by light microscopy. Microencapsulation efficiency was 90 ± 0.2%. The microcapsules showed a spherical shape, the mean size was 4.0 μm, and the powder showed a pink-red color. Feed intake and survival were similar to all diets (p > 0.05). D150 and D300 showed a significant variation in GR, SGR, and WG (p

Rosemary (Rosmarinus officinalis) is a culinary and medicinal plant used in food and pharmaceutical industry. The wide range of biological activities is mainly related to phenolic and terpenic compounds; like carnosic acid (CA), carnosol (CS) and rosmarinic acid (RA), mainly reported in rosemary leaf extracts, and recently described in rosemary callus extracts. The aim of this work was to investigate the chemical profile of rosemary cell lines and evaluate their antiproliferative potential against human HT-29 colorectal cancer cell lines. For this purpose, rosemary leaf explants were dedifferentiated on MS medium and added with 2, 4-D (2, 4-dichlorophenoxyacetic acid; 2 mg/L) and BAP (6-benzylaminopurine; 2 mg/L). Cell aggregates were separated according to colour and three rosemary cell lines cultures were established: green (RoG), yellow (RoY) and white (RoW). The chemical profile of rosemary cell lines extracts was characterized by combining HPLC and GC platforms coupled to HR-MS/MS. The antiproliferative activity against HT-29 cell line was analyzed with MTT assay. A total of 71 compounds, including hydroxycinnamic acid and hydroxybenzoic acid derivatives, flavonoids, phenolic di- and triterpenes, as well as relevant unsaturated fatty acids and their esters, phytosterols, and carotenoids were tentatively identified in the extract of the target cell lines. The antiproliferative activity test against HT-29 cell using the MTT assay revealed that the viability of HT-29 colon cancer cells was affected after treatment with the RoW extract (IC50 of 49.63 μg/mL) at 48 h. These results showed that rosemary cell lines can also accumulate other bioactive phytochemicals with demonstrated antiproliferative potential., Araceli Urquiza-López acknowledges a study grant from CONACYT and mobility grant to CIAL Madrid. This work was supported by Instituto Politécnico Nacional, México (IPN/SIP 20200706) and Ministerio de Ciencia, Innovación y Universidades, Spain (Project AGL2017-89417-R).

Ruta graveolens L. is a plant of agricultural interest due to its biological activity against phytopathogenic microorganism. This activity has been attributed to the presence of alkaloids, furanocoumarins, terpenes and fatty acids. The accumulation of these compounds is affected by environmental factors. The aim of this work was to analyze the seasonal variation the chemical profile of rue and to evaluate the biological activity against phytopathogenic fungi. Extracts were obtained in January, April, June and September 2017, chemically characterized by GC-MS. The antifungal activity was determined by absorbance, four concentrations were evaluated: 4, 8, 16 and 32 mg/mL. In extracts from the aerial parts, fatty acids were the most abundant group in the January sample (29.7%), and alkaloids in the April and June (27.7%) samples. In extracts from the roots, alkaloids were the most abundant group in the four collections. The highest percentage of growth inhibition on Fusarium and Stemphylium was obtained with the extracts of the January sample. Ruta graveolens can be considered an alternative for the management of phytopathogenic fungi of crops of economic importance., L.R.V. acknowledges CONACYT for the doctoral grant 449402. M.B. acknowledges MINECO for the “Juan de La Cierva-Formación” FJCI-2016-30902 and MICIU for the “Juan de La Cierva-Incorporación” IJC 2018-037830-I postdoctoral grants. Authors acknowledge to SIBE and EDI, IPN.

