Callus culture development from two varieties of Tagetes erecta and carotenoids production

AbstractBackgroundThe properties of natural pigments, such as antioxidants, functional, medical, and nutraceutical, have demonstrated the advantages of these natural compounds over synthetic ones. Some products are accepted only when they are pigmented with natural, food-quality colorants: for example poultry products (manly marigold flower extracts). Carotenoids such as β-carotene, β-criptoxanthin and lutein are very attractive as natural food colorants due to their antioxidant and pro-vitamin activities which provide additional value to the target products. Marigold (Tagetes erecta) is an Asteraceous ornamental plant native to Mexico, and it is also important as a carotenoid source for industrial and medicinal purposes but nowadays its production is destined mainly for ornamental purposes.ResultsFriable callus of T. erecta yellow flower (YF) and white flower (WF) varieties was induced from leaf explants on Murashige and Skoog (MS) medium supplemented with 9.0μM 4-dichlorophenoxyacetic acid (2,4-D) and 8.8μM 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 callus were lutein and zeaxanthin, whereas in the extracts of the WF callus the main pigments were lutein, zeaxanthin, β-cryptoxanthin and β-carotene. Callus cultures of T. erecta accumulated pigments even after several rounds of subculture.ConclusionsWF callus appeared to be a suitable candidate as a source of different carotenoids, and tested varieties could represent an alternative for further studies about in vitro pigment production

Assessment of the differences in the phenolic composition and color characteristics of new strawberry (Fragaria x ananassa Duch.) cultivars by HPLC–MS and Imaging Tristimulus Colorimetry

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 cultivar

Morphostructural Characterization of Rice Grain (Oryza sativa L.) Variety Morelos A-98 during Filling Stages

The morphostructure of grain rice Morelos A-98 was characterized in five stages of physiological maturation, in order to generate morphometric information during the filling process. Micrographic images from optical and scanning electron microscopy coupled to a digital capture system were used. Images were digitally processed to measure different descriptors such as shape, fractal dimension, and surface texture. Results showed that, two weeks after anthesis, an accelerated grain filling was observed, particularly on those grains positioned in the distal panicle zone, compared to those located in the base of this one. As deposition of assimilates in the grain increased, the area and perimeter of the transversal cut of the grains also increased (P ≤ 0.05); meanwhile, the rounded shape factor tended to increase as well (P ≤ 0.05), while the elliptic shape factor decreased. As the dehydrated endosperm passed from “milky” to “doughy” stages, values of fractal dimension area and endosperm perimeter as well as surface texture values showed that grain borders tended to become smoother and that there was a greater structured endosperm area (P ≤ 0.05)

PLASTID ANALYSIS OF PIGMENTED UNDIFFERENTIATED CELLS OF MARIGOLD TAGETES ERECTA L. BY TRANSMISSION ELECTRON MICROSCOPY.

http://link.springer.com/article/10.1007%2Fs11627-011-9401-

Metabolite profiling of rosemary cell lines with antiproliferative potential against human HT-29 colon cancer cells

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).Peer reviewe

Chemical analysis of callus extracts from toxic and non-toxic varieties of Jatropha curcas L.

Jatropha curcas L. belongs to Euphorbiaceae family, and it synthesizes flavonoid and diterpene compounds that have showed antioxidant, anti-inflammatory, anticancer, antiviral, antimicrobial, antifungal and insecticide activity. Seeds of this plant accumulate phorbol esters, which are tigliane type diterpenes, reported as toxic and, depending on its concentration, toxic and non-toxic varieties has been identified. The aim of this work was to characterize the chemical profile of the extracts from seeds, leaves and callus of both varieties (toxic and non-toxic) of Jatropha curcas, to verify the presence of important compounds in dedifferentiated cells and consider the possibility of using these cultures for the massive production of metabolites. Callus induction was obtained using NAA (1.5 mg L−1) and BAP (1.5 mg L−1) after 21 d for both varieties. Thin layer chromatography analysis showed differences in compounds accumulation in callus from non-toxic variety throughout the time of culture, diterpenes showed an increase along the time, in contrast with flavonoids which decreased. Based on the results obtained through microQTOF-QII spectrometer it is suggested a higher accumulation of phorbol esters, derived from 12-deoxy-16-hydroxy-phorbol (m/z 365 [M+H]+), in callus of 38 d than those of 14 d culture, from both varieties. Unlike flavonoids accumulation, the MS chromatograms analysis allowed to suggest lower accumulation of flavonoids as the culture time progresses, in callus from both varieties. The presence of six glycosylated flavonoids is also suggested in leaf and callus extracts derived from both varieties (toxic and non-toxic), including: apigenin 6-C-α-L-arabinopyranosyl-8-C-β-D-xylopyranoside (m/z 535 [M+H]+), apigenin 4′-O-rhamnoside (m/z 417 [M+H]+), vitexin (m/z 433 [M+H]+), vitexin 4′-O-glucoside-2″-O-rhamnoside (m/z 741 [M+H]+), vicenin-2 (m/z 595 [M+H]+), and vicenin-2,6″-O-glucoside (m/z 757 [M+H]+)

Characterization of PLE extracts from Kalanchoe daigremontiana by LC-UV/MS and GC-Q-TOF analysis. Evaluation of bioactive properties

Resumen del trabajo presentado a las 15as Jornadas de Análisis Instrumental, celebradas en Barcelona (España) del 3 al 5 de octubre de 2017.This work was supported by the projects AGL2014-53609-P (Ministerio de Economía y Competitividad, Spain) and S2013/ABI-2728 (Comunidad de Madrid). G.A.-R. would like to acknowledge the Ministry of Economy and Competitiveness for a Juan de La Cierva-Formación postdoctoral grant.Peer reviewe

