Mandal B B, Ahuja-Ghosh Sangeeta, Srivastava P S
Tissue Culture and Cryopreservation Unit, National Bureau of Plant Genetic Resources, New Delhi, India.
Cryo Letters. 2008 Sep-Oct;29(5):399-408.
The aim of this study was to develop a cryopreservation protocol for Dioscorea rotundata with maintenance of genetic stability of regenerated plants after cryopreservation. In vitro shoot tips were cryopreserved using vitrification and encapsulation-dehydration to compare the efficacy of the two methods. Both methods produced high levels of plant regeneration from cryopreserved shoot tips. The regeneration level obtained using vitrification (71%) was not significantly different from that obtained using encapsulation-dehydration (67%). Genetic stability of plants derived from cryopreserved shoot tips was evaluated using RAPD markers. Analysis of 50 cryopreserved-derived and 20 in vitro- maintained (control) plantlets showed that 10 primers produced 77 clear, reproducible bands, with the amplification products being monomorphic for all the plantlets tested. A total of 5,390 bands obtained from this study exhibited no aberration in RAPD banding. Thus, the present study showed that both vitrification and encapsulation-dehydration methods are equally applicable to D. rotundata for cryopreservation. The in vitro plantlets derived from cryopreservation were genetically stable at the molecular level tested.
本研究的目的是开发一种用于甘薯的冷冻保存方案，以在冷冻保存后维持再生植株的遗传稳定性。使用玻璃化法和包埋脱水法对离体茎尖进行冷冻保存，以比较这两种方法的效果。两种方法都能使冷冻保存的茎尖产生高水平的植株再生。使用玻璃化法获得的再生率（71%）与使用包埋脱水法获得的再生率（67%）没有显著差异。使用随机扩增多态性DNA（RAPD）标记评估了源自冷冻保存茎尖的植株的遗传稳定性。对50株冷冻保存衍生的和20株离体保存（对照）的植株进行分析表明，10条引物产生了77条清晰、可重复的条带，所有测试植株的扩增产物均为单态性。本研究共获得5390条条带，其RAPD条带没有异常。因此，本研究表明，玻璃化法和包埋脱水法同样适用于甘薯的冷冻保存。在测试的分子水平上，源自冷冻保存的离体植株具有遗传稳定性。
