Isolation of genetically modified potato plant containing the gene of defensive peptide from Amaranthus]

The plants of potato (Solanum tuberosum L., var. Desire) have been transformed with a pH22Kneo vector carrying the gene ac2, encoding the fungicidal peptide (defensin) from the seed of amaranth (Amaranthus caudatus L.). The transformation involved co-cultivation of potato stem explants (excised from aseptically grown plants) and Agrobacterium tumefaciens on solid MS medium. Factors affecting in vitro regeneration of the explants and the transformation efficiency were optimized. Regenerated potato plants harboring the amaranth defensin gene were selected by two traits, growth and ability to form roots on kanamycin-supplemented MS medium. The transgenic state was confirmed PCR analysis of ac2 in tissues of the kanamycin-resistant plants. The transgenic organisms thus obtained differed from the original ambiol-treated plants in growth patterns and proton translocation across the plasma membrane of the tuber cells.     

The effect of melamine salt of bis(oxymethyl)phosphinic acid (melafen) on the growth processes and cytoplasmic membrane function in potato tuber cells

The effects of a new synthetic growth regulator, preparation melafen, on the growth processes in potato plant tubers and the H+ -ATPase activity in cell plasmalemma were studied. It was demonstrated that melafen could both stimulate and inhibit the growth of potato tubers depending on its concentration and the physiological state of the tubers. It is likely that one of the manifestations of melafen action is its influence on the division and extension of apical meristem cells. The growth stimulation caused by melafen is connected with modifications of the plasmalemma of potato tuber cells, namely, the activation of H+ -ATPase and increase in the membrane proton permeability.     

Protocol of a prospective study on the diagnostic value of transcranial duplex scanning of the substantia nigra in patients with parkinsonian symptoms

Background  

Parkinson's disease (PD) is the second most common neurodegenerative disorder. As there is no definitive diagnostic test, its diagnosis is based on clinical criteria. Recently transcranial duplex scanning (TCD) of the substantia nigra in the brainstem has been proposed as an instrument to diagnose PD. We and others have found that TCD scanning of substantia nigra duplex is a relatively accurate diagnostic instrument in patients with parkinsonian symptoms. However, all studies on TCD so far have involved well-defined, later-stage PD patients, which will obviously lead to an overestimate of the diagnostic accuracy of TCD.     

The effect of thaumatin gene overexpression on the properties of H(+)-ATPase from the plasmalemma of potato tuber cells

The introduction of the thaumatin gene into potato plants was accompanied by a decrease in the activity of H(+)-ATPase in the plasmalemma (PL) of tuber cells. When tubers were released from dormancy, the enzyme was activated in the tuber cells of both the original and transgenic plants. Experiments performed in vitro demonstrated that sensitivities to ambiol (AM) and jasmonic acid (JA) of H(+)-ATPase in the PL of tubers from the original plants were lower after the release from a period of deep dormancy. In preparations from the tubers of transgenic plants, the situation was reversed. The differences between the activities of H(+)-ATPase in the PL preparations produced from the original and transgenic tubers that sprouted under the action of AM and JA were detected. Thus, the overexpression of the thaumatin gene in potato plants changed the properties of H(+)-ATPase from PL.     

The conduction of protons in different stereoisomers of dioxolane-linked gramicidin A channels

Two different stereoisomers of the dioxolane-linked gramicidin A (gA) channels were individually synthesized (the SS and RR dimers;. Science. 244:813-817). The structural differences between these dimers arise from different chiralities within the dioxolane linker. The SS dimer mimics the helicity and the inter- and intramolecular hydrogen bonding of the monomer-monomer association of gA's. In contrast, there is a significant disruption of the helicity and hydrogen bonding pattern of the ion channel in the RR dimer. Single ion channels formed by the SS and RR dimers in planar lipid bilayers have different proton transport properties. The lipid environment in which the different dimers are reconstituted also has significant effects on single-channel proton conductance (g(H)). g(H) in the SS dimer is about 2-4 times as large as in the RR. In phospholipid bilayers with 1 M [H(+)](bulk), the current-voltage (I-V) relationship of the SS dimer is sublinear. Under identical experimental conditions, the I-V plot of the RR dimer is supralinear (S-shaped). In glycerylmonooleate bilayers with 1 M [H(+)](bulk), both the SS and RR dimers have a supralinear I-V plot. Consistent with results previously published (. Biophys. J. 73:2489-2502), the SS dimer is stable in lipid bilayers and has fast closures. In contrast, the open state of the RR channel has closed states that can last a few seconds, and the channel eventually inactivates into a closed state in either phospholipid or glycerylmonooleate bilayers. It is concluded that the water dynamics inside the pore as related to proton wire transfer is significantly different in the RR and SS dimers. Different physical mechanisms that could account for this hypothesis are discussed. The gating of the synthetic gA dimers seems to depend on the conformation of the dioxolane link between gA's. The experimental results provide an important framework for a detailed investigation at the atomic level of proton conduction in different and relatively simple ion channel structures.