Wheat (T. aestivum L.) is the second most important staple food crop consumed in the form of various end-use products across the world. However, it contains lower concentrations of Fe and Zn leading to micronutrient deficiency in human beings where wheat is the sole diet. Therefore, increasing grain Fe/Zn content in wheat has become priority in wheat breeding programmes across the world. Understanding the molecular mechanism of Fe/Zn transport and accumulation in grains is required to expedite the breeding process. For this purpose, whole seedling transcriptome analysis was conducted in four wheat genotypes (CRP 1660, Sonora 64, Vinata, : high, and DBW17: low) differing in grain Fe/Zn content under controlled and Fe/Zn deficient conditions. Twenty eight key transcripts involved in phytosiderophore biosynthesis, Fe/Zn uptake and transport were identified. Expression analysis of 12 of the transcripts using qPCR was conducted in seedling stage and flag leaf which exhibited greater differential accumulation in CRP 1660 followed by Vinata, Sonora 64 and DBW 17 in both flag leaf and seedling. However, there was significantly higher differential accumulation of the transcripts in flag leaf as compared to seedling. In CRP 1660, transcripts pertaining to phytosiderophore biosynthesis like DMAS1-B, NRAMP2 and NAAT2-D showed greater accumulation. Additionally, corresponding miRNAs were also identified for these 28 transcripts. The findings will help in better understanding of molecular basis of Fe/Zn transport and accumulation in grain and subsequent utilization in breeding to improve Fe/Zn content in wheat grain., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)