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Item Genome sequencing efforts in minor millets: Current knowledge and emerging insights(Springer Nature Publishing AG, 2025) Singh, Roshan Kumar; Panchal, Anurag; Muthamilarasan, Mehanathan; Prasad, ManojSmall millets (or minor millets) include finger millet (Eleusine coracana), foxtail millet (Setaria italica), proso millet (Panicum miliaceum), barnyard millet (Echinochloa crus-galli), kodo millet (Paspalum scrobiculatum), little millet (Panicum sumatrense), teff (Eragrostis tef), fonio (Digitaria exilis), job’s tears (Coix lacryma-jobi), guinea millet (Brachiaria deflexa), and browntop millet (Urochloa ramosa). These millets are highly nutritious and climate-resilient but marginally cultivated for the production and consumption of particular communities. Though called “poor men’s crops,” minor millets possess the potential to ensure food and nutritional security amid the threat of global climate change. Thus, scope exists to improve the agronomic traits of these minor millets for commercial cultivation; however, lack of genomic resources remains a bottleneck to this advancement. Genome sequencing not only provides an opportunity to decode the genes encoded by the genome, but also provides avenue for the development of genomic resources. The success of genome sequencing for resource development and further implementation of these resources have been proven in other crop plants. Among minor millets, genomes of a few species have been sequenced, including finger millet, foxtail millet, proso millet, barnyard millet, teff, fonio, and job’s tears. However, the genomes of kodo millet, little millet, guinea millet, and browntop millet remains to be sequenced. In this context, the chapter summarizes the outcomes of sequencing efforts and the application of genome sequence information in accelerating genomics studies in minor millets. The chapter also enumerates the status of transcriptome sequencing and its application in dissecting the genes underlying important traits.Item Multi-environment GWAS identifies genomic regions underlying grain nutrient traits in foxtail millet (Setaria italica)(Springer Nature Publishing AG, 2024) Jaiswal, Vandana; Bandyopadhyay, Tirthankar; Singh, Roshan Kumar; Gahlaut, Vijay; Muthamilarasan, Mehanathan; Prasad, ManojA total of 104 foxtail millet accessions were evaluated for 11 nutrients in three environments and 67 high-confidence marker-trait associations (MTAs) were identified. Six SNPs showed pleiotropic effect and associated with two or more nutrients, whereas 24 candidate genes were identified for 28 MTAs involving seven traits. Millets are known for their better nutritional profiles compared to major cereals. Foxtail millet (Setaria italica) is rich in nutrients essential to circumvent malnutrition and hidden hunger. However, the genetic determinants underlying this trait remain elusive. In this context, we evaluated 104 diverse foxtail millet accessions in three different environments (E1, E2, and E3) for 11 nutrients and genotyped with 30K SNPs. The genome-wide association study showed 67 high-confidence (Bonferroni-corrected) marker-trait associations (MTAs) for the nutrients except for phosphorus. Six pleiotropic SNPs were also identified, which were associated with two or more nutrients. Around 24 candidate genes (CGs) were identified for 28 MTAs involving seven nutrients. A total of 17 associated SNPs were present within the gene region, and five (5) were mapped in the exon of the CGs. Significant SNPs, desirable alleles and CGs identified in the present study will be useful in breeding programmes for trait improvement.Item Histone deacetylase 9 interacts with SiHAT3.1 and SiHDA19 to repress dehydration responses through H3K9 deacetylation in foxtail millet(Oxford University Press, 2024) Kumar, Verandra; Singh, Babita; Singh, Roshan Kumar; Sharma, Namisha; Muthamilarasan, Mehanathan; Sawant, Samir V; Prasad, ManojClimate change inflicts several stresses on plants, of which dehydration stress severely affects growth and productivity. C4 plants possess better adaptability to dehydration stress; however, the role of epigenetic modifications underlying this trait is unclear. Particularly, the molecular links between histone modifiers and their regulation remain elusive. In this study, genome-wide H3K9 acetylation (H3K9ac) enrichment using ChIP-seq was performed in two foxtail millet cultivars contrastingly differing in dehydration tolerance (IC403579; cv. IC4 – tolerant, and IC480117; cv. IC41 – sensitive). It revealed that a histone deacetylase, SiHDA9, was significantly up-regulated in the sensitive cultivar. Further characterization indicated that SiHDA9 interacts with SiHAT3.1 and SiHDA19 to form a repressor complex. SiHDA9 might be recruited through the SiHAT3.1 recognition sequence onto the upstream of dehydration-responsive genes to decrease H3K9 acetylation levels. The silencing of SiHDA9 resulted in the up-regulation of crucial genes, namely, SiRAB18, SiRAP2.4, SiP5CS2, SiRD22, SiPIP1;4 and SiLHCB2.3, which imparted dehydration tolerance in the sensitive cultivar (IC41). Overall, the study provides mechanistic insights into SiHDA9-mediated regulation of dehydration stress response in foxtail millet.Item Editorial: Dissecting antinutrient traits using omics approaches(Frontiers Media S.A., 2023) Muthamilarasan, Mehanathan; Prasad, ManojThe human population predominantly depends on plants and plant-based products for their food and nutrition. While plants are a major source of nutrients and health-promoting compounds, they also possess antinutrient factors that affect health and well-being. Global research focuses on improving important traits in plants, particularly food and nutritional security. However, few research groups work towards identifying and characterizing the antinutrient factors in food grains. These factors include cyanogen and cyanogenic glucosides, enzyme inhibitors, erucic acid, exorphins, goitrogens, lectins/hemoglutinins, lipoxygenases, nitrates, oxalate and oxalic acid, phytates, raffinose oligosaccharides, saponins, tannins, and other contextual antinutrients. Though these molecules are vital for the growth and development of plants, they pose health concerns to humans when consumed. In this context, this Research Topic was edited to collate the knowledge available on antinutrient research. The Research Topic received two research and three review articles. A comprehensive review of different antinutrients, their biosynthetic pathways, and physical and chemical methods to remove/reduce the antinutrients was provided by Duraiswamy et al.. The article provides an excellent timeline of research on this aspect and a compendium of genetic and genomic factors that underlie antinutrient biosynthesis. Also, the article provides a roadmap for using advanced genetic and genomic tools to reduce antinutrient content in food grains.Item Comparative transcriptome profiling of two contrasting foxtail millet cultivars provides insights into molecular mechanisms underlying dehydration stress response(Springer Nature Publishing AG, 2023) Muthamilarasan, Mehanathan; Suresh, Bonthala Venkata; Singh, Roshan Kumar; Choudhary, Pooja; Aggarwal, Pooja Rani; Prasad, ManojFoxtail millet (Setaria italica L.) has emerged as a model system to understand its adaptation to environmental stresses in the past decade. However, studies on understanding the molecular mechanism underlying the adaptation to dehydration stress and the regulatory network involved in the process remain elusive. In the present study, RNA-seq was performed during dehydration stress in the tolerant (IC4) and sensitive (IC41) cultivars at different time points (0, 6, and 12 h). A total of 2467 and 3318 differentially expressed genes (DEGs) were identified in IC4, and 2535 and 5572 in IC41 at 6 h and 12 h compared to control (0 h), respectively. Gene ontology (GO) analysis revealed that the DEGs were enriched in water transport, response to water deprivation, oxidative stress, amino acid and sugar transport, lipid biosynthesis, and regulation of stomatal opening. Pathway analysis suggested a significant modulation of genes involved in the metabolism of glutathione and tryptophan and biosynthesis of flavonoid, ascorbate, arginine, and proline in IC4 compared to IC41. Genes encoding for DIVARICATA, SBP family protein (teosinte glume architecture 1), and SRS family proteins (LATERAL ROOT PRIMORDIUM 1 and SHI-RELATED SEQUENCE 1) were found to be exclusively upregulated in IC4 during dehydration stress. Gene co-expression networks constructed based on the expression data showed the key modules and hubs that play critical roles during dehydration