Publications of NIPGR Scientists

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Author: search.filters.author.Mathew, Iny Elizebeth

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    Decoding rice seed storage proteins: From gene identification to structural prediction
    (Oxford University Press, 2026) Yadav, Antima; Jaiswal, Priya; Mathew, Iny Elizebeth; Panwar, Akanksha; Agarwal, Pinky
    Background and Aims: Rice seed storage proteins (SSPs) are major determinants of grain nutritional quality, serving as primary sources of dietary protein, energy, and essential nutrients. However, limited understanding of their diversity, evolution, and regulation constrains efforts to improve grain quality. This study aimed to perform a comprehensive genome-wide characterization of SSPs in rice. Methods: A combined homology- and domain-based approach was employed to identify SSP-encoding genes in the rice genome. These proteins were further analysed through phylogenetic reconstruction, domain and motif characterization, promoter cis-element analysis, expression profiling across seed developmental stages, and three-dimensional structural modelling. Key Results: A total of 65 SSP genes were identified, including 19 previously uncharacterized members. Phylogenetic and domain analyses revealed evolutionary relationships between albumins and prolamins, and between globulins and glutelins. Tandem clustering of albumins, glutelins, and prolamins suggested gene duplication as a major driver of SSP family expansion. Expression profiling indicated that albumins, globulins, and glutelins were transcriptionally active from the S2 stage, whereas prolamins were predominantly expressed from the S3 stage onwards. Promoter analysis identified several seed-specific cis-regulatory elements, including CAATBOX1, EBOXBNNAPA, and DOFCOREZM. Structural modelling showed that albumins and prolamins are primarily composed of α-helices, while globulins and glutelins are enriched in β-strands and coils. Conclusions: This integrative analysis provides comprehensive insights into the classification, evolution, regulatory mechanisms, and structural features of rice SSPs. The findings establish a valuable resource for future functional studies and offer a foundation for strategies aimed at improving grain nutritional quality.
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    SUPER STARCHY1/ONAC025 participates in rice grain filling
    (American Society of Plant Biologists, 2020) Mathew, Iny Elizebeth; Priyadarshini, Richa; Mahto, Arunima; Jaiswal, Priya; Parida, Swarup K.; Agarwal, Pinky
    NAC transcription factors (TFs) are known for their role in development and stress. This article attempts to functionally validate the role of rice SS1/ ONAC025 (LOC_ Os11g31330) during seed development. The gene is seed-specific and its promoter directs reporter expression in the developing endosperm and embryo in rice transgenic plants. Furthermore, rice transgenic plants ectopically expressing SS1/ ONAC025 have a plantlet lethal phenotype with hampered vegetative growth, but increased tillers and an altered shoot apical meristem structure. The vegetative cells of these plantlets are filled with distinct starch granules. RNAseq analysis of two independent plantlets reveals the differential expression of reproductive and photosynthetic genes. A comparison with seed development transcriptome indicates differential regulation of many seed-related genes by SS1/ ONAC025. Genes involved in starch biosynthesis, especially amylopectin and those encoding seed storage proteins, and regulating seed size are also differentially expressed. In conjunction, SS1/ ONAC025 shows highest expression in japonica rice. As a TF, SS1/ ONAC025 is a transcriptional repressor localized to endoplasmic reticulum and nucleus. The article shows that SS1/ ONAC025 is a seed-specific gene promoting grain filling in rice, and negatively affecting vegetative growth.
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    Analysis of rice proteins with DLN repressor Motif/S
    (MDPI AG, 2019) Singh, Purnima; Mathew, Iny Elizebeth; Verma, Ankit; Tyagi, Akhilesh K.; Agarwal, Pinky
    Transcriptional regulation includes both activation and repression of downstream genes. In plants, a well-established class of repressors are proteins with an ERF-associated amphiphilic repression/EAR domain. They contain either DLNxxP or LxLxL as the identifying hexapeptide motif. In rice (Oryza sativa), we have identified a total of 266 DLN repressor proteins, with the former motif and its modifications thereof comprising 227 transcription factors and 39 transcriptional regulators. Apart from DLNxxP motif conservation, DLNxP and DLNxxxP motifs with variable numbers/positions of proline and those without any proline conservation have been identified. Most of the DLN repressome proteins have a single DLN motif, with higher relative percentage in the C-terminal region. We have designed a simple yeast-based experiment wherein a DLN motif can successfully cause strong repression of downstream reporter genes, when fused to a transcriptional activator of rice or yeast. The DLN hexapeptide motif is essential for repression, and at least two “DLN” residues cause maximal repression. Comparatively, rice has more DLN repressor encoding genes than Arabidopsis, and DLNSPP motif from rice is 40% stronger than the known Arabidopsis SRDX motif. The study reports a straightforward assay to analyze repressor activity, along with the identification of a strong DLN repressor from rice.
