Why do flowers close at night? Experiments with the Lesser celandine Ficaria verna Huds (Ranunculaceae)

Although the mechanisms of flower closure have been widely studied, the evolutionary processes underlying this phenomenon are largely unknown. Open flowers may face a trade‐off between successful pollination/pollen transfer on the one hand and destruction of reproductive organs caused by exogenous factors (e.g. rain) on the other. The cost of flower opening was investigated by treating flowers of the Lesser celandine both in the field and under laboratory conditions. No differences were found in anthesis and reproductive success between treated and untreated flowers in any of the experiments. However, treated flowers in the field experiment suffered from significantly higher predation rates, in all probability caused by slugs and roe deer. It is suggested that flower closure may decrease the rate of being eaten by some herbivores, particularly in early‐spring plants that flower at times of insufficient food availability.     

Methods for obtaining more complete species lists in surveys of lichen biodiversity

We tested two methods to obtain more complete species inventories in surveys of lichen biodiversity. The first was to employ eight lichenologists (all experienced, some specialists) acting as individuals in parallel in a competitive survey. The second was to organize those lichenologists into two competing teams. We show that overall recorded biodiversity is distinctly higher than the part of lichen biodiversity recorded by each single lichenologist (45–66%) or team (79–83%). Use of these methods in a survey of epiphytic and epixylic lichens resulted in a list containing 112 species in 1 ha, 192 species in 12.5 ha and 212 species for 30 km² of lowland floodplain old‐growth forest in southeastern Czech Republic. Eleven recorded species are new to the country; four are rediscovered after more than 50 years. In comparison, few previous surveys of mixed montane forests in the same region have yielded more than 200 species, even though it is certain that those forests have greater lichen diversity than our lowland forest.     

Local adaptation of annual weed populations to habitats differing in disturbance regime

Plants have evolved several strategies to cope with disturbance, and one strategy is tolerance. In tolerance, plants store resources (meristems, carbohydrates) so that they can resprout after disturbance and thereby compensate to some degree for losses. Because tolerance is costly (it occurs at the expense of current growth), we can expect adaptation to the local disturbance regime. In this study, we determined whether populations of a common European annual weed, Euphorbia peplus, are adapted to the local disturbance regime. We hypothesized that the tolerance and hence compensation for losses in seed and biomass production after experimental damage are greater in plants from more severely disturbed than from less severely disturbed populations. We also hypothesized that transgenerational effects can alter adaptation. We found that compensation for biomass loss to damage was greater for plants from more severely disturbed habitats than for plants from less severely disturbed habitats. This, however, was not at the expense of growth before damage because plants from both disturbance regimes did not show differences when not damaged. Transgenerational effects played a positive role in adaptation to disturbance during germination and maturity. We conclude that local adaptation together with transgenerational effects have evolved in more severely disturbed populations but not in less severely disturbed populations of E. peplus.     

Tertiary structure stabilization promotes hairpin ribozyme ligation.

The hairpin ribozyme catalyzes a reversible RNA cleavage reaction that participates in processing intermediates of viral satellite RNA replication in plants. A minimal hairpin ribozyme consists of two helix-loop-helix segments. These segments associate noncoaxially in the active folded structure in a way that brings catalytically important loop nucleotides into close proximity. The hairpin ribozyme in the satellite RNA of Tobacco Ringspot Virus assembles in the context of a four-way helical junction. Recent physical characterization of hairpin ribozyme structures using fluorescence resonance energy transfer demonstrated enhanced stability of the folded structure in the context of a four-way helical junction compared to minimal hairpin ribozyme variants. Analysis of the functional consequences of this modification of the helical junction has revealed two changes in the hairpin ribozyme kinetic mechanism. First, ribozymes with a four-way helical junction bind 3' cleavage products with much higher affinity than minimal hairpin ribozymes, evidence that tertiary interactions within the folded structure contribute to product binding energy. Second, the balance between ligation and cleavage shifts in favor of ligation. The enhanced ligation activity of hairpin ribozymes that contain a four-way helical junction supports the notion that tertiary structure stability is a major determinant of the hairpin ribozyme proficiency as a ligase and illustrates the link between RNA structure and biological function.     

