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Samper del Salz, Spain

Roman B.,IFAPA Centro Alameda del Obispo | Gonzalez-Verdejo C.I.,IFAPA Centro Alameda del Obispo | Pena F.,IFAPA Centro Alameda del Obispo | Nadal S.,IFAPA Centro Alameda del Obispo | Gomez P.,IFAPA Centro la Mojonera
Phytochemical Analysis | Year: 2012

Introduction Quality and integrity of RNA are critical for transcription studies in plant molecular biology. In squash fruit and other high water content crops, the grinding of tissue with mortar and pestle in liquid nitrogen fails to produce a homogeneous and fine powered sample desirable to ensure a good penetration of the extraction reagent. Objective To develop an improved pulverisation method to facilitate the homogenisation process of squash fruit tissue prior to RNA extraction without reducing quality and yield of the extracted RNA. Methodology Three methods of pulverisation, each followed by the same extraction protocol, were compared. The first approach consisted of the lyophilisation of the sample in order to remove the excess of water before grinding, the second one used a cryogenic mill and the control one a mortar grinding of frozen tissue. The quality of the isolated RNA was tested by carrying out a quantitative real time downstream amplification. Results In the three situations considered, mean values for A260/A280 indicated minimal interference by proteins and RNA quality indicator (RQI) values were considered appropriate for quantitative real-time polymerase chain reaction (qRT-PCR) amplification. Successful qRT-PCR amplifications were obtained with cDNA isolated with the three protocols. Conclusion Both apparatus can improve and facilitate the grinding step in the RNA extraction process in zucchini, resulting in isolated RNA of high quality and integrity as revealed by qRT-PCR downstream application. This is apparently the first time that a cryogenic mill has been used to prepare fruit samples for RNA extraction, thereby improving the sampling strategy because the fine powder obtained represents a homogeneous mix of the organ tissue. Copyright © 2012 John Wiley & Sons, Ltd. Source

Gonzalez-Verdejo C.I.,IFAPA Centro Alameda del Obispo | Obrero A.,IFAPA Centro Alameda del Obispo | Roman B.,IFAPA Centro Alameda del Obispo | Gomez P.,IFAPA Centro la Mojonera
Plant Foods for Human Nutrition | Year: 2015

Carotenoids are important dietary components that can be found in vegetable crops. The accumulation of these compounds in fruit and vegetables is altered by the activity of carotenoid cleavage dioxygenases (CCDs) enzymes that produce their degradation. The aim of this work was to study the possible implication of CCD genes in preventing carotenoid storage in the horticultural crop summer squash (Cucurbita pepo L.). The relationship between the presence of these compounds and gene expression for CCDs was studied in three varieties showing different peel and flesh colour. Expression analysis for the CCD genes CpNCED1, CpNCED2, CpNCED3, CpNCED9, CpCCD1, CpCCD4a, CpCCD4b and CpCCD8 was carried out on different organs and at several fruit developmental stages. The results showed that the CpCCD4a and CpCCD4b genes were highly expressed in the variety with lowest carotenoid content suggesting a putative role in carotenoid accumulation pattern in summer squash fruit. © 2015, Springer Science+Business Media New York. Source

Villatoro-Pulido M.,IFAPA Centro Alameda del Obispo | Priego-Capote F.,University of Cordoba, Spain | Alvarez-Sanchez B.,University of Cordoba, Spain | Saha S.,UK Institute of Food Research | And 5 more authors.
Journal of the Science of Food and Agriculture | Year: 2013

