Chromosome number and DNA content of tobacco cells adapted to NaCl

Cultured tobacco (Nicotiana tabacum L. cv. Wisconsin 38) cells were found to have altered DNA contents and chromosome numbers after adaptation to NaCl. Cells adapted to 428 mM NaCl were predominately hexaploid compared to the normal tetraploid 2N(2C)=4X=48 chromosome number of unadapted cells. Enrichment of the cell population for hexaploid cells occurred only after exposure to higher NaCl (428 mM), not lower levels of NaCl (171 mM). The majority of adapted cells remain hexaploid for at least 25 cell generations after removal from NaCl exposure. Adapted cell populations were found to have fewer cells with highly polyploid (2N96) nuclei. Salt tolerance of hexaploid cells was not found to be significantly greater than that of tetraploid cells. Cells with higher ploidy levels were less salt tolerant. It is suggested that high levels of NaCl induce polyploidization and that exposure to NaCl selects against cells with very high ploidy levels.     

Activities of DNA polymerases and RNA polymerases detected in situ in growing and differentiating cells of root cortex

Summary Activities of DNA polymerases and RNA polymerases were studied by autoradiographic methods in growing and differentiating root cortex cells ofZea mays — a species in which endomitosis occurs — andTulipa kaufmanniana — in which this process does not occur. InTulipa kaufmanniana, the highest activity of DNA polymerase appears in the nuclei of meristematic zone during the S phase of the cell cycle. InZea mays, endomitotic replication of DNA occurs in all growth and differentiation zones and the activity of DNA polymerase in the nuclei is similar to that in the meristematic zone. In both species, nuclear RNA synthesis, measured with³H uridine incorporation, is highest in the meristematic zone and declines steadily with development. Activity of nuclear RNA polymerase is present in all developmental zones in both species and is similar to that in the meristematic zone.³H uridine incorporation into nucleoli decreases markedly in both species, whereas the activity of nucleolar RNA polymerase remains at a high level in all root segments inZea mays and decreases slightly inTulipa kaufmanniana.It is argued that the differences between the incorporation of³H uridine and that or³H UMP may be caused by a reduction of the pool of endogenous ribonucleoside triphosphates. Marked activities of DNA polymerase and RNA polymerase in cytoplasm are possibly related to the growth and division of plastids and mitochondria.     

Cis-regulatory elements used to control gene expression in plants

Plant biotechnology is a dynamically developing science, which comprises many fields of knowledge. Novel plant genetic engineering findings highly influence the improvement of industrial production. These findings mostly concern cis-regulatory elements, which are sequences controlling gene expression at all developmental stages. They comprise of promoters, enhancers, insulators and silencers, which are used to construct synthetic expression cassettes. Examples of most important cis-regulatory elements are reviewed in the present paper. Variability among core promoters content and distal promoter regions impedes evaluation of interactions between them during the artificial promoters construction. Synthetic promoters and artificial expression cassettes trigger a significant increase in gene expression level, better properties and quality of a product. Accumulating knowledge about gene promoters, cis sequences and their cooperating factors allows uniform expression systems and highly predictable results.     

Fight to the death: Arabidopsis thaliana defense response to fungal necrotrophic pathogens

Necrotrophic fungi, being the largest class of fungal plant pathogens, pose a serious economic problem to crop production. They are the cause of heavy losses in agriculture worldwide. Understanding the process of plant infection by necrotrophic fungi, including subtle interaction networks connecting such evolutionarily distinct organisms has recently been given high research priority. Such studies are now possible mainly because of the utility of the model plant Arabidopsis thaliana. A. thaliana has a sequenced genome and thousands of mutants available, allowing investigation of virtually all aspects of plant pathogenesis. This review focuses on morphological and molecular changes in A. thaliana, which occur during response to infection by necrotrophic fungi. These responses in relation to resistance and susceptibility of the plant will be discussed.     

Flower bud dipping or vacuum infiltration—two methods of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> transformation

The purpose of this work was to evaluate two methods (floral dip and vacuum infiltration) of in planta transformation of Arabidopsis thaliana. The key issue of this work is the identification of the developmental stages of A. thaliana flower buds subjected to agroinfection, optimal for the successful transformation. Histological tests performed after agroinfection made it possible to establish the patterns of a GUSPlus reporter gene expression in the examined plants and thus precisely define the range of flower developmental stages most appropriate for efficient transformation. Two plasmids, CAMBIA 1305.1 and CAMBIA 2301, were used. Verification of the transgenic nature of plants was carried out by detection of CaMV::gusA and CaMV::GUSPlus transgenes and their expression in transgenic plants by appropriate molecular and histochemical methods. For the flower dip transformation, three concentrations of Silwet L-77 surfactant and two inoculation times were tested. The most efficient treatment appeared to be 2-min-long flower bud inoculation and 400 μl/l surfactant (pCAMBIA 1305.1 − 1.73%; pCAMBIA 2301 − 2.01%). In the case of vacuum infiltration method, the highest efficiency of the transformation occurred when the inoculation time was 4 min (pCAMBIA 2301 − 1.55%; pCAMBIA 1305.1 − 1.37%). Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 619–628. This text was submitted by the authors in English.     

Elastin-like polypeptides as a promising family of genetically-engineered protein based polymers

Elastin-like polypeptides (ELP) are artificial, genetically encodable biopolymers, belonging to elastomeric proteins, which are widespread in a wide range of living organisms. They are composed of a repeating pentapeptide sequence Val–Pro–Gly–Xaa–Gly, where the guest residue (Xaa) can be any naturally occurring amino acid except proline. These polymers undergo reversible phase transition that can be triggered by various environmental stimuli, such as temperature, pH or ionic strength. This behavior depends greatly on the molecular weight, concentration of ELP in the solution and composition of the amino acids constituting ELPs. At a temperature below the inverse transition temperature (T_t), ELPs are soluble, but insoluble when the temperature exceeds T_t. Furthermore, this feature is retained even when ELP is fused to the protein of interest. These unique properties make ELP very useful for a wide variety of biomedical applications (e.g. protein purification, drug delivery etc.) and it can be expected that smart biopolymers will play a significant role in the development of most new materials and technologies. Here we present the structure and properties of thermally responsive elastin-like polypeptides with a particular emphasis on biomedical and biotechnological application.