Established In Vitro Cell Lines From Brassica Oleracea L Var,Regulated Metabolism And Optimizated Extraction Process Of Sulforaphane

Sulforaphane derived from glucoraphanin degradation broccoli, has the strongestanti-cancer activity, molecular weight is177.29, formula is C6H11S2NO.Using broccoli callus and seeds as experimental material, combined plant tissue culturetechnology, studied the sulforaphane content in different varieties and callus, and theinfluences of supplementary materials, carbon sources and harvest time on the growth of thecallus from broccoli and its sulforaphane accumulation. The main conclusions of this study asfollowed:1. Broccoli varieties of the highest sulforaphane content were942.8μg·g^-1（yuhuang）, the lowest172.8μg·g^-1（zhongqing No.9F1）; The sulforaphane content of call-usinduced form different explants from high to low were cotyledon(840.5μg·g^-1), hypocotyl(517.1μg·g^-1), radicle part(440.3μg·g^-1).2. The best medium of callus proliferation were MS+1.0mg·L^-1NAA+1.5mg·L^-16-BA,MS+1.5mg·L^-1NAA+1.5mg·L^-16-BA and MS+1.0mg·L^-1NAA+2.0mg·L^-16-BA.3. In the growth cycle of broccoli callus, multiplication growth of callus showed S-typecurve and the content of sulforaphane was bimodal curve, which reached the maximum982.3μg·g^-1at18d. According to the gross production, the best harvest time was27d, thesulforaphane production reached the maximum865.4μg·g^-1.4. The content of sulforaphene of broccoli callus were significantly effected by theexogenous additions. In this investigation, when the concentrations of AgNO3, SA、SAS、sucrose and glucose were respectively10mg·L^-1,12mg·g^-1,0.4mg·L^-1,25g·L^-1and35g·L^-1,the sulforaphene were promoted obviously, with the increase of the concentration of NaCland maltose, were inversely proportional and all lower than control; when the concentrationsof SA, SAS and sucrose were respectively6mg·L^-1,0.4g·L^-1and25g·L^-1, the contents ofsulforaphane in broccoli callus were promoted obviously;and the metabolism and synthesis ofsulforaphane were inhibited by other factors.5. By response surface test design: HPLC method with determination, with methylenechloride for extracting agent, sulforaphane for the best technical parameters: the optimumextraction conditions were the extraction temperature25.7℃, the extraction time2.8h, the liquid solid ratio1g:30mL, the extraction times3. The test value was6.7%.