Relative importance of seed drying rate, desiccation tolerance, and cryotolerance for the conservation of Ardisia elliptica, A. brunnescens and A. virens.

BACKGROUND The pan-tropical genus Ardisia has more than 400 species and is of high horticultural and medicinal value. Due to overexploitation it is important to conserve the germplasm of this genus. OBJECTIVE To investigate the feasibility and methods of cryopreservation for long-term seed storage of three Ardisia species: A. elliptica Thunb., A. brunnescens Walker, and A. virens Kurz. METHODS We tested whether rapid desiccation can increase desiccation tolerance and cryotolerance, and whether the thawing rate can affect cryopreservation success. Seeds were subjected to three desiccation treatments: 1) activated silica gel at 25 +/- 2 degree C, and 4% relative humidity (RH); 2) saturated NaCl solution in closed jars in 25 +/- 2 degree C and 75% RH; and 3) air-drying at room conditions at 27 +/- 2 degree C and RH 60% for different desiccation durations (12h, 24h, 48h, 96h, and 196h). Seeds were then assessed for desiccation tolerance and cryotolerance after rapid thawing (direct immersion in 36 degree C water bath for 2 min) or slow thawing (at 27°C for 1 h). RESULTS For all three species, desiccation method and duration significantly affected cryotolerance (P < 0.0001). Fast desiccation did not improve germination compared to slower desiccation (P < 0.01). Whereas A. elliptica germination was unaffected by desiccation duration, drying time significantly (P < 0.0001) affected germination percentage in the other two species especially after 48h. Although slow thawing improved cryotolerance of A. brunnescens seeds (P < 0.05), there was no significant effect of thawing rate on A. elliptica. A. virens seed did not survive cryopreservation. CONCLUSIONS Cryopreservation protocols of Ardisia species may be species-specific and should be established for each species in the genus so that cryopreservation can be used as a successful conservation strategy.