Tamara I. Muzarok

Panax ginseng natural populations: their past, current state and perspectives.

AbstractAim:The mating system of Panax ginseng, genetics and ontogenetic structure of its natural populations of Primorye (Russia) were investigated.Methods:Genetic diversity was assessed using allozyme and the fluorescently based automated amplified fragment length polymorphism (AFLP) and simple sequence repeats (SSR) markers.Results:Total genetic diversity at species level is low with allozyme assay (0.023), and high with AFLP (0.255) and SSR (0.259) methods. It is observed within populations according to allozyme (>99%), AFLP (>85%), and SSR (>73%) assays. The indices of genetic variability distribution point out the re-colonization of the Sikhote-Alin by ginseng plants from southern refuges during the warming period in the early Holocene. The capability of ginseng plants to cross- and self-pollinate was shown and the assumption that Panax ginseng is a facultative apomictic plant was confirmed. The reproductive system of ginseng possesses high plasticity and stability of the fertilization process that help the species to survive in stress conditions. Disturbances caused by external or internal factors can be reduced due to the morphogenetic potential of ginseng ovule or apomictic embryo development. Analysis of life stages structure of ginseng populations demonstrates that all of them are not full-constituents because some life stages are absent or occur rarely.Conclusion:In all 3 populations, virgin and young generative individuals are predominant. This means that populations studied are viable and the reintroduction of natural ginseng population is possible yet.

Genetic variability and population structure of endangered Panax ginseng in the Russian Primorye.

BackgroundThe natural habitat of wild P. ginseng is currently found only in the Russian Primorye and the populations are extremely exhausted and require restoration. Analysis of the genetic diversity and population structure of an endangered species is a prerequisite for conservation. The present study aims to investigate the patterns and levels of genetic polymorphism and population structures of wild P. ginseng with the AFLP method to (1) estimate the level of genetic diversity in the P. ginseng populations in the Russian Primorsky Krai, (2) calculate the distribution of variability within a population and among populations and (3) examine the genetic relationship between the populations.MethodsGenetic variability and population structure of ten P. ginseng populations were investigated with Amplified Fragment Length Polymorphism (AFLP) markers. The genetic relationships among P. ginseng plants and populations were delineated.ResultsThe mean genetic variability within populations was high. The mean level of polymorphisms was 55.68% at the population level and 99.65% at the species level. The Shannons index ranged between 0.1602 and 0.3222 with an average of 0.2626 at the population level and 0.3967 at the species level. The analysis of molecular variances (AMOVA) showed a significant population structure in P. ginseng. The partition of genetic diversity with AMOVA suggested that the majority of the genetic variation (64.5%) was within populations of P. ginseng. The inter-population variability was approximately 36% of the total variability. The genetic relationships among P. ginseng plants and populations were reconstructed by Minimum Spanning tree (MS-tree) on the basis of Euclidean distances with ARLEQUIN and NTSYS, respectively. The MS-trees suggest that the southern Uss, Part and Nad populations may have promoted P. ginseng distribution throughout the Russian Primorye.ConclusionThe P. ginseng populations in the Russian Primorye are significant in genetic diversity. The high variability demonstrates that the current genetic resources of P. ginseng populations have not been exposed to depletion.

The production of class III plant peroxidases in transgenic callus cultures transformed with the rolB gene of Agrobacterium rhizogenes

The production of plant peroxidases by plant cell cultures is of great interest because of the potential for industrial applications. We used plant cell cultures overexpressing the rolB gene to produce increased amounts of plant class III peroxidases. The rolB gene ensured the stable and permanent activation of peroxidase activity in the transformed callus cultures of different plants. In particular, the total peroxidase activity in transformed Rubia cordifolia cells was increased 23-86-fold, and the abundance of the major peroxidase gene transcripts was increased 17-125-fold (depending on the level of rolB expression) compared with non-transformed control calli. The peroxidase-activating effect of rolB was greater than that of other peroxidase inducers, such as external stresses and methyl jasmonate.

Characterization of the Variability of Nucleoli in the Cells of Panax ginseng Meyer In Vivo and In Vitro

Results of karyological study of intact plants and some callus lines of Panax ginseng are presented. In the native plants of P. ginseng the nucleus with 1 nucleolus (90%) dominate, and nucleus with 2 nucleoli is rare. One nucleolar nucleus also dominate in interphase nuclei of cells of cultivated P. ginseng (from 2006), but we also found nucleus with 2 to 3 nucleoli in the same cell lines. Interphase nuclei of P. ginseng in long cultivated lines (from 1988) contain 1 to 9 nucleoli, with a predominance of nuclei containing from 3 to 4 nucleoli. It was shown that long-time cells (cultivated since 1988) had cytogenetic changes such as increase level of polyploid and aneuploid cells, increase of nucleoli number into interphase nucleus and decrease of nuclei/nucleoli ratio. These long-time cultivated cells had very low ginsenoside content.

Catechin production in cultured cells of Taxus cuspidata and Taxus baccata

The main polyphenols in callus and cell suspension cultures of Taxus cuspidata and T. baccata were (+)-catechin and (−)-epicatechin, while lignans, such as (+)-taxiresinol, (+)-isotaxiresinol, (+)-isolariciresinol and (−)-secoisolariciresinol, were present in trace amounts. T. cuspidata cells contained 1.7% (+)-catechin and 2.4% (−)-epicatechin on dry wt basis but when stimulated with methyl jasmonate produced 3.4% catechin and 5.2% epicatechin. These are the highest levels of these metabolites obtained in plant cell cultures.

