IFAPA Centro La Mojonera

La Mojonera, Spain

IFAPA Centro La Mojonera

La Mojonera, Spain
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PubMed | IFAPA Centro La Mojonera, IFAPA Centro Alameda del Obispo, Polytechnic University of Valencia and French National Institute for Agricultural Research
Type: Journal Article | Journal: PloS one | Year: 2014

Although the availability of genetic and genomic resources for Cucurbita pepo has increased significantly, functional genomic resources are still limited for this crop. In this direction, we have developed a high throughput reverse genetic tool: the first TILLING (Targeting Induced Local Lesions IN Genomes) resource for this species. Additionally, we have used this resource to demonstrate that the previous EMS mutant population we developed has the highest mutation density compared with other cucurbits mutant populations. The overall mutation density in this first C. pepo TILLING platform was estimated to be 1/133 Kb by screening five additional genes. In total, 58 mutations confirmed by sequencing were identified in the five targeted genes, thirteen of which were predicted to have an impact on the function of the protein. The genotype/phenotype correlation was studied in a peroxidase gene, revealing that the phenotype of seedling homozygous for one of the isolated mutant alleles was albino. These results indicate that the TILLING approach in this species was successful at providing new mutations and can address the major challenge of linking sequence information to biological function and also the identification of novel variation for crop breeding.

Perez-Hernandez A.,IFAPA Centro La Mojonera | Serrano-Alonso Y.,IFAPA Centro La Mojonera | Aguilar-Perez M.I.,Laboratorio Of Sanidad Vegetal | Gomez-Uroz R.,Laboratorio Of Sanidad Vegetal | Gomez-Vazquez J.,IFAPA Centro La Mojonera
Plant Disease | Year: 2014

Bell pepper (Capsicum annuum L.) is intensively cropped in ~9,920 ha of plastic houses in southern Spain. In summer 2013, pepper seedlings cv. Melchor, Acorde, Galena, Prometeo, and Souleria, with 4 to 8 leaves, grown in a nursery greenhouse near El Ejido, Almería Province, exhibited root rot and stunting. Incidence of symptomatic plants was ~35% among over 10 million. Fusarium sp. was consistently isolated on potato dextrose agar (PDA) from primary and secondary roots of symptomatic plants. Eight single spore isolates (FC1, FC2, FC3, FC4, FC5, FC6, FC11, and FC12) were identified on PDA, carnation leaf-piece agar medium, and Spezieller Nährstoffarmer agar medium as Fusarium oxysporum because of their production of macroconidia (20.5 to 38.2 × 3.9 to 5.8 μm) containing mostly three or rarely four septa, with foot-shaped basal cells. Microconidia (5.9 to 16.5 × 2.6 to 4.7 μm) with 0 to 1 septa formed on false heads on short monophialides and chlamydospores. DNA was extracted from various isolates used in the pathogenicity test, and a portion of the elongation translation factor 1-alpha using primers EF-1 and EF-2 was amplified and sequenced. All the pathogenic isolates were identical, and they differed from the non-pathogenic in 6 to 8 base pairs. The isolates had 99% homology with several isolates of F. oxysporum corresponding to different specialized forms (vasinfectum, lilii, lycopersici, and radicis-lycopersici) at the Fusarium-ID database (1) and GenBank. The sequences of two isolates, FC-6 and FC-12, were deposited in GenBank with accession nos. KF928930 and KF928931, respectively. The pathogenicity of these eight isolates was tested on pepper cv. Melchor in 1-liter containers filled with vermiculite in August and October. Seedlings were inoculated at sowing. PDA plates fully covered with the colony of each isolate were separately blended and homogenized with 300 ml of sterile distilled water. Inocula (5.0 × 105 to 9.4 × 106 conidia/ml) were poured at 50 ml per container. Each experiment had four replicates and 5 to 6 plants per replicate. Treatments with different isolates were arranged in a randomized complete block design. In both experiments, the same number of uninoculated seedlings served as controls. The plants were maintained for 40 days following inoculation in a greenhouse with mean temperatures of 24.0 to 32.4°C and 23.6 to 31.20°C for August and October experiments, respectively. In both experiments, all control plants and those inoculated with FC2, FC3, and FC4 remained asymptomatic. The first wilting occurred 11 days after inoculation. At the end of the August experiment, plants inoculated respectively with FC1, FC5, FC6, FC11, and FC12 showed symptoms in 60, 70, 65, 80, and 90% and 25, 0, 15, 40, and 25% died. At the October experiment, plants showed symptoms in 91.7, 95.8, 100.0, 91.7, and 87.5% and 83.3, 75, 62.5, 83.3, and 79.2% died. Symptomatic plants exhibited damping-off, necrosis of the primary and secondary roots, and sometimes necrotic streaks on the stem. F. oxysporum was consistently recovered from the primary root of symptomatic plants in both experiments and 10 of these isolates were inoculated in a third pathogenicity test, being all pathogens, thus fulfilling Koch's postulates. Although F. oxysporum was reported in peppers (2), to our knowledge, this is the first report of F. oxysporum as the causal agent of damping-off and root rot in pepper seedlings in Almería Province. © 2014 The American Phytopathological Society.

