Towards the identification of ZmNAC24 transcription factor targets using DNA affinity purification sequencing in maize

Details

Supervisors

Chaumont, François ; Teirlinckx, Estelle

Faculty

Faculté des bioingénieurs

Degree label

Master [120] : bioingénieur en chimie et bioindustries, à finalité spécialisée

Abstract

Maize is one of the world’s most important crops used for animal feed, human food and non-food applications. As climate change progresses, we are observing an increase in abiotic stresses on ecosystems and organisms. Regulation of water uptake and movement in plants are essentiaI for their physiology, especially in case of drought stress, and these processes depend on the activity of aquaporins. The abundance of these water channels in the cell membranes is primarily dependent on gene expression and therefore on transcription factors (TFs). The NAC TF family play roles in plant responses to abiotic stresses but also in development and growth. We previously identified ZmNAC24 as a TF controlling the expression of ZmPIP2;5 and ZmPIP2;9 aquaporin genes, but little more is known about it. In order to identify other NAC24 target genes and its binding motifs, we set up a DNA Affinity Purification sequencing approach using NAC24 protein and maize root gDNA fragments. We adapted and optimized different steps of the protocol, and succeeded in obtaining gDNA fragments that bind to NAC24. These fragments were sent for sequencing. In addition, we prepare genetic constructs to express tagged NAC24 together with the fluorescent mCherry marker in maize BMS suspension cells. These cells were transformed and fluorescent calli were obtained. These cells lines will be used for a NAC24 ChIP-seq analysis and other functional studies.