An efficient immunodetection method for histone modifications in plants

Plant Methods

Clelia De-la-Peña²  Víctor M Loyola-Vargas¹  Angela Kú-González¹  Sara Hernández-Castellano²  Geovanny Nic-Can¹ 

[1] Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán, Calle 43 No. 130, Col. Chuburná de Hidalgo, Mérida CP 97200, Yucatán, México;Unidad de Biotecnología, Centro de Investigación Científica de Yucatán, Calle 43 No. 130, Col. Chuburná de Hidalgo, Mérida CP 97200, Yucatán, México

关键词: Immunodetection;    H3K9me2;    H3K4me3;    Histone H3;    Epigenetic mechanism;    Coffea canephora;    Cedrela odorata;    Capsicum chinense;    Arabidopsis thaliana;    Agave tequilana;   

Others  :  805384
DOI  :  10.1186/1746-4811-9-47

 received in 2013-08-19, accepted in 2013-12-02,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

   
2013 Nic-Can et al.; licensee BioMed Central Ltd.

【 图 表 】

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