Comparative assessment of PLE and SFE for the selective extraction of compounds of interest from Ruta graveolens

Trabajo presentado al 17th European Meeting on Supercritical Fluids (EMSF) y al 7th European Meeting on High Pressure Technology, celebrados en Ciudad Real (España) del 8 al 11 de abril de 2019.[Introduction]: Ruta graveolens, commonly known as rue, is an herbaceous perennial plant with bluish-green leaves that emits a powerful odor and has a bitter taste. This plant has shown different pharmacological activities and is employed in folk medicine for treatment of rheumatism, dermatitis, pain and many inflammatory diseases. The main components reported in Ruta graveolens are quinoline, furoquinoline, acridone alkaloids, flavonoids, and coumarins, mainly furanocoumarins. Although traditional extraction of these compounds is mainly done by maceration, nowadays the use of green extraction techniques provides an efficient, fast, selective and environmentally friendly alternative. In this regard, technologies based on compressed fluids play an important role, being supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE) the most widely employed. The aim of this work was to evaluate the ability of PLE and SFE to selectively extract families of compounds of interest from the aerial part and root of Ruta graveolens. PLE was carried out at 100 bar for 20 min. Three different temperatures were studied (40 °, 105 ° and 170 ° C) and two green solvents (d-limonene and ethyl acetate) and its mixture (50:50 v/v) were used. In SFE, extraction with supercritical CO2 (scCO2) was carried out, increasing the pressure every 60 min (up to 100, 200 and 350 bars) at a temperature of 40 °C. Two conditions were tested: dry sample or moistened sample before reaching the pressure of 350 bar. SFE extracts were collected in ethanol and ethyl acetate. Rue extracts obtained by SFE and PLE were chemically characterized by GC–q-TOF-MS.[Results and discussion]: The extraction yield and the relative abundance of the different groups of metabolites (alkaloids, furanocoumarins, terpenes, amines, fatty acids and other compounds) identified by GC-q-TOF-MS were evaluated. Alkaloids, terpenes and furanocoumarins were identified in all extracts, and differences in abundance and selectivity according to the compressed fluid used and the conditions tested were found. Furthermore, arborinine is an alkaloid that was only identified in the aerial part of the plant, while furanocoumarins -isopimpinelina and xantotoxina- along with some other coumarins and amides were only identified in the root. Concerning PLE, the extraction of alkaloids from the root was favored with any of the solvents used. However, in the aerial part, alkaloids’ extraction was promoted by ethyl acetate while the extraction of terpenes improved with d-limonene and mixture d-limonene – ethyl acetate (50:50 v/v). Regarding SFE, when the extracts were collected in ethanol, terpenes abundance increase, while collecting them in ethyl acetate, increased the alkaloids content while, at the same time, terpenes decreased, with a difference of 20%. Moistening the sample at 350 bar pressure and collecting the extract in ethyl acetate facilitates the extraction and recovery of alkaloids.[Conclusions]: In this study, two different compressed fluid technologies, PLE and SFE, were described for the first time to extract phytochemicals from the aerial part and the roots of Ruta graveolens using GRAS – generally recognized as safe – solvents. Specific compounds were identified for each part of the plant and selectivity of different families could be reached in different conditions.Authors thank CONACYT, COFAA, SIP-IPN and Laboratory of Foodomics. M.B. and G. A-R acknowledge MINECO for the “Juan de La Cierva-Formación” postdoctoral grants FJCI-2016-30902 and FJCI-2015-25504, respectively.Peer reviewe

Phytochemical profiling of hydroalcoholic extract from aerial parts and roots of Ruta graveolens from Morelos (Mexico)

Resumen del trabajo presentado al XVIII Scientific Meeting of the Spanish Society of Chromatography and Related Techniques (SECyTA), celebrado en Granada del 2 al 4 de octubre de 2018.Ruta graveolens, commonly known as rue, is an herbaceous perennial plant with bluish-green leaves that emits a powerful odor and have a bitter taste. This species is original from the Mediterranean region and has been cultivated in several countries all around the world. This plant has shown different pharmacological activities and is employed in folk medicine for treatment of rheumatism, dermatitis, pain and many inflammatory diseases. The main components reported in Ruta graveolens are quinoline, furoquinoline, acridone alkaloids, flavonoids, and coumarins, mainly furanocoumarins. Furanocoumarins have been detected in different parts of the plant (i.e. leaves, stems and roots), whereas acridone alkaloids have been identified in roots and furoquinoline alkaloids were mainly reported in leaves. Since several works in literature report the tissue-specific accumulation of phytochemicals in Ruta graveolens, in this work, a comparative phytochemical profiling of metabolites from the aerial part and the root was proposed to verify the distribution of metabolites in the Mexican variety. For the isolation of target metabolites, aerial part and roots were separated, washed, dehydrated at room temperature and submitted to an ultrasound-assisted extraction procedure using ethanol and water mixtures (85:15 v/v) as solvents. The obtained extracts were analyzed and characterized by GC-MS. A similar number of metabolites were identified in both parts of the plant. Acridone and furoquinoline alkaloids, furanocoumarins, terpenes and ketones, were found in both extracts, although at different concentration levels. Roots extracts showed the highest number of alkaloids and furanocoumarins, and the metabolites most abundant were dictammnine, rutacridona and isomaturnina; whereas terpenes and ketones were mainly abundant in aerial part extracts, where the principal metabolites were kokusaginine, arborinine, bergapten, isomartunine and circimaritin. In this work, acridone alkaloid arborinine was only identified in the aerial part, whereas furanocoumarins as isopimpinellin and kokusaginin were exclusively detected in the roots, which reveal that this variety exhibit a different distribution of metabolites from the reported in literature. Arborinine has been reported anticancer activity.Peer reviewe