System Is Established On Cinnamomum Camphora Chvar. Borneol Cell Suspension Culture And Optimize

Natural D-borneol is the most important secondary metabolite in Cinnamomumcamphora chvar. Borneol. The traditional extraction methods can not meet marketdemand.Cell suspension culture way of production of secondary metabolites is animportant technology method in the current Chinese medicine production.In this study, Cinnamomum camphora chvar. Borneol tissue culture as a materialwere used to carry out the good callus. The good callus after several times subculturewas transferred to liquid medium, followed by suspension culture conditions whilemonitoring D-borneol content of the target product, so Cinnamomum camphora chvar.Borneol callus suspension culture system were established and optimized, and thestudy provides a theoretical basis for the future industrial production of secondarymetabolites by cell culture.The contents and results of this thesis is as follows:1. The excellent Cinnamomum camphora chvar. Borneol callus induction andsubculture In this study, callus induction and subculture, the best explants forinducing callus: young stem of Cinnamomum camphora chvar. Borneol. The bestculture medium: MS+2,4-D2mg/L+6-BA0.6mg/L+sucrose30g/L+Carrageenan6.8g/L, pH value of5.8; Outside regulation as follows:25℃dark pretreatmentmaterial3days, at25℃in the dark for30days, then harvest.The content ofD-borneol is2.7453μg/g DW.The subculture medium means mixing LH100mg/L ininduction medium.After subculture four times can be transferred to suspension culture.The content of D-borneol is2.6397μg/g DW.2. Cinnamomum camphora chvar. Borneol cell suspension culture system isestablished and its physiological and biochemical are regulated The best cellsuspension culture medium: MS+2,4-D2mg/L+6-BA0.6mg/L+LH100mg/L+CH500mg/L+sucrose30mg/L,pH value of5.8; External regulation and controlconditions, light intensity2000Lux by white light continuous irradiation,5days laterunder the low-temperature environment of16℃for3days,take80ml medium in250ml flask, inoculum40g/L, shaking speed is110r/min, sealed with a breathable membrane and cultured for21days, then harvest. The content of D-borneol is3.8159μg/g DW. About39%more than the initial callus the content of D-borneol(2.7453μg/g DW).3. Cinnamomum camphora chvar. Borneol cell culture extraction anddetermination of D-borneol The relatively better extraction method of activeingredient D-borneol is reagent for anhydrous ethanol, the solid-liquid ratio1:10(g/ml), ultrasonic extraction, extraction temperature of40℃,2times extraction,ultrasonic time is120min. Determined by GC.In summary, in the study the plant cell suspension culture technique was firstlyused in medicinal plants Cinnamomum camphora chvar. Borneol, and its cellsuspension culture system was initially established and optimized.