24-Epibrassinolide and methyl jasmonate can encourage cell growth and the production of secondary metabolites in immobilized cells of Hyoscyamus niger

In Vitro Cellular & Developmental Biology - Plant - This study was carried out to determine the effects of 24-epibrassinolide (24-eBL) and methyl jasmonate (MeJA) on cell growth and...     

The N-acetyl-D-glucosaminylphosphatidylinositol De-N-acetylase of glycosylphosphatidylinositol biosynthesis is a zinc metalloenzyme

The de-N-acetylation of N-acetyl-D-glucosaminylphosphatidylinositol (GlcNAc-PI) is the second step of mammalian and trypanosomal glycosylphosphatidylinositol biosynthesis. Glycosylphosphatidylinositol biosynthesis is essential for Trypanosoma brucei, the causative agent of African sleeping sickness, and GlcNAc-PI de-N-acetylase has previously been validated as a drug target. Inhibition of the trypanosome cell-free system and recombinant rat GlcNAc-PI de-N-acetylase by divalent metal cation chelators demonstrates that a tightly bound divalent metal cation is essential for activity. Reconstitution of metal-free GlcNAc-PI de-N-acetylase with divalent metal cations restores activity in the order Zn(2+) > Cu(2+) > Ni(2+) > Co(2+) > Mg(2+). Site-directed mutagenesis and homology modeling were used to identify active site residues and postulate a mechanism of action. The characterization of GlcNAc-PI de-N-acetylase as a zinc metalloenzyme will facilitate the rational design of anti-protozoan parasite drugs.     

Optimization of inoculum density to support root growth and secondary metabolite accumulation in root cultures of endangered Gentiana species: Gentiana lutea and Gentiana boissieri

In Vitro Cellular & Developmental Biology - Plant - Gentiana species belonging to the Gentianaceae family are medicinal plants rich in glycosides and phenolics. Gentiana lutea L. is a highly...     

Physical and biochemical differences in Agrobacterium rhizogenes-mediated transgenic hairy root lines of Echinacea purpurea

In Vitro Cellular & Developmental Biology - Plant - Echinacea (Echinacea purpurea L.), also known as purple coneflower, is one of the important medicinal plants commonly used for respiratory...     

Brassinosteroids Modify Yield,Quality, and Antioxidant Components in Grapes (Vitis vinifera cv. Alphonse Lavallée)

Brassinosteroids (BRs) are a class of polyhydroxysteroids that have been recognized as a “sixth class of plant hormones.” Exogenous application of BRs affects a broad spectrum of physiological responses, secondary metabolite accumulation, and resistance against stress factors in plants. In this study, the effects of pre-harvest 24-epibrassinolide (24-eBL) applications on the accumulation of antioxidant compound, yield, and quality properties of Alphonse Lavallée grape cultivar were examined. 24-eBL was applied to vines with 13 combinations including different application times and concentrations of 24-eBL (0.2, 0.4, 0.6, and 0.8 mg L⁻¹) and control. It was found that 24-eBL increased yield, quality, and antioxidant compounds in grapes when it was applied at appropriate concentration and application time. The concentration 0.2 mg L⁻¹ of 24-eBL applied to vines at three times (7 days after berry set + veraison + 30 days after veraison) was the most suitable application providing the highest yield and some quality properties such as cluster weight, berry weight, and specific gravity. Concentrations 0.6 and 0.8 mg L⁻¹ of 24-eBL applied at veraison to vines can be suggested for total phenolics, β-carotene, ascorbic acid, and trans-resveratrol accumulation for Alphonse Lavallée as a practical application, while the application consists of 0.6 mg L⁻¹ of 24-eBL and 7 days after berry set + veraison + 30 days after veraison seems to be the best treatment for anthocyanin content.

     

Cloning of Trypanosoma brucei and Leishmania major genes encoding the GlcNAc-phosphatidylinositol de-N-acetylase of glycosylphosphatidylinositol biosynthesis that is essential to the African sleeping sickness parasite

The second step of glycosylphosphatidylinositol anchor biosynthesis in all eukaryotes is the conversion of D-GlcNAcalpha1-6-d-myo-inositol-1-HPO(4)-sn-1,2-diacylglycerol (GlcNAc-PI) to d-GlcNalpha1-6-d-myo-inositol-1-HPO(4)-sn-1,2-diacylglycerol by GlcNAc-PI de-N-acetylase. The genes encoding this activity are PIG-L and GPI12 in mammals and yeast, respectively. Fragments of putative GlcNAc-PI de-N-acetylase genes from Trypanosoma brucei and Leishmania major were identified in the respective genome project data bases. The full-length genes TbGPI12 and LmGPI12 were subsequently cloned, sequenced, and shown to complement a PIG-L-deficient Chinese hamster ovary cell line and restore surface expression of GPI-anchored proteins. A tetracycline-inducible bloodstream form T. brucei TbGPI12 conditional null mutant cell line was created and analyzed under nonpermissive conditions. TbGPI12 mRNA levels were reduced to undetectable levels within 8 h of tetracycline removal, and the cells died after 3-4 days. This demonstrates that TbGPI12 is an essential gene for the tsetse-transmitted parasite that causes Nagana in cattle and African sleeping sickness in humans. It also validates GlcNAc-PI de-N-acetylase as a potential drug target against these diseases. Washed parasite membranes were prepared from the conditional null mutant parasites after 48 h without tetracycline. These membranes were shown to be greatly reduced in GlcNAc-PI de-N-acetylase activity, but they retained their ability to make GlcNAc-PI and to process d-GlcNalpha1-6-d-myo-inositol-1-HPO(4)-sn-1,2-diacylglycerol to later glycosylphosphatidylinositol intermediates. These results suggest that the stabilities of other glycosylphosphatidylinositol pathway enzymes are not dependent on GlcNAc-PI de-N-acetylase levels.