Green wild plants (dirctly before flowering) and seeds of Hyoscyamus aureus were collected from natural habitat at Al Qalamon region in Syria. Seeds were surface sterilized and cultured in vitro, after 21 days from germination stem-derived callus was induced on two different nutrient media. Tropane alkaloids were extracted from wild plants and 30 days old in vitro plants and callus, and then analyzed using GC-MS. Genetic variation was also studied between the wild and in vitro plants and the callus culture lines using twenty ISSR markers. The results showed that there were significant variations in tropane alkaloids contents between the wild plants, the in vitro plants and the callus culture lines. The highest content of hyoscyamine was in callus on line A medium, but the highest content of scopolamine was in the wild plants. However, the lowest content of tropane alkaloids was in callus on line B medium. Also the ISSR analyses showed that there was genetic variation between the wild and in vitro plants and the callus culture lines.
The experiment has been carried out in the Syrian National Commission of Biotechnology, during the growing season 2018/2019, to study the effect of abiotic stresses (salinity and osmotic stresses) on the activity of some antioxidant enzymes and biochemical traits in Catharanthus roseus. The experiment has been laid according to (CRD) with three replications. The seeds have been sterilized by NaOCl solution (0.5% v/v), then planted on MS medium. Plantlets have been moved to MS medium enriched with NAA (1 mg.L-1) and BA (2 mg.L-1). The callus has been initiated from leaves using MS medium containing NAA (1 mg L-1) and KIN (2 mg.L-1). After 60 days, callus has been transferred to MS medium supplemented with different concentrations of PEG 6000 (-0.2,-0.3,-0.4,-0.5 MPa), and NaCl (25, 50, 75, 100 mM) in succession as stimulating agents. The results show that the top value of solutes leakage has been in the salt and osmotic treatments (28.04 and 26.98% respectively) compared with the control (8.563%). MDA content has significantly been higher in salt stress (102.3 µmol.g-1 FW) followed by the osmotic stress treatment (79.41 µmol.g-1 FW), while it was significantly lower in the non-stressed treatment (37.76 µmol.g-1 FW). An increase in the proline content occurred in both the stress treatments (4.623, 4.243 mmol.g-1 FW, respectively) compared with the control (2.477 mmol.g-1 FW). The activity of antioxidant enzymes (CAT, APX, and SOD) have significantly been higher in salt stress treatment (506.9, 12270.02 mol.min-1 .mg-1 protein, 191.4 U.mg-1 protein respectively), followed by osmotic stress (259.4, 7106.22 mol.min-1 .mg-1 protein, 65.60 U.mg-1 protein, respectively), while it has been significantly lower in control (126.9, 1800.38 mol.min-1 .mg-1 protein, 36.03 U.mg-1 protein, respectively).
This study investigates the effects of nutrient media on the genetic stability of cell lines of callus produced by Digitalis purpurea leaves explants. Four cell lines of D. purpurea callus were selected after inducing 80 sub-cultures of callus in four different nutrient media: Vollosovich (5C01) medium, Murashige and Skoog (MS), Linsmaier and Skoog (LS), and Gamborg (B5) were compared with control samples of seeds and in vitro grown plant (micropropagation). DNA was isolated from the callus by cetyltrimethylammonium bromide, and molecular biomarkers were quantified by inter-simple sequence repeat (ISSR)-polymerase chain reaction (PCR). The purity of the extracted DNA from cell lines and the in vitro has grown plant ranged from 1.94 to 1.98 compared to seeds (1.74) at wavelength 260/280 nm. Out of 360 ISSR-PCR reactions conducted for amplifying 18 samples with 20 primers, 19 primers provided 232 bands with polymorphism and 25 were unique bands with band sizes ranging from 75 to 3500 Bas pair. Cluster analysis and the Jaccard similarity coefficient showed that the medium 5C01 was the most stable compared to the MS, B5, and LS media. The genetic stability of D. purpurea callus was affected by the type and concentration of electrolyte and cytokinin in the nutrient media.
This study aimed to investigate the effects of certain nutrient medium types enhanced with plant-growth regulators on the direct inductionof somatic embryogenesis in vegetative strains produced and propagated from a tissue culture of Digitalis purpurea. The results showthat MS medium was superior to other tested nutrient media-B5, LS, and 5C01—in terms of the days required to initiate direct somaticembryogenesis induction and induction rate. In contrast, the LS medium resulted in the highest number of embryos. Moreover, 2 mg/LBA (benzyl adenine) was superior to similar concentrations of BAP (benzyl aminopurine) and Kin (kinetin) in the MS medium, whereasBAP performed better in LS and B5. On the other hand, auxin 2.4D (dichlorophenoxy acetic acid) inhibited somatic embryogenesis, as didNAA (naphthalene acetic acid) and IAA (indole acetic acid) when added individually. Regarding the type of explant, the stems showedthe induction rates and numbers of direct-somatic embryos produced in all the basic media.Keywords: Digitalis purpurea; Vegetative strains; Direct-somatic embryogenesis; Explant; Basic media
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