Descripción: |
Background: Epigenetic mechanisms can be highly dynamic, but the cross-talk among them and with the genome
is still poorly understood. Many of these mechanisms work at different places in the cell and at different times of
organism development. Covalent histone modifications are one of the most complex and studied epigenetic
mechanisms involved in cellular reprogramming and development in plants. Therefore, the knowledge of the spatial
distribution of histone methylation in different tissues is important to understand their behavior on specific cells.
Results: Based on the importance of epigenetic marks for biology, we present a simplified, inexpensive and
efficient protocol for in situ immunolocalization on different tissues such as flowers, buds, callus, somatic embryo
and meristematic tissue from several plants of agronomical and biological importance. Here, we fully describe all
the steps to perform the localization of histone modifications. Using this method, we were able to visualize the
distribution of H3K4me3 and H3K9me2 without loss of histological integrity of tissues from several plants, including
Agave tequilana, Capsicum chinense, Coffea canephora and Cedrela odorata, as well as Arabidopsis thaliana.
Conclusions: There are many protocols to study chromatin modifications; however, most of them are expensive,
difficult and require sophisticated equipment. Here, we provide an efficient protocol for in situ localization of
histone methylation that dispenses with the use of expensive and sensitive enzymes. The present method can be
used to investigate the cellular distribution and localization of a wide array of proteins, which could help to clarify
the biological role that they play at specific times and places in different tissues of various plant species |