Comparative physiological, biochemical and transcriptomic analysis of hexaploid wheat (T. aestivum L.) roots and shoots identifies potential pathways and their molecular regulatory network during Fe and Zn starvation.

The combined effect of iron (Fe) and zinc (Zn) starvation on their uptake and transportation and the molecular regulatory networks is poorly understood in wheat. To fill this gap, we performed a comprehensive physiological, biochemical and transcriptome analysis in two bread wheat genotypes, i.e. Narmada 195 and PBW 502, differing in inherent Fe and Zn content. Compared to PBW 502, Narmada 195 exhibited increased tolerance to Fe and Zn withdrawal by significantly modulating the critical physiological and biochemical parameters. We identified 25 core genes associated with four key pathways, i.e. methionine cycle, phytosiderophore biosynthesis, antioxidant and transport system, that exhibited significant up-regulation in both the genotypes with a maximum in Narmada 195. We also identified 26 microRNAs targeting 14 core genes across the four pathways. Together, core genes identified can serve as valuable resources for further functional research for genetic improvement of Fe and Zn content in wheat grain.

Transcriptomic dataset reveals the molecular basis of genotypic variation in hexaploid wheat (T. aestivum L.) in response to Fe/Zn deficiency.

The datasets depicted in the paper are related to the original article entitled "Identifying transcripts associated with efficient transport and accumulation of Fe and Zn in hexaploid wheat (T. aestivum L.)" [1]. Four wheat genotypes i.e. Sonora 64, CRP 1660, Vinata, and DBW 17 were selected for RNA sequencing using Illumina HiSeq4000 platform. These genotypes were grown in Fe/Zn sufficient and deficient conditions in sand pot culture with intermittent administration of Hoagland solution. Pooled assembly was carried out for all of the four varieties subsequent to discarding low-quality reads, adaptor sequences and contamination resulting in approximately 315,904 clean transcripts of around 937 bp lengths and N50 of 1,294 bp. For the functional annotation of the identified transcripts databases like Pfam, KEGG pathway, Uniprot, PlnTFDB and wheat proteins were utilized. Differential expression calculation of transcripts was carried out by DESeq, an R package and real-time PCR study of 12 Fe/Zn metabolic pathway related transcripts was utilized for further revalidation of data. Elemental analysis of grain Fe and Zn was performed using Flame Atomic Absorption Spectrometry (FAAS). The RNA-seq data of all the four wheat genotypes was uploaded on Sequence Read Archive (SRA: SUB6961770 and BioProject: PRJNA605909), enabling easy access to the researchers worldwide.

Identifying transcripts associated with efficient transport and accumulation of Fe and Zn in hexaploid wheat (T. aestivum L.).

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.