The rolC gene increases caffeoylquinic acid production in transformed artichoke cells.

Caffeoylquinic acids are found in artichokes, and they are currently considered important therapeutic or preventive agents for treating Alzheimer's disease and diabetes. We transformed artichoke [the cultivated cardoon or Cynara cardunculus var. altilis DC (Asteraceae)] with the rolC gene, which is a known inducer of secondary metabolism. High-performance liquid chromatography with UV and high-resolution mass spectrometry (HPLC-UV-HRMS) revealed that the predominant metabolites synthesized in the transgenic calli were 1,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, and chlorogenic acid. The rolC-transformed calli contained 1.5% caffeoylquinic acids by dry weight. The overall production of these metabolites was three times higher than that of the corresponding control calli. The enhancing effect of rolC remained stable over long-term cultivation.

[Genetic variation of the relict species Acanthopanax sessiliflorus (Rupr. et Maxim.) Seem. (Araliaceae) in Primorsky Krai].

Based on the analysis of 17 genes encoding the allozyme diversity of 12 enzyme systems, data were obtained on the genetic variation of a relict of the Tertiary flora, a valuable medicinal plant Acanthopanax sessiliflorus (Rupr. et Maxim.) Seem. (Araliaceae) in the Russian area of its habitat. Indicators of polymorphism for populations had rather high values on average (P95 = 42.4%, A = 1.55, H(o) = 0.211, and H(e) = 0.168), which are comparable with the known data for populations of A. sessiliflorus from the peninsula of Korea. The level of genetic diversity and its distribution among populations reflects the interaction of several factors, among which the most important are the historical past of the species, genetic drift, and the plasticity of the reproduction system. The obtained data can serve as a basis for the conservation of genetic resources of Far Eastern Araliaceae species.

[Genetic variation in six species of the genus Oxytropis DC. (Fabaceae) from Kamchatka Peninsula].

Using the isozyme analysis, genetic variations in six species of the genus Oxytropis DC. (Fabaceae) from Kamchatka was assessed. It was demonstrated that diploid species from the section Arctobia were characterized by a low level of variations typical of endemic plant species. At the same time, polyploid species from the Orobia section demonstrated very high values of the heterozygosity parameters (H(o) varied from 0.200 to 0.274). It has been suggested that the level of polymorphism of the oxytropes from Kamchatka was shaped as a result of the interaction of a number of factors, among which the most important are the ecological confinedness of the species, the specific features of the reproductive system, and gene drift. In the species of Orobia section, it is also the presence of the polyploid genome.

[Low level of allozyme polymorphism in relict aquatic plants of the Far East Nelumbo komarovii Grossh. and Euryale ferox Salisb].

Using allozyme analysis, genetic variation of two relict aquatic plants from Primorsky krai, Komarov lotus (Neliumbo komarovii Grossh.) and Gorgon plant (Euryale ferox Salisb.), was examined. The absence of allozyme variation in the Primorye populations of Neliumbo komarovii along with low polymorphism level in the population of Euryale ferox (P95 = 7.69; A = 1.07; Ho = 0.072; He = 0.038) was demonstrated. Since the data for the species examined are reported for the first time ever, the pheonotypes and genetic interpretation of the enzyme systems tested are presented. The izoenzyme profiles of N. komarovii were compared with the data reported for N. nucifera from China. The absence ofallozyme variation in N. komarovii, along with extremely low level of variation revealed for E. ferox, is discussed in association with the evolutionary histories of these species, their dispersal after the Pleistocene-Holocene cooling, and survival on this territory in range boundaries.

[Genetic diversity of a rare species Aristolochia contorta bungae (aristolochiaceae) in Primorsky Krai].

The herbaceous vine, twisted birthwort Aristolochia contorta Bunge, is a rare species listed in the Red Data Book of Primorsky krai (2008). On the northern boundary of its range (south of the Russian Far East), the species is represented by small isolated populations confined to the river drainages. Using allozyme analysis, genetic variation of nine natural populations ofA. contorta (247 accessions), which represented the main part of the species range in Russia, was examined. The values of genetic variation indices (P = 22.7%; A = 1.28; Ho = 0.129; He = 0.101) were low and comparable with the data obtained for other rare plants. The proportion of unique genotypes (G/N) and Simpson's genotypic diversity index (D) ranged from 0.32 to 0.64 and from 0.60 to 0.98, respectively. This means that A. contorta is characterized by sexual and asexual reproduction. Moreover, the ratio between these types of reproduction varied among the populations. Complete absence of inbreeding and excess of heterozygotes (F(IS) = -0.282), which was low probable in case of free mating, was observed. Evidently, clonal growth and (or) apomixis enables the species to maintain certain level of heterozygosity despite of small population sizes and non-regular gene exchange.

