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Licorice is commonly used in Chinese medicine , is an important medicinal plants. In addition to medicinal value, licorice in food, beverages , cigarettes , cosmetics and other industries are also widely used. In recent years , as people destructive excavation of wild licorice , so wild almost extinct. Therefore, to carry out licorice tissue culture work, especially large-scale suspension culture is important. Licorice flavonoids are a large class of biologically active ingredients , which have anti-ulcer , anti-bacterial , anti-inflammatory , anti-oxidation , anti- HIV, anti-tumor effect. Chalcone synthase flavonoid synthesis of all the key enzyme . It catalyzes three molecules of malonyl coenzyme A molecule with an acrylic -hydroxybenzoate coenzyme A reduced synthesis naringenin chalcone , as a stent to other flavonoids derived . Chalcone synthase activity accumulation and accumulation of flavonoids are closely related . This experiment was cut Glycyrrhiza aseptic cotyledons , hypocotyl , root segment three kinds of explants callus induction and subculture , the establishment of liquorice suspension cell culture system , suspension cultured cells were measured growth curve ; and studied the inoculum size , conditioned medium and exogenous hormones on suspension cells. The results show that the source cotyledon callus suspension culture the most suitable ; suspension culture the optimum inoculum 35g / L; conditioned medium of the optimum amount of the total culture volume of 1/ 3 ; most suitable Glycyrrhiza suspension culture of medium was added NAA 1.0mg / L KT 0.5mg / L MS medium hormone combination . By RT-PCR method , callus from Glycyrrhiza cloned chalcone synthase gene cDNA fragment and sequenced . Sequencing results suggest that we have cloned a cDNA fragment obtained contains an open reading frame , the 1170bp nucleotides encoding a 389 -amino acid polypeptide . And alfalfa, soybeans, peas and other legumes CHS several nucleotide homology of 80% or more amino acid homology rate of 90 % or more. The sequence has been submitted to GenBank / EMBL / DDBJ registration , serial number EU706287.
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