Determination of the adenine-specific methylase recognition site of Shigella sonnei 47, using hydrazinolysis with subsequent separation of purine oligonucleotides by thin layer chromatography on DEAE- cellulose

A procedure for separation of oligopurine blocks of different length and composition by two-dimensional thin layer chromatography on DEAE-cellulose plates has been developed. This method allows a comparative analysis of the purine isostich content in the DNAs of various origin. In case of methylated DNA, the method permits to compare the substrate specificity of different enzymes responsible for the adenine residue methylation in the DNA. In combination with enzymatic treatment of labeled methylated isostichs, the method described can be used for the deciphering of the methylated sequences as well as for constructing, in a number of cases, the recognition site of adenine-specific methylases. Thus, it was demonstrated that methylase SsoI recognizes the 5...G-A-A-T-T-C ... 3' sequence and methylates its adenine residue nearest to the 5'-end.     

Is intensity of plant root mycorrhizal colonization a good proxy for plant growth rate,dominance and decomposition in nutrient poor conditions?

Questions

Mycorrhizae may be a key element of plant nutritional strategies and of carbon and nutrient cycling. Recent research suggests that in natural conditions, intensity of mycorrhizal colonization should be considered an important plant feature. How are inter‐specific variations in mycorrhizal colonization rate, plant relative growth rate (RGR ) and leaf litter decomposability related? Is (arbuscular) mycorrhizal colonization linked to the dominance of plant species in nutrient‐stressed ecosystems?

Location

Teberda State Biosphere Reserve, northwest Caucasus, Russia.

Methods

We measured plant RGR under mycorrhizal limitation and under natural nutrition conditions, together with leaf litter decomposability and field intensity of mycorrhizal colonization across a wide range of plant species, typical for alpine communities of European mountains. We applied regression analysis to test whether the intensity of mycorrhizal colonization is a good predictor of RGR and decomposition rate, and tested how these traits predict plant dominance in communities.

Results

Forb species with a high level of field mycorrhizal colonization had lower RGR under nutritional and mycorrhizal limitation, while grasses were unaffected. Litter decomposition rate was not related to the intensity of mycorrhizal colonization. Dominant species mostly had a higher level of mycorrhizal colonization and lower RGR without mycorrhizal colonization than subordinate species, implying that they were more dependent on mycorrhizal symbionts. There were no differences in litter decomposability.

Conclusions

In alpine herbaceous plant communities dominated by arbuscular mycorrhizae, nutrient dynamics are to a large extent controlled by mycorrhizal symbiosis. Intensity of mycorrhizal colonization is a negative predictor for whole plant RGR . Our study highlights the importance of mycorrhizal colonization as a key trait underpinning the role of plant species in carbon and nutrient dynamics in nutrient‐limited herbaceous plant communities.

     

Temperature and Photoperiodic Control of Development in the Green Shield Bug Palomena prasina (L.) (Heteroptera,Pentatomidae) in Leningrad Province

Thermal reaction norms for development (the lower temperature threshold, temperature sensitivity, and sum of degree-days) can show phenotypic plasticity in response to a combination of ecological factors. The goal of this study was to evaluate the degree of plasticity of the thermal reaction norms for development under different photoperiodic conditions in the green shield bug Palomena prasina. Experiments were conducted in 2015 and 2016. In 2015, two photoperiodic regimens (12 and 22 h of light per day) and five constant temperatures (20, 22, 24, 26, and 28°C) were used; a lower temperature of 16°C was added in 2016. There were no differences in the egg developmental time between the two photoperiodic conditions and across the two experiments. Under the short-day photoperiodic regimen, nymphal development was faster at all the temperatures and was characterized by greater thermal sensitivity and a higher temperature threshold than under the long-day photoperiod. Besides, the relationship between the developmental rate and temperature deviated from linearity at 26 and 28°C under short-day conditions. The adaptive nature of the observed nymphal response to photoperiodic conditions was confirmed by our phenological observations and an outdoor cage experiment. The adult body mass slightly increased with rising temperature under short-day conditions but did not depend on the temperature under long-day ones. Females were larger than males, and both sexes had a greater body mass under long-day conditions than under short-day ones. In 2015, the eggs for experiments were collected before mid-July, almost a month later than in 2016. Nymphs that hatched from the later eggs (in 2015) had significantly higher relative growth rates than the early-season nymphs (in 2016) at 20, 22, and 24°C under both photoperiods. This discrepancy between years was probably related to the maternal effect, namely, the difference in the female physiological age.