[Genetic variation of Oplopanax elatus (Nakai) Nakai (Araliaceae)].

Using allozyme analysis, genetic and genotypic variation of rare relict species, a member of one of ancient angiosperm families, Araliaceae, the clone-forming plant Oplopanax elatus (Nakai) Nakai, was evaluated. Electrophoretic separation of the enzymes is described, and genetic interpretation of the enzymes variation patterns is presented. The values of genetic variation indices obtained were low (P = 25%; A = 1.45: Ho = 0.131; He = 0.113) and comparable with the data reported for the rare plant species and the representatives of the family Araliaceae. The main factors responsible for the polymorphism level observed might be the evolutionary history of the species and gene drift. The level of genotypic diversity (G/N = 0.76; D = 0.97) was substantially higher, compared to the values reported for species with vegetative reproduction (D = 0.62). These results suggest certain contribution of the propagation by seeds in the formation of the species of interest.

[Characteristics of gene pools of three Pinus pumila (Pall.) Regel populations at the range margins].

Genetic variation of Siberian dwarf pine Pinus pumila (Pall.) Regel was characterized in three marginal populations in southwestern, southern and eastern parts of the natural species range (Transbaikalia, Primorye, Kamchatka) using isozyme analysis. Analysis involving 16 isozyme loci encoding ten enzyme systems was conducted. Our results confirm that P. pumila is among the most polymorphic species in the family Pinus. Three marginal populations exhibited high genetic variation (P95 = 68.8%, Ho = 0.247, He = 0.291). Populations heterogeneity and significantly high level of divergence in coniferous (F(ST) = 0.050, D(N) = 0.044) reflect their genetic originality. In summary, it was shown that the level of genetic variation characteristic for P. pumila in other parts of the not only is reproduced in the populations examined but even is close to maximum there.

[Genetic structure of natural populations of the relict species Aristolochia manshuriensis (Aristolochiaceae) in disturbed and intact habitats].

Genetic structure of natural populations of the rare relict plant Manchurian birthwort (Aristolochia manshuriensis Kom.) in the Russian part of its area was analyzed using allozyme markers. The studied A. manshuriensis populations differed in the degree of their intrapopulation differentiation. The populations Nezhinka and Anan'evka were more differentiated (F(ST) = 0.1209 and 0.0576, respectively); these populations are located close in the regions of intense economic activity and are exposed to the strongest anthropogenic impact. A low degree of differentiation was detected in the population Malaya Borisovka (F(ST) = 0.0393), localized to intact habitats. The overall heterogeneity test has demonstrated that the population Malaya Anan'evka, exposed at present to small anthropogenic stress yet growing in disturbed habitats, displays no differentiation. These results suggest that at least three populations are influenced by genetic drift connected with a decrease in the reproductive and effective population sizes, which is caused, in particular, by anthropogenic impact. A high level of genetic similarity between the A. manshuriensis populations in Primor'e is discussed in connection with the evolutionary history of this species.

[Genetic structure and differentiation of populations of the tetraploid species Oxytropis chankaensis (Fabaceae)].

The population genetic variation of the tetraploid species Oxytropis chankaensis Jurtz. (Fabaceae), a local endemic of the western coast of Khanka Lake (Primorye), was examined. Five populations were analyzed using 28 isozyme loci encoding 16 enzyme systems. Significant allelic heterogeneity among the populations was found for six out of twelve polymorphic loci. The heterozygosity of the samples (total sample size 294 plants) He = 0.301 was considerable higher than the mean values in populations of endemic species (0.076). Based on the results of this study, we identified two groups of O. chankaensis populations (southern and northern), in spite of the absence of marked hiatus between them. Of special interest is the population from Przhewalski Spit, which is a natural reserve of genetic diversity of the species and the putative center of formation of the autotetraploid O. chankaensis.

[Specific features of pollination in the Manchurian birthwort Aristolochia manshuriensis].

An analysis of pollination system in Aristolochia manshuriensis has shown that flower structure in this species is strictly adapted to cross-pollination, but the possibility of an autogamous or geitonogamous type of self-pollination with the involvement of insects is not excluded. The flowers of A. manshuriensis are most frequently visited by flies of the family Anthomyiidae, which markedly contribute to their pollination. Males account for 65% of pollinator insects collected from the flowers.

[Genetic variation and differentiation in population of Japanese emperor oak (Quercus dentata Thunb.) and Mongolian oak (quercus mongolica fisch. ex ledeb.) in the south of the Russian far east].

