Morphometric and molecular analysis of a pink-berried mutant within the population of grape cultivar 'Plavac mali'

This study reports characteristics of pink-berried mutants found in the clone population of the Croatian red wine cultivar 'Plavac mali' on the basis of comparative ampelographic and DNA marker (SSR and Gret1) analysis. The pink-berried accession, also called 'Plavac mali sivi' (Croatian: sivi = English: grey or French: gris), along with the other 58 'Plavac mali' accessions of standard blue-black berry skin color, has been characterized for the first time using OIV descriptors and molecular markers. Using a set of 9 simple sequence repeat (SSR) markers, an identical SSR profile for all the analyzed accessions was revealed, indicating their monozygotic status. The analysis of Gret1 insertion within the VvMYBA1 locus revealed no DNA polymorphism responsible for the pink-berried phenotype. Surface color of the berry skin was measured with the CIELab technique using a reflectance spectrophotometer at full ripeness. The results of colorimetric variables (L*, a* and b*) suggest a significantly lower accumulation of anthocyanins in the pink-berried accession compared to the standard blue-black berries. The pink-berried accession shares all assessed morphological and genetic traits of 'Plavac mali', with the only difference being the color of the berry skin. This suggests that the pinkberried genotype is the result of a spontaneous mutation of a standard 'Plavac mali' genotype

Genetic diversity and differentiation within and between cultivated (Vitis vinifera L. ssp. sativa) and wild (Vitis vinifera L. ssp. sylvestris) grapes

Genetic characterization of 502 diverse grape accessions including 342 cultivated (V. vinifera ssp. sativa) and 160 wild (V. vinifera ssp. sylvestris) grapes showed considerable genetic diversity among accessions. A total of 117 alleles were detected across eight SSR loci with the average of 14 alleles per locus. The genetic diversity of wild grapes was slightly lower than that observed in the cultivated grapes probably due to small populations and severe natural selection leading to drift and loss of alleles and heterozygosity in wild grapes. The distance cluster analysis (CA) supported the classical ecogeographic groups with moderate genetic differentiation among them. There was a greater affinity of Occidentalis grape to wild grape from the Caucasus than other groups. However, a number of low to moderate frequency alleles that are present in the cultivated grape are not represented in the wild grape.

Cluster architecture of old, neglected Croatian grapevine varieties (Vitis vinifera L.)

Grape cluster architecture is an important trait affecting fruit composition and yield. Damage caused by gray mold (Botrytis cinerea) is directly related to cluster density and some other environmental factors. The aim of this study was to quantify characteristics of fully mature clusters of eleven old Croatian grapevine varieties (Vitis vinifera L.) which so far have not been phenotypically described in detail, and to examine Botrytis cinerea disease severity on their grapes under field conditions. Eight variables that describe the appearance of clusters were quantified in three consecutive years including cluster weight, length, width, volume, compactness, rachis weight, number of berries and single berry weight. Two-factor analysis of variance provided estimates of varietal and annual differences. Significant differences among eleven old Croatian varieties for all cluster traits were determined. Results suggest diverse cluster morphology within the old Croatian varieties revealing three variety groups by PCA analysis and their varying degrees of sensitivity to Botrytis. These phenotypic data on cluster structure represent a basis for future research, such as QTL analysis or assessment of productivity of these varieties

Guiding principles for identification, evaluation and conservation of Vitis vinifera L. subsp. sylvestris

Conservation of grapevine genetic resources is an important and long lasting task. Here, partners of the InWiGrape Activity of the European Cooperative Programme for Plant Genetic Resources have proposed a set of descriptors that will assist in identification, conservation and study of genetic resources of Vitis vinifera L. subsp. sylvestris. A distribution map of Vitis vinifera L. subsp. sylvestris populations in Europe was produced, with on-line access through the European Vitis Database. The several different aspects of conservation of Vitis vinifera L. subsp. sylvestris including bibliographical references, identification in the wild, in situ and ex situ conservation have been discussed. The descriptors and the map will assist different stakeholders, working on biodiversity and ecosystems in more effective conservation of wild grapevine genetic resources

Differentiation between Croatian dessert wine Prošek and dry wines based on phenolic composition

The phenolic composition of the Croatian dessert wine Prošek and dry wines Plavac mali and Pošip produced from the same autochthonous cultivars was investigated to determine which phenolic compounds best discriminate between these wine types. The wines were analyzed by the targeted metabolomic method using UPLC/QqQ-MS/MS. Forty-five (45) phenolic compounds were identified and classified into five groups based on chemical structure: benzoic acid derivatives, cinnamic acid derivatives, flavan-3-ols, stilbenes and flavonols. ANOVA indicated that the grape-drying process heavily influences the complex phenolic composition of Prošek dessert wine, which differs significantly from dry wines produced from the same cultivars. The data was grouped by principal component analysis and linear discriminant analysis to derive a classification function that distinguished dry and dessert wines with 98% accuracy. Principal component analysis separated the samples and showed that 23 phenolic compounds depending to phenolic acids, phenolic aldehydes, flavan-3-ols and flavonols were the compounds that best differentiated the Prošek from the dry wines

True-to-type confirmation of 20 unknown grapevine accessions at a grape germplasm collection in Split, Croatia