Variation in the number of nucleoli and incomplete homogenization of 18S ribosomal DNA sequences in leaf cells of the cultivated Oriental ginseng (Panax ginseng Meyer)

Background Wild ginseng, Panax ginseng Meyer, is an endangered species of medicinal plants. In the present study, we analyzed variations within the ribosomal DNA (rDNA) cluster to gain insight into the genetic diversity of the Oriental ginseng, P. ginseng, at artificial plant cultivation. Methods The roots of wild P. ginseng plants were sampled from a nonprotected natural population of the Russian Far East. The slides were prepared from leaf tissues using the squash technique for cytogenetic analysis. The 18S rDNA sequences were cloned and sequenced. The distribution of nucleotide diversity, recombination events, and interspecific phylogenies for the total 18S rDNA sequence data set was also examined. Results In mesophyll cells, mononucleolar nuclei were estimated to be dominant (75.7%), while the remaining nuclei contained two to four nucleoli. Among the analyzed 18S rDNA clones, 20% were identical to the 18S rDNA sequence of P. ginseng from Japan, and other clones differed in one to six substitutions. The nucleotide polymorphism was more expressed at the positions 440–640 bp, and distributed in variable regions, expansion segments, and conservative elements of core structure. The phylogenetic analysis confirmed conspecificity of ginseng plants cultivated in different regions, with two fixed mutations between P. ginseng and other species. Conclusion This study identified the evidences of the intragenomic nucleotide polymorphism in the 18S rDNA sequences of P. ginseng. These data suggest that, in cultivated plants, the observed genome instability may influence the synthesis of biologically active compounds, which are widely used in traditional medicine.

The Identification of Araliaceae Species by ITS2 Genetic Barcoding and Pollen Morphology

The genetic barcode ITS2 (ITS: internal transcribed spacer) and pollen morphology were used for the identification of the pharmacologically valuable wild Araliaceae species Panax ginseng, Oplopanax elatus, Aralia elata, Aralia continentalis, Eleutherococcus senticosus, and Eleutherococcus sessiliflorus inhabiting the natural forests of Primorye, Russia. The ITS2 locus successfully identified all six species, which supports the use of ITS2 as a standard barcode for medicinal plants. However, the ITS2 locus was insufficient for intra-specific discrimination in these species, neither within Primorye nor from other world representatives within GenBank. Araliaceae pollen was confirmed to undergo size-reducing metamorphosis. The final morphotypes were species-specific for each of the six species but could not discriminate intra-species geographic localities within Primorye. The morphologies of the final pollen morphotypes from homologous species inhabiting other parts of the world are not yet known. Therefore, whether pollen is applicable for Araliaceae intra-species discrimination between Primorye and other world localities could not be established. Based on these findings, we propose that the ITS2 genetic barcode and the final pollen morphotypes are suitable for the identification of Araliaceae species. However, further studies will be needed to determine the suitability of genetic and pollen traits for Araliaceae geographic authentication.

State of antioxidant systems and ginsenoside contents in the leaves of Panax ginseng in a natural habitat and an artificial plantation

Native Panax ginseng Meyer plants are now quite rare in their natural environment, and thus, artificial plantations are used for commercial purposes. However, ginseng plantations are frequently exposed to abiotic and biotic stress stimuli, which can decrease biomass accumulation and cause plants to wither and die. The antioxidant systems of the plants play a vital role in their defense mechanism against adverse stresses through maintaining the balance between reactive oxygen species generation and detoxification. Here, the adaptability of wild and cultivated P. ginseng was compared with respect to the antioxidant enzyme activities and gene expression, lipid peroxidation, ratio of reduced-to-oxidized glutathione, and ginsenoside content. Several new genes for antioxidant enzymes, including PgApx1, PgApx2, PgCSD2, PgCSD3, PgMSD1, PgGR1, PgPrx2, and PgPrx3, were identified, and their relative expression levels were determined together with previously characterized genes (PgCSD1, PgCat1, and PgPrx1). The relative transcription of PgMSD1 was higher in wild ginseng, whereas PgApx2 was overexpressed in cultivated plants. Expression of other antioxidant genes remained constant. The activity of superoxide dismutase, class III peroxidase, and glutathione reductase was significantly decreased in cultivated P. ginseng, whereas the activity of ascorbate peroxidase and catalase was not changed. Moreover, oxidative stress markers such as malondialdehyde concentration, the ratio of reduced-to-oxidized glutathione, and Rg-type ginsenosides content were elevated in cultivated ginseng plants. Our results indicate that P. ginseng plants grown in their natural habitat or artificial plantations have different antioxidative statuses. The process of domestication appears to have reduced the antioxidant defense system of ginseng.

Анатомо-морфологические особенности организации побега Filipendula camtschatica (Rosaceae) в культуре in vitro

The results of the study morphogenetic processes in plants of Filipendula camtschatica (Pall.) Maxim. under propagation in vitro are presented. ½MS medium supplemented Kn 1 mg/l, indolyl acetic acid 0,05 mg/l and Gb 2 mg/l provide a high multiplication rate. Micro plants formed a rosette shoots. Nutritional medium is selected for cultivation vegetative rosette shoots and further transfer micro plants into the ground.