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选择高粱正交F_1杂种和它们纯系亲本作为试验材料,用He-Ne激光器在低强度下处理其萌发种子,用甲基化敏感扩增片断多态性(MSAP)分子标记技术在全基因组范围内检测分析高粱各种基因型DNA甲基化水平和变异模式,旨在探讨激光辐射和杂交对基因组DNA甲基化的交互作用机制。结果表明,激光辐射诱导高粱F_1杂种和相应亲本在DNA甲基化水平和模式上发生一定变化,再次证明激光辐射诱导是一种表观遗传诱变手段。从甲基化水平上看,激光辐射诱导使高粱3种基因型整体表现为去甲基化。从甲基化变异模式看,CG位点甲基化变异模式要高于CNG位点甲基化变异模式。比较激光辐射对不同基因型的影响,发现F_1杂交种的甲基化总体水平要低于两纯系亲本甲基化总体水平的中间值。总之,通过试验证明激光辐射是一种直接的表观遗传诱变手段,激光辐射和杂交联用诱导高粱产生更复杂的甲基化变异,这两种手段联用可能是一个重要改良作物的手段,值得深入研究。
Orthogonal F_1 hybrids of Sorghum and their pure line parents were selected as experimental materials, their germinating seeds were treated with He-Ne laser at low intensity, and the whole genomes were screened by methylation sensitive amplified fragment polymorphism (MSAP) The detection and analysis of DNA methylation and variation patterns of various genotypes in sorghum were carried out in-house to investigate the interaction mechanism of laser radiation and hybridization on genomic DNA methylation. The results showed that there was a certain change in DNA methylation level and pattern in F_1 hybrids and corresponding parents induced by laser radiation, which proved once again that laser induced radiation was an epigenetic mutagenesis method. From the perspective of methylation level, three types of sorghum genotypes were demethylated by laser radiation. From the methylation mutation pattern, the methylation variation pattern of CG locus is higher than that of CNG locus methylation mutation pattern. Comparing the effects of laser radiation on different genotypes, we found that the overall level of methylation in F_1 hybrids was lower than the median of the overall level of methylation in two pure lines. In conclusion, laser irradiation was proved to be a direct epigenetic mutagenesis experiment by means of experiments. The combination of laser irradiation and hybridization induced more complicated methylation variation in sorghum. The combination of these two methods may be an important crop improvement method , Worth in-depth study.