stress. Altogether, the present study has identified key genes, pathways, and regulatory modules that would serve as a base for further studies to gain insights into the dehydration-responsive molecular circuitry in foxtail millet.Item SiHSFA2e regulated expression of SisHSP21.9 maintains chloroplast proteome integrity under high temperature stress(Springer Nature Publishing AG, 2022) Singh, Roshan Kumar; Muthamilarasan, Mehanathan; Prasad, ManojHigh temperature-induced crop failures are prominent nowadays in major staples, including rice, wheat, and maize; however, crops such as foxtail millet (Setaria italica) are resilient to temperature stress. In this study, a novel small heat shock protein of foxtail millet, SisHSP21.9, is identified and characterized for its role in conferring tolerance to high-temperature stress. SisHSP21.9 is a panicoid-specific gene, which is highly upregulated during high-temperature in leaves, and the protein is localized in the chloroplast. Its expression is directly regulated by heat shock factor, SiHSFA2e, during temperature stress. Further, overexpression of SiHSP21.9 in rice enhanced the survival of transgenics during high-temperature stress (> 80% survival frequency), and the transgenic lines showed improved plant architecture and overall grain yield. Compared to WT plants, transgenic lines maintained optimal photosynthesis rates with higher photosystem efficiencies at high temperatures, and this is conferred through protecting the components of photosystems, chlorophyll-binding proteins, and chloroplast-localized functional proteins by SisHSP21.9. Prolonged high-temperature stress showed minimal damage to chloroplast proteins resulting in comparatively lower yield loss (35–37%) in transgenic lines. Altogether, the study suggests that SisHSP21.9 is a potential candidate for designing thermotolerant crops for climate-resilient agriculture; however, further research is needed because tolerance to abiotic stresses is polygenic.Item De novo transcriptome analysis identifies key genes involved in dehydration stress response in kodo millet (Paspalum scrobiculatum L.)(Elsevier B.V., 2022) Suresh, Bonthala Venkata; Choudhary, Pooja; Aggarwal, Pooja Rani; Rana, Sumi; Singh, Roshan Kumar; Ravikesavan, Rajasekaran; Prasad, Manoj; Muthamilarasan, MehanathanKodo millet (Paspalum scrobiculatum L.) is a small millet species known for its excellent nutritional and climate-resilient traits. To understand the genes and pathways underlying dehydration stress tolerance of kodo millet, the transcriptome of cultivar ‘CO3’ subjected to dehydration stress (0 h, 3 h, and 6 h) was sequenced. The study generated 239.1 million clean reads that identified 9201, 9814, and 2346 differentially expressed genes (DEGs) in 0 h vs. 3 h, 0 h vs. 6 h, and 3 h vs. 6 h libraries, respectively. The DEGs were found to be associated with vital molecular pathways, including hormone metabolism and signaling, antioxidant scavenging, photosynthesis, and cellular metabolism, and were validated using qRT-PCR. Also, a higher abundance of uncharacterized genes expressed during stress warrants further studies to characterize this class of genes to understand their role in dehydration stress response. Altogether, the study provides insights into the transcriptomic response of kodo millet during dehydration stress.Item Genetics and genomics interventions for promoting millets as functional foods(Bentham Science, 2021) Dhaka, Annvi; Singh, Roshan Kumar; Muthamilarasan, Mehanathan; Prasad, ManojSeveral crops, including millets with immense nutritional and therapeutic values, were once a part of our regular diet. However, due to domestication and selection pressures, many of them have become marginally cultivated crops confined to a particular region, race, or locality. Millets are a perfect example of neglected species that have the potential to address both food and nutritional insecurities prevalent among the ever-growing global population. Starvation and malnutrition contribute to a large number of health-related issues, being the main reason behind the occurrence of most of the severe diseases worldwide. These constraints are repeatedly disturbing both the social and economic health of global society. Naturally, millets are rich in minerals, nutrients, and bioactive compounds, and these crops are less dependent on