Thymus algeriensis Boiss. et Reut., a north African endemic plant species: genetic diversity and population structure as assessed by molecular markers, a pioneer step for conservation implications.

BACKGROUND: Thymus algeriensis Boiss. et Reut. is one of the most widespread North African species of the genus Thymus L. The species is subshrub growing primarily in subtropical biome of Morocco, Algeria, Tunisia and Libya. In Tunisia, the plant species is under high pressure of anthropogenic activities including over-collecting. The assessment of genetic diversity and population structure of T. algeriensis is a pioneer step to retrace its evolutionary history and to perform appropriate conservation strategies of the plant species. METHODS AND RESULTS: Seven wild populations growing, widely, in different bioclimatic zones were selected and analysed using two molecular markers systems. Fifteen Simple Sequence Repeats (SSRs) and fifteen Inter-Simple Sequence Repeats (ISSRs) were used to characterize genetically 140 different genotypes. The results showed a high molecular variation within populations and among the studied genotypes. The intra-populations genetic diversity revealed by SSRs was higher (P = 80.95%, Na = 2.143 and He = 0.364) than that based on ISSRs (P = 78.12%, Na = 1.632, He = 0.265 and I = 0.398). As demonstrated by inbreeding coefficients, a significant level of differentiation and a low level of gene flow were detected among studied populations (FST = 0.161 for SSRs and ΦST = 0.197 for ISSRs). Furthermore, the results of ISSRs marker suggest land strips as barriers in population genetic structure. While SSRs marker reflects a relatively structured bioclimatic patterns of studied populations. The Bayesian analysis showed a specific adaptation of populations to local environments. CONCLUSIONS: The used molecular markers (ISSRs and SSRs) seem to be effective in deciphering genetic polymorphism of Tunisian genotypes of T. algeriensis. Therefore, the genetic structure of the studied genotypes could constitute a starting point for further conservation, characterization and breeding programs.

Variation in Phenolic Content, Antioxidant Activity and Alpha-amylase and Acetylcholinesterase Inhibitory Capacities of Different Extracts from Tunisian Satureja barceloi (Willk) L.

The antioxidant activity and the anti-α-amylase and anti-acetylcholinesterase capacities of secondary metabolites from different organs (roots, stems, leaves and flowers) of Tunisian Satureja barceloi were determined. The variation in the distribution of phenolic metabolites among roots, stems, leaves and flowers extracts of S. barceloi with various solvent systems (methanol, ethyl acetate, hexane and distilled water) has not been characterized before. Significant variation of phenolic compounds was observed according to organs rather than to extracting solvents. The analyzed organs show a high level of phenolic compounds although the stems contains the highest total polyphenols (132.53±0.48 mg AGE/g Ex), flavonoids (48.99±0.65 mg RE/g Ex) and flavonols (34.93±0.29 mg QE/g Ex) contents. The phenolic fraction was dominated by sagerinic acid, caffeic acid glucoside and epigallocatechin, detected using HPLC-PDA/ESI-MS. The antioxidant activity of all extracts, evaluated by four in vitro tests, was high and varied significantly according to the type of solvent used and the plant organ. The aqueous extracts of leaves exhibited the greatest inhibitory effect on alpha-amylase while the methanolic extract of leaves and stems revealed the most important acetylcholinesterase inhibitory effect. Hence, S. barceloi extracts could be used as a source of various bioactive molecules in pharmaceutical industry.

Insights for the control of dried-fruit beetle Carpophilus hemipterus (Nitidulidae) using rosemary essential oil loaded in chitosan nanoparticles.

Natural insecticidal products, essentially essential oils and their bioactive compounds are available as an excellent alternative method to control insect pests as well as they are less toxic to human health. However, due to their volatile constituents, encapsulation could be considered as the most suitable solution for their practical application. Therefore, this study aims to explore the insecticidal toxicity of chitosan nanoparticles loaded with Rosmarinus officinalis essential oil against adults of Carpophilus hemipterus. Chitosan nanoparticles were prepared by ionic gelation method with an encapsulation efficiency 41.543.1% and loading capacity 5.24G0.28%. Results revealed that chitosan nanoparticles loaded rosemary oil exhibited interesting insecticidal toxicity towards C. hemipterus adults in stored dates with 50.73% mortality. Overall, this work pointed out that the innovative design method based on chitosan-nanoparticles loaded rosemary oil can be promoted in integrated pest management program for stored date pests.

Screening for insecticidal toxicity of Urginea maritima (L.) bulbs and Asphodelus microcarpus (L.) tubers for the control of Tribolium castaneum (Tenebrionidae).

This research was conducted to determine the effect of geographical distribution on morphological characteristics of plants and some chemical compounds such as polyphenols. Moreover, we explore the contact toxicity of ethanolic and methanolic extracts from Urginea maritima bulbs and Asphodelus microcarpus tubers collected from three localities in the North West of Tunisia against Tribolium castaneum adults. The toxicity results demonstrated that both of the studied plant extracts had an effective control against T. castaneum at concentration 7.5 µL/L air, thus total mortality reached. In addition, results showed plant extracts with methanol exhibited high mortality percentage of T. castaneum. On the other hand, results revealed that chemical composition of U. maritime bulbs and A. microcarpus tuber extract depends on the soil components and geographical distribution of plants. The present investigation confirmed that A. microcarpus and U. maritima may be recommended as an eco-friendly alternative to synthetic insecticide against T. castaneum.

