168 research outputs found
Efficient production of saffron crocins and picrocrocin in Nicotiana benthamiana using a virus-driven system
[EN] Crocins and picrocrocin are glycosylated apocarotenoids responsible, respectively, for the color and the unique taste of the saffron spice, known as red gold due to its high price. Several studies have also shown the health-promoting properties of these compounds. However, their high costs hamper the wide use of these metabolites in the pharmaceutical sector. We have developed a virus-driven system to produce remarkable amounts of crocins and picrocrocin in adult Nicotiana benthamiana plants in only two weeks. The system consists of viral clones derived from tobacco etch potyvirus that express specific carotenoid cleavage dioxygenase (CCD) enzymes from Crocus sativus and Buddleja davidii. Metabolic analyses of infected tissues demonstrated that the sole virus driven expression of C. sativus CsCCD2L or B. davidii BdCCD4.1 resulted in the production of crocins, picrocrocin and safranal. Using the recombinant virus that expressed CsCCD2L, accumulations of 0.2% of crocins and 0.8% of picrocrocin in leaf dry weight were reached in only two weeks. In an attempt to improve apocarotenoid content in N. benthamiana, co-expression of CsCCD2L with other carotenogenic enzymes, such as Pantoea ananatis phytoene synthase (PaCrtB) and saffron beta-carotene hydroxylase 2 (BCH2), was performed using the same viral system. This combinatorial approach led to an additional crocin increase up to 0.35% in leaves in which CsCCD2L and PaCrtB were co-expressed. Considering that saffron apocarotenoids are costly harvested from flower stigma once a year, and that Buddleja spp. flowers accumulate lower amounts, this system may be an attractive alternative for the sustainable production of these appreciated metabolites.We thank K. Schreiber and C. Mares (IBMCP, CSIC-UPV, Valencia, Spain) for technical assistance during plant transformation. We thank M. Gasc.on and M.D. G.omez-Jim.enez (IBMCP, CSIC-UPV, Valencia, Spain) for helpful assistance with LSCM analyses. We thank D. Dubbala (IBMCP, CSIC-UPV, Valencia, Spain) for English revision. This work was supported by grants BIO2016-77000-R and BIO2017-83184-R from the Spanish Ministerio de Ciencia e Innovacion (co-financed European Union ERDF), and SBPLY/17/180501/000234 from Junta de Comunidades de Castilla-La Mancha. M.M. was the recipient of a predoctoral fellowship from the Spanish Ministerio de Educacion, Cultura y Deporte (FPU16/05294). G.D. and L.G.G. are participants of the European COST action CA15136 (EUROCAROTEN). L.G.G. is a participant of the CARNET network (BIO2015-71703-REDT and BIO2017-90877-RED).Martí, M.; Diretto, G.; Aragones, V.; Frusciante, S.; Ahrazem, O.; Gómez-Gómez, L.; Daròs, J. (2020). Efficient production of saffron crocins and picrocrocin in Nicotiana benthamiana using a virus-driven system. Metabolic Engineering. 61:238-250. https://doi.org/10.1016/j.ymben.2020.06.009S23825061Ahrazem, O., Argandoña, J., Fiore, A., Aguado, C., Luján, R., Rubio-Moraga, Á., … Gómez-Gómez, L. (2018). Transcriptome analysis in tissue sectors with contrasting crocins accumulation provides novel insights into apocarotenoid biosynthesis and regulation during chromoplast biogenesis. Scientific Reports, 8(1). doi:10.1038/s41598-018-21225-zAhrazem, O., Diretto, G., Argandoña, J., Rubio-Moraga, Á., Julve, J. M., Orzáez, D., … Gómez-Gómez, L. (2017). Evolutionarily distinct carotenoid cleavage dioxygenases are responsible for crocetin production in Buddleja davidii. Journal of Experimental Botany, 68(16), 4663-4677. doi:10.1093/jxb/erx277Ahrazem, O., Gómez-Gómez, L., Rodrigo, M., Avalos, J., & Limón, M. (2016). Carotenoid Cleavage Oxygenases from Microbes and Photosynthetic Organisms: Features and Functions. International Journal of Molecular Sciences, 17(11), 1781. doi:10.3390/ijms17111781Ahrazem, O., Rubio-Moraga, A., Argandoña-Picazo, J., Castillo, R., & Gómez-Gómez, L. (2016). Intron retention and rhythmic diel pattern regulation of carotenoid cleavage dioxygenase 2 during crocetin biosynthesis in saffron. Plant Molecular Biology, 91(3), 355-374. doi:10.1007/s11103-016-0473-8Ahrazem, O., Rubio‐Moraga, A., Berman, J., Capell, T., Christou, P., Zhu, C., & Gómez‐Gómez, L. (2015). The carotenoid cleavage dioxygenase
CCD
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Biochemical features of native red wines and genetic diversity of the corresponding grape varieties from Campania Region
Campania region has always been considered one of the most appreciated Italian districts for wine production. Wine distinctiveness arises from their native grapevines. To better define the chemical profile of Campania autochthonous red grape varieties, we analysed the phenolic composition of Aglianico di Taurasi, Aglianico del Vulture, Aglianico del Taburno, Piedirosso wines, and a minor native variety, Lingua di Femmina in comparison with Merlot and Cabernet Sauvignon, as reference cultivars. A genetic profiling was also carried out using microsatellite molecular markers with high polymorphic and unambiguous profiles. Principal component analysis applied to 72 wines based on the 18 biochemical parameters, explained 77.6% of the total variance and highlighted important biological entities providing insightful patterns. Moreover, comparison of SSR-based data with phenylpropanoid molecules exhibited a statistically significant correlation. Our approach might be reasonably adopted for future characterisations and traceability of grapevines and corresponding wines
