Embryonic axes of a tropical recalcitrant-seeded species: successful elimination of micro-organisms and potential for zygotic synthetic seed (synseed) production

作者：Berjak P^*; Cherian J; Makhathini A P; Sershen; Pammenter N W

来源：Seed Science and Technology, 2014, 42(2): 150-160.

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 (R) bags were stored for up to 42 days with undiminished viability, while rapid loss of viability occurred when the capsules were stored in Magenta (TM) vessels, Microtubes (R) 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.

• 出版日期2014