No abstract

Neem is a medicinal plant used as antimalarial, antibacterial, antiviral, insecticide, and antimicrobial drug. This study aimed to investigate and predict the effect of yeast extract and sampling time on cell growth, secondary metabolites synthesis, SQS1 and MOF1 genes expression by response surface methodology. The highest fresh and dry cell weights were 580.25 g/L and 21.01 g/L, respectively obtained 6 days after using 100 mg/L yeast extract. The highest azadirachtin accumulation and production were 16.08 mg/g DW and 219.78 mg/L obtained 2 and 4 days, respectively after using 25 mg/L yeast extract. Maximum mevalonic acid accumulation (1.75 mg/g DW) and production (23.77 mg/L) were observed 2 days after application of 50 mg/L yeast extract. The highest amount of squalene accumulation (0.22 mg/g DW) and production (4.53 mg/L) were achieved 4 days after using 50 mg/L yeast extract. Prediction results exhibited the highest azadirachtin accumulation (13.61 mg/g DW) and production (190.50 mg/L), mevalonic acid accumulation (0.50 mg/g DW) and production (5.57 mg/L), and squalene accumulation (0.30 mg/g DW) by using 245 mg/L yeast extract for 2 days, 71 mg/L yeast extract for 2 days, 200 mg/L yeast extract for 4.96 days, without yeast extract for 6.54 days and 4 days, respectively. Also, it was predicted that the highest squalene production is achieved by long-term exposure to high concentrations of yeast extract. The qRT-PCR analysis displayed the maximum relative gene expression of SQS1 and MOF1 by using 150 and 25 mg/L yeast extract for 4 and 2 days treatment.

In this study, the effect of methyl jasmonate (MJ) and ultrasound (US), individually and in combination with L-tyrosine, on the stimulation of thebaine production in Papaver bracteatum cell suspension cultures was studied. The addition of L-tyrosine did not significantly affect the cell biomass, but significantly increased the thebaine yield of cells compared with the control. The synergistic effects of MJ and L-tyrosine in the combined treatment of 100 μM MJ and 2 mM L-tyrosine increased the thebaine yield of cells up to 84.62 mg L(- 1) at 6 days after treatment. Sonication of the cells for 20 s caused a significant decrease in cell growth and biomass, whereas the thebaine yield increased up to 39.60 mg L(- 1) at 6 days after treatment. The combination of US (10 s) and L-tyrosine feeding (2 mM) significantly increased the production of thebaine in comparison to individual utilisation of 2 mM L-tyrosine and US (10 s).

The seeds were surface-sterilized with 70% (v/v) ethanol for 45 s and 2.5% (w/v) sodium hypochlorite solution for 12 min, and then rinsed 3 times in sterile distilled water. The seeds were then germinated on MS medium (Murashige and Skoog, 1962) solidified with 8 g/L plant agar, and maintained in a growth chamber at 25 ± 2 °C with a 16-h photoperiod of cool white fluorescent light (400-500 lx).

No abstract

Neem is the medicinal plant used as antimalarial, antibacterial, antiviral, insecticide, and antimicrobial. This study aimed to investigate and prediction the effect of yeast extract and sampling time on cell growth, secondary metabolites synthesis, SQS1 and MOF1 genes expression by response surface methodology. The highest fresh and dry cell weights were 580.25 g/L and 21.01 g/L obtained 6 days after using 100 mg/L yeast extract. The highest azadirachtin accumulation and production were 16.08 mg/g DW and 219.78 mg/L obtained 2 and 4 days after using 25 mg/L yeast extract. Maximum mevalonic acid accumulation and production were 1.75 mg/g DW and 23.77 mg/L observed 2 days after application of 50 mg/L yeast extract. The highest amount of squalene accumulation (0.22 mg/g DW) and production (4.53 mg/L) were achieved 4 days after using 50 mg/L yeast extract. Prediction results showed that the highest azadirachtin accumulation (13.61 mg/g DW) and production (190.50 mg/L), mevalonic acid accumulation (0.50 mg/g DW) and production (5.57 mg/L), and squalene accumulation (0.30 mg/g DW) obtained by using 245 mg/L yeast extract for 2 days, 71 mg/L yeast extract for 2 days, 200 mg/L yeast extract for 4.96 days, without yeast extract for 6.54 days and 4 days, respectively. Also, predicted that the highest squalene production is achieved by long-term exposure to high concentrations of yeast extract. The qRT-PCR analysis showed the maximum relative gene expression of SQS1 and MOF1 obtained by using 150 mg/L yeast extract for 4 days and 25 mg/L yeast extract for 2 days, respectively.

No abstract

Optimizing of the in vitro culture condition, somatic embryogenesis, and plant regeneration is the first step in transformation. In this study, the response surface methodology was used to optimize and predict the concentrations of TDZ, NAA, BAP, and kinetin for callus formation, and regeneration by indirect somatic embryogenesis of Hyoscyamus niger. The results proved that TDZ is more efficient than NAA for the regeneration of H. niger. By using different concentrations of TDZ, BAP, and kinetin, the highest percentage of callusing (100.00%), callus fresh weight (1207.75 mg/explant), percentage of somatic embryogenesis (92.66%) and shooting (93.30%), and the number of the shoot (7.75 per callus) was recorded from petiole explants on MS medium containing 0.25 mg/L TDZ and 1 mg/L BAP, leaf explants by using 0.5 mg/L TDZ and 1 mg/L kinetin, petiole explants with addition 0.5 mg/L TDZ, leaf explants by using 0.50 mg/L TDZ and 1 mg/L kinetin, and petiole explants with addition 0.25 mg/L TDZ and 1 mg/L BAP, respectively. Also, the highest percentage of callusing (100.00%), callus fresh weight (3273.64 mg/explant), percentage of somatic embryogenesis (100%), shooting (100.00%), and the number of the shoot (8.61 per callus) were predicted by culturing of petiole explant on the MS medium containing 0.11 mg/L TDZ and 1.253 mg/L BAP, leaf explant by using 0.52 mg/L TDZ, petiole explant by using 0.533 mg/L TDZ and 1 mg/L BAP, leaf explant with addition 0.18 mg/L TDZ and 2.89 mg/L BAP and petiole explant by using 0.1 mg/L TDZ and 3 mg/L BAP.