Abstract

As efforts are made to increase food security, millets are gaining increasing importance due to their excellent nutritional credentials. Among the millets, pearl millet is the predominant species possessing several health benefiting nutritional traits in its grain that are helpful in mitigating chronic illnesses such as type−2 diabetes and obesity. In this paper, we conducted metabolomic fingerprinting of 197 pearl millet inbred lines drawn randomly from within the world collection of pearl millet germplasm and report the extent of genetic variation for health benefitting metabolites in these genotypes. Metabolites were extracted from seeds and assessed using flow infusion high-resolution mass spectrometry (FIE-HRMS). Metabolite features (m/z), whose levels significantly differed among the germplasm inbred lines, were identified by ANOVA corrected for FDR and subjected to functional pathway analysis. A number of health-benefiting metabolites linked to dietary starch, antioxidants, vitamins, and lipid metabolism-related compounds were identified. Metabolic genome-wide association analysis (mGWAS) performed using the 396 m/z as phenotypic traits and the 76 K SNP as genotypic variants identified a total of 897 SNPs associated with health benefiting nutritional metabolite at the -log p-value ≤ 4.0. From these associations, 738 probable candidate genes were predicted to have an important role in starch, antioxidants, vitamins, and lipid metabolism. The mGWAS analysis focused on genes involved in starch branching (α-amylase, β-amylase), vitamin-K reductase, UDP-glucuronosyl, and UDP-glucosyl transferase (UGTs), L-ascorbate oxidase, and isoflavone 2′-monooxygenase genes, which are known to be linked to increases in human health benefiting metabolites. We demonstrate how metabolomic, genomic, and statistical approaches can be utilized to pinpoint genetic variations and their functions linked to key nutritional properties in pearl millet, which in turn can be bred into millets and other cereals crops using plant breeding methods.

Highlights

  • More than half of all deaths are due to malnutrition, and this is driving the need to improve the nutritional content of staple food crops that are consumed on a daily basis

  • Using a metabolic genome-wide association study approach, we report candidate genes associated with a diversity of plant metabolites in this germplasm set

  • Metabolite fingerprinting was performed using FIE-HRMS for 197 pearl millet accessions originating from different parts of the world

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Summary

Introduction

More than half of all deaths are due to malnutrition, and this is driving the need to improve the nutritional content of staple food crops that are consumed on a daily basis. Most of the millets, including pearl millet, have high protein, folic acid, vitamins, carotenoid contents [4,5], possess non-glutinous and non-acid forming properties, and are easy to digest They are rich in nutritionally important minerals like iron, calcium, zinc, magnesium, phosphorous, and potassium [6,7,8], and dietary fiber and several vitamins (β-Carotene, niacin, vitamin B6 and folic acid) [9,10]. Millets are rich in polyphenols, tannins, phytosterols and are a good source of antioxidants Despite these positive attributes, the use of millets as a food is restricted to the conventional consumers, predominantly the subsistence farming families who grow them. This can be attributed to the non-availability of user-friendly, ready to use/ready to eat millet-based food products

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