山药愈伤组织EMS诱变及其再生苗变异研究

EMS Mutagenesis in Callus of Dioscorea opposita and It's Regeneration Plantlets Variation

摘要:

为了加速创造山药新种质，以“明淮2号”山药愈伤组织为材料，研究了不同浓度EMS[0（ck），0.05%，0.1%，0.2%，0.3%]、不同时间[0(ck)，6h，12h，24h，48h]处理对山药愈伤组织致死率的影响，以确定半致死浓度和时间；比较了在半致死处理后愈伤组织不同继代次数的影响情况；并对再生植株进行形态学比较和分子鉴定；结果表明，EMS诱变浓度和时间都对山药愈伤组织的致死率有显著影响，且EMS诱变浓度影响更显著，适宜的诱变方法为0.1%EMS浓度处理48h，0.2％EMS处理12h；诱变后愈伤组织增殖系数和分化率随继代次数增加而增加，但诱变率随继代次数增加而显著降低，诱变后愈伤组织经过一次继代后转入分化有利诱变体的筛选，再生植株主要发生叶色、叶型变异，通过RAPD标记鉴定结果显示部分植株条带确实出现了差异。

Abstract:

To accelerate the creation of new germplasm of Dioscorea opposita, with the callus of “Minghuai 2” as material, the effects of different concentrations of EMS [0(ck)，0.05%，0.1%，0.2%，0.3%] and different time [0(ck)，6h，12h，24h，48h] on the mortality of callus were studied, so as to determine the semi lethal concentration and time; The effect of callus in different subculture times was compared after semi lethal treatment; And the morphological comparison and molecular identification of regeneration plantlets were carried out; The results show that: The concentration and time of EMS had a significant effect on the death rate of the callus; And the effect of EMS concentration on the death rate of callus was more significant than that of the treatment time; The suitable EMS mutation method was: 0.1%EMS concentration was treated by 48h, 0.2%EMS was treated by 12h; The proliferation coefficient and regeneration rate of callus increased with the increase of subculture times; However, the mutation rate decreased significantly with the increase of subculture; So after mutation, the callus was subcultured once and then transferred to regeneration is the most favorable screening of the mutant; The variation of regenerated plants mainly occurred in leaf color and leaf shape; By RAPD molecular markers, the results showed that the DNA bands of some regeneration plantlets did appear to differ.