Application of the small-angle x-ray scattering technique for the study of two-step equilibrium enzyme-substrate interactions

The small-angle x-ray scattering (SAXS) technique is used for the investigation of two-stage equilibrium macromolecular interactions of the enzyme-substrate type in solution. Experimental procedures and methods of analyzing the data obtained from SAXS have been elaborated. The algorithm for the data analysis allows one to determine the stoichiometric, equilibrium, and structural parameters of the enzyme-substrate complexes obtained. The thermodynamic characteristics for the formation of complexes of double-stranded oligonucleotide with Eco dam methyltransferase (MTase) have been determined and demonstrate a high cooperativity of MTase binding when the ternary complex containing the dimeric enzyme is formed. The structural parameters (R_g, R_c, semiaxes) have been determined for free enzyme and polynucleotides and of enzyme-substrate complexes, indicating structural rearrangements of the enzyme in the interaction with substrates. © 1996 John Wiley & Sons, Inc.     

Melatonin promotes and synchronizes protein synthesis in hepatocyte culture from old rats

The effect of 1 to 1000 nM melatonin was studied on daily cultures of rat hepatocytes on slides in serum-free medium. The minimum melatonin concentration (1 nM) proved to synchronize protein synthesis in asynchronous sparse cultures of hepatocytes from rats of different age, and a circahoralian rhythm of protein synthesis was revealed in them. In dense weekly synchronous hepatocytes from old rats (2.5 years old with the weight of about 600 g), melatonin improved cell synchronization to the level of young animals. Melatonin treatment increased the mean rate of protein synthesis in rats of different age.     

Signaling factors of self-organization of protein synthesis rhythm in hepatocyte cultures—gangliosides and catecholamines function independently

Considering the data on the low level of self-organization (self-synchronization) of protein synthesis rhythm in aging, we studied the possible interference of the signaling factors of self-organization, gangliosides and catecholamines, as well as catecholamine reception. Experiments were carried out on primary cultures of rat hepatocytes on slides. Inhibited ganglioside synthesis did not prevent the organization of protein synthesis rhythm by the α-adrenomimetic agent phenylephrine. Upon the blockade of α-receptors by prazosin, the protein synthesis rhythm was observed after the exposure to gangliosides. α-Adrenolytic agents prazosin and benoxathian abolished the synchronizing effect of the β-adrenomimetic isoproterenol. A mixture of α-and β-adrenomimetic agents inhibited the protein synthesis rhythm-organizing effect of noradrenaline. Thus, the signaling molecules of self-organization of protein synthesis function independently via specific receptors.     

Survival of Bifidobacterium animalis DN-173 010 in the faecal microbiota after administration in lyophilised form or in fermented product - a randomised study in healthy adults

The survival of Bifidobacterium animalis strain DN-173 010 was assessed after its ingestion in a fermented product or in a lyophilised form. Twelve healthy subjects were included in a randomised, open study with 2 parallel groups. The composition and activities of the faecal microbiota were monitored before (10-day baseline step), during (1-week product administration step) and after (10-day follow-up step) the ingestion of 1 of the 2 products. A colony immunoblotting method, fluorescent in situ hybridisation with group-specific DNA probes, and temporal temperature gradient gel electrophoresis using group-specific primers were carried out to compare survival of B. animalis strain DN-173 010 after ingestion of the 2 products, together with analyses of enzyme activities and faecal metabolites. At the end of the supplementation step, the mean number of B. animalis DN-173 010 quantified by immunodetection in the faeces of 5 of 6 subjects in each treatment group was >/=10(8) colony-forming units/g faeces. These numbers corresponded to an average survival of 22% for the lyophilised form and 20% for the fermented product. At the same step, the PCR temporal temperature gradient gel electrophoresis profiles showed a double band corresponding to the B. animalis DN-173 010 pattern for 11 subjects. No major modification was observed during the trial in either the dominant members of the faecal microbiota assessed by fluorescent in situ hybridisation or their activities. In conclusion, we show that the lyophilised form of B. animalis DN-173 010 survives transit and could represent a more convenient form to administer for long-term clinical trials.     

