Embryogenic cultures of plants are exposed to various stress factors both in vitro and during cryostorage. In order to safely include the plant material obtained by somatic embryogenesis in combination with cryopreservation for breeding programs, it is necessary to monitor its genetic stability. The aim of the present study was the assessment of somaclonal variation in plant material obtained from embryogenic cultures of Picea abies (L.) Karst. and P. omorika (Pancic) Purk. maintained in vitro or stored in liquid nitrogen by the pregrowth-dehydration method. The analysis of genetic confoimity with using microsatellite markers was performed on cotyledonary somatic embryos (CSE), germinating somatic embryos (GSE) and somatic seedlings (SS), obtained from tissues maintained in vitro or from recovered embryogenic tissues (ETc) and CSE obtained after cryopreservation. The analysis revealed changes in the DNA of somatic embryogenesis-derived plant material of both Picea spp. They were found in plant material from 8 out of 10 tested embryogenic lines of P abies and in 10 out of 19 embryogenic lines of P. omorika after in vitro culture. Changes were also detected in plant material obtained after cryopreservation. Somaclonal variation was observed in ETc and CSE of P omorika and at ETv stage of P abies. However, most of the changes were induced at the stage of somatic embryogenesis initiation. These results confirm the need for monitoring the genetic stability of plants obtained by somatic embryogenesis and after cryopreservation for both spruce species.

In vitro embryogenic callus is a critical factor for genetic transformation of rice, especially for indica varieties. In this study, we investigated the relationship between polyamines, including putrescine (Put), spermidine (Spd) and spermine (Spm), and callus browning, and we studied the effect of exogenous Put on callus regeneration and on the content of endogenous polyamines. In addition, the expression levels of arginine decarboxylase gene (Adcl) and S-adenosylmethionine decarboxylase gene (Samdc) in embryogenic callus were studied by quantitative Real-time PCR analysis. The results showed that the contents of endogenous Put and Spd in the browning callus were significantly lower than those in normal callus. Exogenous Put could effectively improve the growing state of callus of indica rice and enhance the development of embryogenic callus. The content of endogenous polyamines in embryogenic callus, especially Spd and Spm, was increased after addition of exogenous Put. Additionally, exogenous Put also had an obvious impact on the expression levels of Adcl but partial effect on the expression levels of Samdc gene. This study could increase the knowledge of both embryogenic callus induction and polyamine catabolism in callus in indica rice.

Ascorbic acid is a well-known antioxidant found in plants. The content of ascorbic acid was assayed using a normal phase European Pharmacopoeia HPLC method for ascorbic acid in medicinal products. The content of ascorbic acid in herbs was calculated in % for absolutely dry drug. Ascorbic acid was not detected in the roots of Primula ueris, in aerial parts it was detected in flowers (0.43 +/- 0.034%), in blades (1.43 +/- 0.11%) and petioles (1.56 +/- 0.12%). In fresh leaves collected at weekly intervals the content of ascorbic acid varied from 1.19 to 2.39%, being highest from mid-May to mid-June. The fresh leaves contained 2.35 +/- 0.18% of ascorbic acid and when frozen its content was quite stable for one year. The content of ascorbic acid in dried leaves decreased more than ten times in three months, in twelve months it was less than 1/20th of the initial level. Compared to the analyzed common fresh fruits and salads (n = 10) the fresh leaves of common cowslip contained considerably more ascorbic acid. Commercial orange juices could be recommended as the most convenient source of ascorbic acid (8.6-50.4 mg/100 ml); 1-5 glasses of orange juice could fulfill the recommended daily intake of vitamin C (60 mg).

Until recently, Festuca arietina was practically an unknown species in the flora of Eastern Europe. Such a situation can be treated as a consequence of insufficient studying of Festuca valesiaca group species in Eastern Europe and misinterpretation of the volume of some taxa. As a result of a complex study of F arietina populations from the territory of Ukraine (including the material from locus classicus), Belarus and Lithuania, original anatomy, morphology and molecular data were obtained. These data confirmed the taxonomical status of F arietina as a separate species. Eleven morphological and 12 anatomical characters, ITS1-5.8S-ITS2 cluster of nuclear ribosomal genes, as well as the models of secondary structure of ITS1 and ITS2 transcripts were studied in this approach. It was found for the first time that F arietina is hexaploid (6x = 42), which is distinguished from all the other narrow-leaved fescues by specific leaf anatomy as well as in ITS1-5.8S-ITS2 sequences. Molecular data indicating possible hybridogenous origin of F arietina, fall in line with the anatomical-morphological data and explain the tendency toward sclerenchyma strands fusion with formation of a continuous ring in F arietina, as well as E arietina ecological confinement to psammophyte biotopes.

