DNA-methylation changes in grapevine somaclones following in vitro culture and thermotherapy
2010
Baránek, Miroslav | Křižan, Břetislav | Ondrušíková, Eva | Pidra, Miroslav
The methylation-sensitive amplified polymorphism (MSAP) technique using HpaII and MspI isoschizomers was used to analyse DNA-methylation alterations in stressed grapevine plants. The stress used was in vitro propagation via nodal segments and in vitro thermotherapy for virus elimination. A set of pertinent grapevine plants derived from two cultivars (18 plants each for Müller Thurgau and Riesling) was used as stressed variants for analyses. A total of 695 and 700 MSAP bands were recognised and evaluated as present/absent for all analysed variants derived from both cvs. Müller Thurgau and Riesling. Average computed similarity of MSAP banding between analysed variants (Dice/Nei and Li coefficient) was 0.935 for both cultivars. Clustering of variants within resulting dendrograms showed significant differences between woody cuttings despite originating from the one plant. Further, there was a strong ‘donor' effect of maternal plants on future arrays of DNA methylation in their regenerants. The ‘donor' effect even seemed to prevail in the effect of stress on final DNA-methylation state in stressed regenerants. Additional MSAP evaluation suggests that thermotherapy induced an additional array of methylation changes when compared with stress caused by in vitro cultivation. From the viewpoint of whether methylation of CCGG loci increased/decreased due to stress, the results showed moderate prevalence for decreasing CCGG loci methylation.
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