Construction of aseptic cultural system of Salvia miltiorrhiza and determination of active components by HPLC analysis / 中草药

Objective: The regeneration system of Salvia miltiorrhiza was constructed and contents of active components of the sterile materials were determined, so as to provide the basis for the rapid propagation and secondary metabolism regulation of S. miltiorrhiza. Methods: The optimum hormone ratio for inducing callus was screened by orthogonal test. The buds and roots induction were conducted to estabilish the tissue culture system. The contents of effective components were evaluated by HPLC analysis. Results: The suitable medium for callus induction was MS+6-BA (2.0 mg/L)+NAA (1.0 mg/L)+2,4-D (0.5 mg/L). The preferred enrichment medium of adventitious bud induction was MS+6-BA (2.0 mg/L)+NAA (1.0 mg/L). And rooting medium was 1/2 MS+NAA (0.5 mg/L). Seven active components in aseptic seedlings, callus, and regenerated seedlings could be detected with significant differences in different aseptic materials (P < 0.05). The contents of rosmarinic acid and salvianolic acid B were higher than the others. Conclusion The culture system of S. miltiorrhiza was successfully established, and vigor regenerated seedlings were also obtained. The accumulation of active components in the three sterile materials showed difference, laying a foundation for further study in S. miltiorrhiza.

Regenerated plantlet from salt-tolerant callus of Glycyrrhiza uralensis / 中草药

Objective To develop the technique of artificial cultivation of Glycyrrhiza uralensis on salina by exploring regenerated plantlet from the salt-tolerant callus.Methods The experiments of dedifferentiation were successful in inducing cotyledon sections and embryonic axle sections from sterilized shoot,as explants,on salty media.The induced callus was to be multiplied and adventitious buds tobe produced when the salt concentration of medium increased gradually.Then the buds were rooting and a lot of plantlet occurring.Results Dedifferentiation of cotyledon sections and embryonic axle sections was preferable on MS+BA 1.5 mg/L+2.4-D 1.2 mg/L+NaCl 100 mg/L.Callus was hazel and just a little translucent.Growth of callus became weaker and weaker after increasing concentration of NaCl to 250 mg/L,the color got darker,and much callus died.A lot of adventitious buds came into being on MS+BA(0.5 mg/L+)KT 1.0 mg/L+NAA 1.0 mg/L+NaCl 200 mg/L after inoculating the callus on non-salty media for multiplying twice.It proved that the salt-tolerant character of the buds was induced by reason of genetics.Decreasing temperature(18—22 ℃) at natural light and adding an appropriate quantity of GA_3 in medium were helpful for the formation of strong sprouts.The rooting rate was 66.26% with rooting 55 adventitious buds among 83,and medium of 1/2 MS+IAA 1.0 mg/L+NAA 1.0 mg/L+NaCl 200 mg/L adapted to root. Conclusion A lot of salt-tolerant regenerated plantlets of G.uralensis occur after inducing the salt-tolerant callus to redifferentiate for getting the regenerated plantlet.

Establishment of asexual multiplication system of Arnebia euchroma / 中草药

Object In order to promote the artificial cultivation of Arnebia euchroma (Royle) Johnst , and save the wild herb from relieving the lack of resources, the asexual multiplication system of A euchroma was established by tissue culture technique Methods Dedifferentiation in many kinds of explants of A euchroma by inducement succeeded After the embryos were multiplied, there were a lot of germinated transplants in media After callus were multiplied, many adventitious buds had formed in the media, then they were rooting; the transplants formed by inducement and the plants survived after being cultured in soil Results During the dedifferentiation, 2,4 D was essential, the effect of KT was better than BA The shoot sections were dedifferentiated in the most rapid way among the explants The callus from them were in very good quality It is of benefit to the callus multiplication and redifferentiation after reducing the concentration of 2,4 D Both KT and BA can induce adventitious buds, KT was also suitable for the formation of globular shaped embryos In initial period of sphere embryos culture, it was necessary to add the plant growth regulators in media for multiplying sphere embryos and promoting their development Proper temperature difference between day and night (10 ℃-28 ℃) and natural light were helpful for the formation of strong sprouts and survival of transplants The survival rate was 13 2% with 31 plantlets survived among 236 transplants cultured in soil Conclusion The asexual multiplication system of A euchroma established through tissue culture is stable and available