Cryopreservation of protonemata of Ditrichum cornubicum (Paton) comparing the effectiveness of four cryoprotectant pretreatments

If an ex situ programme for the conservation of rare and threatened bryophytes is to succeed, a method of long-term, basal storage is required. The aim of this study was to determine the effectiveness of sucrose, ABA and sucrose/ABA combined, as cryoprotectant pretreatments, for the long-term storage of protonemata of the endangered Cornish path moss, Ditrichum cornubicum. This work is a prelude to the in vitro conservation of rare and endangered UK bryophytes. The addition of sucrose and/or ABA in the pre-culture medium, reduced protonemal growth rate. However, pretreatment afforded a high level of protection against tissue damage during dehydration and freezing, with 100 % regeneration of pretreated plants after thawing, compared to 53 % regeneration of controls. Sucrose combined with ABA in the pre-culture medium gave the most effective cryoprotection, with protonemata having fewer necrotic tissues and rapid recovery of growth.