Metapontum Agrobios


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Minoia S.,Metapontum Agrobios | Minoia S.,ENEA | Petrozza A.,Metapontum Agrobios | D'Onofrio O.,Metapontum Agrobios | And 6 more authors.
BMC Research Notes | Year: 2010

Background. In the last decade, the availability of gene sequences of many plant species, including tomato, has encouraged the development of strategies that do not rely on genetic transformation techniques (GMOs) for imparting desired traits in crops. One of these new emerging technology is TILLING (Targeting Induced Local Lesions In Genomes), a reverse genetics tool, which is proving to be very valuable in creating new traits in different crop species. Results. To apply TILLING to tomato, a new mutant collection was generated in the genetic background of the processing tomato cultivar Red Setter by treating seeds with two different ethylemethane sulfonate doses (0.7% and 1%). An associated phenotype database, LycoTILL, was developed and a TILLING platform was also established. The interactive and evolving database is available online to the community for phenotypic alteration inquiries. To validate the Red Setter TILLING platform, induced point mutations were searched in 7 tomato genes with the mismatch-specific ENDO1 nuclease. In total 9.5 kb of tomato genome were screened and 66 nucleotide substitutions were identified. The overall mutation density was estimated and it resulted to be 1/322 kb and 1/574 kb for the 1% EMS and 0.7% EMS treatment respectively. Conclusions. The mutation density estimated in our collection and its comparison with other TILLING populations demonstrate that the Red Setter genetic resource is suitable for use in high-throughput mutation discovery. The Red Setter TILLING platform is open to the research community and is publicly available via web for requesting mutation screening services. © 2010 Carriero et al; licensee BioMed Central Ltd.

Belviso C.,CNR Institute of Methodologies for Environmental analysis | Cavalcante F.,CNR Institute of Methodologies for Environmental analysis | Di Gennaro S.,Metapontum Agrobios | Lettino A.,CNR Institute of Methodologies for Environmental analysis | And 3 more authors.
Journal of Environmental Management | Year: 2014

A number of water purification processes have been developed in recent years based on the utilisation of low-cost materials with high pollutant removal efficiency. Among these materials, fly ash and zeolite synthesised from fly ash are two examples of high-efficiency adsorbents. Column absorption tests were performed in order to compare the manganese sorption behaviour of an Italian coal fly ash and zeolite synthesised from it. Different masses of both materials (10-60g) were exposed to solutions containing a total metal concentration of 10mg/L. Batch adsorption studies were also conducted to determine the effect of time on the removal on Mn sequestration. The results indicate that both materials are effective for the removal of Mn from aqueous solution by precipitation due to the high pH of the solid/liquid mixtures. However, the leaching tests reveal that the amount of Mn removed from the fly ash was greater than that leached from the zeolite, thereby indicating that the metal is partially sequestrated by zeolite. © 2014 Elsevier Ltd.

Bianco G.,University of Basilicata | Zianni R.,University of Bari | Anzillotta G.,Metapontum Agrobios | Palma A.,Metapontum Agrobios | And 3 more authors.
Analytical and Bioanalytical Chemistry | Year: 2013

We report on the content of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in 15 breast milk samples of nursing women living in the city of Taranto (Southern, Italy) or nearby. Breast milk samples were collected over the 2008-2009 period and analyzed by gas chromatography coupled with high-resolution mass spectrometry (GC-HRMS) upon accelerated solvent extraction (ASE) using acetone/n-hexane mixture 1:1 (v/v). The method was validated demonstrating good performing features. Profiles of PCDD/PCDF congeners in breast milk samples exhibited a prevalence of PCDFs compared to PCDDs. Toxic equivalents (TEQs in picogram per gram fat) of four breast milk were far above the legal limit for human consumption of 3.0 pg/g; their estimated daily and weekly dietary intake were almost 5-20 and 10-40 times higher, respectively, than the tolerable intake values established by the World Health Organization. © 2013 Springer-Verlag Berlin Heidelberg.

Di Matteo A.,University of Naples Federico II | Ruggieri V.,University of Naples Federico II | Sacco A.,University of Naples Federico II | Rigano M.M.,University of Naples Federico II | And 6 more authors.
Plant Science | Year: 2013

Phenolics are antioxidants present in tomato fruit that confer healthy benefits and exhibit crucial roles for plant metabolism and response to environmental stimuli. An approach based on two genomics platforms was undertaken to identify candidate genes associated to higher phenolics content in tomato fruit. A comparative transcriptomic analysis between the S. pennellii Introgression Line 7-3, which produced an average higher level of fruit phenolics, and the cultivated variety M82, revealed that their differences are attributed to genes involved in phenolics accumulation into the vacuole. The up-regulation of genes coding for one MATE-transporter, one vacuolar sorting protein and three GSTs supported this hypothesis. The observed balancing effect between two ethylene responsive factors (ERF1 and ERF4) was also hypothesized to drive the transcriptional regulation of these transport genes. In order to confirm such model a TILLING platform was explored. A mutant was isolated harbouring a point mutation in the ERF1 cds that affects the protein sequence and its expected function. Fruits of the mutant exhibited a significant reduced level of phenolics than the control variety. Changes in the expression of genes involved in sequestration of phenolics in vacuole also supported the hypothesized key-role of ERF1 in orchestrating these genes. © 2013 Elsevier Ireland Ltd.

