Salicylic acid improves podophyllotoxin production in cell cultures of Linum album by increasing the expression of genes related with its biosynthesis

Treatment of Linum album cell cultures with 10 μM salicylic acid (SA) for 3 days improved podophyllotoxin (PTOX) production up to 333 μg/g dry weight (DW): over three times that of the control cultures. qPCR analyses showed that in SA-treated cells, the expression of the genes coding for phenylalanine ammonia-lyase (PAL), cinnamoyl-CoA reductase (CCR) and cinnamyl-alcohol dehydrogenase (CAD), all involved in the first steps of PTOX biosynthesis, also increased reaching a peak 8–12 h after the treatment. Expression of the pinoresinol-lariciresinol reductase gene (PLR), which is involved in one of the last biosynthetic steps, was not affected by SA. The selective action of SA on these genes can be applied to control the biotechnological production of this anticancer agent.     

Preparation of New Spiropyrazole,Pyrazole and Hydantoin Derivatives and Investigation of Their Antioxidant and Antibacterial Activities.

In this study, the synthesis of new spiropyrazoles, pyrazole and hydantoin heterocycles is reported by three component reactions of parabanic acids, hydrazine derivatives, and phenacyl bromides in the presence of triphenylphosphine as a nucleophile and triethylamine as a base in good to high yields (69–91 %). Evaluation of the synthesized compounds revealed a good to excellent antioxidant activities (37.6–96.2 %) using DPPH inhibitory potency. Among these compounds, hydantoin derivatives displayed higher antioxidant activities (93.7–96.2 %) comparing with spiropyrazoles and pyrazoles. The obtained results showed that Cl and Br substituents on the phenyl ring increased antioxidant activities of the related heterocycles. The antibacterial activities of the synthesized compounds were examined against two Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and two Gram-positive (Staphylococcus aureus and Bacillus subtilis) bacteria. Among the synthesized heterocycles, 2-[1,3-dimethyl-2,5-dioxo-4-(2-oxo-2-phenylethyl)imidazolidin-4-yl]hydrazine-1-carbothioamide exhibited the excellent antibacterial activity against both Gram-positive and Gram-negative bacteria.     

Up-regulation of matrix metalloproteinase-9 in primary bone tumors and its association with tumor aggressiveness

Molecular Biology Reports - Understanding the molecular mechanism underlying the pathophysiology of primary skeletal tumors is crucial due to the tumor-related complications, incidence at...     

The influence of Agrobacterium rhizogenes on induction of hairy roots and ß-carboline alkaloids production in Tribulus terrestris L.

We have developed an efficient transformation system for Tribulus terrestris L., an important medicinal plant, using Agrobacterium rhizogenes strains AR15834 and GMI9534 to generate hairy roots. Hairy roots were formed directly from the cut edges of leaf explants 10–14 days after inoculation with the Agrobacterium with highest frequency transformation being 49 %, which was achieved using Agrobacterium rhizogenes AR15834 on hormone-free MS medium after 28 days inoculation. PCR analysis showed that rolB genes of Ri plasmid of A. rhizogenes were integrated and expressed into the genome of transformed hairy roots. Isolated transgenic hairy roots grew rapidly on MS medium supplemented with indole-3-butyric acid. They showed characteristics of transformed roots such as fast growth and high lateral branching in comparison with untransformed roots. Isolated control and transgenic hairy roots grown in liquid medium containing IBA were analyzed to detect ß-carboline alkaloids by High Performance Thin Layer Chromatograghy (HPTLC). Harmine content was estimated to be 1.7 μg g⁻¹ of the dried weight of transgenic hairy root cultures at the end of 50 days of culturing. The transformed roots induced by AR15834 strain, spontaneously, dedifferentiated as callus on MS medium without hormone. Optimum callus induction and shoot regeneration of transformed roots in vitro was achieved on MS medium containing 0.4 mg L⁻¹ naphthaleneacetic acid and 2 mg L⁻¹ 6-benzylaminopurine (BAP) after 50 days. The main objective of this investigation was to establish hairy roots in this plant by using A. rhizogenes to synthesize secondary products at levels comparable to the wild-type roots.     

