as mean ± standard deviation of triplicate experiments. In order to compare the four organs and to prove that the differences between these organs are not an accident or due to a chance, we utilised the Kruskal-Wallis test. This test rejects the hypothesis that the organs are from identical populations. Moreover, we used ANOVA (at a significant level of p < 0.05) coupled with multiple comparisons of means (Tukey Contrasts) to investigate: the differences between different organ assays and to compare selected M. minima populations. In addition, we completed discriminative analysis, namely PCA, for each organ by the using TPC as a matrix. We evaluated the association between variables by the Pearson correlation method. The levels and the statistical analysis (ANOVA, Kruskal-Wallis test and Tukey test) are shown in Supplementary Tables S1, S2. We used R (version 3.5.1) for all statistical analyses. The utilised packages were: Agricolae, Rcmdr, car, RcmdrMisc, corrplot, tidyverse, hrbrthemes, ggplot2 and RColorBrewer 68-71 .
Amplified fragment length polymorphism (AFLP) markers were used to characterize the genetic diversity within and among natural populations and cultivars of Hedysarum coronarium. Twelve populations within Tunisia were evaluated with three AFLP primer combinations. A total of 207 reproducible bands was detected of which 178 (86%) were polymorphic. The great discriminative power of AFLP markers and their ability to represent genetic relationships among Hedysarum plants was demonstrated. Genetic diversity within and among populations was assessed through Principal Component Analysis (PCA) and cluster analysis by using the Neighbor-joining clustering algorithm. AFLP technology has provided evidence of a high degree of intra-and inter-population genetic diversity in H. coronarium. AFLP banding patterns provided molecular markers correlated with the plants' geotropism. In addition, AFLP markers can differentiate wild accessions from cultivars. Moreover, geographical origins did not correspond to population clustering.
This study aims to evaluate the bio-morphological and biochemical variability of three Tunisian wild populations and one growing cultivar of purslane (Portulaca oleracea L.). The studied varieties should be easily distinguished by the color and the habitus of the plant as mentioned in literature, but the various analyses have shown a strong morphological heterogeneity within and among the wild and cultivated accessions as presented by the variance analysis test (ANOVA) and the PCA (Principal component analysis). We found high intrapopulation variability through the wild populations that make it hard to differentiate them only on the base of morphology. We analyzed the biochemical profile of those populations based on the analysis of freeze-dried samples of leaves and stems. We identified and quantified twelve different phenolic compounds by the HPLC-diode array detector (DAD) technique. Six phenolic acids and flavonoids were identified in the leaves and stems of the wild and cultivated populations. Sinapic acid and myricetin are the majors identified compounds through our samples. The results were significantly different in relation to the plant organs and to the geographic origin for most of the compounds. The obtained results highlighted the importance of Portulaca as a medicinal plant by showing its richness in phenols and flavonoids that have multi-medicinal effects besides their antioxidant power.
Sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA were analysed to precise their length (637-643 bp) and resolve phylogenetic relationships among eight Mediterranean species of the genus Hedysarum (Fabaceae). The infra-specific variability levels of the ITS sequences of spontaneous population of H. coronarium proved a lack of polymorphism both in the length and in the sequences examined in this species. Hence, a consensus ITS sequence characterising each Hedysarum species has been investigated for analysis of inter-specific polymorphisms. The level of variation of ITS sequence was high enough to make the ITS1 and ITS2 a useful tool for phylogenetic reconstruction. However, ITS2 seems to be relatively more polymorphic and more informative than ITS1 regarding length or GC percent. The phylogenic relationships in the genus Hedysarum based on ITS1 and ITS2 sequences taken independently or together, are discussed in the context of current work in molecular biosystematics. Results exhibited the distinctiveness of the two H. spinosissimum subspecies (i.e. H. spinosissimum ssp. capitatum and H. spinosissimum ssp. spinosissimum). In addition, the great similarity of the ITS sequences between H. coronarium (the only cultivated species of the genus) and H. carnosum suggests the usefulness of the latter in selection programmes to improve pastoral production in semi-arid areas.
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