Musto M.,University of Basilicata | Faraone D.,University of Basilicata | Cellini F.,Metapontum Agrobios
Journal of Applied Animal Welfare Science | Year: 2014

Given the increasing importance of exploring consumers' concerns about the welfare of farmed animals, a survey questionnaire was designed to investigate the role of cognitive styles along with sociodemographic characteristics in consumers' perceptions about nonhuman animal welfare (AW) and their willingness to pay for animal-friendly products. The results revealed that the survey respondents were concerned about AW and had negative perceptions of the way animals were treated. They showed positive attitudes toward some actions to be taken for improving AW and strongly agreed to pay more for animal-friendly products. Consistent with previous studies, results revealed significant associations between sociodemographics and concern toward AW. However, some observed differences were highlighted by cognitive styles rather than by sociodemographic characteristics. These results indicate a significant link between cognitive styles and perceptions and attitudes toward AW, which may outweigh previously found sociodemographic differences and fuel the contemporary debate on AW. © Taylor & Francis.

Bianco G.,University of Basilicata | Novario G.,University of Basilicata | Anzilotta G.,Metapontum Agrobios | Palma A.,Metapontum Agrobios | And 2 more authors.
Journal of Mass Spectrometry | Year: 2010

Samples of Mytilus galloprovincialis were examined to investigate the levels of polybrominated diphenyl ethers (PBDEs) in the Apulia's marine environment, a region in the South of Italy. The levels of nine PBDE congeners were measured in 40 mussel samples taken from the aquaculture farms in the South of Adriatic Sea and North Ionian Sea along the Apulia coast. While accelerated solvent extraction (ASE) was the extraction technique adopted using acetone/n-hexane (1:1, v/v), the content of PBDEs was evaluated by using gas chromatography coupled with high-resolution mass spectrometry (GC-HRMS) via electron ionization (EI) in the multiple ion monitoring (MIM) mode (two ions for each compound). The 13C mass-labeled compounds were used to establish the levels of PBDEs in M. galloprovincialis samples. The total concentration of PBDEs (ΣPBDEs) ranged from 0.2 to 6.9 ng/g dry mass, with the highest concentrations found around coastal areas of Salento. With regard to the composition of PBDE congeners, BDE-47, BDE-99 and BDE-100 were the predominant congeners in most of the samples. The present study is the first to report levels of PBDEs occurring in aquatic organisms living in aquaculture farms of Apulia region. Copyright © 2010 John Wiley & Sons, Ltd.

Stigliani A.L.,Metapontum Agrobios | Giorio G.,Metapontum Agrobios | D'Ambrosio C.,Metapontum Agrobios
Plant and Cell Physiology | Year: 2011

The pathway of carotenoids starts with the synthesis of phytoene and proceeds along a single path up to lycopene which can be transformed to β-carotene by the action of lycopene β-cyclase or to α-carotene through the sequential action of lycopene -cyclase and lycopene β-cyclase. All xanthophylls are produced from these two cyclic precursors following two hydroxylation steps. β,β-Xanthophyll biosynthesis requires hydroxylases belonging to the so-called 'non-heme di-iron' group while the biosynthesis of lutein involves enzymes belonging to the vast group of P450 monooxygenases with different enzymatic specificity due to the distinct rings of α-carotene. Here we report on the isolation and functional characterization of tomato CYP97A29 and CYP97C11 genes encoding the P450 carotenoid β- and -hydroxylases. Through a reverse transcription- quantitative real-time PCR analysis of the two P450 and nine other carotenoid biosynthetic genes it was possible to highlight the transcriptional patterns of the 11 genes in root, leaf, petal and fruit at three stages of development and ripening. Finally, the characterization of the two P450 carotenoid (A29 and C11) hydroxylases was complemented by an in planta analysis through the use of transgenic plants. Results of this study have permitted us to model the lutein synthesis in leaf and in fruit of tomato. © 2011 The Author.

