본 실험은 국화의 바이로이드 제거에 이용되는 초저온처리 시 국화 품종 'White ND'을 적합한 처리조건을 확립하기 위해 초저온처리의 단계별 요인을 실험하였다. 그 결과 생장점의 크기는 1 mm(엽원기 2~3매 포함)에서 높은 생존율과 신초 재생율을 나타내었고, vitirification 처리시 PVS3가 효과적이었으며, 처리 시간은 60분 처리 하였을 때 높은 생존율 및 정 상 신초 재생율을 보였다. 또한 vitrification을 위한 전처리 조건은 sucrose 농도를 88 mM 24시간, sucrose 0.3 M 16시간, sucrose 0.5 M 6시간, sucrose 0.7 M 3시간으로 처리하는 것이 초저온 처리 후 생존율 및 신초 재생율을 높이는데 효과적이었으며, 재생된 정상 식물체는 모본과 비교하여 ploidy level이 동일한 것으로 보아 식물체의 유전적 변이가 일어나지 않았다.

The preservation of pear germplasm, like that of other clonal germplasms, is difficult because it requires conservation of whole plants or their tissues. Among the currently available methods for long-term conservation of clonal germplasm, cryopreservation of shoot tips is the most reliable and cost- and space-effective option. Alginate-coated axillary shoot tips from in vitro−grown pear were conserved successfully in liquid nitrogen (LN) following dehydration. Shoot recovery from cryopreserved shoot tips was improved greatly after 8 weeks of cold acclimation, but recovery decreased slightly after then. The highest regeneration rate was observed when in vitro shoot tips were preincubated in MS (Murashige and Skoog) medium with 0.3 M sucrose for 48 h, and when alginate-coated shoot tips were precultured in MS medium with increasing sucrose concentrations (0.5 M and 0.7 M) for 8 and 16 h, respectively. When the encapsulated beads were dehydrated for up to 7 h [25% water content (fresh weight basis)] under laminar flow, the highest regeneration rate was observed in “BaeYun No. 3” (55.7%) and “Whanggeum” (43.3%) after warming from LN. This technique is useful as a practical procedure to cryopreserve plant material that is sensitive to freezing of the surrounding cryoprotectant medium. Therefore, this technique appears to be promising for the cryopreservation of shoot tips from in vitro−grown plantlets of pear germplasm.

A simplified technique which cryoprotects zygotic embryos by encapsulation-dehydration was developed for the germplasm conservation of herbaceous peony (Paeonia lactiflora Pall.). The highest survival rate (85~%) was obtained from embryos treated by encapsulation-dehydration. The zygotic embryos were precultured on MS medium containing 0.3mg/L GA3 for 1 day. The precultured embryos were encapsulated in 3~% (w/v) alginate beads and immersed for 1 h in MS medium containing 2 M glycerol and 0.5 M sucrose. The encapsulated embryos were dehydrated for 5h by air drying prior to direct immersion in liquid nitrogen. This encapsulation-dehydration method appears to be a promising technique for germplasm cryopreservation of a herbaceous peony.

The rol e of osm otol erance and dehydr ation, in terms of both medium com posi tion and exposur e duration, on sur vi val ofem bryos of Citrus m adurensi s after cryopr eser vation usi ng the encapsul ation-dehydr ation and modified encapsul ation-dehydr