1,721,005 research outputs found
The effect of methyl mercury on cytoskeleton and glutathion-dependent enzymes of CHO cultured cells
No full textValidation of a simplified small-scale DNA extraction protocol from wine by quantitative real-time PCR
Get PDFIn the present study, we compared a simplified small-scale purification protocol to obtain DNA admixtures out of wine, with our large-scale published method. The extraction methods must provide DNA free of PCR inhibitors, that can interfere with DNA amplification. To evaluate the efficiency of grapevine’s nuclear DNA extraction from wine, the new protocol was also compared in terms of purity and yield to the DNA obtained out of grapevine’s (Vitis vinifera) leaf tissue, using a commercial kit. Two single-copy nuclear genes, nine-cis-epoxy carotenoid dioxygenase 2 (NCED2), and prefoldin subunit 5-like (PS5) were amplified in DNA extracted from wine and grapevine by real-time TaqMan PCR to determine the presence of inhibitors in relation to the diversity of starting biological matrix. This study showed that the small-scale, simpler method for extracting DNA from wine produced effective results in terms of inhibitor presence and purity. Furthermore, even though the initial biological matrix was more complicated, the grapevine nuclear DNA that was removed from wine was qualitatively equivalent to the DNA that was isolated from the leaves
Characterization of genetic biodiversity with Vitis vinifera L. Sangiovese and Colorino genotypes by AFLP and ISTR DNA marker technology
No full textA comparison between two recently developed, PCR-based DNA marker technologies (amplified fragment length polymorphism, AFLP; inverse sequence-tagged repeat analysis, ISTR) was carried out in a group of 19 Vitis vinifera L. accessions, including 13 putative Sangiovese-related grapevines and 6 "coloured" ecotypes whose fruits are of importance for conferring intense red colour to the wine. A large amount of polymorphic DNA fragments was revealed by both molecular techniques: 8 different AFLP and 5 ISTR primer combinations generated 264 and 249 polymorphic markers, respectively. Similarity relationships among the accessions were described by cluster analysis. The AFLP analysis revealed the existence of a uniform group for the Sangiovese (SG) ecotypes showing a high degree of genetic relatedness for the members of this cultivar. Among the coloured ecotypes (CLR), variability was more evident. Only the so called Colorino americano ecotype significantly diverged from both groups. ISTR analysis confirmed the genetic dissimilarity of Colorino americano and the existence of the SG and CLR groups, but in addition detected a higher proportion of polymorphism among the Sangiovese accessions compared to AFLP analysis. Sangiovese forte and Saragiolo apparently differed from the other SG-related grapevines in agreement with AFLP results. It is possible that the observed genetic dissimilarity between Sangiovese forte, Saragiolo and other SG-related types could be interpreted by the putative polyclonal origin of many grapevine cultivars, a concept which is generally accepted by the grapevine research community. Both AFLP and ISTR appear to represent innovative, efficient and sensitive molecular tools for investigating genetic diversity among Vitis vinifera ecotypes and for the eventual identification of clones
Palaeobotanical evidence and SSR DNA analysis of archaeological apple seeds found in first century winery of Roman farm on Elba Island (Italy)
No full textIn the late republican Roman period (2nd-1st century BC) a Roman farm was
established at San Giovanni, on the Island of Elba. About 6th century BC the area had
hosted intense metallurgical activity (reduction of ion ore). In 1st century AD the farm
was destroyed by fire. Since 2012, the remains of walls defining five rectangular areas
have been uncovered. The first contained eight broken amphorae used to store,
conserve and transport beverages. Palaeobotanical analysis of set layers of the
archaeological site were performed. Palynological observations showed little pollen
and major presence of fungal spores and non-pollen objects typical of moist soil,
stagnant water and heavy human impact. Anthracological analysis revealed plants
useful for reconstructing the palaeoecological characteristics and human attitudes of
the period. In the bottom of amphorae protected by a crust of burnt soil, two sublayers
of deposition and a compact layer of mud and organic material, we found about 500
well conserved apple seeds, which when compared with contemporary seeds showed
morphological affinities with Malus sylvestris. Phenotypic study and genetic analysis of
genomic DNA with SSR molecular markers were performed against seeds of known
contemporary varieties of Malus sylvestris and Malus domestica as controls
Post-translational modifications of α-tubulin in Zea mays L. are highly tissue specific
No full textTo further understand post-translational modifications (PTMs) of plant α-tubulin, post-translationally modified α-tubulin isoforms from selected tissues of Zea mays L. were examined using two-dimensional electrophoresis and immunoblotting. Except for polyglycylated tubulin, tyrosinated, detyrosinated, acetylated and polyglutamylated α-tubulin isoforms were all present in maize tissues. Tyrosinated α-tubulin was the predominant variant in all cases, with isoforms α1-α4 (α5) being the most common components. Leaves exhibited a striking difference in PTM patterns of α-tubulin isoforms compared to other tissues examined. In leaves, several major specific isoforms were highly modified by detyrosination, acetylation and polyglutamylation. In pollen and anthers, only the most abundant isoform α3 was acetylated to an appreciable extent, and no acetylated isoform was found in roots. Similarly, in pollen, anthers and roots, only α3 was appreciably polyglutamylated. Additionally, a detyrosinated isoform α6 was present in anthers and in leaves, while the tyrosinated isoform α6 seemed to be pollen specific. These results indicate that certain types of PTM of plant α-tubulin preferentially occur in a tissue-specific way
Vineyards genetic monitoring and Vernaccia di San Gimignano wine molecular fingerprinting
Get PDFThe definition of the genetic profile of Vernaccia di San Gimignano (VSG) in the areas of production is an essential step for both the implementation of a plan of analytical traceability and the evaluation of the biological future potential of the same grape variety in relation to any environmental change. The genetic variability of the VSG was monitored by use of SSRs genotyping of a representative portion of individuals belonging to both the productive vineyards and the germplasm collections that represent the “mother plants” reservoir for future vineyards. 74% of the individuals have been shown to be identical to the grapevine genotype reported in databases as VSG truetype. In order to determine the wine varietal composition by DNA analysis, four wine types commercialized as VSG were DNA-tested at 14 loci SSRs. The molecular data obtained demonstrate the presence as prevalent component of the VSG in the four wine types. All the wines revealed the presence of minor varieties, whose presence/absence was estimated by extrapolating the allele configuration that best matched to a standard genotype. Molecular data allow us to exclude the presence of three aromatic white grapevines that are not allowed by the actual production rules (Disciplinare di Produzione
DNA extracted with optimized protocols can be genotyped to reconstruct the varietal composition of monovarietal wines
No full textA new DNA extraction protocol has been established to reconstruct the genetic identity of grapevine cultivars used for the production of experimental and commercial monovarietal wines. Seven wines originating from Merlot, Pinot noir, Zinfandel, Riesling, Sauvignon blanc, Sangiovese, and Alicante were identified by simple sequence repeat (SSR) analysis. The average quantity of Vitis vinifera DNA that can be extracted from wine, estimated by RT-PCR, was between 2,199 and 87,995 gc/mL. This is the first study that reports source vine identity reconstructed with a significant statistical correspondence (PI ⤠10-6) in both experimental and commercial wines. © 2012 by the American Society for Enology and Viticulture. All rights reserved
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