High cytokinin concentration and nutrient starvation trigger DNA methylation changes in somaclonal variants of Agave angustifolia Haw

Abstract

Somaclonal variation (SV) is observed during micropropagation, especially in organogenic processes when differentiated somatic cells are induced into a pluripotent state. However, the clones obtained show differences in phenotype from each other and from the mother plants, which could be an agricultural problem or an ornamental advantage. Few of the mechanisms associated with how a somatic cell generates phenotypic variation have been elucidated. Recently, our group demonstrated that an albino phenotype produced in Agave in in vitro culture was linked to changes in DNA methylation through the micropropagation process, suggesting a new way to investigate the mechanism. Therefore, here we evaluate the effect of stressful conditions in the culture media on three different phenotypes (green, variegated and albino) of Agaveangustifolia Haw. to determine the changes in DNA methylation linked to SV in the new formed shoots. The effects of nutrient availability and exogen cytokinin concentration in the culture media were evaluated for five months in each phenotype. We observed that a reduction of 25 % in the nutrients in the culture media affected the coloration of green plantlets. On the other hand, a high 6-benzyladenine (BA) concentration exposure (88.8 μM) in the variegated phenotype enhanced organogenic capability, increased phenotypic variation, and changed DNA methylation levels. We have demonstrated that long periods of time under nutrient-limited conditions or high cytokinin concentration provoke changes in DNA methylation levels that increase SV frequency in the newly formed shoots. Epigenetic changes may thus be an overlooked factor in SV induced by culture media stress.