D'Ambrosio C.,Metapontum Agrobios | Stigliani A.L.,Metapontum Agrobios | Giorio G.,Metapontum Agrobios
Transgenic Research | Year: 2011

Plant chloroplasts are enriched in xanthophylls which participate in photosynthesis as light-absorbing pigments and as dissipaters of excess light. In comparison, chromoplasts have evolved the capacity to synthesize and store brightly coloured carotenoid pigments to give flowers and fruits the power to attract pollinators and fruit dispersers. The best performing accumulator of xanthophylls in tomato is the petal chromoplast in contrast to the fruit chromoplast which only seems able to store carotenes. We have generated genetically engineered tomato lines carrying the tomato CrtR-b2 transgene with the aim of forcing the fruit to accumulate beta-xanthophylls. Both chloroplast and chromoplast-containing tissues of hemizygous transgenic plants were found to contain elevated xanthophyll contents as a direct consequence of the increased number of CrtR-b2 transcripts. Hemizygous transgenic leaves contained fourfold more violaxanthin than control leaves. Developing fruits were yellow instead of green since they lacked chlorophyll a, and their violaxanthin and neoxanthin contents were seven- and threefold higher, respectively, than those of the control. Ripe fruits of hemizygous transgenic plants contained free violaxanthin and significant amounts of esterified xanthophylls. Esterified xanthophylls were present also in ripe fruits of control and homozygous plants. However, in transgenic homozygous plants, we observed a reduction in transcript content in most tissues, particularly in petals, due to a post-transcriptional gene silencing process. These findings demonstrate that tomato fruit chromoplasts can accumulate xanthophylls with the same sequestration mechanism (esterification) as that exploited by chromoplasts of the tomato petal and pepper fruit. This study on transgenic plants overexpressing an important carotenoid gene (CrtR-b2) provides an interesting model for future investigations on perturbations in beta-carotene-derived xanthophyll synthesis which in turn may provide insights into the molecular mechanisms controlling carotenoid metabolism in tomato. © 2010 Springer Science+Business Media B.V.

Mishra K.B.,Academy of Sciences of the Czech Republic | Iannacone R.,Metapontum Agrobios | Petrozza A.,Metapontum Agrobios | Mishra A.,Academy of Sciences of the Czech Republic | And 6 more authors.
Plant Science | Year: 2012

Drought stress is one of the most important factors that limit crop productivity worldwide. In order to obtain tomato plants with enhanced drought tolerance, we inserted the transcription factor gene ATHB-7 into the tomato genome. This gene was demonstrated earlier to be up-regulated during drought stress in Arabidopsis thaliana thus acting as a negative regulator of growth. We compared the performance of wild type and transgenic tomato line DTL-20, carrying ATHB-7 gene, under well-irrigated and water limited conditions. We found that transgenic plants had reduced stomatal density and stomatal pore size and exhibited an enhanced resistance to soil water deficit. We used the transgenic plants to investigate the potential of chlorophyll fluorescence to report drought tolerance in a simulated high-throughput screening procedure. Wild type and transgenic tomato plants were exposed to drought stress lasting 18 days. The stress was then terminated by rehydration after which recovery was studied for another 2 days. Plant growth, leaf water potential, and chlorophyll fluorescence were measured during the entire experimental period. We found that water potential in wild type and drought tolerant transgenic plants diverged around day 11 of induced drought stress. The chlorophyll fluorescence parameters: the non-photochemical quenching, effective quantum efficiency of PSII, and the maximum quantum yield of PSII photochemistry yielded a good contrast between wild type and transgenic plants from day 7, day 12, and day 14 of induced stress, respectively. We propose that chlorophyll fluorescence emission reports well on the level of water stress and, thus, can be used to identify elevated drought tolerance in high-throughput screens for selection of resistant genotypes. © 2011 Elsevier Ireland Ltd.

Silletti M.F.,Metapontum Agrobios | Petrozza A.,Metapontum Agrobios | Stigliani A.L.,Metapontum Agrobios | Giorio G.,Metapontum Agrobios | And 3 more authors.
Molecular Breeding | Year: 2013

Tomato Cyc-B gene encodes a chromoplast-specific lycopene β-cyclase that converts lycopene to β-carotene during ripening of the fruit. By screening the tomato Red Setter mutant population with the TILLING method, we identified eight new alleles at the Cyc-B locus. Results of greenhouse phenotypic analysis revealed that the novel A949G Cyc-B allele produced modifications in the carotenoid profile and content of tomato petals and fruit. The cyc-b7 genotype, carrying the A949G Cyc-B allele, was therefore evaluated in an open field trial for standard agronomic traits as well as carotenoid content of the fruit. Results of the field trial confirmed that the induced A949G missense mutation favored the accumulation of lycopene in the fruit with no detrimental effects on the yield or on other agronomic and technological properties such as fruit firmness and Brix degree of fruit juice. On the basis of these results, it can be affirmed that the A949G Cyc-B allele constitutes a useful new genetic variant which can be used for improving carotenoid content in tomato fruit and for the development of new tomato commercial lines. Finally, the results presented here furthermore demonstrate that TILLING is a powerful methodology not only as a confirmatory system for gene functional analysis but also for selecting new gene variants useful for genetic improvement of important crops. © 2012 Springer Science+Business Media Dordrecht.

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