Sana Ghaffari, Nejib Hasnaoui[show abstract] [hide abstract]
ABSTRACT: Microsatellites (SSRs) are the most informative and popular class of molecular markers used for diverse purposes, particularly in plants: genetic diversity study, marker assisted selection, breeding, mapping, phylogenetics and phylogeography, systematics, etc. They have become a routine technique practically in each laboratory for studying molecular plant genetics. Despite their wide utilization, however, setup and optimization of various conditions involved in PCR amplification is a prerequisite for reliable inference of results. In this chapter, we describe optimization of SSR-PCR conditions and give ranges of concentrations for different parameters. The protocol provided here is inspired from bench work on the use of microsatellite to study diversity of Vitis vinifera germplasm.Methods in molecular biology (Clifton, N.J.) 01/2013; 1006:139-46.
Article: Rapid, High Quality DNA Isolation from Tunisian Grapevine (Vitis vinifera L.) Cultivars and Optimization of the RAPD Marker Technique[show abstract] [hide abstract]
ABSTRACT: 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.Romanian Biotechnological Letters. 01/2011; 16.
Article: Seed and juice characterization of pomegranate fruits grown in Tunisia: Comparison between sour and sweet cultivars revealed interesting properties for prospective industrial applications[show abstract] [hide abstract]
ABSTRACT: Tunisian pomegranate 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 pomegranate cultivars. Results showed that citric acid was predominant in sour pomegranates, 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 pomegranate 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 pomegranate showed several characteristics that could be of great interest for food and nutraceutical industries.Industrial Crops and Products 01/2011; 33(2):374-381. · 2.47 Impact Factor
Néjib Hasnaoui, Rania Jbir, Messaoud Mars, Mokhtar Trifi, Afaf Kamal-Eldin, Pablo Melgarejo, Francisca Hernandez[show abstract] [hide abstract]
ABSTRACT: 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.International Journal of Food Properties 07/2011; 14(4):741-757. · 0.67 Impact Factor
Article: Molecular Polymorphisms in Tunisian Pomegranate (Punica granatum L.) as Revealed by RAPD Fingerprints[show abstract] [hide abstract]
ABSTRACT: The genetic diversity among Tunisian pomegranate cultivars has been investigated. Using universal primers, the random amplified polymorphic DNA (RAPD) method was used to generate banding profiles from a set of twelve cultivars. Data was then computed with appropriate programs to construct a dendrogram illustrating the relationships between the studied cultivars. Our data proved the efficiency of the designed method to examine the DNA polymorphism in this crop since the tested primers are characterized by a collective resolving power of 12.83. In addition, the cluster analysis has exhibited a parsimonious tree branching independent from the geographic origin of the cultivars. In spite of the relatively low number of primers and cultivars, RAPD constitutes an appropriate procedure to assess the genetic diversity and to survey the phylogenetic relationships in this crop.Diversity. 01/2010;