Gomez J.,IFAPA Centro La Mojonera | Perez A.,IFAPA Centro La Mojonera | Serrano Y.,IFAPA Centro La Mojonera | Aguilar M.I.,Laboratorio Of Sanidad Vegetal | Gomez R.,Laboratorio Of Sanidad Vegetal
Plant Disease | Year: 2013

Zucchini (Cucurbita pepo) is intensively cropped in approximately 4,500 ha of plastic houses in southern Spain. In 2008 to 2009, Consul, Cronos, and Tosca zucchini plants showed symptoms of leaf wilting, basal stem necrosis, and plant death. Incidences of dead plants were 20 to 30% and these plants were distributed in clusters. Phytophthora capsici Leonian was isolated from the basal stems of symptomatic plants, using PDA and cornmeal agar amended with a pimaricin, ampicillin, and rifampicin. Five resultant isolates (PCl-211, PCl-221, PCl-611, PCl-612, and PCl-811) on lima beans agar (LBA) produced white mycelia with lemon-shaped and papillate sporangia borne on long pedicels, but no oospores or chlamydospores. These isolates had an identical ribosomal DNA ITS sequence, matching with that of P. capsici in GenBank. The sequences of two representative isolates, PCl-211 and PCl-811, were deposited in GenBank with accession nos. KC662328 and KC688317, respectively. The pathogenicity of these five isolates was tested on zucchini cv. Consul in 1-liter containers filled with vermiculite in May and September of 2009. Plants were inoculated at the 2 to 3 true-leaf stage. Plates with LBA fully covered with colony of each isolate were separately blended and homogenized with 300 ml of sterile distilled water. Inocula were poured around stem at 50 ml per plant. Each experiment had three replicates and four plants per replicate. Treatments with different isolates were arranged in a randomized complete block design. In both experiments, 12 uninoculated plants served as controls. Test plants were maintained for a month following inoculation in a greenhouse with mean temperatures ranging from 21.9 to 27.9°C and from 20.7 to 24.6°C for the May and September experiments, respectively. The first wilting occurred 5 days after inoculation. At the end of the May experiment, all control plants and those inoculated with PCl-221 remained asymptomatic while 83.3% of those inoculated with PCl-211 and 100% of those with the other isolates were dead. Inoculated plants exhibited crown and root rots, excluding the secondary roots. In the September experiment, 83.3% and 33.3% of plants inoculated with PCl-211 and PCl-221, respectively, were symptomatic, while all plants inoculated with the other isolates were dead. The control plants remained healthy. The pathogen was consistently recovered from symptomatic plants in both experiments. Although P. capsici was reported in peppers (Capsicum annuum) in several provinces of Spain (1), to our knowledge, this is the first report of P. capsici as the causal agent of crown rot in zucchini plants in plastic houses in the Almería Province of Spain, one of the world's largest concentrations of greenhouses. © The American Phytopathological Society.

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.

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.

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.

Medranoa E.,IFAPA Centro La Mojonera | Alonso F.J.,IFAPA Centro La Mojonera | Lorenzo P.,IFAPA Centro La Mojonera | Sanchez-Guerrero M.C.,IFAPA Centro La Mojonera
Acta Horticulturae | Year: 2011

