Chemical constituents from Rhodiola sachalinensis / 中草药

Objective: To investigate the chemical constituents from the roots and rhizomes of Rhodiola sachalinensis. Methods: The chemical constituents were isolated by repeated silica gel chromatography, medium pressure column chromatography, and semi-preparative liquid chromatography, and their structures were elucidated by chemical properties and spectroscopic analyses. Results: Eighteen compounds were isolated and identified to be gallic acid (1), p-hydroxybenzoic acid (2), salidroside (3), benzyl-O-β-D-glucopyranodide (4), phenylethyl-8-O-β-D-glucopyranodide (5), cinnamyl-β-D-glucopyranoside (6), sachalinol (7), quercetin (8), quercetin-3-O-β-D-glucopyranoside (9), kaempferol (10), kaemferol-7-O-α-L-rhamnopyranoside (11), kaempferol- 7-O-β-D-glucopyranoside (12), kaemnpferol-3-O-α-L-rhamnoside (13), kaempferol-3-O-β-D-glucopyranoside-7-O-α-L-rhamnoside (14), tricin (15), tamarixetin (16), herbacetin-7-O-α-L-rhamnoside (17), and herbacetin-3-O-β-D-glucopyranoside-7-O-α-L- rhamnoside (18). Conclusion: Compounds 9, 12, and 16 are obtained from the plants in Rhodiola L. for the first time. Compounds 2, 7, 8, 14, and 18 are obtained from this plant for the first time.

Autotetraploid induction and identification of Rhodiola sachalinensis / 中草药

Objective: To obtain autotetraploid clones of Rhodiola sachalinensis. Methods: Germplasm with doubled chromosomes was acquired according to colchicine soaking seeds, and ploidy was identified according to stomata size and density measurements, chromosome counting and flow cytometry analysis. Chimera was purified after several times of regenerations. Results: Colchicine concentration and treatment duration had significant effect on seed germination rate, seedling death rate and ploidy mutation rates, and they were 18.2%, 73.6%, and 68.7%, respectively after the seeds were soaked in 0.2% colchicine solution for 72 h. Stomata diameter was promoted and density was declined significantly in morphological variation plantlets; normal diploid chromosome number was 2n = 2x = 26, and tetraploid chromosome number was 2n = 4x = 52. Meanwhile, aneuploidy was found in the population of plantlets after colchicine soaking, leaves of ploidy mutation plantlets were used as expiants to regenerate to tube plants, and homozygous clones were obained after three regeneration and purification cycles. There was no chimera in the homozygous clones according to flow cytometry analysis. Conclusion: It is feasible to obtain autotetraploid according to colchicine treatment on seeds of R. sachalinensis.

Protoplast isolation and plant regeneration from leaves of Rhodiola sachalinensis / 中草药

Objective To isolate protoplasts from tube plant leaves of Rhodiola sachalinensis and regenerate plantlets after protoplast culture.Methods Preculture treatment and size of explants,enzyme concentration,mannitol concentration in enzyme mixture related with protoplasts isolation were studied to determine the superior optimized conditions.Results Explants could be used to isolate protoplast without dark preculture,and leaf length should be longer than 1.5 cm.The best incubating enzyme solution contains 1.0% cellulase Onzuka R-10,0.5% Macerozyme R-10,10 mmol/L CaCl2?2H2O,0.1% MES,0.7 mmol/L KH2PO4,and 0.5 mol/L mannitol.The enzyme and explants mixture were shaken for 4 h at 25 ℃.The protoplasts yield and viability were 39.43?106/g fresh weight and 78.6%,respectively.Purified protoplasts were cultured in medium 1/2 MS+1 mg/L 2,4-D+0.5 mg/L ZT+0.5 mol/L mannitol +500 mg/L hydrolysis of casein initially with shallow liquid layers,and calli formed within 40 d.After calli were transfered to MS+1 mg/L 6-BA+0.1 mg/L NAA,adventitious buds were induced from calli.Shoots longer than 2 cm rooted within 30 d when they were transfered to 1/2 MS medium.Conclusion The study provides the scientific base for protoplast fusion in polyploidy breeding of R.sachalinensis.