Embryonic axes of a tropical recalcitrant-seeded species: successful elimination of micro-organisms and potential for zygotic synthetic seed (synseed) production
A major problem encountered with embryos/embryonic axes excised from tropical recalcitrant seeds as explants for cryopreservation is contamination by micro-organisms, making it essential that effective, non-injurious decontamination procedures are developed a priori. The effects of commonly-used hypochlorites, mercuric chloride and sodium dichloroisocyanurate (NaDCC) were evaluated on excised embryonic axes of Trichilia dregeana and their associated microflora. Only exposure of the axes to 0.2% NaDCC for 20 minutes or 0.3% NADCC for 10 and 20 minutes facilitated full viability retention by the explants and completely eliminated all axis-associated contaminants. Axes pre-treated with 0.3% NaDCC for 20 minutes were encapsulated in alginate gel incorporating 0.5% sucrose to form zygotic synseeds. Those alginate capsules sealed in polythene derived from Ziploc® bags were stored for up to 42 days with undiminished viability, while rapid loss of viability occurred when the capsules were stored in Magenta™ vessels, Microtubes® or metallised pouches. This study using recalcitrant embryonic axes of T. dregeana has demonstrated the efficacy of decontamination with NaDCC, the unequivocal potential of synseed technology for short-term storage and dissemination (ultimately after retrieval from cryopreservation), and the importance of containment which prevents/curtails water loss while facilitating gaseous exchange.
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Document Type: Research Article
Publication date: August 1, 2014
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