Abstract:
To explore whether abscisic acid (ABA) and cold adaptation can improve the cryopreservation efficiency of
Porphyra sensu lato, the free living conchocelis of
Pyropia haitanensis were treated with ABA (100 μmol/L) for 48h at 20℃, light intensity of 40 μmol·photons/(m
2·s) with photoperiod 12L﹕12D or cultured in dark at 4℃ for 6 days before cryopreservation at –80℃. The effects of cryopreservation were evaluated by continuous measurement of algal survival rate after frozen, the maximum light quantum efficiency (
Fv/
Fm) of PSⅡ and the expression of cold adaptative related genes, such as
P(
H+)
-ATPase, ald, Mn-sod A
, Cu/Zn-sod 1,
apx-2,
hsp70-4, psy-2,
gapdh, and
gpxha-1. The results showed that: (1) Both ABA treatment and cold adaptation at 4℃ increased the survival rate of
P. haitanensis conchocelis after cryopreservation, 59% in the ABA groups and 89% in the cold adapt groups; (2) The
Fv/
Fm of the conchocelis after cold adaptation recovered significantly faster than those of the other groups (
P<0.05), with the optimal level after 35 day culture; (3) ABA treatment and 4℃ cold adaptation significantly up-regulated genes like
P(H
+)
-ATPase,
ald,
Mn-sod A
, Cu/Zn-sod 1
, gapdh (
P<0.01) and
psy-2 (
P<0.05) in the algae, but only ABA induced
apx-2 and
hsp70-4 (
P<0.01). In summary, ABA treatment and cold adaptation can improve the frost resistance of
P. haitanensis and improve the efficiency of cryopreservation. The cold adaptation method at 4℃ is simple and effective, and has greater application potential.