Optimization of PCR Conditions to Amplify Microsatellite Loci in Grapevine (Vitis vinifera L.) Genomic DNA

A total of three different primer pairs were optimized for polymerase chain reaction (PCR) to amplify microsatellite loci in total genomic DNA of grapevine (Vitis vinifera L.). Different concentrations of MgCl2, DNA and different regimes of annealing temperature were optimized. For all the primer pairs, 2.5 mM MgCl2 concentration was found optimum. For DNA concentration, 100 ng in the final reaction volume was suitable for good amplification. Annealing temperatures 56°C, 61°C and 58°C were found optimum to amplify with primer pairs VVMD5, scu04vv and VMC8E6, respectively. The other reagents used in PCR and temperature regimes (denaturation and extension temperature) were kept constant. The protocol has been successfully applied producing scorable and clear amplicons in all cultivars studied. These loci can be used to evaluate the genetic variability and cultivar relatedness in autochthonous Vitis vinifera cultivars from Tunisia

Organic Acids, Sugars, and Anthocyanins Contents in Juices of Tunisian Pomegranate Fruits

Juices made from fruits of 30 Tunisian accessions of pomegranate were studied for their organic acids, sugars, and anthocyanin contents, using high performance liquid chromatography. Among the detected organic acids, malic acid was the major one (>50%) followed by citric acid (>22%), while among sugars, fructose and glucose were most present in pomegranate juice contributing 53.9 and 43.4% of the total sugar content, respectively. The total anthocyanin content ranged from 9-115 mg per litre of juice with the following ranges of the six compounds found: cyanidin-3,5-diglucoside (3.1-74.4 mg/L), delphinidin-3-glucoside (0.7-22.0 mg/L), cyanidin-3-glucoside (0.8-21.0 mg/L), pelargonidin-3-glucoside (0.5-16.1 mg/L), pelargonidin-3,5-diglucoside (0.0-11.8 mg/L), and delphinidin-3,5-diglucoside (0.0-5.4 mg/L). Based on the analyzed parameters, cluster analysis allowed grouping cultivars into two main clusters. One was made of sour cultivars and the second of the sweet ones. Principle component and cluster analyses suggested that the composition of the pomegranate fruits is determined by cultivar rather than cultivation location

Amplification des Microsatellites chez les Plantes: Optimisation des composés de PCR

peer reviewe

Tunisian South Cultivated Grapevine (Vitis vinifera L.) -Foliar Composition on Major Mineral Elements-

peer reviewedThe diversity of the local grapevine varietal set of Tunisia was remarkably noted in areas of Sfax, Kerkhennah, Gabes, Tozeur, Nafta, Degache, Zarzis and Djerba. Some varieties showed requested characteristics at the level of the production quality, its lateness and its fitness to take on tree, and its adaptation to various Tunisian pedoclimatic conditions. In an attempt to explore the genetic diversity of 25 autochthones cultivars of grapevine (Vitis vinifera L.) in South East of Tunisia, the determination of leaves composition on major mineral elements (Na, K, Ca, Mg, Cu, Zn, Fe and Mn) have permitted to appreciate the genetic variability inter-cultivars and its structuring in this collection. Data (PCA plot, cluster analysis) provide that some of these parameters were designed suitable to discriminate significantly few cultivars (ARCz, KORg, BAKz, MGB, BAK, TON, SAKj…). In fact, the recorded differences could be due to sever environmental conditions such as: nature of the soil, culture and climatic conditions, irrigation, salinity, position of leaves on the branches, rootstock etc

Rapid, High Quality DNA Isolation from Tunisian Grapevine (Vitis vinifera L.) Cultivars and Optimization of the RAPD Marker Technique

Various problems are encountered during DNA extraction from Vitis species, harbouring high levels of secondary metabolites and polysaccharides. A simple and highly efficient protocol for isolating large quantities (0.5±0.3 mg.g-1 of leaf tissue) of high-quality DNA, from dry young Vitis vinifera leaves, is described in the present study. Thus, three different DNA extraction protocols were examined. The isolated DNA is essentially free of polysaccharides, polyphenols, and other major contaminants as judged by viscosity, clear color, A260/280 ratio, and RAPD suitability. Moreover, the RAPD profiling from the isolated DNA was optimized to produce scorable and clear amplicons in all studied cultivars

Tunisian South Cultivated Grapevine (Vitis vinifera L.) -Foliar Composition on Major Mineral Elements