Genetic diversity and population structure of Tunisian wild Kermes oak (Quercus coccifera L.): Assessment by ISSR molecular markers and implication for conservation.

BACKGROUND: In Tunisia, Kermes oak (Quercus coccifera L.) populations are severely destroyed due to deforestation. Nowadays, no preservation programs are attempted, yet, to conserve and promote the potential value of this resource. In this work, we assessed the genetic diversity of seven natural Tunisian populations of Q. coccifera from different bioclimates using Inter-Simple Sequence Repeats molecular markers. The distribution of the genetic diversity of Q. coccifera constitutes the pioneer step in the process of the conservation of the species. METHODS AND RESULTS: Nine selected ISSR markers were analyzed to characterize the genetic profiles of 70 different genotypes. The ISSR primers produced 64 loci ranging from 6 (UBC809 and UBC810) to 9 (UBC873) with an average of 7.11 at the species level. The average percentage of the polymorphic loci varied from 64.06% (Tabarka) to 76.56% (El Haouaria). The analyzed genotypes (70 individuals) revealed a high level of genetic diversity at species level (Na = 1.697; Ne = 1.517; He = 0.289; I = 0.418). The major proportion of the variation was attributable to individual differences within populations (76.07%). Analysis of molecular variance revealed also significant differentiation among all populations (ΦST = 0.245) and among populations within bioclimates (ΦSC = 0.233), even at a low scale space. The UPGMA and the PCoA analyses showed that most populations clustered independently to bioclimate or geographical distance indicating that genetic differentiation mainly occurs at local space scale due to genetic drift. CONCLUSIONS: The in-situ conservation of the species should be maintained on natural populations as a forest genetic resources. Moreover, ex-situ conservation should involve the selection of genotypes with extensive collection of seeds and cuttings from different populations of the target area.

Effect of processing technology on chemical, sensory, and consumers' hedonic rating of seven olive oil varieties.

This study established physicochemical and sensory characteristics of virgin olive oils (VOOs) and linked them to consumers' liking using external preference mapping. We used five Tunisian and two foreign VOO varieties produced by two processing systems: discontinuous (sp) and continuous three-phase decanter (3p). The samples were analyzed and evaluated by a panel of 274 consumers. The external preference mapping revealed five VOO clusters with a consumer preference scores rating from 40% to 65%. Consumers highly appreciated the foreign Coratina cultivar's olive oil; the main drivers being richness in polyphenols (markers of bitterness and pungency), mainly the oleuropein aglycone, and volatile compounds (markers of green fruity, green leaves, green apple, cut grassy almond, and bitterness), particularly the trans-2-hexenol. The Tunisian Chemlali (3p) oil was second highly preferred (scoring 55%). The positive drivers for olive oil preference (a profile of almond fruity green and low bitterness and pungency) are the richness in hexanal compounds. Arbequina (sp and 3p) and Chemlali (sp) were the least appreciated due to the fact that Arbequina VOO is not in the tradition of Tunisian consumers, whereas Chemchali VOO is a minor variety representing only 2% of olive oil production in Tunisia and consumed mostly in blends. The differentiation between the two processing systems depends on the variety of cultivar; consumers are able to identify the two processing system in the case of Chetoui, Leguim, and Chemchali.

Essential Oils of Myrtaceae Species Growing Wild in Tunisia: Chemical Variability and Antifungal Activity Against Biscogniauxia mediterranea, the Causative Agent of Charcoal Canker.

The chemical composition of five Eucalyptus species and five Myrtus communis L. populations was investigated using GC/MS and GC-FID. For Eucalyptus essential oils, 32 compounds, representing 88.56 - 96.83% of the total oil according to species, were identified. The main compounds were 1,8-cineole, α-pinene, p-cymene, γ-gurjunene, α-aromadendrene, and ß-phellandrene. For Myrtle essential oils, 26 compounds, representing 93.13 - 98.91% of the total oil were identified. α-Pinene, 1,8-cineole, linalool, and myrtenyl acetate were found to be the major compounds. Principal component analysis (PCA) showed chemical differentiation between Eucalyptus species and between Myrtle populations. Biscogniauxia mediterranea, the causative agent of charcoal canker, was identified according to its morphological and molecular characteristics. Essential oils of the investigated Eucalyptus species and Myrtle populations were tested for their antifungal capacity against this fungus. The antifungal activity varied according to the essential oil composition. Biscogniauxia mediterranea exhibited powerful resistance to some essential oils including them of Eucalyptus lehmannii and Eucalyptus sideroxylon but it was very sensitive to Eucalyptus camaldulensis oil (IC50  = 3.83 mg/ml) and M. communis oil from Zaghouan (IC50  = 1 mg/ml). This sensitivity was found to be correlated to some essential oil compounds such as p-cymene, carvacrol, cuminaldehyde, and linalool.