Optimizing Pueraria candollei var. mirifica cell suspension culture for prolonged maintenance and decreased variation of isoflavonoid from single cell lines
2018
Rani, Dolly | Meelaph, Thitirat | Kobtrakul, Khwanlada | Vimolmangkang, Sornkanok
Isoflavonoids are naturally occurring polyphenolic compounds that have various biochemical and pharmacological properties. The tuberous roots of Pueraria candollei var. mirifica contain isoflavonoids and are widely used as dietary supplements and in therapeutics. The present study was aimed at producing reliable sources of isoflavonoids from single Pueraria cells and at optimizing a culture medium with increased biomass and isoflavonoid content. The medium optimized for prolonged maintenance of callus was MS medium supplemented with 200 mg/L KH₂PO₄, 1 mg/L thiamine HCl, 100 mg/L of myo-inositol with 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and subculturing for 30 days for the highest growth. The amount of isoflavonoids varied among calli obtained from different explants. The highest amounts of puerarin and daidzein were quantified as 0.63 and 1.80 mg/g of dry weight (DW) in the examined suspension cell culture of single clones, whereas daidzein (0.50 mg/g) and genistein (0.03 mg/g) were present in the suspension cell culture obtained from a mixture of cells. The suspension of cell cultures from the same origin gave comparable amounts of isoflavonoids, but cells from different sources showed varied amounts of isoflavonoids. The amount of isoflavonoid also varied depending on the cell age. The present study provided the first evidence of Pueraria cell suspension from single cells and highlighted the importance of screening the starting cells from a single cell line and the cell age prior to the scale-up process to select desired sources of isoflavonoid compounds.
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