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Measuring Gene Expression in Bombarded Barley Aleurone Layers with Increased Throughput
Published on: March 30, 2018
Expression GWAS and Differential Methylome Profiling Reveal Upstream Regulators of Drought Memory Genes in
Debankona Marik1, Rishabh Kumar1, Ayan Sadhukhan1
1Department of Bioscience and Bioengineering, Indian Institute of Technology Jodhpur, Jodhpur, Rajasthan, India.
Abstract:
Plants adapt to recurrent drought through transcriptional memory, yet the upstream regulators remain largely unknown. This study integrated expression genome-wide association study (eGWAS) across 115 Arabidopsis thaliana ecotypes with differential methylome profiling to identify these regulators. Focusing on the memory genes LKR, HIS1-3, and DREB1A, eGWAS identified signaling and epigenetic loci involved in ABA/JA responses and DNA methylation. Methylome profiling by whole-genome bisulfite sequencing of ecotypes contrasting in drought tolerance, and in the superinduction of memory genes revealed significantly greater methylation variation during the second drought (D2) than during the first (D1), highlighting the role of epigenetic reprogramming in memory maintenance. Functional validation using T-DNA mutants demonstrated specific modulation of the D2/D1 induction ratio without affecting initial drought responses. Mutants of LKR eGWAS-delineated genes AT1G56660, AT2G19120, AT4G16490, and DEG3, those of HIS1-3 eGWAS genes AT1G14220, AT2G24960, AT3G10845, AT3G19340, CNGC10, EMB2770, GRF7, and RPP2A, and DREB1A eGWAS genes AT1G67000, AT3G61610, AT5G62110, HK2, JMJ12, and LUP5 abolished respective memory gene induction. The eGWAS and methylome approaches converged on DNA repair, chromatin modification, vesicular transport, and proteostasis as core memory hubs. These findings reveal a genetic-epigenetic interplay that coordinates transcriptional memory, priming plants for rapid reactivation of stress pathways during recurrent drought.
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