Endopolyploidy is a condition of a cell containing reduplicated genetic material in its nucleus. Cells with the nuclei of different ploidy levels are often present within a single polysomatic organism. Endoreduplication is thus a modified cell cycle that omits cytokinesis and leads to chromatin replication in the endopolyploid cells. This study aimed to research the effect of salinity on endopolyploidy of Trifolium pratense and T. repens. Both species are important pasture legumes and belong to the genus Fabaceae with the well documented endopolyploidy occurence. Endopolyploidy levels in the seedlings treated with 0, 30, 60, 90 and 120 mM NaCl were investigated by flow cytometry. The seedling organs were evaluated during three ontogeny stages. The cytometric data plotted on a histogram showed the presence of 2C-16C nuclei in T. pratense and 2C-8C in T. repens. The hypothesis that salinity induces additional endocycles was not confirmed. Our results show that the distribution of nuclei among ploidy levels does not differ markedly between the treatment groups and the control ones. Additionally, only minor changes were observed among the endoreduplication indexes (EI) of plant organs after exposure to various salt concentrations. Endopolyploidy patterns within the salt-treated seedlings during ontogeny are similar to the controls. We suggest that endopolyploidy in Trifolium species is a conserved genetic trait, rather than an adaptation to salinity stress. The analyses of the roots of T'. pratense at stage III show that with the increased concentrations of NaCl the length of roots decreased, but no evident changes in endopolyploidy occured.

Girdling was applied to 5-year-old potted beech individuals of early, intermediate and late phenological forms to block assimilate export from leaves. Phloem severance caused accumulation of soluble carbohydrates and starch in leaves and increased the C/N ratio. While the hexose content increased continuously until the end of the experiment, the sucrose and starch contents peaked earlier, depending on the plant's phenological features. Different rates of chlorophyll degradation and H2O2 and TBARS (thiobarbituric acid-reactive substances) production in different phenological forms implied that phloem girdling was the source of oxidative stress and, depending on the phenological form, accelerated leaf senescence to different degrees. The variable rate of the increase in soluble carbohydrate and starch content, characteristic of the different phenological forms, had different modifying effects on the antioxidant activity in leaves. Compared with the early phenological form, the late form was characterized by a smaller increase in H2O2 and TBARS content and delayed and slowed chlorophyll and carotenoid degradation. In conjunction with the larger increase in the activity of antioxidant enzymes (catalase, ascorbate peroxidase and superoxide clismutase) induced by carbohydrate accumulation and slower carotenoid degradation, these changes led to the late form having greater resistance to oxidative stress and slower senescence.

Oilseed rape (Brassica napus L. ssp. oleifera Metzg) was the subject of the study in two forms: winter cv. 'Muller' (at the rosette stage - the first internode BBCH 30-31) and spring cv. `Feliks' (at the yellow bud stage BBCH 59). The main gas-exchange parameters, net photosynthetic rate (P-N) transpiration rate (E), stomatal conductance (g(s)), and intercellular CO2 concentration (Ci) were measured on leaves prior to the piercing and immediately after the short-term piercing. The effect of mechanical wounding revealed different progress of the gas exchange process for the two forms. Piecewise linear regression with the breakpoint estimation showed that the plants at the same age but at a different vegetal stage, manage mechanical leaf-piercing differently. The differences concerned the stomatal conductance and transpiration changes since for rosette leaves the process consisted of five intervals with a uniform direction, while for stem leaves-of five intervals with a fluctuating direction. These parameters got stabilized within a similar time (220 mins) for both forms. The process of net photosynthetic rate was altered by the plant stages. 'Muller' plants at the rosette stage demonstrated dependence of P-N on time in log-linear progression: y (P-N) = 8.01+ 2.73 log(10) (x t(2)); 7 < t(2) < 220; R-2 = 0.96. For stem leaves of Teliks' plants the process of transpiration, in terms of directions, was convergent with the process of photosynthesis. Those two processes were synchronized from 1st to 114th min of the test (r = 0.85; p < 0.001) in plants at the rosette stage and from 26th to 148th min in stem leaves (r = 0.95; p < 0.001).

Several parasitic plants are known to have acquired mitochondrial genes via a horizontal transfer from their hosts. However, mitochondrial gene transfer in this direction has not yet been found in the parasite-rich family Orobanchaceae. Based on a phylogenetic analysis of the mitochondrial atp6 gene in selected species of Orobanche s.l., we provide evidence of a host-to-parasite transfer of this gene in O. coerulescens, which is a Eurasiatic species that parasitises Artemisia (Asteraceae). We did not find the original Orobanche atp6 gene in this species, which suggests that it has been replaced by a gene that was acquired from Asteraceae. In addition, our data suggest the occurrence of a second HGT event in the atp6 sequence – from Asteraceae to Phelipanche. Our results support the view that the transfer of genetic material from hosts to parasites influences the mitochondrial genome evolution in the latter.

Genetic diversity is often considered a major determinant of long term population persistence and its potential to adapt to variable environmental conditions. The ability of populations to maintain their genetic diversity across generations seems to be a major prerequisite for their sustainability, which is particularly important for keystone forest tree species. However, little is known about genetic consequences of demographic alterations occurring during natural processes of ecological succession involving changes in the species composition. Using microsatellites, we investigated genetic diversity of adult and offspring generations in beech (Fagus sylvatica L.) and oak (Quercus robur L.) populations coexisting in a naturally established old-growth forest stand, showing some symptoms of ongoing ecological succession from oak- to beech- dominated forest. In general, adult generations of both species exhibited high levels of genetic diversity (0.657 for beech; 0.821 for oak), which, however, depended on the sets of selected genetic markers. Nevertheless, several symptoms such as differences in genetic diversity indices between generations, significant levels of inbreeding (up to 0.029) and low estimates of effective population size (48-80) confirmed the declining status of the oak population. On the other hand, the uniform distribution of genetic diversity indices across generations, low levels of inbreeding (0.004), low genetic differentiation among adults and offspring and, most importantly, large estimates of effective population size (119-716), all supported beech as a successive and successful tree species in the studied forest stand.