OBJECTIVETo study the cause of the seeds dormancy of Glehnia littoralis in vitro and to establish plant regeneration methods via somatic embryos.

METHODThe effects of endosperm and exogenous hormone on the seed dormancy breaking of G. littoralis and the effect of hormone concentration on embryonic callus induction and plant regeneration via somatic embryos were observed,

RESULTSThe germination rate of the seeds with 1/3 endosperm was the highest which achieved 31%. TDZ, 6-BA and GA3 treatment could not break seed dormancy but easily lead to abnormal seedlings. Embryogenic callus induction rates was up to 57% on MS supplemented with 1.0 mg x L(-1) 2,4-D. After 20 days culture, embryogenic calli were transferred to MS medium and cotyledonary embryos were formed in 40 days. The regenerated plants were obtained in 20 days.

CONCLUSIONAn effective system of plant regeneration of G. littoralis was established in this study.

Apiaceae ; physiology ; Endangered Species ; Plant Somatic Embryogenesis Techniques ; Plants, Medicinal ; physiology ; Regeneration ; Seeds ; physiology

OBJECTIVETo establish HPLC characteristic fingerprints of the saponins in Pulsatilla medicinal plants, and provide the basis for authentication and classification of Pulsatilla species.

METHODThe HPLC profiles were determined at 35 degrees C on a Symmetry C18 column (4.6 mm x 250 mm,5 microm) eluted with water (A) and acetonitrile (B) as mobile phases in a linear gradient elution with the flowrate of 0.5 mL x min(-1). The elution program was as follows: 0-8 min, 90% A to 77% A, 8-25 min, changed to 71% A, 25-40 min, to 60% A, 40-50 min, to 50% A, 50-75 min, to 10% A, 75-80 min, to 0% A. The detection wavelength was set at 210 nm.

RESULTThe different species of Pulsatilla showed different HPLC fingerprints, but with 10 common peaks. A cluster analysis of 14 accessions indicated that they were divided into four groups: all accessions from P. koreana were classified into group I, P. ambigua in group II, P. dahurica and P. turczaninovii in group III, and P. chinensis in group IV, respectively. The significant differences between P. koreana and P. dahurica, and between P. turczaninovii and P. ambigua were observed.

CONCLUSIONThe results obtained were in agreement with the traditional taxonomic study. The method was rapid and precise, not only can be used to classify and authenticate Pulsatilla species, but also provides important references for HPLC fingerprints and quality control of Pulsatilla medicinal plants.

Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Plants, Medicinal ; chemistry ; classification ; Pulsatilla ; chemistry ; classification ; Quality Control

OBJECTIVETo develop a HPLC quantitative method for determination of schisandrin, schisandrol B, schisantherin A, deoxyschisandrin and gamma-schisandrin in Fructus Schisandrae.

METHODA symmetry C18 column (4.6 mm x 250 mm, 5 microm) was used with methanol (A) and water (B) as mobile phases, in gradient elution. The gradient program was as follows: 0-36 min, changed from 60% A to 66% A, 36-65 min, to 80% A, 65-70 min, kept for 80% A, 70-75 min, to 100% A. The flow rate was 0.5 mL x min(-1) and detection wavelength was set at 254 nm.

RESULTThe linearities of schisandrin, schisandrol B, schisantherin A, deoxyschisandrin and gamma-schisandrin were in the ranges of 0.0214. 160 (r = 0.9999), 0.020-4.000 (r = 0.9999), 0.021-4.240 (r = 0.9999), 0.020-3.960 (r = 0.9999) and 0.021-4.200 (r = 0.9999). The average recoveries were 104.8%, 104.2%, 102.7%, 104.6%, 104.5%, respectively.

CONCLUSIONThe method developed in this study was reliable, and can be used for the quality control of the fruits of S. chinensis.

Chromatography, High Pressure Liquid ; methods ; Lignans ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Schisandra ; chemistry