Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC

Casas de Fernando Alonso, Spain

Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC

Casas de Fernando Alonso, Spain
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Contreras-Gutierrez P.K.,University of Granada | Hurtado-Fernandez E.,University of Granada | Gomez-Romero M.,Imperial College London | Hormaza J.I.,Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC | And 2 more authors.
Electrophoresis | Year: 2013

A CZE method with two different MS detection conditions (MRM and Full Scan) was developed to determine qualitative and quantitative changes in the metabolic profile of avocado fruits (Persea americana). LODs in MRM approach were found between 20.1 and 203.0 ppb for abscisic acid and perseitol, respectively, whilst in Full Scan, varied within the range 0.22-1.90 ppm for the same metabolites. The RSDs for reproducibility test did not exceed 11.45%. The two MS approaches were used to quantify 10 metabolites (phenolic acids, flavonoids, a carbohydrate, an organic acid, a vitamin and a phytohormone) in 18 samples of avocado at different ripening states, and the achieved results were compared. Perseitol, quinic, chlorogenic, trans-cinnamic, pantothenic and abscisic acids, as well as epicatechin and catechin decreased during the ripening process, whereas ferulic and p-coumaric acids showed the opposite trend. Moreover, some other unknown compounds whose concentration changed largely during ripening were also studied by MS/MS and QTOF MS to get a tentative identification. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Madhou M.,Mauritius Research Council | Normand F.,CIRAD | Bahorun T.,University of Mauritius | Hormaza J.I.,Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC
Tree Genetics and Genomes | Year: 2013

Litchi (Litchi chinensis Sonn.) is a fruit crop with a clear niche for expansion in countries with subtropical climates. One of the main limitations for breeding purposes and optimum germplasm management in this species is the confusion in cultivar denomination among different producing countries and germplasm collections worldwide. Litchi cultivar identification is still mainly based on morphological characters, and homonymies and synonymies are very frequent. To address this gap, a molecular study was conducted to characterize litchi accessions from two of the main litchi-producing regions for the export market, Mauritius and Réunion, and to compare them to those obtained from litchi cultivars from different origins conserved in a germplasm collection in Spain. Eleven simple sequence repeat (SSR) loci were used to characterize molecular polymorphisms among 88 litchi accessions conserved in Mauritius, Réunion, and Spain. A total of 67 amplification fragments were detected with those 11 SSRs, with an average of 6. 1 bands/SSR. Three primer pairs seemed to amplify more than one locus. The mean expected and observed heterozygosities over the eight single locus SSRs averaged 0. 53 and 0. 61, respectively. The total value for the probability of identity was 9. 78 × 10-4. Molecular characterization revealed the existence of 42 different genetic profiles. Several synonymies and homonymies in litchi cultivar nomenclature both within and across geographical regions were found. This comparative study provides the basis for the standardization of litchi cultivar nomenclature over the studied regions and in other litchi-producing countries. © 2012 Springer-Verlag.

Merchante C.,Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC | Vallarino J.G.,Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC | Osorio S.,Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC | Araguez I.,Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC | And 10 more authors.
Journal of Experimental Botany | Year: 2013

The fruit of the strawberry Fragaria×ananassa has traditionally been classified as non-climacteric because its ripening process is not governed by ethylene. However, previous studies have reported the timely endogenous production of minor amounts of ethylene by the fruit as well as the differential expression of genes of the ethylene synthesis, reception, and signalling pathways during fruit development. Mining of the Fragaria vesca genome allowed for the identification of the two main ethylene biosynthetic genes, 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase. Their expression pattern during fruit ripening was found to be stage and organ (achene or receptacle) specific. Strawberry plants with altered sensitivity to ethylene could be employed to unravel the role of ethylene in the ripening process of the strawberry fruit. To this end, independent lines of transgenic strawberry plants were generated that overexpress the Arabidopsis etr1-1 mutant ethylene receptor, which is a dominant negative allele, causing diminished sensitivity to ethylene. Genes involved in ethylene perception as well as in its related downstream processes, such as flavonoid biosynthesis, pectin metabolism, and volatile biosynthesis, were differently expressed in two transgenic tissues, the achene and the receptacle. The different transcriptional responsiveness of the achene and the receptacle to ethylene was also revealed by the metabolic profiling of the primary metabolites in these two organs. The free amino acid content was higher in the transgenic lines compared with the control in the mature achene, while glucose and fructose, and citric and malic acids were at lower levels. In the receptacle, the most conspicuous change in the transgenic lines was the depletion of the tricarboxylic acid cycle intermediates at the white stage of development, most probably as a consequence of diminished respiration. The results are discussed in the context of the importance of ethylene during strawberry fruit ripening. © The Author 2013.

Amaya I.,Instituto Andaluz Of Investigacion Y Formacion Agraria Y Pesquera Ifapa | Osorio S.,Max Planck Institute of Molecular Plant Physiology | Osorio S.,Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC | Martinez-Ferri E.,Instituto Andaluz Of Investigacion Y Formacion Agraria Y Pesquera Ifapa | And 6 more authors.
Biotechnology Journal | Year: 2015

Increasing L-ascorbic acid (AsA, vitamin C) content in fruits is a common goal in current breeding programs due to its beneficial effect on human health. Attempts to increase AsA content by genetic engineering have resulted in variable success likely due to AsA's complex regulation. Here, we report the effect of ectopically expressing in tomato the D-galacturonate reductase (FaGalUR) gene from strawberry, involved in AsA biosynthesis, either under the control of the constitutive 35S or the tomato fruit-specific polygalucturonase (PG) promoters. Although transgenic lines showed a moderate increase on AsA content, complex changes in metabolites were found in transgenic fruits. Metabolomic analyses of ripe fruits identified a decrease in citrate, glutamate, asparagine, glucose, and fructose, accompanied by an increase of sucrose, galactinol, and chlorogenic acid. Significant metabolic changes also occurred in leaves of 35S-FaGalUR lines, which showed higher non-photochemical fluorescence quenching (NPQ), indicative of a higher constitutive photo-protective capacity. Overall, overexpression of FaGalUR increased total antioxidant capacity in fruits and the results suggest a tight control of AsA content, probably linked to a complex regulation of cellular redox state and metabolic adjustment. Engineering increased vitamin C content in fruits, particularly in tomato due to its high consumption, has been a goal of many laboratories. Here, the authors report the ectopic expression in tomato fruits of a strawberry D-galacturonic acid reductase (GalUR) involves in Vitamin C biosynthesis causes moderate increases of this compound. However, the expression of GalUR alters the content of several metabolites in the transgenic fruits and causes a general increase of antioxidant capacity. This study indicates that the homeostasis of Vitamin C is under a tight control likely because of a complex regulation of the cellular redox state. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

PubMed | University of British Columbia, Center for Research in Agricultural Genomics IRTA UAB UB and Institute Hortofruticultura Subtropical y Mediterranea IHSM UMA CSIC
Type: | Journal: Methods in molecular biology (Clifton, N.J.) | Year: 2015

C2 domains (C2s) are regulatory protein modules identified in eukaryotic proteins targeted to cell membranes. C2s were initially characterized as independently folded Ca(2+)-dependent phospholipids binding domains; however, later studies have shown that C2s have evolutionarily diverged into Ca(2+)-dependent and Ca(2+)-independent forms. These forms interact and regulate their affinity to diverse lipid species using different binding mechanisms. In this protocol we describe a biochemical approach to produce, purify, and solubilize functional C2 domains bound to GST for the identification of their putative Ca(2+)-dependent and Ca(2+)-independent lipid-binding partners.

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