Molecular phylogenetic identification of Artemisia l. Species from Mongolia

The Artemisia L. genus, one of the largest genera in the Asteraceae family,   consists of many medicinally important and phylogenetically unresolved species. To define the phylogenetic relationship of Artemisia species, nucleotide sequences of the nuclear ITS (Intergenic spacer DNA) region, chloroplast trnL-trnF intergenic spacer, partial sequences of plastid rbcL gene were identified from medicinally important 12 species included in 3 subgenera. The phylogenetic tree was constructed through the Neighbor-Joining and Maximum Parsimony analysis, respectively. The results of study revealed that the combination of the nucleotide sequences from the ITS and rbcL region was more efficient in determining the phylogenic relationship of species

Determination of triterpenoid saponin and polysaccharide content from in vitro cultures of Astragalus mongholicus Bunge

In this study, the efficient micropropagation protocol of Astragalus mongholicus Bunge was established and also triterpenoid saponin and polysaccharide content in ethanol, methanol and aqueous extracts of different samples were determined by using spectrophotometric methods to investigate whether the content of biologically active compounds depends on the stage of development of the plant during in vitro culture. The content of total saponins and polysaccharides in different cultures of A. mongholicus grown in vitro was higher (990 and 505 μg/ml) in ethanol extracted 14-day-old young shoot samples than in 28-day-old propagated shoot samples and rooted shoots

Determination of triterpenoid saponin and polysaccharide content from in vitro cultures of Astragalus mongholicus Bunge

In this study, the efficient micropropagation protocol of Astragalus mongholicus Bunge was established and also triterpenoid saponin and polysaccharide content in ethanol, methanol and aqueous extracts of different samples were determined by using spectrophotometric methods to investigate whether the content of biologically active compounds depends on the stage of development of the plant during in vitro culture. The content of total saponins and polysaccharides in different cultures of A. mongholicus grown in vitro was higher (990 and 505 μg/ml) in ethanol extracted 14-day-old young shoot samples than in 28-day-old propagated shoot samples and rooted shoots

Cryopreservation of PLBs of Brassidium Fly Away Using Encapsulation-Dehydration Techniqu

In vitro grown protocorm-like bodies (PLBs) of Brassidium Fly Away orchid hybrid were cryopreserved using encapsulation- dehydration technique. The viability of the cryopreserved cells was determined by 2,3,5-triphenyltetrazolium chloride (TTC) assay. For the preculture treatment, the PLBs were excised into two standard sizes of 1-2 and 4-5 mm and were precultured on half-strength Murashige and Skoog (MS) semi solid medium supplemented with diff erent concentrations of sucrose (0, 0.2, 0.4, 0.6, 0.8 and 1.0M). The PLBs size 4-5 mm and 0.6 M sucrose concentration was selected based on highest viability obtained in TTC assay. The PLBs were encapsulated for 30 minutes using 3% (w/v) liquid sodium alginate medium supplemented with 0.4M sucrose and 0.1M calcium chloride and osmoprotected in 0.75M sucrose solution for 24 hours at 25°C. The beads were then dehydrated using 50g heat-sterilised silica gel for four hours, cryopreserved for 24 hours, thawed in a 40±2°C water bath for 90 seconds, and regenerated in semi-solid half-strength. Biochemical analyses were conducted and the cryopreserved PLBs had produced lower content of chlorophyll while the highest specifi c peroxidase activity was observed in cryopreserved PLBs

-transformed roots of

In this study the results of agrobacterial transformation of Astragalus membranaceus (Fisch. Ex Link) Bunge, a medicinal plant from Mongolia and China with a wide spectrum of pharmacological action were presented. As explants for transformation, in vitro derived microshoots as well as hypocotyls, cotyledons and primary shoots isolated from seedlings were used. The most effective from two tested Agrobacterium rhizogenes strains (A4-RT, 15834 SWISS) was 15834 SWISS, causing the transformation of various types of explants, including primary shoots as a more responsive type. The optimal co-cultivation time of explants with bacterial suspension (48 hours) and the highest transformation rate of primary shoots (36.4%) were established. PCR analysis confirmed the transgenic nature of the roots transformed with strain 15834 SWISS and the absence of bacterial contamination

Characterization of the Second Generation Cryopreserved Dendrobium Bobby Messina Using Histological and RAPD Analyses

This study was conducted to detect the morphological, histological and molecular diff erences in the second generation of the PVS2 cryopreserved Dendrobium Bobby Messina [DBM] (18 months old culture) plantlets. Morphological analyses indicated that similarities and diff erences in cryopreserved DBM plantlets comparing to control stock culture based on selected morphological criteria. Morphological criteria, such as root length, number of shoot per explant and shoot length displayed diff erences, while the other three criteria, leaf diameter, leaf length and PLBs size were similar in cryopreserved compared to the control stock culture plant. Higher amount of homogenous cell population and denser cytoplasm were observed in cryopreserved PLBs compared to control stock culture PLBs based on histological analysis. This suggests the existance of somatic embryogenesis development mechanism taking place during the recovery and regeneration of the cryopreserved PLBs. However, RAPD analyses based on 10 primers indicated that cryopreserved DBM regenerated from vitrifi cation method generated a total of 20 to 39.9% polymorphic bands as compared to stock culture indicating potential somaclonal variation. Hence, an increase percentage of polymorphics bands in cryopreserved plantlets 18 months post cryopreservation as compared to previous report of 10% polymorphic bands in cryopreserved DBM 3 months post cryopreservation

Metabolite Profiling of In Vitro Cultured and Field Grown Rhizomes of Acorus calamus from Mongolia Using GC-MS

Acorus calamus (sweet flag) is used in the traditional Chinese and Indian medicines for various ailments. Due to its extensive use in herbal medicine, natural resources from the world's forests are being depleted at an alarming rate. In the present study, an in vitro cell culture technique is being explored as an alternative to field grown A. calamus with respect to the metabolite profile, antioxidant properties, total phenol, and total flavonoid content. Gas chromatography mass spectrometry (GC-MS) was utilized to compare the metabolite profiling between methanolic extracts of in vitro and field grown rhizome tissues of A. calamus. A statistical analysis indicated an upregulation of a-selinene, which is representative of sesquiterpene ketones, and a cyclic polyol, D-pinitol, which has an insulin mimicking effect in the in vitro cultivated rhizome tissue when compared to field grown rhizomes. Significantly higher free-radical scavenging activity (IC50 69.32 mu g mL(-1)), total phenolic content (71.60 mg GAE g(-1)), and total flavonoid content (42.34 mg CE g(-1)) were observed in in vitro rhizome tissues compared with those from field grown rhizomes. These observations suggest that the in vitro cultivation of Acorus rhizomes could be exploited as an alternative to field grown A. calamus, as it is an endangered medicinal plant. The production of useful metabolites by the in vitro cultured rhizomes can be explored successfully for utilization by various food and drug industries