New Spherically Symmetric Solutions in f(R)-gravity by Noether Symmetries
Spherical symmetry for f(R)-gravity is discussed by searching for Noether
symmetries. The method consists in selecting conserved quantities in form of
currents that reduce dynamics of f(R)-models compatible with symmetries. In
this way we get a general method to obtain constants of motion without setting
a priori the form of f(R). In this sense, the Noether symmetry results a
physical criterium. Relevant cases are discussed.Comment: 9 pages, accepted for publication in General Relativity and
Gravitatio
Serum thyroglobulin and 131I whole body scan after recombinant human TSH stimulation in the follow-up of low-risk patients with differentiated thyroid cancer
OBJECTIVE: The 'standard' postoperative follow-up of patients with differentiated thyroid cancer (DTC) has been based upon serum thyroglobulin (Tg) measurement and (131)I whole body scan ((131)I-WBS) after thyroid hormone (T(4)) treatment withdrawal. However, (131)I-WBS sensitivity has been reported to be low. Thyroid hormone withdrawal, often associated with hypothyroidism-related side effects, may now be replaced by recombinant human thyroid stimulating hormone (rhTSH). The aim of our study was to evaluate the diagnostic accuracy of (131)I-WBS and serum Tg measurement obtained after rhTSH stimulation and of neck ultrasonography in the first follow-up of DTC patients. DESIGN: Ninety-nine consecutive patients previously treated with total thyroidectomy and (131)I ablation, with no uptake outside the thyroid bed on the post-ablative (131)I-WBS (low-risk patients) were enrolled. METHODS: Measurement of serum Tg and (131)I-WBS after rhTSH stimulation, and ultrasound examination (US) of the neck. RESULTS: rhTSH-stimulated Tg was 1 ng/ml (Tg+) in 21 patients, including 6 patients with Tg levels >5 ng/ml. (131)I-WBS was negative for persistent or recurrent disease in all patients (i.e. sensitivity = 0%). US identified lymph-node metastases (confirmed at surgery) in 4/6 (67%) patients with stimulated Tg levels >5 ng/ml, in 2/15 (13%) with Tg>1<5 ng/ml, and in 2/78 (3%) who were Tg-negative. CONCLUSIONS: (i) diagnostic (131)I-WBS performed after rhTSH stimulation is useless in the first follow-up of DTC patients; (ii) US may identify lymph node metastases even in patients with low or undetectable serum Tg levels
Improvements in cosmological constraints from breaking growth degeneracy
The key probes of the growth of a large-scale structure are its rate f and amplitude σ8. Redshift space distortions in the galaxy power spectrum allow us to measure only the combination fσ8, which can be used to constrain the standard cosmological model or alternatives. By using measurements of the galaxy-galaxy lensing cross-correlation spectrum or of the galaxy bispectrum, it is possible to break the fσ8 degeneracy and obtain separate estimates of f and σ8 from the same galaxy sample. Currently there are very few such separate measurements, but even this allows for improved constraints on cosmological models
Use of SSR and retrotransposon-based markers to interpret the population structure of native grapevines from Southern Italy
Native grapevines are the quintessential ele- ments of Southern Italy winemaking, and genomic char- acterization plays a role of primary importance for preservation and sustainable use of these unexploited genetic resources. Among the various molecular techniques available, SSR and retrotransposons-based markers result to be the most valuable for cultivars and biotypes distinc- tiveness. A total of 62 accessions including 38 local grape cultivars were analyzed with 30 SSR, four REMAP and one IRAP markers to assess their genetic diversity and obtain a complete genomic profiling. The use of VrZAG79, VrZAG112, VVS2, VVMD25 and VVMD5 combined with retrotransposon-based markers proved to be the most dis- criminating and polymorphic markers for the rapid and unambiguous identification of minority grapevines from Campania region, which is considered one of the most appreciated Italian districts for wine production. Results revealed 58 SSR marker-specific alleles, 22 genotype- specific SSR alleles, and four REMAP and IRAP private bands. Cases of synonymy and homonymy were discov- ered. In conclusion, we provided evidences that the inte- grating SSR and retrotransposon-based markers is an effective strategy to assess the genetic diversity of autochthonous grapes, allowing their easy identification
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