Targeting polyketide synthase gene pool within actinomycetes: new degenerate primers

Natural products provide a unique element of molecular diversity and biological functionality and they are still indispensable for drug discovery. The polyketides, comprising a large and structurally diverse family of bioactive natural products, have been isolated from a group of mycelia-forming Gram-positive microorganisms, the actinomycetes. Relatively high amino acid sequence identity of the actinomycetes type I polyketide synthases (PKSs) was used to design three degenerate primer pairs for homology-based PCR detection of novel PKS genes, with particular interest into PKSs involved in biosynthesis of immunosuppressive-like metabolites. The stepdown PCR method, described here, enables fast insight into the PKS arsenal within actinomycetes. Designed primers and stepdown PCR were applied for the analysis of two natural isolates, Streptomyces sp. strains NP13 and MS405. Sequence analysis of chosen clones revealed the presence of two distinctive sequences in strain Streptomyces sp. NP13, but only one of these showed homology to PKS-related sequences. On analysing PCR amplicons derived from Streptomyces sp. strain MS405, three different PKS-related sequences were identified demonstrating a potential of designed primers to target PKS gene pool within single organism.     

Effect of inorganic nutrition on the formation of lateral roots in Dracaena fragrans Ker-Gawl cultured in vitro

The effect of inorganic nutrition on the formation of lateral roots has been studied in Dracaena fragrans Ker-Gawl cultured in vitro. MS macronutrient salts imposed inhibition of lateral root formation in 95% of primary adventitious roots which could be overcome by:     

Seasonal dynamics of foliar antioxidative enzymes and total anthocyanins in natural populations of Iris pumila L.

Aims Plants in their natural habitats frequently cope with a multitude of abiotic stresses, such as high light intensity, extreme temperatures and water deficit, which often co-occur during periods of drought, especially in semi-arid and arid regions. Exposure of plants to stressful environmental conditions usually induce overproduction of reactive oxygen species (ROS) that, as highly toxic derivatives of O 2, can assault all cell macromolecules, leading to the disruption of cellular homeostasis and, consequently, the uncoupling of major metabolic processes, the photosynthesis and photorespiration. In order to minimize ROS-mediated cellular damage, plants have evolved highly efficient antioxidative defense systems that include both enzymatic and non-enzymatic components. Since abiotic stress can also operate as a strong evolutionary force that shapes adaptations in natural plant populations, the aim of this study was to examine the seasonal variation patterns of two distinct antioxidative systems, ROS-scavenging enzymes and anthocyanin pigments, in the leaf tissue of a steppe plant, Iris pumila, as expressed under contrasting light conditions that the species regularly experiences in the wild.Methods We selected two natural populations of I.pumila inhabiting the alternative radiation environments in the Deliblato Sands, a sun-exposed dune site and a woodland understory. The specific activity of three antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) and the content of total anthocyanins were examined in leaves of I.pumila plants collected from each of the 31 Iris clones (17 in the exposed population and 14 in the shaded population) once during each of the three seasons, spring, summer and autumn in 2004. Specifically, a fully expanded leaf was cut from each clonal plant between 15:00 and 16:00 h, immediately frozen in liquid nitrogen and stored at ?70°C until preparation.Important findings Generally, all three antioxidative enzymes were up-regulated in summer-harvested leaves compared to their spring or autumn counterparts, as was observed for the concentration of foliar anthocyanins, indicating that strengthening of antioxidant systems was the key mechanism for long-term acclimatization of I.pumila plants to stressful environmental conditions within their natural ecological niches. When plants from contrasting radiation environments were compared, SOD and CAT activities appeared to be greater in shade-exposed than in sun-exposed leaves. Conversely, POD activity and the content of foliar anthocyanins were notably higher in foliage experiencing full sunlight relative to those developed under vegetation canopy, suggesting the synergistic function of these two molecules in protecting leaf cells against photoinhibitory and photooxidative effects of strong light.     

Linking oxidative and salinity stress tolerance in barley: can root antioxidant enzyme activity be used as a measure of stress tolerance?

Aims

A causal relationship between salinity and oxidative stress tolerance and a suitability of using root antioxidant activity as a biochemical marker for salinity tolerance in barley was investigated.

Methods

Net ion fluxes were measured from the mature zone of excised roots of two barley varieties contrasting in their salinity tolerance using non-invasive MIFE technique in response to acute and prolonged salinity treatment. These changes were correlated with activity of major antioxidant enzymes; ascorbate peroxidase, catalase, and superoxide dismutase.

Results

It was found that genotypic difference in salinity tolerance was largely independent of root integrity, and observed not only for short-term but also long-term NaCl exposures. Higher K⁺ retention ability (and, hence, salinity tolerance) positively correlated with oxidative stress tolerance. At the same time, antioxidant activities were constitutively higher in a sensitive but not tolerant variety, and no correlation was found between SOD activity and salinity tolerance index during large-scale screening.

Conclusion

Although salinity tolerance in barley correlates with its oxidative stress tolerance, higher antioxidant activity at one particular time does not correlate with salinity tolerance and, as such, cannot be used as a biochemical marker in barley screening programs.     

Morphological features and isoenzyme characterization of endosymbiotic algae from green hydra

Symbiotic associations are of a wide significance in evolution and biodiversity. Green hydra is a typical example of endosymbiosis. In its gastrodermal myoepithelial cells, it harbors individuals of unicellular green algae. Morphological characteristics of isolated algae determined by light and electron microscopy are presented. Cytological morphometric parameters (cell area, cell radius, chloroplast area) of isolated algae from green hydra (Cx), as well as from reference species Chlorella kessleri (Ck) and Chlorella vulgaris (Cv), revealed similarity between the isolated endosymbiont and C. kessleri. Isoenzyme patterns of esterase (EST), peroxidase (POX), and catalase (CAT) were used for the investigation of genetic variability in endosymbiotic algae isolated from green hydra. Out of 14 EST isoenzymes observed in Cx species, 9 were expressed in the Cx sample. Results of the EST isoenzyme analysis indicated a higher degree of similarity between Cx and Cv than between Cx and Ck. Due to much higher heterogeneity, EST isoenzymes seem to be more suitable genetic markers for identification of different Chlorella species than CAT and POX isoenzymes. Results obtained suggest that symbiogenesis in green hydra has probably not been terminated yet.