Chemical Composition and Antimicrobial Activity of Essential Oils from Three Mediterranean Plants against Eighteen Pathogenic Bacteria and Fungi.

The chemical composition and antimicrobial activity of essential oils (EOs) obtained from three medicinal plants of the Moroccan flora were evaluated. The chemical composition of EOs of Thymus leptobotrys, Laurus nobilis and Syzygium aromaticum was determined using a gas chromatograph coupled with mass spectrometry. Carvacrol (75.05%) was the main constituent of T. leptobotrys EOs, while 1,8-cineole (31.48%) and eugenol (82.16%) were the predominant components of L. nobilis and S. aromaticum EOs, respectively. The antimicrobial activity of the EOs was evaluated qualitatively and quantitatively against 18 microbial strains pathogenic to humans by using the disc diffusion method, and by measuring the minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC). The EOs of T. leptobotrys were the most active against the strains tested, with inhibitory zone values ranging from 7.00 to 45.00 mm, and MIC and MMC values ranging from 0.312 to 80.00 mg/mL. In many cases, these EOs exhibited higher antibacterial and antifungal activities than the chemical compounds ciprofloxacin and fluconazole, respectively. This high antimicrobial activity can be ascribed to their richness in carvacrol. The EOs of T. leptobotrys, L. nobilis, and S. aromaticum could be considered a promising alternative to replace chemical antimicrobials, and a readily available natural source of bioactive compounds.

In Vitro Root Induction from Argan (Argania spinosa (L.) Skeels) Adventitious Shoots: Influence of Ammonium Nitrate, Auxins, Silver Nitrate and Putrescine, and Evaluation of Plantlet Acclimatization.

Argania spinosa (L.) Skeels is an endangered plant species endemic to Morocco. In recent years, attempts to develop in vitro regeneration systems for this species were made. However, rooting and acclimatization of in vitro plants have been a bottleneck for successful propagation. In the present study, the effects of different concentrations of auxins, putrescine, silver nitrate (AgNO3) and ammonium nitrate on the in vitro rooting of adventitious shoots of two argan genotypes "Mejji" and "R'zwa", were evaluated. The highest rooting percentages (86.6% in "Mejji" and 84.4% in "R'zwa") were observed on Murashige and Skoog (MS) medium modified by reducing the ammonium nitrate concentration and supplemented with 1.5 mg L-1 indole-3-butyric acid (IBA), 0.5 mg L-1 1-naphthalene acetic acid (NAA), 2 mg L-1 AgNO3 and 160 mg L-1 putrescine. This medium resulted in the development of a good root system after only 10 days of culture. Plantlet acclimatization was carried out using different substrate mixtures, and high survival rates (100%) were observed when the substrate contained either peat alone or a sand-peat mixture (1:1, w/w). The high percentages of rooting and acclimatization reported in the present study are of high importance for rapid and large-scale propagation of this endangered species.

Maturation and Conversion of Somatic Embryos Derived from Seeds of Olive (Olea europaea L.) cv. Dahbia: Occurrence of Secondary Embryogenesis and Adventitious Bud Formation.

Maturation and conversion of somatic embryos are two crucial steps that hamper the development of efficient somatic embryogenesis systems in olive. Herein, a simple and efficient protocol for the maturation and conversion of olive somatic embryos is reported. Globular somatic embryos derived from seeds of cv. Dahbia were cultured on either half-strength olive (OM) or olive cyclic embryogenesis (ECO) media, with and without plant growth regulators (PGRs). The embryos reached the cotyledonary stage in 9 weeks, but those cultured on ECO medium containing 0.1 mg·L-1 6-(dimethylallylamino)purine (2iP), 0.1 mg·L-1 6-benzyladenine (BA) and 0.05 mg·L-1 indole-3-butyric acid (IBA) exhibited the largest sizes, with an average of 4.7 mm. Somatic embryo conversion into plantlets was evaluated using different culture media (half-strength OM or one-third strength Murashige and Skoog (MS)), light conditions (light or dark) and desiccation pretreatments. The highest rate of somatic embryo conversion (45%) was observed under a 16 h photoperiod on half strength OM medium containing 0.1 mg·L-1 gibberellic acid (GA3) and 0.1 mg·L-1 1-naphthalene acetic acid (NAA). The embryos that failed to germinate showed either necrosis, cotyledon greening with no further conversion, adventitious bud formation or secondary embryogenesis. The findings of this study will be beneficial for biotechnological applications in olive.

A combined pathway of organogenesis and somatic embryogenesis for an efficient large-scale propagation in date palm (Phoenix dactylifera L.) cv. Mejhoul.

