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采用扩增片段长度多态性(AFLP)和甲基化敏感扩增多态性(MSAP)技术分析红豆杉脱分化前后基因组DNA和DNA甲基化状态的变化。选用32个AFLP引物组合从红豆杉植株及其愈伤组织分别扩增出1834个片段,无多态性片段产生。这说明红豆杉植株在诱导形成愈伤组织的过程中基因组DNA保持高度的遗传稳定性。另用32个MSAP引物组合从红豆杉植株及其愈伤组织分别扩增出1197个片段,总扩增位点的甲基化水平由脱分化前的12.4%上升为16.2%,表明红豆杉在脱分化过程中的某些位点发生了甲基化。红豆杉脱分化前后的DNA甲基化模式也存在较大差异,说明DNA甲基化对愈伤组织形成有调控作用。
The genomic DNA and DNA methylation status of Taxus chinensis before and after dedifferentiation were analyzed by amplified fragment length polymorphism (AFLP) and methylation-sensitive amplified polymorphism (MSAP). Twenty-two AFLP primers were used to amplify 1834 fragments from the plants and their callus, respectively. No polymorphic fragments were produced. This shows that the genomic DNA of the yew plants maintained a high degree of genetic stability during induction of callus formation. In addition, 1197 fragments were amplified from the yew plants and their callus using 32 MSAP primer combinations respectively. The methylation level of total amplification sites increased from 12.4% before dedifferentiation to 16.2% Methylation occurs at some sites during dedifferentiation. DNA methylation patterns before and after the dedifferentiation of Taxus also have a large difference, indicating that DNA methylation on callus formation has a regulatory role.