Diversity of life strategies and population structure of Kamchatka mykiss <Emphasis Type="Italic">Parasalmo mykiss</Emphasis> in the ecosystems of small salmon rivers of various types

The diversity of life strategies and population structure of Kamchatka mykiss Parasalmo mykiss in the ecosystems of small salmon rivers of various types are considered. Hydrogeomorphological differences of the model rivers Kol and Kekhta are found. The preferred habitats are investigated, and their area is determined. It is shown that, in the Kol River, the mykiss having a resident life strategy predominated, and in the Kekhta River—the mykiss with migratory strategy. The key parameter controlling the prevalence of life strategies in each river is the ratio of the area of spawning grounds to the area of feeding grounds and their productivity. The hypothesis is confirmed that, in complex river systems, due to the diversity of biotopes and a higher productivity, the food resources are sufficient for maturation of mykiss and for realization of the resident life strategy (without migration to the sea for feeding). In small rivers of the channel type with their insufficient food resources, the specimens having a migratory life strategy prevail.     

Micropropagation of Wild Service Tree (Sorbus torminalis [L.] Crantz): The Regulative Role of Different Aromatic Cytokinins During Organogenesis

The influences of three different aromatic cytokinin derivatives [6-benzylaminopurine, meta-topolin, and 6-(3-methoxybenzylamino)purine-9-ß-D-ribofuranoside (MeOBAPR)] on in vitro multiplication and rhizogenesis of the wild service tree (Sorbus torminalis [L.] Crantz) were compared. The highest micropropagation rate (24 new shoots per explant after 3 months of cultivation) was achieved on media containing BAP. On the other hand, the best rooting microcuttings were those multiplied on a medium containing MeoBAPR. To compare these results with the levels of endogenous cytokinins in multiplied explants, a newly developed UPLC-ESI(+)-MS/MS method was used to determine levels of 50 cytokinin metabolites in explants cultivated 12 weeks on media supplemented by BAP and of the two other aromatic cytokinin analogs used. Several significant differences among the levels of endogenous cytokinins, extracted from the explants, were found. The concentration of BAP9G, an important metabolite suspected to be responsible for inhibition of rooting and acclimatization problems of newly formed plantlets, was found to be the highest in microcuttings grown on media supplemented with BAP. This agrees well with the results of our rooting experiments; the lowest percentages of rooted plantlets 6 weeks after transferring shoots on rooting medium were present on explants multiplied on BAP. In contrast, BAP was still the most effective for the induction of bud formation on primary explants. Levels of the most active endogenous isoprenoid cytokinins, tZ, tZR, and iPR, as well as O-glucosides were also suppressed in explants grown on BAP compared with those of explants treated with other cytokinin derivatives. This may be the result of a very high BAP uptake into the explants grown on this cytokinin. On the other hand, endogenous concentrations of cis-zeatin derivatives as well as dihydrozeatin derivatives were not affected. Differences in the production of another plant hormone, ethylene, that plays an important role in controlling organogenesis in tissue culture, were also observed among S. torminalis plantlets grown in vitro on media containing different cytokinins tested. The highest ethylene levels were detected in the vessels containing media supplemented with mT. They were two to four times higher compared with the production by the S. torminalis explants cultivated on other media used. Finally, the levels of free IAA were also determined in the explants. S. torminalis plantlets grown on media containing BAP contained the lowest level of auxin, which is again in good agreement with their loss of rooting capacity. The results found in this study about optimal plant hormone concentrations may be used to improve in vitro rooting efficiency of the wild service tree and possibly also of other plant species.