Production of Anthocyanins by Habituated Cultured Cells of Nyoho Strawberry (Fragaria ananassa Duch.)

Cultured leaf cells of Nyoho strawberry (Fragaria ananassa Duch.) produced 4 times more anthocyanins than fruit, with additions of 2.0 mg/l of 1-naphthalenacetic acid (NAA) and 0.2 mg/l of benzyladenine (BA) for 2 weeks. They were habituated after 6 passages at NAA 1.25 and BA 0.125 μg/l in 107 cultures with stepwise decrease of the additions at 2-week intervals, and supervention of a gradual decrease in anthocyanin contents of 80%. They fully recovered through the following 43 passages without the additions. Finally, 3 types of cell line rich in cyanidin 3-glucoside (62%), peonidin 3-glucoside (55%) or both (49 and 36%, respectively) were obtained. They also produced pelargonidin 3-glucoside (0.6 to 1.8%), which was the main component in the fruit. The fruit produced peonidin 3-glucoside (0.4%), as well as pelargonidin 3-O-(6-O-malonyl-β-D-glucoside) and cyanidin 3-glucoside. The habituation seems to have caused some activation or inhibition of the cell’s anthocyanin formation, which originated from the mother plant.