以留兰香为材料,研究了土霉素处理对留兰香生长发育和生理生化反应的影响,以及不同浓度土霉素处理对留兰香基因组DNA的甲基化水平和模式的影响。在无菌条件下,将留兰香茎段接种到含5种不同浓度0(对照)、5、10、15、20μmol/L的土霉素处理培养基上,连续处理14 d。基因组DNA用混合取样法,取100株组培苗嫩叶,混合并提取。用AFLP、MSAP的方法进行相关的分析。结果表明,土霉素处理14 d后,叶绿素含量随着土霉素浓度的升高逐渐降低,丙二醛( MDA)的合成也呈下降趋势。 AFLP分析表明,土霉素处理组与对照组之间未发现特异性片段。 MSAP分析表明,5、10、15、20μmol/L 土霉素处理后,留兰香全基因组 DNA 甲基化水平分别为14.16%、13.92%、18.84%、8.95%,均低于对照(22.75%)；与对照相比,土霉素处理后DNA甲基化率分别为10.31%、10.87%、10.94%、11.34%,去甲基化率分别为11.86%、11.41%、15.63%、17.53%。土霉素影响了留兰香的生长发育,并引起各项生理生化指标的改变。土霉素未造成留兰香基因序列的变异,却能引起基因组DNA甲基化水平降低。
Effects of Oxytetracycline Treatment on Genomic DNA and Its Methylation of Menthae spicatae L.
The objectives of this research were to assess the effect of oxytetracycline treatment on the de-velopment as well as genomic DNA methylation levels and patterns of Menthae spicatae L.. Spearmint stems were cultured on MS medium supplied with oxytetracycline(0,5,10,15,20 μmol/L,respectively) for 14 d. Genomic DNA was extracted from leaves. The amplified fragment length polymorphisms( AFLP) and methylation sensitive amplification polymorphism( MSAP) were used to test the genetic and epigene-tic stability. When Menthae spicatae L. were treated by oxytetracycline for 14 d, the chlorophyll content were significantly reduced,whereas the MDA content increased as the concentration of oxytetracycline in-creased. AFLP technique did not detect any sequence change. MSAP technique showed that the genomic DNA methylation levels of Menthae spicatae L. were 14. 16%,13. 92%,18. 84%,8. 95% under 5,10, 15,20 μmol/L oxytetracycline stress,which were lower than that of the untreated plants with 22. 75%;the ratio of methylation and demethylation of Menthae spicatae L. genomic DNA at oxytetracycline concen-trations of 5 ,10 ,15 ,20 μmol/L were 10 . 31%,10. 87%,10. 94%,11. 34% and 11. 86%,11. 41%,15. 63%, 17. 53%. The growth and physiological and biochemical reactionis of Menthae spicatae L. had changed un-der oxytetracycline treatment. Oxytetracycline could not impact the genomic DNA sequence, however, it could result in the reduction of DNA methylation.
Menthae spicatae L.methylationoxytetracyclinegenomic DNA
国家科技期刊平台
NSTL
万方数据
维普