synthetic fertilizers, systemic irrigation, and pest/weed control. Given this, the review emphasizes the nutritional values, health benefits, processing techniques, and genomic advancements of millets. In addition, it proposes a roadmap for enhancing the utility and commercialization of millets.Item The Sw5a gene confers resistance to ToLCNDV and triggers an HR response after direct AC4 effector recognition(PNAS, USA, 2021) Sharma, Namisha; Sahu, Pranav Pankaj; Prasad, Ashish; Muthamilarasan, Mehanathan; Waseem, Mohd; Khan, Yusuf; Thakur, Jitendra K.; Chakraborty, Supriya; Prasad, ManojSeveral attempts have been made to identify antiviral genes against Tomato leaf curl New Delhi virus (ToLCNDV) and related viruses. This has led to the recognition of Ty genes (Ty1-Ty6), which have been successful in developing virus-resistant crops to some extent. Owing to the regular appearance of resistance-breaking strains of these viruses, it is important to identify genes related to resistance. In the present study, we identified a ToLCNDV resistance (R) gene, SlSw5a, in a ToLCNDV-resistant tomato cultivar, H-88-78-1, which lacks the known Ty genes. The expression of SlSw5a is controlled by the transcription factor SlMyb33, which in turn is regulated by microRNA159 (sly-miR159). Virus-induced gene silencing of either SlSw5a or SlMyb33 severely increases the disease symptoms and viral titer in leaves of resistant cultivar. Moreover, in SlMyb33-silenced plants, the relative messenger RNA level of SlSw5a was reduced, suggesting SlSw5a is downstream of the sly-miR159-SlMyb33 module. We also demonstrate that SlSw5a interacts physically with ToLCNDV-AC4 (viral suppressor of RNA silencing) to trigger a hypersensitive response (HR) and generate reactive oxygen species at infection sites to limit the spread of the virus. The "RTSK" motif in the AC4 C terminus is important for the interaction, and its mutation completely abolishes the interaction with Sw5a and HR elicitation. Overall, our research reports an R gene against ToLCNDV and establishes a connection between the upstream miR159-Myb33 module and its downstream target Sw5a to activate HR in the tomato, resulting in geminivirus resistance.Item Genetic determinants of micronutrient traits in graminaceous crops to combat hidden hunger(Springer Nature Publishing AG, 2021) Sushree, Shyamli P; Rana, Sumi; Suranjika, Sandhya; Muthamilarasan, Mehanathan; Parida, Ajay; Prasad, ManojKey message: Improving the nutritional content of graminaceous crops is imperative to ensure nutritional security, wherein omics approaches play pivotal roles in dissecting this complex trait and contributing to trait improvement. Abstract: Micronutrients regulate the metabolic processes to ensure the normal functioning of the biological system in all living organisms. Micronutrient defciency, thereby, can be detrimental that can result in serious health issues. Grains of graminaceous crops serve as an important source of micronutrients to the human population; however, the rise in hidden hunger and malnutrition indicates an insufciency in meeting the nutritional requirements. Improving the elemental composition and nutritional value of the graminaceous crops using conventional and biotechnological approaches is imperative to address this issue. Identifying the genetic determinants underlying the micronutrient biosynthesis and accumulation is the frst step toward achieving this goal. Genetic and genomic dissection of this complex trait has been accomplished in major cereals, and several genes, alleles, and QTLs underlying grain micronutrient content were identifed and characterized. However, no comprehensive study has been reported on minor cereals such as small millets, which are rich in micronutrients and other bioactive compounds. A comparative narrative on the reports available in major and minor Graminaceae species will illustrate the knowledge gained from studying the micronutrient traits in major cereals and provides a roadmap for dissecting this trait in other minor species, including millets. In this context, this review explains the progress made in studying micronutrient traits in major cereals and millets using omics approaches. Moreover, it provides insights into deploying integrated omics approaches and strategies for genetic improvement in micronutrient traits in graminaceous crops.