Growth inhibitor effects on protoplasmic drought tolerance and protein synthesis in leaf cells of the resurrection grass, Sporobolus stapfianus

Hydrated leaves of the resurrection grass S.stapfianus Gandoger are not desiccation tolerant, but tolerance can be induced in them by moderate to severe drought stress. When brassinolide (BR) and methyljasmonic acid (MJA) were applied separately, each improved PDT by approximately 6 MPa. Exogenous abscisic acid (ABA) improved the protoplasmic drought tolerance (PDT) of suspended cells from hydrated leaves of S. stapfianus only slightly (about 1 MPa).BR, MJA or ABA treatment of leaves on fully hydrated S. stapfianus plants induced changes in the leaf protein complement (partitioned by 2-D PAG electrophoresis), with the induction of apparently novel proteins and increased and decreased abundances of other proteins. Most of the changes that were induced by MJA differed from those produced by ABA and also by BR. Two proteins increased in abundance after treatment of leaves with MJA, BR or ABA.     

Contents of sugars in leaves of drying desiccation tolerant flowering plants, particularly grasses

Sugar complements were analysed in extracts from leaves of desiccation tolerant species in the angiosperm families Cyperaceae, Gesneriaceae, Liliaceae, Poaceae and Velloziaceae. Total sugar content was higher in live air-dry leaves of all desiccation tolerant species (except the grass Eragrostiella nardoides; 22 µmoles/g dw) than in the dead air-dry leaves of the desiccation sensitive grass Sporobolus pyramidalis (36 µmoles/g dw). Sucrose contents rose to high levels (40–98 µmoles/g dw) in live air-dry leaves of all species (except the grass Eragrostiella nardoides in which it rose to only 11 µmoles/g dw) to become the predominant sugar. Glucose and/or fructose contents frequently were lower after leaf drying but usually these were the sugars of next highest contents in live air-dry leaves. Contents of raffinose (that has been postulated to reduce sucrose crystallization) rose to c. 10% of sucrose contents in air-dry leaves of most desiccation tolerant species (but only c. 4% in Tripogon jacquemontii) compared with c. 2% of sucrose contents in the sensitive grass S. pyramidalis. Trehalose (a rare sugar in seed-plants) was present in all but one desiccation tolerant species (Xerophyta villosa) but only in minor amounts. The results are consistent with the views that sugars play a protective role during drying of desiccation tolerant plants in general but that other factors are also involved indesiccation tolerance, that in desiccation tolerant angiospermae sucrose is generally the predominant protective sugar and that raffinose and trehalose may supplement the role of sucrose.     

Improved Taxol production by combination of inducing factors in suspension cell culture of Taxus baccata