An efficient regeneration system via a combined pathway of organogenesis and somatic embryogenesis was developed for date palm (Phoenix dactylifera L.) cv. Mejhoul. Adventitious buds were obtained from shoot-tip explants with a frequency of 53.3% after 9 months of culture: 6 months on half-strength Murashige and Skoog (MS/2) medium containing 14.2 µM indole-3-acetic acid (IAA), 13.4 µM 1-naphthaleneacetic acid (NAA) and 0.5 µM 6-(dimethylallylamino) purine (2iP), and 3 months on MS/2 medium supplemented with 1.1 µM IAA, 1.1 µM NAA, 0.5 µM 2iP, 2.2 µM 6-benzyladenine (BA) and 0.4 µM kinetin. Adventitious bud segments were used as explants to induce somatic embryogenesis, and the effects of different concentrations (22.5, 45, 90, 225 or 450 µM) of 3,6-dichloro-o-anisic acid (dicamba) and 4-amino-3,5,6-trichloropicolinic acid (picloram) were evaluated. The optimal medium for somatic embryogenesis induction was MS medium supplemented with 45 µM picloram and 5 µM 2iP, in which the somatic embryogenesis rate was 70%. For somatic embryo maturation, the effects of sorbitol, mannitol, polyethylene glycol (PEG) and abscisic acid (ABA) were tested. The highest maturation rate (88.6 mature somatic embryos per 100 mg fresh weight callus) was observed on liquid MS medium supplemented with 20 g L-1 PEG. Subsequent somatic embryo germination was achieved with up to 52.0% in MS medium containing 2.5 µM NAA and 2.5 µM BA. The regenerated plantlets were transferred to the glasshouse where 76.0% of them survived.

Somatic embryogenesis from bud and leaf explants of date palm (Phoenix dactylifera L.) cv. Najda.

An efficient regeneration system through somatic embryogenesis was developed for date palm cv. Najda. Adventitious bud and proximal leaf segments cultured on Murashige and Skoog (MS) medium supplemented with various combinations of auxins and cytokinins induced embryogenesis after at least 6 months of culture. Somatic embryogenesis induction seemed correlated with the type of the explant, the induction period and the auxin used. The highest rate of somatic embryogenesis (86.0%) was obtained on bud explants cultured on MS medium supplemented with 45.0 µM 2,4-dichlorophenoxyacetic acid (2,4-D), and 4.5 µM kinetin or 4.5 µM 6-(dimethylallylamino) purine (2iP). Whereas, low levels of embryogenesis were obtained on media supplemented with 1-naphthalene acetic acid (NAA) or 2-naphthoxyacetic acid (NOA). Proximal leaf segments showed somatic embryogenesis only when cultured on media supplemented with 2,4-D or picloram. Statistical analysis revealed significant effects of explant type and plant growth regulators (PGRs) combination on somatic embryogenesis. Somatic embryos were germinated successfully on PGR-free MS medium with or without activated charcoal (50.0-60.0 and 26.6-36.6%, respectively), and 80.0% of plantlets survived after transferring to a glasshouse for 6 months. Our results will be useful for large-scale propagation of date palm cv. Najda, characterized by high fruit quality and bayoud disease resistance.

Optimization of medium composition for in vitro shoot proliferation and growth of date palm cv. Mejhoul.

The effects of major mineral salts, L-glutamine, myo-inositol and carbon source on shoot bud proliferation of date palm (Phoenix dactylifera L.) cv. Mejhoul were evaluated. Different concentrations of ammonium nitrate (NH4NO3; 550, 825 or 1650 mg/L), potassium nitrate (KNO3; 633.3, 950 or 1900 mg/L), calcium chloride dehydrate (CaCl2·2H2O; 147, 220 or 440 mg/L), potassium dihydrogen phosphate (KH2PO4; 57, 85 or 170 mg/L), magnesium sulfate heptahydrate (MgSO4·7H2O; 123, 185 or 370 mg/L), L-glutamine and myo-inositol (0.5, 1, 1.5 and 2 g/L), sucrose, sorbitol, mannitol or commercial granulated sugar (10, 20, 30, 40 or 50 g/L) were tested. The highest number of shoot buds per explant (18.7) occurred on the medium containing 825 mg/L NH4NO3, 1900 mg/L KNO3, 220 mg/L CaCl2·2H2O, 170 mg/L KH2PO4, 370 mg/L MgSO4·7H2O as well as 1 g/L L-glutamine, 2 g/L myo-inositol and 30 g/L sucrose. The results showed that the frequency of hyperhydricity significantly increased in media containing 1650 mg/L NH4NO3. The concentrations of L-glutamine, myo-inositol and carbon source significantly affected the number of shoot buds per explant. However, they had no effect on hyperhydricity, tissue browning and precocious rooting. Shoots of 4.5-6.0 cm in length were isolated and transferred onto hormone-free media for elongation and rooting. After 3 months, the developed plantlets were successfully transplanted in a glasshouse and over 90 % survived acclimatization.