Tagetes erecta is an asteraceous plant of industrial, ornamental and medicinal importance; its inflorescences have been used as a pigment source for food coloring, mainly for poultry skin and eggs. Nevertheless, there are few reports on plant regeneration or micropropagation, because unsuccesfull results in the plant's reaction to the growth regulators, developing embryogenesis on Tagetes erecta. In this study, somatic embryogenesis was induced and plantlets of Tagetes erecta were regenerated. For induction of globular structures MS medium supplemented with 2,4-D (4.5 µM) and BAP (8.8 µM) was used; globular structures were transferred to MS medium with 45 g l-1 sucrose until the embryos maturation. Transmission electron microscopy showed characteristic subcellular structures of embryogenic callus. Somatic embryos were transferred to MS medium without plant growth regulators and whole plantlets were obtained. In vitro plants were successfully transplanted into a mixture of peat moss and vermiculite (1 : 1 v/v) under greenhouse conditions. In this study, somatic embryogenesis and plant regeneration system from foliar explants were established, an important requirement for performing genetic transformation events on Tagetes erecta.

Roselle is a plant that accumulates anthocyanins significantly, hence its importance as food coloring and as a source of antioxidant compounds for human health. This study was aimed to determine phenolic composition and antioxidant capacity of methanolic extracts, and beverages obtained from native roselle cultivars in Mexico (Negra, Sudan, Rosa and Blanca) with different degrees of pigmentation, and to establish the color-composition relationship. Chromatographic methods were used to determine phenolic compounds: flavanols, flavonols, benzoic, hibiscus and phenolic acids as well as two main anthocyanins (cyanidin 3-sambubioside and delphinidin 3-sambubioside). The antioxidant capacity was evaluated by ABTS and FRAP assays. Tristimulus colorimetry showed to be a useful technique to determine the color-composition relationship, leading to equations that allowed to predict anthocyanin content of roselle (R > 0.84). Also, a stepwise linear discriminant analysis (SLDA) was developed in order to classify roselle cultivars. The obtained mathematical model could be an important tool to be used in colorimetric characterization of functional compounds used in food processing.

Tagetes erecta is an asteraceous plant of industrial, ornamental and medicinal importance; its inflorescences have been used as a pigment source for food coloring, mainly for poultry skin and eggs. Nevertheless, there are few reports on plant regeneration or micropropagation, because unsuccesfull results in the plant’s reaction to the growth regulators, developing embryogenesis on Tagetes erecta. In this study, somatic embryogenesis was induced and plantlets of Tagetes erecta were regenerated. For induction of globular structures MS medium supplemented with 2,4-D (4.5 µM) and BAP (8.8 µM) was used; globular structures were transferred to MS medium with 45 g l(−1) sucrose until the embryos maturation. Transmission electron microscopy showed characteristic subcellular structures of embryogenic callus. Somatic embryos were transferred to MS medium without plant growth regulators and whole plantlets were obtained. In vitro plants were successfully transplanted into a mixture of peat moss and vermiculite (1 : 1 v/v) under greenhouse conditions. In this study, somatic embryogenesis and plant regeneration system from foliar explants were established, an important requirement for performing genetic transformation events on Tagetes erecta.

BACKGROUND: Digital image analysis has an important role in geographical provenance of grains, as it can provide parameters of size, shape and color, which are important quality parameters for the design of engineering processes such as drying and milling of grains. In this study, digital image analysis was used to classify nine rice cultivars based on different morphometric parameters using the three sides of the grain (lateral, ventral and axial), Feret diameter, and 10 different form factors and color parameters (CIE L*, a* and b*). RESULTS: Result of principal component analyisis was an equation with seven variables (area, perimeter, length, width, thickness, sphericity and color), which was useful for distinguishing between nine different cultivars. The morphometric and color parameters for the Mor A-98 and Mor A-92 varieties showed they had 88% similarity. The variability was expressed with a confidence of 95%. CONCLUSION: Multivariate analysis indicated that the lateral side is the most sensitive for the classification of Mexican rice grains because of its color and morphometric characteristics. These results showed the application of image analysis for the future classifications of grains. Copyright © 2012 Society of Chemical Industry

