Mohamed Boussaid

Essential oils composition in two Rosmarinus officinalis L. varieties and incidence for antimicrobial and antioxidant activities

The essential oil composition of Rosmarinus officinalis var. typicus and var. troglodytorum endemic to Tunisia, and growing wild in different bioclimates, was determined by GC and GC-MS. Oils were assessed for their antimicrobial and antioxidant activity. A variation of the chemical composition attributed to varieties rather than to bioclimates was revealed. 1.8-Cineole (47.2-27.5%) and camphor (12.9-27.9%) were identified as the main constituents of var. typicus and var. troglodytorum, respectively. The principal component analysis performed on oil constituents for all the populations allowed the distinction of two distinct population groups in accordance to the varietal subdivision. Based on the determination of the diameter of inhibition and the determination of the minimum inhibitory concentration, a low to moderate antimicrobial activity according to oils was revealed against eight bacteria tested. However, oils from var. troglodytorum showed higher bactericidal effect than those from var. typicus. The oils antioxidant activity, determined by 1,1-diphenyl-1-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and β-carotene bleaching test, was relatively high. The highest activity was found in oils from var. troglodytorum and in one population of var. typicus from the upper semi-arid bioclimate.

Myrtus communis L. Infusions: The Effect of Infusion Time on Phytochemical Composition, Antioxidant, and Antimicrobial Activities

In traditional medicine, myrtle (Myrtus communis L.) is frequently consumed as an infusion and decoction. In this study, we investigate the phenolic and volatile compositions and antioxidant and antibacterial activities of leaf infusions prepared during 3 different times. The total phenolics contents (146.74 to 179.55 mg GAE/g DM) varied significantly between infusions. Eleven phenolic compounds were identified by reversed-phase high-performance liquid chromatography. Phenolic acids (7.64 to 14.28 μmol/g DM) and flavonol glycosides (7.05 to 12.11 μmol/g DM) were the major phenolic fractions of infusions. Significant quantitative variation in 6 phenolic components was observed between infusions. Sixteen volatile components were identified by gas chromatography (GC) and GC mass spectrometry analyses. The main constituents were 1,8-cineole (42.58% to 51.39%), α-terpineol (9.45% to 9.72%), methyl eugenol (6.69% to 7.11%), and linalool (5.91% to 6.06%). Quantitative variations of the volatile components of the analyzed oils in relation to the infusion time were observed. The antioxidant properties of infusions, assayed through DPPH (2,2- diphenyl-1-picrylhydrazyl) method, β-carotene bleaching test, chelating effect on ferrous ions, and ferric reducing power method, were considerable and varied according to the infusion time. Myrtle infusions exhibited a substantial antimicrobial activity against 6 tested bacteria.

Chemical composition and antioxidant activities of essential oils and methanol extracts of three wild Lavandula L. species

A comparative study of essential oil composition, polyphenol content and antioxidant activities of Lavandula coronopifolia, Lavandula multifida and Lavandula stoechas subsp. stoechas were reported. Qualitative and quantitative variations in the composition of oils according to species were shown. Lavandula coronopifolias oil was characterised by high proportions of trans-β-ocimene (26.9%), carvacrol (18.5%), β-bisabolene (13.1%) and myrcene (7.5%). The main components of L. multifida oil are carvacrol (65.1%) and β-bisabolene (24.7%). Lavandula stoechas oil is rich in fenchone (34.3%) and comphor (27.4%). The total phenolic and flavonoid contents also significantly varied among species. Lavandula coronopifolia exhibits the highest phenolic and flavonoid contents (31.3u2009mg GAEu2009g−1 and 16.3u2009mg REu2009g−1, respectively), followed by L. multifida (30.8u2009mg GAEu2009g−1 and 12.3u2009mg REu2009g−1). Methanolic extracts and essential oils displayed significant antioxidant activities. The level of antioxidant capacity varied according to extracts and species.

Plant regeneration via somatic embryogenesis from in vitro tissue culture in two Tunisian Cucumis melo cultivars Maazoun and Beji

Hypocotyl, cotyledon and zygotic embryo explants from two Tunisian Cucumis melo L. cultivars Beji and Maazoun, cultured on the MS medium added with 2,4-D (0.25–1xa0mgxa0l−1) and BA (0.10–0.50xa0mgxa0l−1), produce calluses with somatic embryos after 3xa0weeks of culture. For Beji c.v. the highest percentage (62.50%) of embryogenesis was observed for cotyledons. The average embryo number per callus was 10.40. Embryogenesis induction for zygotic embryos reached 33.50% with 29 embryos per callus. The embryogenesis ability of hypocotyls did not exceed 12.50% (2.50 embryos per callus). Somatic embryogenesis for Maazoun c.v. explants was less efficient. Embryos formation was observed only for cotyledons (29%) and zygotic embryos (25%). Cotyledonary staged embryos, when transferred to hormone free MS medium, germinated. The maximum germination rates were 51.50 and 44.50%, respectively for Maazoun and Beji c.v. The highest percentage (36.50%) of survival plants was noted for Beji c.v. Regenerants were diploids (2nu2009=u20092xu2009=u200924) and morphologically similar to their parents issued from seeds.

