Evaluation of DNA methylation using methylation-sensitive amplification polymorphism in plant tissues grown in vivo and in vitro

Abstract

In present study, methylation-sensitive AFLP (MSAP) markers were employed to assess DNA methylation, degree of alterations in DNA methylation and methylation polymorphism in plant tissues growing in vivo and in vitro. The leaf tissues of six plants growing in vivo and in vitro were subjected to MSAP profiling. A total of 717 MSAP markers in Salvadora persica, 801 in Commiphora wightii, 874 in male (M) and 845 in female (F) genotype of Simmondsia chinensis, 719 in Jatropha curcas and 880 in Withania coagulans were obtained with seventeen MSAP primer combinations. Percentage methylation in genome obtained was higher in in vivo-grown tissues of S. persica (39.47 %), S. chinensis—M (61.71 %) and W. coagulans (71.59 %); and in in vitro-grown tissues of C. wightii (65.17 %), S. chinensis—F (60.83 %) and J. curcas (68.29 %). The percentage polymorphism in methylated DNA obtained was 8.71 % in S. persica, 9.81 % in J. curcas, 10.10 % in S. chinensis—F, 10.26 % in W. coagulans, 10.66 % in S. chinensis—M and 13.98 % in C. wightii. The difference in DNA methylation and polymorphism in genomes reflect the plasticity in genomes of the plants growing under two different environments. Different pattern of DNA methylation of the homologous nucleotide sequences and polymorphism in the methylated DNA in tissues under in vitro and in vivo conditions suggest possibility of involvement of these fragments in the dynamic processes regulating plant growth and development under prevailing growth conditions.