Cryopreservation techniques of important vegetable seed germplasm and some recalcitrant seed of economic fruit
1997
Panie Tongpamnak | Monthar Wongmaneeroj | Bualuang Joypoy (Kasetsart Univ., Bangkok (Thailand). Kamphaeng Saen Campus. Central Laboratory and Greenhouse Complex)
Methodology of seed cryopreservation techniques was set up from the study on vegetables and lequmes. The experiment found out that, it was necessary to adjust moisture content of the seed to equilibrium point or lower, then they were filled up in cryotube and placed directly in liquid nitrogen tank at -196 deg C. The seeds could be kept as long as liquid nitrogen available. When the seeds are needed to be grown, they could be removed from liquid nitrogen tank without any special treatment for thawing. Then they could be planted directly in the field and handling them as if they were normal seeds. Several kinds of seed being survival after storage under liquid nitrogen such as sweet basil, chilli, tomato, sweet corn, okra, cucumber, chinese kale, mung bean, yard long bean and soybean. Except green soybean, its survival percentage was lower than control seeds. In order to save space for storage in 2.5 ml cryotube, the big seed size such as mung bean, yard long bean, soybean and green soybean, were excised embryo during imbibition storage and transfer into cryotube by antiseptic techniques. It was necessary to pass the step of pre-cooling at -40 deg C before deeping embryo into liquid nitrogen. Recovering the freezing embryo, needed thawing process at 40 deg C least 16 hours before transplanting in MS media. Then after acclimatization that seedlings could bring out in the field.
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