     

E. coli multidrug transporter MdfA is a monomer

MdfA is a 410-residue-long secondary multidrug transporter from E. coli. Cells expressing MdfA from a multicopy plasmid exhibit resistance against a diverse group of toxic compounds, including neutral and cationic ones, because of active multidrug export. As a prerequisite for high-resolution structural studies and a better understanding of the mechanism of substrate recognition and translocation by MdfA, we investigated its biochemical properties and overall structural characteristics. To this end, we purified the beta-dodecyl maltopyranoside (DDM)-solubilized protein using a 6-His tag and metal affinity chromatography, and size exclusion chromatography (SE-HPLC). Purified MdfA was analyzed for its DDM and phospholipid (PL) content, and tetraphenylphosphonium (TPP+)-binding activity. The results are consistent with MdfA being an active monomer in DDM solution. Furthermore, an investigation of two-dimensional crystals by electron crystallography and 3D reconstruction lent support to the notion that MdfA may also be monomeric in reconstituted proteoliposomes.     

The influence of temperature transfers on subsequent growth of the European peacock butterfly <Emphasis Type="Italic">Inachis io</Emphasis> (Lepidoptera,Nymphalidae)

The study addresses the effects of higher and lower temperatures experienced during the early stages of ontogeny on the parameters of the subsequent growth and development of the butterfly Inachis io. The caterpillars reared at a temperature of 22°С during instars I–III and then transferred to lower temperatures (16, 18, and 20°C) had higher instantaneous growth rates, larger body mass, and shorter duration of instars IV and V than the caterpillars that developed permanently at these lower temperatures. Vice versa, the caterpillars reared at 16°С during instars I–III and transferred to higher temperatures (18, 20, and 22°C) developed and grew slower than the caterpillars reared at these higher temperatures from the beginning. Possible physiological mechanisms underlying the observed phenomena and their ecological consequences are discussed. Our results are compared with the previously published data on fish—another group of ectothermic organisms, on which similar experiments have been carried out.     

Plant species traits are the predominant control on litter decomposition rates within biomes worldwide

Worldwide decomposition rates depend both on climate and the legacy of plant functional traits as litter quality. To quantify the degree to which functional differentiation among species affects their litter decomposition rates, we brought together leaf trait and litter mass loss data for 818 species from 66 decomposition experiments on six continents. We show that: (i) the magnitude of species-driven differences is much larger than previously thought and greater than climate-driven variation; (ii) the decomposability of a species' litter is consistently correlated with that species' ecological strategy within different ecosystems globally, representing a new connection between whole plant carbon strategy and biogeochemical cycling. This connection between plant strategies and decomposability is crucial for both understanding vegetation-soil feedbacks, and for improving forecasts of the global carbon cycle.     

Trapping of human DNA topoisomerase I by DNA structures mimicking intermediates of DNA repair

In this report we show that human DNA Topoisomerase I (Top1) forms DNA-protein adducts with nicked and gapped DNA structures lacking a conventional Top1 cleavage site. The radioactively labeled crosslinking products were identified by SDS-gel electrophoresis. The chemical structure of the groups at 5' or 3' end of the nick does not have an effect on the formation of these covalent adducts. Therefore, all kinds of nicks, either directly induced by ionizing radiation or reactive oxygen species or indirectly induced in the course of base excision repair (BER) are targets for Top1 that competes with BER proteins and other nick-sensors. Top1-DNA covalent adducts formed in cells exposed to DNA damaging agents can promote genetic instability.     

Photoperiod-temperature interaction--a new form of seasonal control of growth and development in insects and in particular carabid beetle, Amara communis (coleoptera: carabidae)

Amara communis larvae are found to develop significantly faster and have higher growth rate at short-day (12 h) as compared to long-day (22 h) photoperiods under all temperatures (16, 18, 20 and 22 degrees C) used. The coefficient of linear regression of larval development rate on temperature was significantly higher at short days than at long days. At that thermal developmental thresholds appeared similar at both photoperiods. Body weight of young beetles reared under different photoperiods was just the same. Thus, the photoperiodic effect does not simply accelerate or retard insect development, but modifies their thermal reaction norm. Under short days larval development becomes faster and more temperature dependent, which ensures the timely completion of the development at the end of summer. The analysis of data from literature allowed us to find photoperiodic modification of thermal requirements for development in 5 insect orders--Orthoptera, Heteroptera, Coleoptera, Lepidoptera, Diptera. Modification may result in significant changes in the slope of the regression line and hence in the sum of degree-days and thermal developmental threshold. Consequently, during summer under the influence of changing day-length the thermal requirements for development in many insects gradually vary, which may have adaptive significance. Thus, the photoperiodic modification of thermal reaction norm acts as a specific form of seasonal control of insect development.