Variability of Volatiles in Tunisian Mentha pulegium L. (Lamiaceae)

Abstract Fourteen Tunisian Mentha pulegium L. populations were assessed for their extract composition by GC and GC/MS. They were collected in different geographical areas corresponding with their bioclimates. Thirty-four components representing 78.4% of the total extract were identified. Pulegone (41.8%), followed by isomenthone (11.3%) and carvone (6.2%) were the main compounds. The amounts of menthofuran and menthol did not exceed 3.7% and 2.8%, respectively. Differences in the extract compositions were recorded among populations, and three bioclimate related chemotypes were identified.

Genetic variation of Tunisian Myrtus communis L. (Myrtaceae) populations assessed by isozymes and RAPDs

The genetic variation of six Tunisian Myrtus communis L. (Myrtaceae) populations was assessed using nine isozymes coding for 17 putative loci and 79 RAPD markers, amplified by five decamer random primers. The analysed populations belonged to three bioclimatic zones (lower humid, sub-humid and upper semi-arid). A high genetic diversity within populations was detected both by isozymes and RAPDs. The level of variation differed according to bioclimate. Populations collected from sub-humid bioclimate showed more polymorphism than those grown in the upper semi-arid zone. For all populations, the genetic diversity revealed by RAPDs was more pronounced than that detected with isozymes. A high differentiation among populations related to bioclimate and geographic distance was revealed by both methods. Population’s structure based on RAPD markers was more concordant with bioclimatic zones in comparison with isozymes. Differentiation between ecological groups was higher than that revealed within groups. Conservation programs should take into account the level of genetic diversity within population revealed by the two complementary classes of markers according to bioclimate.RésuméLa variabilité génétique de six populations tunisiennes de Myrtus communis L. (Myrtaceae) a été estimée à l’aide de neuf systèmes isoenzymatiques contrôlés par 17 loci et 79 marqueurs RAPD amplifiés par cinq amorces. Les populations analysées appartiennent à trois étages bioclimatiques différents : humide inférieur, sub-humide et semi-aride supérieur. Une diversité génétique intrapopulation importante a été détectée. Le niveau de polymorphisme varie selon le bioclimat. Les populations du sub-humide sont plus polymorphes. Pour l’ensemble des populations, la diversité génétique révélée par les RAPDs est plus importante que celle détectée par les isozymes. Une forte différenciation entre les populations, selon le bioclimat et l’éloignement géographique, a été révélée par les deux méthodes. La structuration des populations selon les marqueurs RAPD concorde mieux avec le bioclimat. La différenciation entre les populations appartenant à des groupes écologiques différents est plus importante que celle entre populations d’un même groupe. Les programmes de conservation de l’espèce doivent tenir compte aussi bien du degré de la diversité génétique intrapopulation révélé par les deux types de marqueurs que du bioclimat.

Genetic Diversity and Population Structure of Teucrium polium (Lamiaceae) in Tunisia

Random amplified polymorphic DNA markers were used to assess the genetic diversity within and among seven Tunisian diploid and polyploid populations of Teucriumpolium L. from five bioclimatic areas. Out of the 141 bands generated from eight selected primers, 124 were polymorphic. The genetic diversity within a population (Shannon’s index) was high and varied according both the ploidal levels and bioclimatic zones. The genetic differentiation among populations assessed by GST and ΦST statistics was high, suggesting a low level of gene flow among them. The major proportion of the variation was attributable to individual differences within populations. The UPGMA analysis based on Nei and Li’s coefficient showed that individuals from each population clustered together. In a dendrogram using the ΦST distance matrix, population grouping is concordant with bioclimates and cytotypes. Conservation strategies should take into account the level of the genetic diversity of the populations according to their bioclimate and ploidal levels.

