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研究马铃薯茎尖超低温保存技术的结果表明,4℃低温下锻炼6d,在添加二甲基亚砜(DMSO)和乙酰胺的培养基中预培养5d,60%PVS2于室温下装载30min,0℃下PVS2脱水40min时,茎尖成活率最高(71.6%),再生植株生长分化正常。进一步对再生植株进行AFLP分析,6对引物组合共扩增出385条带,超低温保存前后的材料之间未见到明显差的异带,但用MSAP技术分析超低温保存前后植株甲基化的结果显示:超低温保存后的材料均有不同程度的甲基化。在扩增的624条带中,处理与否之间完全一致的带型为584条;有变化的带型为40条,处理2(茎尖经过完整的超低温保存过程,区别于处理1,增加了冷冻、解冻和洗涤后恢复培养)有13个位点的甲基化增加,21个位点去甲基化。
The results of cryopreservation techniques of potato shoot tips showed that pre-cultured for 5 days in 4 ℃ low temperature for 6 days, DMSO for 5 days and acetamide, 60% PVS2 for 30 minutes at 0 ℃ Under 40 minutes dehydration of PVS2, the survival rate of shoot tip was the highest (71.6%), and the growth and differentiation of regenerated plants were normal. Further AFLP analysis of regenerated plants showed that 385 bands were amplified by 6 pairs of primers and no significant difference was found between the materials before and after cryopreservation. However, the results of plant methylation before and after cryopreservation were analyzed by MSAP Show: After cryogenic storage of materials have different degrees of methylation. Among the 624 amplified bands, 584 bands were completely consistent between treatments; 40 bands changed, treatment 2 (the tips of the shoots were completely preserved after cryopreservation, different from treatment 1, increased After freezing, thawing and washing, the methylation was increased in 13 loci and demethylation in 21 loci.