Rafael Zárate

The lipid and fatty acid profiles of 14 marine macroalgal species from the Madeira Archipelago, including two green (Ulvales and Dasycladales), three red (Corallinales, Bonnemaisoniales, and Ceramiales) and nine brown (Fucales, Dictyotales, and Sphacelariales) species were characterised in order to determine their potential use for animal and human...

Seaweed beach-casts are a seasonal phenomenon that regularly deposits tons of algae biomass on beaches, which are usually disposed of in landfills. The present work aimed to contribute to the valorization of this biomass by studying bioactivities that reveal its potential in the pharmaceutical and/or cosmeceutical industries. Methanol and ethanol e...

Continuous use of anticoccidial treatments against Eimeria infections has resulted in the development of drug resistance. This study aimed to evaluate the anticoccidial efficacy of a methanolic extract derived from the endemic Canary rue (Ruta pinnata) plant of the Canary Islands, Spain, against Eimeria ninakohlyakimovae using in vitro assays. Fres...

In the last decades, the development of new technologies applied to lipidomics has revitalized the analysis of lipid profile alterations and the understanding of the underlying molecular mechanisms of lipid metabolism, together with their involvement in the occurrence of human disease. Of particular interest is the study of omega-3 and omega-6 long...

Omega-3 long change polyunsaturated fatty acids in higher plants are limited with just a few plant genus showing the accumulation of stearidonic acid (SDA) being also the longest and more unsaturated omega-3 fatty acid present. Echium acanthocarpum has been proven to be an efficient and attractive producer of SDA. Improved production of this fatty...

Marine algae are non-vascular photoautotrophic organisms containing chlorophyll. They are the primary producers in marine food webs, and responsible for the maintenance of life in many ecosystems. Algae are able to produce a wide range of active metabolites, many of which are of great importance to human health, such as the highly valued omega-3 ei...

Hyoscyamine and scopolamine, anti-cholinergic agents widely used in medicine, are typically obtained from plants grown under natural conditions. Since field cultivation entails certain difficulties (changeable weather, pests, etc.), attempts have been made to develop a plant in vitro culture system as an alternative source for the production of the...

The additional information accessible on-line provides data presented in tables and figures of the different parameters recorded in this investigation at the other sampling times of the different culture growth curves, as well as additional data of the statistical analyses conducted and not present in the printed version of the publication.

Fatty acids are of great nutritional, therapeutic, and physiological importance, especially the polyunsaturated n-3 fatty acids, possessing larger carbon chains and abundant double bonds or their immediate precursors. A few higher plant species are able to accumulate these compounds, like those belonging to the Echium genus. Here, the novel E. acan...

Plant natural products are useful compounds, such as pigments, fragrances, pharmaceuticals used for the treatment of several human diseases. Development of plant manipulation techniques, such as particle bombardment, Agrobacterium-mediated transformation, vacuum infiltration, agrodrench, viral vector, protoplasts fusion and ultrasound, as well as r...

The therapeutic and health promoting role of highly unsaturated fatty acids (HUFAs) from fish, i.e. eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) are well known. These same benefits may however be shared by some of their precursors, the polyunsaturated fatty acids (PUFAs), such as stearidonic acid (SDA, 18:4 n-3). In...

Tables (S1-S6) and Images S1a-f. Tables S1-S5 show the different sampling points not included in the main text. In each table the total lipid content and general fatty acid profiles (%) of the two cell lines of Echium acanthocarpum hairy roots at different sampling points are presented. n-6 and n-3 Δ6-Desaturation Indexes were calculated as 18:3n-6...

Figures S1-S3. Figures S1-S3 show plots of PC1 and PC2 factor scores categorized by type of culture and by sampling points.

Rafael Zárate was born on 7 April 1963 in La Orotava, Tenerife, Spain and graduated from The University of La Laguna, Tenerife with a B.Sc. (Biology) degree in 1989. Later, he moved to The University of Edinburgh where he conducted his Ph.D. studies (1994) on plant biotechnology and natural product synthesis in in vitro culture systems under the su...

A new cDNA encoding hyoscyamine 6beta-hydroxylase (H6H, EC 1.14.11.11), a bifunctional enzyme catalyzing the last two steps in the biosynthesis of scopolamine, was isolated from Atropa baetica roots (GenBank accession no. EF442802). The full cDNA sequence showed an ORF of 1035bp, coding for a protein with 344 amino acid residues. Sequence analyses...

The existence of drug-resistant human immunodeficiency virus (HIV) viruses in patients receiving antiretroviral treatment urgently requires the characterization and development of new antiretroviral drugs designed to inhibit resistant viruses and to complement the existing antiretroviral strategies against AIDS. We assayed several natural or semi-s...

This review deals with the application of recombinant DNA technology to investigate and tailor plant secondary metabolism as a tool for the control of the biosynthesis and accumulation of plant natural products. Firstly, it presents the different plant transformation technologies, followed by the description of the various strategies implemented fo...

Elicitation of transgenic Atropa baetica overexpressing the h6h gene with salicylic acid (SA), acetylsalicylic acid (ASA),or methyl jasmonate (MeJ) was conducted to boost tropane alkaloid yields. Scopolamine (1) amounts increased after treatment with ASA and MeJ, but not with SA. The highest enhancement of 1 was achieved with MeJ followed by ASA di...

The usefulness of the solvent mixture dichloromethane/methanol for lipid extraction and the determination of lipid classes and fatty acids in samples of different natures was conducted. Two different extraction methods were compared, one containing chloroform/methanol, another containing dichloromethane/methanol. Total lipid extraction showed some...

Seed germination and micropropagation protocols of the medicinal species Maytenus canariensis (Loes.) G. Kunkel & Sunding were optimized. In vitro seed germination occurred (86 to 94.7 %) only after treatment of the seeds with H2SO4, followed by surface sterilization and culture on solid nutrient medium without any growth regulators. Micropropagati...