Marigold (Tagetes erecta) flowers are primarily used in industry for their high pigment content. Flower color development implies that chloroplast–chromoplast transition is associated with carotenoid biosynthesis. We report the recovery of undifferentiated pigmented marigold cells, various callus tissues, and their analysis by transmission electron microscopy in order to observe accumulating pigment and development of subcellular structures. Callus was generated from leaf explants and after several rounds of recurrent selection. Green-, yellow-, and brown-colored callus were obtained that showed distinct carotenoid profiles. For green material, violaxanthin, lutein, zeaxanthin, and β-carotene were produced, while yellow callus generated mainly lutein, as did the brown callus. Chloroplast–chromoplast transition was followed by measuring plastid size and shape in undifferentiated marigold cells by digital image analysis. Cellular alterations were evident in brown callus. Chloroplasts were the main structure in green callus, while yellow callus clearly showed the formation of plastoglobules, structures that are correlated with chloroplast–chromoplast transition. The high number of plastoglobules observed in yellow callus is possibly directly related to pigment synthesis and accumulation.

The phenolic composition (by HPLC-DAD-MS) and color characteristics (by Imaging Tristimulus Colorimetry) of four strawberry cultivars that have shown good climate adaptation to subtropical area (Nikte, Zamorana, Jacona and Pakal) have been assessed. 24 monomeric phenolics were identified, including 15 anthocyanins, 5 phenolic acids, 1 flavanol and 4 flavonols. Nikte and Zamorana showed the highest phenolic potential mainly due to their higher content of anthocyanins, while Pakal was richer in phenolic acids. Regarding color, Nikte and Zamorana were the more similar cultivars having the lowest values of lightness and hue. On the contrary, the color of Pakal was quite different from all the rest, due to the specific distribution between pelargonidin and cyanidin. The inclusion of both phenolic and colorimetric information in the Linear Discriminant Analysis allowed reaching very good discriminations among cultivars.

Article history:Received 8 January 2013Accepted 17 January 2014Available online 9 May 2014Keywords:2,4-DBACell developmentLuteinMarigold Background:Thepropertiesofnaturalpigments,suchasantioxidants,functional,medical,andnutraceutical,havedemonstratedtheadvantagesofthesenaturalcompoundsoversyntheticones.Someproductsareacceptedonlywhen they are pigmented with natural, food-quality colorants: for example poultry products (manly marigoldflower extracts). Carotenoids such as β-carotene, β-criptoxanthin and lutein are very attractive as natural foodcolorants due to their antioxidant and pro-vitamin activities which provide additional value to the targetproducts.Marigold (Tagetes erecta)isan Asteraceous ornamentalplantnativetoMexico,anditisalsoimportantasa carotenoid source for industrial and medicinalpurposesbut nowadays its production isdestined mainlyforornamental purposes.Results: Friable callus of T. erecta yellow flower (YF) and white flower (WF) varieties was induced from leafexplants on Murashige and Skoog (MS) medium supplemented with 9.0 μM 4-dichlorophenoxyacetic acid(2,4-D) and 8.8 μM benzyladenine (BA). Calluses developed from both varieties were different in pigmentation.Extract characterization from callus cultures was carried out by high-performance liquid chromatography(HPLC). This analytical process detected several carotenoids; the main pigments in extracts from YF calluswereluteinandzeaxanthin,whereasintheextractsoftheWFcallusthemainpigmentswerelutein,zeaxanthin,β-cryptoxanthin and β-carotene. Callus cultures of T. erecta accumulated pigments even after several rounds ofsubculture.Conclusions:WFcallusappearedtobeasuitablecandidateasasou rceofdifferentcarotenoids,andtestedvarietiescould represent an alternative for further studies about in vitro pigment production.© 2014 Pontificia Universidad Catolica de Valparaiso. Production and hosting by Elsevier B.V. All rights reserved.

Digital image analysis has an important role in geographical provenance of grains, as it can provide parameters of size, shape and color, which are important quality parameters for the design of engineering processes such as drying and milling of grains. In this study, digital image analysis was used to classify nine rice cultivars based on different morphometric parameters using the three sides of the grain (lateral, ventral and axial), Feret diameter, and 10 different form factors and color parameters (CIE L*, a* and b*).Result of principal component analyisis was an equation with seven variables (area, perimeter, length, width, thickness, sphericity and color), which was useful for distinguishing between nine different cultivars. The morphometric and color parameters for the Mor A-98 and Mor A-92 varieties showed they had 88% similarity. The variability was expressed with a confidence of 95%.Multivariate analysis indicated that the lateral side is the most sensitive for the classification of Mexican rice grains because of its color and morphometric characteristics. These results showed the application of image analysis for the future classifications of grains.

