Spatial Gene Expression Analysis of Histone Modifications of ChIP-DNA in Durum Wheat Tissues

Histone modifications are associated with gene expression in their ability to condense or relax chromatin, allowing their expression or repression. In acetylation, the chromatin relaxes, allowing transcription factors to reach their target, resulting in gene expression. In methylation, the chromatin condenses, restricting transcription factors from reaching their target, impeding gene expression. Chromatin Immunoprecipitation (ChIP) examines the interactions between protein and DNA within the genome of an organism by studying histone modifications. These interactions may play a major role in combating drought, adjusting to heat stress, and fighting diseases during plant developmental stages. In this study, we seek to optimize ChIP-DNA isolation in Durum wheat and examine ChIP-DNA isolates in four tissue types (leaf, root, head, and two week old seedling) to identify differential histone-DNA interactions among the group using Illumina sequencing. Optimization of the ChIP-DNA isolation protocol in Durum wheat may allow us to better understand histone modifications and the role they play in combating abiotic and biotic stresses. We will also use quantitative PCR to identify chromosome remodeling gene expression differences among tissue types. By knowing where these specific genes are located in the genome we may be able to better understand their spatial importance. The goal of this research is to serve as a blueprint for studying spatial histone methylation and acetylation in wheat using the ChIP assay.