Figure 2.

Effect of activated charcoal on the survival (A) and regrowth (B) of cryopreserved (LN+) chestnut embryonic axes by encapsulation–vitrification. EAs were encapsulated in WPM solution containing 2.5% (w/v) sodium alginate with (AC+) or without (AC−) 0.3% (w/v) activated charcoal in the artificial matrix. Encapsulated EAs were treated for 60 min with LS (2.0 M glycerol, 0.4 M sucrose), followed by treatment with PVS2 at 0 °C for 30–120 min, prior to direct immersion in liquid nitrogen for 1 h. Control EAs received no LS and PVS2 treatments. Within each exposure time, different letters indicate significant differences between AC+ and AC− at p ≤ 0.05 by chi-squared test (bars, SE of means).