"Los carotenoides son pigmentos fotosensibles frente a un exceso de intensidad luminosa durante el proceso de fotosíntesis. El objetivo de este trabajo fue el estudio del efecto en el desarrollo de la planta y la acumulación de carotenoides por la exposición a dos diferentes intensidades lumínicas (con y sin malla de sombra al 50%). Se evaluó el desarrollo de T. erecta en cuanto a la altura de la planta, número de botones florales y el diámetro de las lígulas. Adicionalmente, en las lígulas se cuantificaron e identificaron los carotenoides por HPLC. Los resultados mostraron diferencias significativas (p¿0.05) en cuanto al desarrollo de las plantas expuestas a mayor radiación UV y temperatura, presentaron reducción del diámetro de las lígulas y disminución en el contenido de Xantófilas totales ( luteína y zeaxantina) con respecto a las cultivadas con malla,. La ultraestructura de los plastidios mostró mayor cantidad de membranas tilacoidales y gránulos de almidón más conspicuos en las células de las plantas de T erecta desarrolladas con malla."

Marigold (Tagetes erecta) is an Asteraceous plant of industrial, ornamental and medicinal importance. Tagetes erecta species, popularly known as marigold, is grown as ornamental plant and is adapted to several agro climates. Inflorescences have been utilized as pigment source for food coloring, mainly of poultry skin and eggs. Lutein is the main pigment in marigold flowers. Some carotenoids are well known as provitamin A compounds. There are many reports on carotenoids and their effect on the prevention of certain ocular diseases, ischemic heart disease, strokes, photoprotection, immune response, aging and cancer. Marigold flowers are certainly a good source of carotenoids; they show very different pigmentation levels. This chapter describes the establishment of techniques for plant regeneration, gene expression, pigment extraction and pigment deposition in specific structures of marigold.

El cempasúchil (Tagetes erecta L.) es una planta originaria de México y sus flores acumulan carotenoides, principalmente luteína. Los pigmentos del cempasúchil se utilizan como aditivos en la elaboración de alimentos para aves, peces y crustáceos, y así mejorar el aspecto de éstos para el consumo humano. Existen estudios en los que se relacionan a los carotenoides con la prevención de algunas enfermedades oculares asociadas con la edad, como las cataratas y la degeneración macular, los cuales han producido resultados interesantes y han promovido el avance en otras áreas del conocimiento relacionadas con estos compuestos, como el mejoramiento de los carotenoides para su utilización en alimentos de consumo común, a través del uso de técnicas de ingeniería genética.

This chapter presents architecture to design emotional agents evolving in an artificial 3D environment. The agent behavior and environment emulator are independent of implementation. To achieve this, a Language of Interface for Animations in 3D called LIA-3D, is presented. The agent and environment simulator uses LIA to establish communication with each other.

Marigold (Tagetes erecta) flowers are a good source of carotenoids and can be used as a model studies on pigmentation during flower development. They show different levels of pigmentation caused by lutein. Here we describe the expression of several genes in the carotenoid biosynthetic pathway: phytoene synthase (Psy), phytoene desaturase (Pds), lycopene beta-cyclase (Lcy-b) and lycopene epsilon-cyclase (Lcy-e). cDNA inserts from isolated clones were 1376-1916bp long. The predicted amino acid sequences showed from 66 to 100% homology with other reported sequences (NCBI gene bank). Northern blot analyses of three varieties of marigold showed that most gene transcripts were expressed during flower development. The ultrastructural changes that occurred during plastid differentiation in flower morphogenesis were analyzed, and pigment accumulation among varieties was evaluated. The pigment deposition in specific structures (lipidic vesicles) during flower development was demonstrated.