Preservation of multiple bud bodies of mulberry (Morus alba L.) under low and ultra-low temperatures
1997
Oka, S. (National Inst. of Agrobiological Resources, Tsukuba, Ibaraki (Japan)) | Niino, T.
Multiple bud bodies (MBB) of mulberry (cv. Kenmochi, Hayatesakari, and shin-ichinose) were induced and subcultured using MS liquid medium supplemented with 2 mg/l 4-PU on a rotatory shaker. MBB fragmented into small pieces were preserved at 2.5-10 degrees C either in solid or in liquid medium. Solid medium gave better results than liquid medium in maintaining viability of MBB. MBB preserved in solid medium at 1.5-5 degrees C for up to 90 days resumed proliferation after shake culture (120 rpm) at 25 degrees C. MBB cold-hardened at 5 degrees C for 30 days were treated with PVS-2 solution at 25 degrees C for 80 min and then transferred to liquid nitrogen followed by rapid thawing at 30 degrees C. MBB could be successfully cryopreserved and regenerated shoots through this vitrification method. The cold-hardened MBB were encapsulated in calcium alginate beads and bried with silica gel for 4-24 hr at 25 degrees C. The dried beads were transferred to liquid nitrogen followed by rapid thawing at 254 degrees C. The highest survival and regeneration rates were obtained when the beads were dried for 16 hr, under which condition the water content of the beads was reduced to about 16%. MBB dried for longer time than 20 hr failed to grow. Morphologically normal shoots were recovered from the cryopreserved MBB
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