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    May the fittest protein evolve: favoring the plant-specific origin and expansion of NAC transcription factors
    (John Wiley & Sons, 2018) Mathew, Iny Elizebeth; Agarwal, Pinky
    Plant‐specific NAC transcription factors (TFs) evolve during the transition from aquatic to terrestrial plant life and are amplified to become one of the biggest TF families. This is because they regulate genes involved in water conductance and cell support. They also control flower and fruit formation. The review presented here focuses on various properties, regulatory intricacies, and developmental roles of NAC family members. Processes controlled by NACs depend majorly on their transcriptional properties. NACs can function as both activators and/or repressors. Additionally, their homo/hetero dimerization abilities can also affect DNA binding and activation properties. The active protein levels are dependent on the regulatory cascades. Because NACs regulate both development and stress responses in plants, in‐depth knowledge about them has the potential to help guide future crop improvement studies.
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    Decoding the transcriptome of rice seed during development
    (InTech, 2017) Mahto, Arunima; Mathew, Iny Elizebeth; Agarwal, Pinky
    Rice seed development is a continuous process wherein it undergoes complex molecular and tissue reprogramming. It is a collective effect of embryo and endosperm development, each of which undertakes its own developmental paths, with endosperm development significantly affecting embryo. Understanding the mechanistics of the regulatory networks administrating this process is the building block for any future research on grain yield and quality. High-throughput transcript profiling and small RNA profiling studies have proved useful in providing information about the molecular changes occurring in various tissues associated with seed development. Transcriptome sequencing studies have highlighted the significant genes and pathways that are operating during seed development. The involvement of TFs and hormones has also been implicated in regulating key aspects of seed development, including embryo patterning and seed maturation. This chapter will review the information provided by high-throughput sequencing studies on various aspects of rice seed development, highlighting the developmental complexities of embryo and endosperm.
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    Three rice NAC transcription factors heteromerize and are associated with seed size
    (Frontiers Media S.A., 2016) Mathew, Iny Elizebeth; Das, Sweta; Mahto, Arunima; Agarwal, Pinky
    NACs are plant-specific transcription factors (TFs) involved in multiple aspects of development and stress. In rice, three NAC TF encoding genes, namely ONAC020, ONAC026, and ONAC023 express specifically during seed development, at extremely high levels. They exhibit significantly strong association with seed size/weight with the sequence variations located in the upstream regulatory region. Concomitantly, their expression pattern/levels during seed development vary amongst different accessions with variation in seed size. The alterations in the promoter sequences of the three genes, amongst the five rice accessions, correlate with the expression levels to a certain extent only. In terms of transcriptional properties, the three NAC TFs can activate and/or suppress downstream genes, though to different extents. Only ONAC026 is localized to the nucleus while ONAC020 and ONAC023 are targeted to the ER and cytoplasm, respectively. Interestingly, these two proteins interact with ONAC026 and the dimers localize in the nucleus. Trans-splicing between ONAC020 and ONAC026 results in three additional forms of ONAC020. The transcriptional properties including activation, repression, subcellular localization and heterodimerization of trans-spliced forms of ONAC020 and ONAC026 are different, indicating toward their role as competitors. The analysis presented in this paper helps to conclude that the three NAC genes, which are associated with seed size, have independent as well as overlapping roles during the process and can be exploited as potential targets for crop improvement.
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    Expanding frontiers in plant transcriptomics in aid of functional genomics and molecular breeding
    (John Wiley & Sons Ltd, 2014) Agarwal, Pinky; Parida, Swarup K.; Mahto, Arunima; Das, Sweta; Mathew, Iny Elizebeth; Malik, Naveen; Tyagi, Akhilesh K.
    The transcript pool of a plant part, under any given condition, is a collection of mRNAs that will pave the way for a biochemical reaction of the plant to stimuli. Over the past decades, transcriptome study has advanced from Northern blotting to RNA sequencing (RNA-seq), through other techniques, of which real-time quantitative polymerase chain reaction (PCR) and microarray are the most significant ones. The questions being addressed by such studies have also matured from a solitary process to expression atlas and marker-assisted genetic enhancement. Not only genes and their networks involved in various developmental processes of plant parts have been elucidated, but also stress tolerant genes have been highlighted. The transcriptome of a plant with altered expression of a target gene has given information about the downstream genes. Marker information has been used for breeding improved varieties. Fortunately, the data generated by transcriptome analysis has been made freely available for ample utilization and comparison. The review discusses this wide variety of transcriptome data being generated in plants, which includes developmental stages, abiotic and biotic stress, effect of altered gene expression, as well as comparative transcriptomics, with a special emphasis on microarray and RNA-seq. Such data can be used to determine the regulatory gene networks, which can subsequently be utilized for generating improved plant varieties.