BACKGROUND: Eruca sativa (rocket) contains a wide range of compounds with nutraceutical and organoleptical properties. This research aimed to characterise the nutraceutical interest of four rocket accessions by analysis of glucosinolates, isothiocyanates, phenolics, carotenoids and carbohydrates. Different methods based on chromatographic separation with ultraviolet absorbance or mass spectrometry detection were used. RESULTS: The total content of glucosinolates ranged from 14.02 to 28.24 μmol g-1 of dry weight. Glucoraphanin represented up to 52% of the total glucosinolates in leaves of one accession. Accessions showed differences in the hydrolysis of glucoraphanin to the isothiocyanate sulforaphane. No correlation between these compounds was observed, which insisted differences in the myrosinase activity within accessions. Rocket leaves had variable phenolic profiles represented by quercetin-3-glucoside, rutin, myricetin, quercetin and ferulic and p-coumaric acids. A high variability was observed for the total carotenoids ranged from 16.2 to 275 μg g-1 with lutein as the main carotenoid. Glucose was the predominant sugar, representing >70% of the total soluble carbohydrates. CONCLUSIONS: Some accessions could be candidates for future breeding programmes because of their pattern of beneficial compounds for human health. However, further research is essential to evaluate the biological activity of these accessions before designing functional food. © 2013 Society of Chemical Industry. Source

Villatoro-Pulido M.,IFAPA Centro Alameda del Obispo | Moreno Rojas R.,University of Cordoba, Spain | Munoz-Serrano A.,University of Cordoba, Spain | Cardenosa V.,IFAPA Centro Alameda del Obispo | And 3 more authors.
Journal of the Science of Food and Agriculture | Year: 2012

Background: Minerals are essential for human nutrition and must be obtained from our diet. Crucifer vegetables are a good source of these nutrients. Our objectives were to determine the genetic variability for mineral content and to evaluate the use of near-infrared reflectance spectroscopy (NIRS) for prediction of ashes and minerals among and within the rocket species Eruca vesicaria subsp. sativa and vesicaria. The minerals studied were iron (Fe), copper (Cu), sodium (Na), potassium (K), calcium (Ca), magnesium (Mg), manganese (Mn) and zinc (Zn). Results: The maximum mean values obtained for all the accessions (mean ± SE) were 235.5 ± 1.5 mg ashes kg -1, 273.3 ± 4.2 mg Fe kg -1, 18.1 ± 0.4 mg Cu kg -1, 2.8 ± 0.1 g Na kg -1, 71.6 ± 1.0 g K kg -1, 64.6 ± 1.2 g Ca kg -1, 6.8 ± 0.1 g mg kg -1, 101.6 ± 1.2 mg Mn kg -1, and 67.1 ± 0.4 mg Zn kg -1 of dry weight. Conclusion: The statistical analysis showed significant differences for all the minerals, except Ca, for each accession studied individually and for accessions grouped within countries. The results indicate that NIRS can be used as a rapid screening method for determining total mineral, Fe, Na, K, and Zn in rocket. © 2011 Society of Chemical Industry. Source

Vicente-Dolera N.,IFAPA Centro la Mojonera | Pinillos V.,University of Almeria | Moya M.,IFAPA Centro la Mojonera | Del Rio-Celestino M.,IFAPA Centro la Mojonera | And 3 more authors.
Scientia Horticulturae | Year: 2014

A new genetic resource for Cucurbita pepo has been developed with chemically induced mutagenesis. The seeds of the zucchini cultivar MU-CU16 were treated with 40. mM-80. mM ethyl methanesulfonate (EMS), reaching high germination rates between 70 and 85%. However, most plants of those M1 populations did not produce offspring, and the fertility rates were lower in plants treated with higher concentrations of EMS. Once we established that visual flower abnormality rates were not sufficient to explain low fruit yield, pollen viability was analysed with fluorochromatic reaction. Compared with untreated plants, treatment with EMS produced a substantial decrease in pollen viability, and only the group of plants with pollen viability rates higher than 45% yielded nearly 70% of fruits with seeds. Therefore, the main issues to be addressed for developing mutant lines in this species are to increase the number of mutations in the genome and to increase the number of mutant lines with sufficient fertility. In this case, the early plantlet selection for high pollen viability carried out as part of this work represents a useful tool for use in future breeding programs by mutagenesis, allowing an increase of up to 40% in the production of mutant lines for a dosage of 65. mM EMS. © 2014 Elsevier B.V. Source

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