Allozyme variation of Japanese emperor oak Quercus dentata Thunb. and Mongolian oak Quercus mongolica Fisch. ex Lebed.) was examined in 11 populations of these species from southern Primorye. Analysis of 18 loci showed that in these populations, 66.7% of genes of Mongolian oak are polymorphic, the number of alleles per locus being 2.28. In Japanese emperor oak, these parameters were respectively 59.8 and 2.36%. The observed heterozygosity in both species did not differ from the expected values, constituting 0.198 in Japanese emperor oak and 0.161 in Maongolian oak. The Nei's distance between Quercus mongolica and Q. dentate was 0.053. The Japanese emperor oak and Mongolian oak populations formed separate clusters in an UPGMA dendrogram. Taking into account morphological and ecological isolation of the species, we suggest that the differences in gene frequencies may reflect selection acting in favor of adaptation to different growth conditions and counteracting between-species hybridization.

[Allozyme variation of the relict plant Aristolochia manshuriensis Kom. (Aristolochiaceae)].

Allozyme variation of a rare relict plant, birthwort Aristolochia manshuriensis Kom., was examined. The main parameters of genetic variation in natural populations of A. manshuriensis from the Anan'evka, Nezhinka, and Malaya Borisovka river basins (Primorskii Krai) were inferred from analysis of nine enzyme systems, presumably encoded by 18 loci. At 99% polymorphism criterion, 24.4% of A. manshuriensis loci were shown to be polymorphic. The mean number of alleles per locus was 1.24; the mean observed and expected heterozygosities, 0.12 and 0.10, respectively.

The Agrobacterium rhizogenes rolC-gene-induced somatic embryogenesis and shoot organogenesis in Panax ginseng transformed calluses.

Expression of the Agrobacterium rhizogenes rolC gene in Panax ginseng callus cells results in formation of tumors that are capable to form roots. The selection of non-root forming tumor clusters yielded the embryogenic 2c3 callus line, which formed somatic embryos and shoots independently of external growth factors. Although the 2c3 somatic embryos developed through a typical embryogenesis process, they terminated prematurely and repeatedly formed adventitious shoot meristems and embryo-like structures. A part of the shoots and somatic embryos formed enlarged and fasciated meristems. This is the first indication of the rolC gene embryogenic effect and, to our knowledge, the first indication that a single gene of non-plant origin can induce somatic embryogenesis in plants.

[High level of polymorphism and autotetraploid origin of rare endemic species of Fabaceae, Oxytropis chankaensis Jurtz., inferred from allozyme data]. / Vysokii uroven' polimorfiszma i avtotetrapolidnoe proiskhozhednie redkogo éndemicheskogo vida ostrolodochnika khankaiskogo Oxytropis chankaensis Jurtz. (Fabaceae): dannye allozimnogo analiza.

Using starch electrophoresis, we examined 19 enzyme systems presumably controlled by 35 loci in the rare endemic tetraploid species Oxytropis chankaensis Jurtz. (Fabaceae). Electrophoretic patterns and their genetic interpretation are presented. The isozyme data suggest tetrasomic inheritance in O. chankaensis. Three or four alleles at a particular locus were found in a number of individual plants, which indicate the autotetraploid origin of this species. Seventeen loci were shown to be polymorphic. As reliable gene markers for population systems, we recommend highly active polymorphic systems showing good allozyme separation (Ce-2, Gpi-2, Gpt-2, Idh-2, Lap, Mdh-2, and Mdh-3). Parameters of allozyme variability proved to be very high for a rare species with a restricted range: P = 48.6%, Ap = 3.06, Hob = 0.173.

[RAPD- and allozyme analysis of genetic variability of Panax ginseng C.A. Meyer and P. quinquefolius L]. / RAPD- i allozimnyi geneticheskoi izmenchivosti Panax ginseng S. A. Meyer i P. quinquefolius L.

Inter- and intraspecific variation of two ginseng species Panax ginseng and P. quinquefolius was estimated by studying 159 RAPD and 39 allozyme loci. Parameters of polymorphism and genetic diversity were determined and a tree was constructed to characterize the differences between individual plants, samples, and species. Genetic variation in P. ginseng proved to be lower than in P. quinquefolius. Gene diversity in the total P. ginseng sample was comparable with the mean expected heterozygosity of herbaceous plants. This suggests that wild P. ginseng plants in various areas of the currently fragmented natural habitat and cultivated plants of different origin have retained a significant proportion of their gene pool. The mean heterozygosity calculated per polymorphic locus for the RAPD phenotypes is similar to that of the allozyme loci and may be helpful in estimating gene diversity in populations of rare and endangered plant species.