Twenty unknown accessions maintained in the field collection of the Institute for Adriatic Crops and Karst Reclamation (IAC), Split, Croatia, were identified. Morphological validation was done using standard OIV descriptors encompassing shoot, leaf and cluster characteristics. A standard set of nine simple sequence repeats (SSRs) were amplified and genetic diversity indices were calculated. The allelic data of the 20 unknown genotypes were compared with reference SSR profiles from the European database Vitis International Variety Catalogue (VIVC; http://www.vivc.de/) and grape germplasm collection of the UK. Three accessions proved to be mislabelled and were identified as known European grapevine cultivars (‘Blaufraenkisch’, ‘Chardonnay’ and ‘Merlot’), two unknown accessions showed identical genotypes matching ‘Chasselas blanc’ and ‘Chasselas rose’, while 12 unknown genotypes confirmed their declared status. Finally, three unknown genotypes of interspecific crosses (‘Ljana’, ‘Original’, ‘Nero’) were not found in VIVC database or in published literature, revealing SSR profiles that are novel to the best of our knowledge. This study presents necessary information to establish cultivar status (true to type) of 20 unknown accessions from the IAC Split grape collection

Seasonal variation in biopharmaceutical activity and fatty acid content of endemic Fucus virsoides algae from Adriatic sea

© 2019 Polish Pharmaceutical Society. All rights reserved. Macroalgae from Fucus genus are a valuable source of bioactive components as they are abundant in complex polysaccharides, fatty acids and polyphenols. In this work, the biological activity and chemical composition of extracts and fractions obtained from endemic Fucus virsoides J. Agardh species collected in the summer and the fall were investigated. From dichloromethane:methanol (1: 1) extract three fractions were made: petroleum-ether, ethyl-acetate and n-butanol. The aim of the study was to examine the influence of the seasonal variations on algal composition and activity. The significant seasonal variation in content and biological activity of Fucus virsoides samples was found. Fall extract and fractions exerted higher cytotoxic effects on cancer cell lines in comparison with summer extract and fractions. The examined extracts and fractions showed higher cytotoxic activity towards cancer cells compared to normal fibroblast MRC-5 cells. Morphological evaluation and cell cycle distribution analysis demonstrated their proapoptotic activity in human cervical adenocarcinoma HeLa cells. Fall extract and fractions better suppressed the migration and tube formation of EA.hy926 cells in comparison with summer extract and fractions. Fall extract and fractions were more potent in inhibition of α-glucosidase enzymatic activity. Ethyl-acetate fractions, from both seasons, exhibited the best antibacterial and antifungal activity on all tested bacteria and fungi. In conclusion, the two fall fractions ethyl-acetate and petroleum-ether rich in polyphenols and polyunsaturated fatty acids were the most active and exhibited prominent anticancer and anti-α-glucosidase activities

Usefulness of near infrared spectroscopy and chemometrics in screening of the quality of dessert wine Prošek

NIRS has been applied for the analysis of dessert wine Prošek produced from dried grapes of Pošip and Plavac mali cv. in a semi-scaled fermentation. It is a fast and non-destructive analytical method that in association with chemometrics is becoming more frequently used technique. NIR spectra were measured and the relative density, alcohol content, dry extract, reducing sugars, pH, total acidity, volatile acidity, ash, free sulphur dioxide and total sulphur dioxide were determined. The range of NIRS used in this paper was 904–1699 nm (i.e. 11062–5886 cm−1) because in this range vibrations of C–H, O–H, S–H and N–H bonds can mainly be observed. Using the basic principles of multivariate analysis, principal component analysis (PCA), was used to reduce the input matrix in order to look for significant relationships between the NIR spectra and the observed characteristics of wine Prošek. Chemometrics applied here resulted in further discrimination between samples. The main features of the spectra are absorption bands at 1580 and 1670 nm, which are related to the first overtone of the C–H stretch. Absorption bands around 908 nm, present the stretches related to the 3rd C–H overtone and 2nd overtone of the O–H stretch of H2O and ethanol. PC1 and PC2, the first two components, gave the highest level of classification (≈95 %) based on the grape used for wine production as well as on the added yeast. The potential of NIRS, as a non-destructive method, is screening of basic parameters that are usually determined during the winemaking and ageing of wine. The results of investigations confirm that NIRS combined with chemometric analysis is a promising tool for quality control and for on-line application in the control of the final product, Prošek

ANTIOXIDANT, ANTIPLATELET AND CYTOTOXIC ACTIVITY OF EXTRACT OF CYSTOSEIRA AMENTACEA FROM THE COAST OF MONTENEGRO (SOUTH-EAST ADRIATIC SEA)

Seaweeds are an excellent source of compounds with biological activity. Since algae have been used in traditional medicine for a long time and also some algal substances have shown cytotoxic, anticoagulant and antiplatelet activity, they have been extensively studied. We examined the cytotoxicity and cell cycle distribution of acetone extract of Brown alga Cystoseira amentacea, on different human malignant cell lines. The antioxidant and antiplatelet activity of the extract has been evaluated. The results indicated that extract of Cystoseira amentacea significantly decreased cell survival in all tested cell lines. The IC50 values in the MTT assay in human cervix carcinoma (HeLa), two human breast cancer cell lines (MDA-MB-361 and MDA-MB-453) and a human colon carcinoma (LS174) cells were ranged from 22.87±1.26 to 64.34±2.78µg/ml. In vitro antitumor activites was accompained by an important sub-G1 accumulation of all cells after treatment of tested cell lines with extract. Antioxidant activity of Cystoseira amentacea extract in vitro was shown using chemically and cell-based assays, in erythrocytes and neutrophils. Finally, extract of Cystoseira amentacea showed significant in vitro inhibitory effects on both platelet activation and aggregation with other blood cells in response to agonist in healthy donors