High incident radiation levels during the summer cropping cycles in Mediterranean greenhouses generate microclimates of high temperature and evaporative demand so the incident radiation reduction by white-washing is usually applied. The aim of this work is to evaluate the transpiration and nutrient uptake of a sweet pepper crop (Capsicum annuum L.) in two greenhouses equipped with (i) external mobile shading (M) and (ii) external fixed shading (F). Transpiration was measured every five minutes in both greenhouses with two lysimeter balances. Leaf transpiration and intercepted radiation relationship was decreasing during the crop cycle. A multiple regression analysis, based on the simplified Penman-Monteith formula, was established between the measured transpiration and the radiative and advective components. A light extinction coefficient of 0.45 was determined and the leaf area index (LAI) was estimated in function of the days after transplanting. To get a good adjustment between observed and estimated transpiration it was necessary to change the P-M model coefficients through the crop development. Total nutrient uptake (N, K, Ca, Mg and P) was calculated weekly by the measure of supply and leached nutrient solution and its chemical analysis. It was initially similar in both treatments and finally higher in the F than the M treatment. By contrast, the nutrient uptake efficiency, in terms of fruit fresh weight (g g -1), was higher in the M treatment during the early yield period, although both treatments showed the same efficiency at the end of the crop.

Blanco-Diaz M.T.,IFAPA Centro La Mojonera | Font R.,IFAPA Centro La Mojonera | Martinez-Valdivieso D.,IFAPA Centro La Mojonera | Del Rio-Celestino M.,IFAPA Centro La Mojonera
Scientia Horticulturae | Year: 2015

The accessions variability in crop species is crucial for crop improvement and consequently enables the development of more new valuable agricultural products. This study aimed to evaluate the ascorbic acid (AA), chlorophyll a (Chl-A), chlorophyll b (Chl-B), total phenolic compounds (TPC) as well as dry matter (DM) in different tissues (exocarp and mesocarp) of Cucurbita pepo fruits belonging to 27 morphologically diverse germplasm accessions (14 traditional accessions and 13 commercial hybrids from different countries). DM content ranged from 4.2 to 11.76%, while natural pigments were between 5.85 and 2602.2. μg/g DW (corresponding to Chl-B and Chl-A, respectively). Other antioxidant compounds registered higher values, 0.47-1.29 mg/g DW for AA and 1.98-5.49 mg/g DW for TPC. Additionally, pattern-recognition techniques including distribution, principal component analysis (PCA) and dendogram cluster analysis were used to compare accessions. PCA revealed that the first two components represented 76% and 84% of the total variability in exocarp and mesocarp tissues, respectively; while clusters regrouped those accessions with greater similarity. Exocarp samples exhibited a significantly higher content in all compounds studied, thus positively suggesting the beneficial consumption of fruit peel. This work also highlights the potential contribution of 'zucchini' morphotype to dietary intake requirements, being moderate in 'vegetable marrow' and 'pumpkin'. Accessions rich in AA, Chl-A, Chl-B and TPC like those namely Ac-2, Ac-8, Ac-23 and Ac-27 could be exploited as potential donors for novel variety development in future breeding programmes. © 2015 Elsevier B.V.

Martinez-Valdivieso D.,IFAPA Centro La Mojonera | Font R.,IFAPA Centro La Mojonera | Gomez P.,IFAPA Centro La Mojonera | Blanco-Diaz T.,IFAPA Centro La Mojonera | Del Rio-Celestino M.,IFAPA Centro La Mojonera
Journal of the Science of Food and Agriculture | Year: 2014

BACKGROUND: Efforts through conventional breeding to improve the mineral content in horticultural crops have not always been successful mainly due to the fact that standard analytical methods are both costly and time-consuming. We investigated the feasibility of applying near infrared reflectance spectroscopy (NIRS) to the estimation of essential mineral composition in the skin and flesh of summer squash fruits (Cucurbita pepo subsp. pepo) using a 200-sample set from diverse morphotypes. RESULTS: The coefficients of determination in the external validation (R2 VAL) obtained for the skin and flesh of the fruit were: total mineral content, 0.84 and 0.70; P, 0.74 and 0.62; K, 0.83 and 0.67; Ca, 0.57 and 0.60; Mg, 0.78 and 0.45; Fe, 0.78 and 0.65; Cu, 0.67 and 0.66; Mn, 0.67 and 0.64; Zn, 0.80 and 0.79 and Na, 0.33 and 0.33; respectively. CONCLUSIONS: NIRS combined with different spectral transformations by modified partial least-squares (MPLS) regression has shown to be useful in determining the mineral composition of summer squash fruit, being a fast and low-cost analytical technique. Components such as chlorophyll, starch and lipids were used by MPLS for modelling the predicting equations. The promotion of micronutrient-rich summer squash varieties could have a significant long-term beneficial impact on the health of mineral deficient human populations. © 2014 Society of Chemical Industry.

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.

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