Cryopreservation of Hyoscyamus niger adventitious roots by vitrification

An auxin-independent adventitious root culture of Hyoscyamus niger was esta blished, and the roots were successfully cryopreserved with a high regenera tion rate of 93.3% by vitrification method. The root tips were cultured for 12 to 14 days in phytohormone-free Murashige and Skoog (MS) liquid medium, and were excised and precultured on Woody Plant (WP) solid medium suppleme nted with 0.3 mol/L sucrose at 25 degreesC in the dark. After 1 day, they w ere treated with MS-based loading solution for 10 min, followed by soaking in MS-based PVS2 for 10 min at 0 degreesC. The treated root tips were immer sed in liquid nitrogen (-196 degreesC). For recovery, the root tips were th awed rapidly at 40 degreesC and washed with MS medium containing 1 mol/L su crose prior to plating onto WP solid medium. The regenerated roots were eva luated by their growth and tropane alkaloid production. The growth and alka loid content of regenerated roots analyzed using HPLC were found to be almo st the same as those of non-treated roots.