To date enormous attempts have been devoted to improve Taxol production exploiting various methodologies from bioprocess engineering to biotechnological and synthetic approaches. We have developed a 2-stage suspension cell culture of Taxus baccata L. using modified B5 medium in order to improve cell growth as well as productivity. After callus induction and cell line selection, B5 medium was supplemented with vanadyl sulfate (0.1 mg/l), silver nitrate (0.3 mg/l) and cobalt chloride (0.25 mg/l) at the first day of stage I culture to maximize cell growth. This medium was further supplemented with sucrose (1%) and ammonium citrate (50 mg/l) on day 10 and sucrose (1%) and phenylalanine (0.1 mM) on day 20 (i.e., biomass growth medium). At stage II (day 25), two different concentrations of several elicitors such as methyl jasmonate (10 or 20 mg/l), salicylic acid (50 or 100 mg/l) and fungal elicitor (25 or 50 mg/l) were added to the biomass growth medium with the aim of improving cellular productivity. For morphological analysis, microscopic inspection was carried out during cultivation. Cell-associated and extracellular amount of Taxol were detected and measured using HPLC methodology. At stage I, overall Taxol amount of biomass growth medium was 13.75 mg/l (i.e., 5.6-fold higher than that of untreated B5 control). At stage II, treated cells with methyl jasmonate (10 mg/l), salicylic acid (100 mg/l) and fungal elicitor (25 mg/l) produced the highest amount of Taxol (39.5 mg/l), which is 16-fold higher than that of untreated B5 control (2.45 mg/l). Microscopic analyses of Taxus cells in suspension cultures showed various positional auto-fluorescence showing direct correlation with Taxol production. Our studies revealed that intervallic supplementation of B5 medium with combination of biomass growth factors at stage I and mixture of elicitors at stage II could significantly increase Taxol production. Thus, we suggest that the exploitation of this methodology may improve the production of Taxol since demands for Taxol pharmaceuticals are increasingly growing and resource paucities have limited its direct harvesting from Taxus trees.     

Role of Jasmonic and Salicylic Acid on Enzymatic Changes in the Root of Two Alyssum inflatum Náyr. Populations Exposed to Nickel Toxicity

Phytohormones, including salicylic acid (SA) and jasmonic acid (JA) have the potential to ameliorate plant development and tolerance to deleterious effects of toxic metals like nickel (Ni). Therefore, the current study was carried out to evaluate SA and JA's interactive effect on the root antioxidative response of two Alyssum inflatum Nyár. populations against Ni-toxicity. Two A. inflatum species under different Ni concentrations (0, 100, 200, and 400 μM) were exposed to alone or combined levels of SA (0, 50, and 200 μM) and JA (0, 5, and 10 μM) treatments. Results showed that high Ni concentration (400 μM) reduced roots fresh weight in both populations than in control. However, external application of individual SA and JA or combined SA?+?JA in higher doses had ameliorated roots biomass by mitigating Ni-toxicity, especially in the NM population, in comparison to 400 μM Ni. Under Ni toxicity, SA and JA, especially their combination, induced high Ni accumulation in plants' roots. Moreover, the application of SA and JA alone, as well as combined SA?+?JA, was found to be effective in the scavenging of hydrogen peroxide by improving the activity of superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase in both populations under Ni-toxicity. Overall, our results manifest that SA and JA's external use, especially combined SA?+?JA treatments, ameliorate root biomass and plant tolerance by restricting Ni translocation to the shoot, accumulating in roots, and enhancing antioxidant defense systems.

     

Structure characterization of a methylated ester biosurfactant produced by a newly isolated <Emphasis Type="Italic">Dietzia cinnamea</Emphasis> KA1

Biosurfactans are amphiphilic compounds synthesized by a wide group of microorganisms and tend to interact with surfaces of different polarities. In the present study we purified and characterized a biosurfactant produced by Dietzia cinnamea KA-1 when cultured by n-hexadecane as sole carbon source. The crude biosurfactant was extracted with ethyl acetate and purified by freezing at–20°C and then silica Gel column chromatography. The purified biosurfactant applied for more characterization using Elemental analysis (CHNS), Fourier Transform Infrared Spectroscopy (FTIR), Mass Spectroscopy (MS) and Nuclear Magnetic Resonance (¹H and ¹³C-NMR) analysis. CHNS analysis showed the presence of C (74.92%) and H (11.63%) but not N or S. Functional groups of OH, CH₂, CH₃, C=O and aliphatic C?O revealed by FTIR analysis. The presence and position of these groups were confirmed by NMR analysis, and molecular mass of biosurfactant calculated using MS analysis. Finally, the product characterized as a methylated ester compound with molecular formula of C₂₁H₄₂O₄. This is the first report of biosurfactant of species D. cinnamea identified as ester, furthermore the ester was found to be in the methylated form.