Plant tissue culture techniques are handy tools for obtaining unique
plant materials that are difficult to propagate or important for agriculture.
Homozygous materials derived through in vitro cultures are invaluable and
significantly accelerate the evaluation of new varieties, e.g., cereals. The
induction of somatic embryogenesis/androgenesis and the regeneration and
its efficiency can be influenced by the external conditions of tissue culture,
such as the ingredients present in the induction or regeneration media.
We have developed an approach based on biological system, molecular
markers, Fourier Transform Infrared spectroscopy, and structural equation
modeling technique to establish links between changes in sequence and
DNA methylation at specific symmetric (CG, CHG) and asymmetric (CHH)
sequences, glutathione, and green plant regeneration efficiency in the
presence of variable supplementation of induction medium with copper ions.
The methylation-sensitive Amplified Fragment Length Polymorphism was
used to assess tissue culture-induced variation, Fourier Transform Infrared
spectroscopy to describe the glutathione spectrum, and a structural equation
model to develop the relationship between sequence variation, de novo
DNA methylation within asymmetric sequence contexts, and copper ions in
the induction medium, as well as, glutathione, and green plant efficiency. An
essential aspect of the study is demonstrating the contribution of glutathione
to green plant regeneration efficiency and indicating the critical role of copper
ions in influencing tissue culture-induced variation, glutathione, and obtaining
green regenerants. The model presented here also has practical implications,
showing that manipulating the concentration of copper ions in the induction
medium may influence cell function and increases green plant regeneration
efficiency.