High-efficiency vitrification protocols for cryopreservation of in vitro grown shoot tips of transgenic papaya lines.

Shu-Fen Tsai; Shyi-Dong Yeh; Chin-Feng Chan; Song-Iuan Liaw

Abstract  In vitro grown shoot tips of transgenic papaya lines (Carica papaya L.) were successfully cryopreserved by vitrification. Shoot tips were excised from stock shoots that were preconditioned in vitro for 45–50-day-old and placed on hormone-free MS medium with 0.09 M sucrose. After loading for 60 min with a mixture of 2 M glycerol and 0.4 M sucrose at 25°C, shoot tips were dehydrated with a highly concentrated vitrification solution (PVS2) for 80 min at 0°C and plunged directly into liquid nitrogen. The regeneration rate was approximately 90% after 2 months post-thawing. Successfully vitrified and warmed shoot tips of three non-transgenic varieties and 13 transgenic lines resumed growth within 2 months and developed shoots in the absence of intermediate callus formation. Dehydration with PVS2 was important for the cryopreservation of transgenic papaya lines. This vitrification procedure for cryopreservation appears to be promising as a routine method for cryopreserving shoot tips of transgenic papaya line germplasm.

CRYOPRESERVATION of plant cells & tissues; PLANT shoots; TRANSGENIC plants; PAPAYA; PLANT growing media; REGENERATION (Botany); GERMPLASM conservation; CALLUS (Botany)

Plant Cell, Tissue & Organ Culture, 2009, Vol 98, Issue 2, p157

0167-6857

Academic Journal

10.1007/s11240-009-9548-4