Essential Oil and Phenolic Compounds of Artemisia herba-alba (Asso.): Composition, Antioxidant, Antiacetylcholinesterase, and Antibacterial Activities

Total phenols, flavonoids, flavonols, and flavanols of the methanolic extract of the aerial part of Artemisia herba-alba were determined. The extract was analyzed by liquid chromatography with photodiode array coupled with electrospray ionisation mass spectrometry and allowed to identify of 10 phenolic compounds. Apigenin-6-C-glycosyl flavonoids and caffeoylquinic acids were identified. Chlorogenic acid and 1,4 dicaffeoylquinic acid being the major constituents. The essential oil obtained by hydrodistillation was analyzed by gas chromatography–mass spectrometry. Twenty-three compounds, representing 97.8% of the total oil, were identified. The most abundant components were β-thujone (41.9%), α-thujone (18.4%), and camphor (13.2%). Methanolic extract and essential oil exhibited a considerable antioxidant activity as evaluated by 2,2-diphenyl-pycrilhydrazil hydrate scavenging activity, reducing power, β-carotene bleaching test, and chelating ability. The methanolic extract was found to be more efficient, while the essential oil exhibited the highest acetylcholinesterase inhibitory activity. Analysis of the antibacterial activity showed that A. herba-alba methanolic extract and essential oil are efficient against gram positive and gram negative bacteria.

Genetic Diversity and Structure of Wild Tunisian Myrtus communis L. (Myrtaceae) Populations

Myrtus communis L. (Myrtaceae), in Tunisia, is closely associated with Quercus suber L. forest which stretched continuously from the North to parts of Cap Bon and Tunisian Dorsal. The destruction of the primary oak forests associated to an over-exploitation of Myrtus for its essential oil quality had led to discontinuous populations exhibiting various levels of degradation. Using starch gel electrophoresis, we analyzed the polymorphism of nine isozymes in order to assess genetic diversity and structuring of 17 natural populations prospected in the three geographical regions and coinciding with subhumid, humid inferior and semi-arid superior bioclimates. The analysis of the level and the distribution of the genetic diversity in this species might help in its conservation. Out of the 18 loci detected for all populations and isozymes analyzed, 12 loci were polymorphic. Allelic frequencies differed according to populations and particular alleles characterized ecological groups. A high level of genetic variation within populations was observed. The mean number of alleles per polymorphic locus was Ap xa0=xa01.67, the percentage of polymorphic loci was Pxa0=xa060.3% and the observed (Ho) and the expected (He) heterozygosities were respectively 0.144 and 0.215. Populations belonging to subhumid (Cap Bon) and semi-arid superior (Tunisian Dorsal) climates, located in degraded sites exhibited the highest level of inbreeding (0.425xa0<xa0FISxa0<xa00.450). A high level of differentiation (FSTxa0=xa00.396) and a low gene flow (Nmxa0=xa00.337) among populations, as a result of habitat intermediate destruction, were revealed. The differentiation of populations within the same bioclimate (or geographic) group was substantial and relatively higher for semi-arid superior populations (FSTxa0=xa00.262), which were more distant. The three ecological groups exhibited a high level of structuring (FSTxa0=xa00.401). These differentiations might be due to geographic distances and to the variations of ecological factors between sites, including human activities and environmental factors. Nei’s (1972) genetic distances calculated between pairs of populations were globally low (0.006xa0<xa0Dxa0<xa00.367) with a mean of 0.15. They indicate a high level of similarity between populations. UPGMA clustering, established through Nei’s genetic distances, showed three population aggregates according to their geographic/bioclimatic appartenances. The high differentiation between populations and the low level of their genetic divergence indicated their recent isolation under anthropic pressures. The species conservation (in situ or ex situ) strategies should take into account the genetic diversity level within populations and its variation between geographic groups.

Genetic diversity in wild Tunisian populations of Mentha pulegium L. (Lamiaceae)

The allozyme variation of 15 Tunisian wild populations of Mentha pulegium L. threatened by human activities (clearing, hard-grazing, ploughing, traditional uses) was surveyed by the analysis of 14 isozyme loci using horizontal gel starch electrophoresis. The species exhibited a high level of genetic variation within populations (the mean Ap = 2.20, P = 72%, Ho = 0.349 and He = 0.229), which indicates a predominately outcrossing mating system and the recruitment of new genotypes via dispersal seeds. The genetic structure analysis of the populations using F statistics indicates no inbreeding, and showed an excess of heterozygosity for few loci. The moderate differentiation of populations (FST = 0.110) and the low rate of gene flow between them (Nm = 2.02) might been caused by recent isolation of the populations through biotope disturbances. The value of Neis genetic identity varied from 0.839 to 0.999 reflecting a relatively low genetic divergence between populations. Cluster analysis using UPGMA method and Neis genetic identity values, showed that populations geographically close didnt always cluster together. However, populations within the same bioclimatic stage generally subclustered together indicating that differentiation between bioclimatic regions occurred.