Development of cryopreservation for Loxocarya cinerea---an endemic Australian plant species important for post-mining restoration.
We report the development of a cryopreservation protocol for the endemic Western Australian plant species Loxocarya cinerea (Restionaceae). Shoot tips from two genotypes, SXH404 and SXH804, were cryopreserved using the droplet-vitrification technique. Control explants, which were cryoprotected, but not cooled, showed regeneration for both genotypes (SXH404, 22.1 +/- 5.9%; SXH804, 67.7 +/- 9.6%). Extension of incubation in PVS2 from 30 to 60 min did not lead to survival after cryopreservation. Thermal analysis using differential scanning calorimetry confirmed the beneficial effect of a loading phase but also revealed no or very little ice formation after cryoprotection of shoot tips in other treatments. Regeneration following cryopreservation was obtained for genotype SXH804 (4.3 +/- 2.1%) but not for SXH404. Regenerated explants of L. cinerea SXH804 were morphologically identical to tissue-cultured plants. As an alternative to shoot tips, callus tissues of clone SXH404 were successfully cryopreserved (> 66.7% post LN survival) using the same protocol.
School of Biomedical Sciences, CHIRI Biosciences Research Precinct, Faculty of Health Sciences, Curtin University, GPO Box U1987, Perth WA 6845, Australia.
Anja
A
Kaczmarczyk
School of Biomedical Sciences, CHIRI Biosciences Research Precinct, Faculty of Health Sciences, Curtin University, GPO Box U1987, Perth WA 6845, Australia.
Bryn
B
Funnekotter
Botanic Gardens and Parks Authority, Fraser Avenue, West Perth, WA 6005, Australia.
Shane R
SR
Turner
Botanic Gardens and Parks Authority, Fraser Avenue, West Perth, WA 6005, Australia.
Eric
E
Bunn
Applied Physics, School of Applied Sciences, RMIT University, GPO Box 2476, Melbourne, VIC 3001, Australia.
Gary
G
Bryant
Applied Physics, School of Applied Sciences, RMIT University, GPO Box 2476, Melbourne, VIC 3001, Australia.
Taavi E
TE
Hunt
School of Biomedical Sciences, CHIRI Biosciences Research Precinct, Faculty of Health Sciences, Curtin University, GPO Box U1987, Perth WA 6845, Australia.
Ricardo L
RL
Mancera
D001315:Australia; D002152:Calorimetry, Differential Scanning; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000378:metabolism; D005838:Genotype; D029562:Magnoliaceae / Q000235:genetics / Q000502:physiology*; D018520:Plant Shoots / Q000235:genetics / Q000502:physiology; D046509:Tissue Culture Techniques; D058989:Vitrification
D003451:Cryoprotective Agents