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Fluorescence imaging of cryopreserved Loxocarya cinerea shoot tips. Green indicates living tissue and blue, dead tissue. Photo: B. Funnekotter.View image slideshow

Project title: Advanced cryobanking for propagation-recalcitrant and critically endangered plant species.

Dates: 2009 - 2017

Funding: Australian Research Council (ARC) Linkage Projects schemes

Location: Biodiversity Conservation Centre (Kings Park)

Research themes

  • rare species conservation
  • south-west Western Australian species
  • cryopreservation
  • tissue culture
  • plant physiology
  • plant metabolomics

Project description

This project will advance methods for the repatriation of recalcitrant/threatened plant species to post-mined sites to achieve best practice ecological restoration in areas of high biodiversity. In particular, the nature of freezing and dehydration damage to cell membranes at various stages of tissue culture and cryopreservation will be investigated to try to minimise its deleterious effects.

In addition, metabolic changes that affect cryogenic survival will be assessed to provide further insight into the role of oxidative stress and the toxicity of cryopreservation processes. Ultimately improved cryogenic protocols will be developed to maintain the integrity of long-term cryobanks of key species for post-mining ecological restoration.

Key staff

Prof Ricardo Mancera (Curtin University), Prof Gary Bryant (RMIT), Dr Shane Turner (BGPA) and Dr Eric Bunn (BGPA)


Dr Bryn Funnekotter (Curtin University), Dr Anja Kaczmarczyk (Curtin University), Keran Keys (BGPA)


Bryn Funnekotter (PhD; Curtin University 2011-2015) The Role of Oxidative Stress in Successful Cryopreservation of South-west Western Australian Plant Species.

Michael Cahill (Hons; Curtin University 2015) Extraction of mitochondrial DNA from three Australian plant species assessing DNase1 and Percoll gradient purification protocols.

Sharon Fletcher (Hons; Curtin University 2014) Extraction and sequencing of plastid genome (cpDNA) of potential value for determining post-cryopreservation genetic integrity.

Anamika Sortey (Hons; Curtin University 2013) Influence of abiotic stress factors during preconditioning on post-cryogenic survival, antioxidant enzymes and electrolyte leakage in Lomandra sonderi.

Akshay Menon (Hons; Curtin University 2011) Enhanced cryogenic tolerance by cold acclimation as a means of conserving endangered native Australian species.

Bryn Funnekotter (Hons; Curtin University 2010) Factors Affecting Post-cryogenic Survival of Plant Shoot Tips.


Funnekotter B, Bunn E, Mancera RL (2017) Cryo-mesh: a simple alternative cryopreservation protocol. CryoLetters 38 (2):155-159.

Funnekotter B, Sortey A, Bunn E, Turner S, Mancera R (2016) Influence of abiotic stress preconditioning on antioxidant enzymes in shoot tips of Lomandra sonderi (Asparagaceae) prior to cryostorage. Aust J Bot 64 (3):260-268.

Funnekotter B, Bunn E, Mancera RL (2016) Cryo-mesh: a simple alternative cryopreservation protocol. CryoLetters In Press:1-5. Accepted 3.10.2016.

Funnekotter B, Whiteley SE, Turner SR, Bunn E, Mancera RL (2015) Evaluation of the new vacuum infiltration vitrification (VIV) cryopreservation technique for native Australian plant shoot tips. CryoLetters 36 (2):104-113.

Menon A, Funnekotter B, Kaczmarczyk A, Bunn E, Turner S, Mancera RL (2014) Cold-induced changes affect survival after exposure to vitrification solution during cryopreservation in the south-west Australian Mediterranean climate species Lomandra sonderi (Asparagaceae). Plant Cell Tiss Org Cult 119 (2):347-358.

Hughes ZE, Mancera RL (2014) Molecular Mechanism of the Synergistic Effects of Vitrification Solutions on the Stability of Phospholipid Bilayers. Biophys J 106 (12):2617-2624.

Malajczuk CJ, Hughes ZE, Mancera RL (2013) Molecular dynamics simulations of the interactions of DMSO, mono-and polyhydroxylated cryosolvents with a hydrated phospholipid bilayer. Biochimica et Biophysica Acta (BBA)-Biomembranes 1828 (9):2041-2055.

Kaczmarczyk A, Funnekotter B, Turner SR, Bunn E, Bryant G, Hunt TE, Mancera RL (2013) Development of Cryopreservation for Loxocarya cinerea-An Endemic Australian Plant Species Important for Post-Mining Restoration. CryoLetters 34 (5):508-519.

Hughes ZE, Mancera RL (2013) Molecular dynamics simulations of mixed DOPC−β-sitosterol bilayers and their interactions with DMSO. Soft Matter 9:2920-2935.

Hughes ZE, Malajczuk CJ, Mancera RL (2013) The Effects of Cryosolvents on DOPC− β-Sitosterol Bilayers Determined from Molecular Dynamics Simulations. The Journal of Physical Chemistry B 117 (12):3362-3375.

Funnekotter B, Kaczmarczyk A, Turner SR, Bunn E, Zhou W, Smith S, Flematti G, Mancera RL (2013) Acclimation-induced changes in cell membrane composition and influence on cryotolerance of in vitro shoots of native plant species. Plant Cell Tiss Org Cult 114 (1):83-96.

Menon A, Funnekotter B, Kaczmarczyk A, Bunn E, Turner S, Mancera RL (2012) Cryopreservation of Lomandra sonderi (Asparagaceae) shoot tips using droplet-vitrification. CryoLetters 33 (4):259-270.

Kaczmarczyk A, Funnekotter B, Menon A, Phang PY, Al-Hanbali A, Bunn E, Mancera RL (2012) Current Issues in Plant Cryopreservation. In: Katkov II (ed) Current Frontiers in Cryobiology. InTech, pp 417-438.

Hughes ZE, Mark AE, Mancera RL (2012) Molecular Dynamics Simulations of the Interactions of DMSO with DPPC and DOPC Phospholipid Membranes. The Journal of Physical Chemistry B 116 (39):11911-11923.

Mandumpal JB, Kreck CA, Mancera RL (2011) A molecular mechanism of solvent cryoprotection in aqueous DMSO solutions. Phys Chem Chem Phys 13 (9):3839-3842.

Kreck CA, Mandumpal JB, Mancera RL (2011) Prediction of the glass transition in aqueous solutions of simple amides by molecular dynamics simulations. Chem Phys Lett 501 (4-6):273-277.

Kaczmarczyk A, Turner SR, Bunn E, Mancera RL, Dixon KW (2011) Cryopreservation of threatened native Australian species—what have we learned and where to from here? In Vitro Cell Dev Biol Plant 47 (1):17-25.

Regenerating shoot tips of Anigozanthos viridis after cryopreservation. Photo: B. Funnekotter.

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