Several bis-pyrano-l,4-benzoquinones have been synthesized by a double domino Knoevenagel hetero Diels–Alder reaction. The synthetic approach is highly efficient allowing the construction of complex polycyclic scaffolds with six new σ-bonds. These reactions performed more efficiently and more rapidly using microwave irradiation. The resulting bis-p...

Four new terpenoids (1-4) were isolated from the root bark of Maytenus ilicifolia. Their structures were determined by spectroscopic studies, and complete 1H and 13C NMR assignments were achieved by 2D NMR spectroscopy. We also report an efficient method for the separation of quinonemethide triterpenes based on centrifugal partition chromatography.

This review describes the different plant transformation techniques, including guided infection with Agrobacterium tumefaciens and A. rhizogenes, particle bombardment and protoplast fusion, that have been attempted to create transgenic Catharanthus roseus (L.) G. Don cell cultures, hairy roots and whole plants. The review also focuses on the differ...

Atropa baetica hairy roots, over-expressing cDNA from Hyoscyamus niger encoding the gene for hyoscyamine 6beta-hydroxylase (H6H), were produced by Agrobacterium rhizogenes infection. The transgenic roots over-expressing h6h had an altered alkaloid profile in which hyoscyamine was entirely converted into scopolamine. In the best h6h clone, scopolami...

In order to investigate the production of tropane alkaloids by hairy roots of Atropa baetica, transgenic for the gene h6h encoding the enzyme hyoscyamine 6beta-hydroxylase, solvent extraction with chloroform and with dichloromethane of the metabolites present in the liquid medium and in the root tissue was compared. The extraction of scopolamine fr...

Agrobacterium rhizogenes transformed hairy roots, callus and suspension cultures of Rudbeckia hirta produced pulchelin E, a compound of the sesquiterpene lactone group, which is not present in the intact plant. During the research the level of the investigated lactone was optimised yielding 8.3 mg g−1 in the callus, 9.1 mg g−1 in the suspension cul...

Different transgenic cell lines of Nicotiana tabacum expressing strictosidine β-D-glucosidase cDNA from Catharanthus roseus were established following Agrobacterium tumefaciens infection. The fresh weight and protein levels did not appear to be affected by the gene insertion performed, and in several instances these seemed to be boosted. Regarding...

The terpenoid indole alkaloid content of Catharanthuspusillus was investigated during development from young to old plants. Different plant organs were assessed showing that the new leaves were the main repository site with vindoline (≈4.8 mg/g DW) and catharanthine (≈2.2 mg/g DW) being the major metabolites with the highest yields at the second an...

This review which is concerned with the application of recombinant DNA technology to studies on plant secondary metabolism, presents the more common plant transformation strategies and shows how these genetic approaches are being used in attempts to manipulate and increase the yield of secondary metabolites, both in cultures and in transformed plan...

Atropa baetica hairy root cultures were induced after infecting stem segments with Agrobacterium rhizogenes strain ATCC 15834. Accumulation of the tropane alkaloids atropine and scopolamine by hairy roots cultured in half- and full-strength Murashige and Skoog (MS) medium was high, although this was not growth associated. These alkaloids were also...

In vitro propagation of Catharanthus roseus was achieved using nodal explants. Bud induction was best on medium containing 1.0mg benzyl aminopurine l–1. Hardening of rooted shoots to soil was very successful with 98% survival. Genetically transformed C.roseus plantlets were obtained after bombardment of nodal explants, which were then micropropagat...

Germination of intact seeds, seeds without testa and isolated embryos of two subspecies of Juniperus oxycedrus (oxycedrus and macrocarpa) was compared both in vitro and under greenhouse conditions (for intact and seeds without testa). Intact seeds did not germinate in the greenhouse or in vitro. Seeds without testa did not germinate under greenhous...

chinospartum algibicum (Leguminosae) regeneración de plantas mediante organogénesis adventicia. La germinación in vitro de semillas de E. algibicum, especie endémica de la Sierra de Grazalema, S de España, así como la inducción de yemas múltiples y el desarrollo posterior de brotes, se consiguió en el medio de cultivo de Murashige y Skoog a mitad d...

Fifteen different tropane alkaloids and derivatives were identified by GC-MS in various plant organs of Atropa baetica. The main root and leaves displayed the largest number of tropane alkaloids, most of which were also present in lateral roots. In contrast, only five of these alkaloids appeared in stem tissue. Quantitative analysis by HPLC showed...

In vitro propagation of Atropa baetica was established employing axillary buds. Single buds were cultured through a multiple shoot induction phase, rooting phase, and then followed by acclimatization in soil. For multiple shoot induction, Murashige and Skoog (MS) medium with 3% sucrose, supplemented with either 0.75 or 1.25 mg l-1 of BAP provided t...

The accumulation of [6]gingerol and [6]shogaol (phenolic pungent principles of ginger) was much higher in culture systems of Zingiber officinale where morphological differentiation was apparent. Cultures grown on a callus-inducing medium also accumulated these metabolites but to a lesser extent. There is a positive relationship between product accu...

summaryExamination of cryo-sections from mature rhizomes, immature rhizomes and adventitious roots of ginger, Zingiber officinale Roscoe, showed the presence of yellow-pigmented cells. There is a correlation between the number of pigrnenteci cells and the amount of [6]-gingerol, the main pungent principle in ginger, in these organs. It is also show...

Two rapid techniques of column chromatography, flash chromatography and vacuum chromatography, together with solvent systems differing in polarity, have been used to separate the pungent principles (gingerols and shogaols) of an extract of ginger powder. Gingerols consist of a homologous series of aldols each containing a phenolic group which toget...