Publications of NIPGR Scientists

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Author: search.filters.author.Majee, ManojSubject: search.filters.subject.Germination

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    Molecular intricacies of modulating seed dormancy through CRISPR/Cas9 technology
    (Oxford University Press, 2026) Gautam, Shikha; Kamble, Nitin Uttam; Majee, Manoj
    The processes of seed development, maturation, and dormancy acquisition are complex and tightly regulated, and play a critical role in plant survival and propagation. Over the past decades, significant advances have been made in elucidating the molecular mechanisms that govern these intricate processes. The interplay among hormone signaling, epigenetic regulation, reactive oxygen species (ROS), and environmental cues has been recognized as central to determining seed fate. Despite these advancements, many molecular components remain to be fully discovered. Recent developments in CRISPR-based gene-editing technologies have provided promising tools for the precise regulation of seed dormancy without compromising other seed traits. Although CRISPR has been effectively utilized to modify genes controlling physiological characteristics in a wide range of crops, its application in regulating dormancy remains at an early stage. This review synthesizes current knowledge on the molecular and genetic mechanisms controlling seed maturation, dormancy acquisition and germination, with particular emphasis on emerging CRISPR-based strategies. Realizing this potential, however, requires a deeper understanding of the complex regulatory networks orchestrating seed dormancy acquisition and germination. Identifying optimal gene targets and refining editing strategies will be crucial for developing reliable and sustainable dormancy-control systems. Therefore, we highlight key gene targets, summarize their functional relevance, and discuss how genome editing could be leveraged to fine-tune dormancy and germination behaviour. The studies discussed herein underscore the transformative potential of CRISPR/Cas9 and related genome-editing platforms in advancing seed biology and crop improvement, paving the way for next-generation seed technologies.
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    A misannotated locus positively influencing Arabidopsis seed germination is deconvoluted using multiple methods, including surrogate splicing
    (Elsevier B.V., 2017) Majee, Manoj; Wu, Shuiqin; Salaita, Louai; Gingerich, Derek; Dirk, Lynnette M.A.; Chappell, Joseph; Hunt, Art G.; Vierstra, Richard; Downie, A. Bruce
    A screen of activation tagged lines of Arabidopsis thaliana retrieved COLD TEMPERATURE GERMINATING10-D(tag) (CTG10-D(tag)) seeds, capable of radicle protrusion in advance of wild type (WT) at suboptimal- and optimal-temperatures. Genomic walking revealed T-DNA in the intragenic region that upregulates expression of the At4g19330 locus previously predicted to encode an F-BOX protein. A combination of surrogate splicing, primer scanning, RACE, and Illumina PolyAdenylation Tag (PAT) sequencing in petunia (Petunia X hybrida) and Arabidopsis were required to demonstrate that the region around At4g19330 was misannotated and is actually compromised of two separate genes. Even though homologous regions nearby and elsewhere on chromosome 4 are confounding elements in the molecular characterization of the locus, we could determine that the 5′ entity encodes a ribonucleoprotein of unknown function whereas the 3′ gene includes the promoter and full coding region of an F-BOX protein. Although both genes were upregulated, only independently-transformed lines over-expressing the F-Box exhibited enhanced completion of seed germination. We named it CTG10, and a single, poorly penetrant, mutant line of ctg10 manifested the expected reduced completion of seed germination. Whereas CTG10-OE lines are hyposensitive to the gibberellin biosynthetic inhibitor paclobutrazol, the ctg10 mutant line is hypersensitive; a phenotype which could be alleviated when transgenically rescued with CTG10. The F-Box moiety promoted association of CTG10 with ASK proteins in yeast two hybrid assays, indicating that it likely assembles into an SCF-type ubiquitin ligase to promote the ubiquitination of one or more substrates. In this capacity CTG10 might target a protein repressing seed germination for polyubiquitination and subsequent proteasomal degradation.