Abstract: The diversity of the local grapevine varietal set of Tunisia was remarkably noted in areas of Sfax, Kerkhennah, Gabes, Tozeur, Nafta, Degache, Zarzis and Djerba. Some varieties showed requested characteristics at the level of the production quality, its lateness and its fitness to take on tree, and its adaptation to various Tunisian pedoclimatic conditions. In an attempt to explore the genetic diversity of 25 autochthones cultivars of grapevine (Vitis vinifera L.) in South East of Tunisia, the determination of leaves composition on major mineral elements (Na, K, Ca, Mg, Cu, Zn, Fe and Mn) have permitted to appreciate the genetic variability inter-cultivars and its structuring in this collection. Data (PCA plot, cluster analysis) provide that some of these parameters were designed suitable to discriminate significantly few cultivars (ARCz, KORg, BAKz, MGB, BAK, TON, SAKj…). In fact, the recorded differences could be due to sever environmental conditions such as: nature of the soil, culture and climatic conditions, irrigation, salinity, position of leaves on the branches, rootstock etc

Seed and juice characterization of pomegranatenext term fruits grown in Tunisia: Comparison between sour and sweet cultivars revealed interesting properties for prospective industrial applications

Tunisian previous termpomegranatenext term genetic resources consist of sweet and sour cultivars, showing large morphometric variability. In the present work we characterized seeds and juice contents of sugars and organic acids of 5 sour and 7 sweet previous termpomegranatenext term cultivars. Results showed that citric acid was predominant in sour previous termpomegranatesnext term, while malic acid was the most prevalent in sweet ones. Paradoxically, sour cultivars have higher sugar content than the sweet ones. A strong correlation was found between sourness and citric acid content, which is assumed to be the major factor that determines sour taste in previous termpomegranatenext term fruits. Besides, some of the seed parameters showed a significant positive correlation with acidity. Sweet cultivars were appropriate for fresh consumption and juice production due to several attributes in addition to their sweetness. Equally, sour previous termpomegranatenext term showed several characteristics that could be of great interest for food and nutraceutical industries

Etude des protéines totales et de l’ADN issus des feuilles de Moricandia arvensis collectées de quatre régions du sud tunisien

A partir des protéines et de l'ADN extraits des feuilles de Moricandia arvensis, collectées de quatre régions du sud tunisien, nous avons étudié la variabilité de cette plante reconnue pour ses vertus médicinales. Nous avons analysé d’une part les protéines totales extraites puis séparées par la technique SDS-PAGE. Cette méthode a révélé un degré de similitude très remarquable entre les plantes issues des quatre régions et qui varie entre 0.77 et 0.91. Nous avons effectué d’autre part, une analyse de la variabilité génétique par l’amplification aléatoire de l'ADN polymorphe par la technique PCR-RAPD en utilisant trois amorces différentes. Nous avons remarqué que la similitude est plus accentuée entre les plantes collectées des régions du Sud Ouest, avec un degré de similitude pouvant atteindre 100%. Celles issues du Sud Est présentent un degré de similitude de 66%

Characterization of Tunisian pomegranatenext term (Punica granatum L.) cultivars using amplified fragment length polymorphism analysis

The amplified fragment length polymorphism (AFLP) analysis of DNA was used to characterize 34 previous termpomegranatenext term cultivars. By using a combination of six primers, a total of 327 markers were scored with a mean of 57.5. The high percentage of polymorphic bands (ppb) of 94.7 and the resolving power (Rp) collective rate value of 129.14 were scored. Data proved that the tested primers were informative to discriminate among cultivars and to survey the genetic diversity in this fruit crop. It has been assumed that the local previous termpomegranatenext term germplasm is characterized by a typically continuous genetic diversity. The derived dendrogram proved that cultivars are clustered independently from their geographical origin and their denomination. In addition, AFLP permitted the generation of a nearly unlimited number of molecular markers that are reliable in differentiating the cultivars and/or the polyclonal varieties

Molecular genetic diversity of Punica granatum L. (pomegranate) as revealed by microsatellite DNA markers (SSR)

Pomegranate (Punica granatum L.) is one of the oldest known edible fruits and more and more it arouse interest of scientific community given its numerous biological activities. However, information about its genetic resources and characterization using reliable molecular markers are still scarce. In the present study, we report the development of 4 new polymorphic SSR markers. They have been used in addition to 11 SSRs previously published to investigate molecular diversity of 33 Punica granatum ecotypes. Based on the multi-locus profiles, twenty-two distinctive genotypes were identified. Globally, quite low genetic diversity has been revealed, as measured by allele richness (2.83 per locus) and heterozygosity (He = 0.245; Ho = 0.243), reflecting the narrow genetic background of the plant material. Four synonymous groups could be detected involving 15 accessions. Results of ordination and cluster analysis suggested that almost all the Tunisian cultivars share similar genetic background, and are likely derived from a small number of introductions in ancient times. Results issued from this study provide essential information to project a previous termpomegranatenext term core-collection without plant material duplication and for sustainable management of previous termpomegranatenext term landraces at national and international level. Furthermore, these SSR markers are powerful tool for marker assisted selection (MAS) program and for QTL studies