Publications of NIPGR Scientists

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Author: search.filters.author.Datta, AsisSubject: search.filters.subject.Ethylene

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    Ethylene-mediated regulation of a ripening-specific N-glycan-processing enzyme β-D-N-Acetylhexosaminidase
    (Springer Nature Publishing AG, 2025) Irfan, Mohammad; Kumar, Pankaj; Datta, Asis
    N-glycoproteins in plants play a critical role in various biological processes such as plant stress response, seed development, and fruit ripening. Further, N-glycans and N-glycan-processing enzymes have been identified as critical regulators of fruit ripening in various fruit crop including tomato. Fruit ripening, a multifaceted process influenced by diverse factors is controlled by the upregulation of cell wall degradation enzymes. Among these, N-glycan-processing enzymes, specifically β-D-N-acetylhexosaminidase (β-Hex), are involved in ripening-associated fruit softening, and its suppression enhances fruit shelf life and firmness in both climacteric and non-climacteric fruits. Ethylene is among the various factors that regulates the ripening-specific expression of β-Hex. Notably, ethylene response factor 6 (SlERF6), also known as SlERF.E4, has been identified as a potential transcriptional regulator of β-Hex expression. However, in order to get deeper insights, a thorough understanding of the specific ethylene-mediated control of β-Hex during ripening is crucial. Thus, we present a protocol for investigating the ethylene-mediated regulation of this crucial ripening-related enzyme β-D-N-acetylhexosaminidase.
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    Fruit ripening specific expression of β-D-N-acetylhexosaminidase (β-Hex) gene in tomato is transcriptionally regulated by Ethylene Response Factor SlERF.E4
    (Elsevier B.V., 2022) Irfan, Mohammad; Kumar, Pankaj; Kumar, Vinay; Datta, Asis
    N-glycans and N-glycan processing enzymes are key players in regulating the ripening of tomato (Solanum lycopersicum) fruits, a model for fleshy fruit ripening. β-D-N-acetylhexosaminidase (β-Hex) is a N-glycan processing enzyme involved in fruit ripening. The suppression of β-Hex results in enhanced fruit shelf life and firmness in both climacteric and non-climacteric fruits. Previously, we have shown that ripening specific expression of β-Hex is regulated by RIPENING INHIBITOR (RIN), ABSCISIC ACID STRESS RIPENING 1 (SlASR1) and ethylene. However, the precise mechanism of ethylene-mediated regulation of β-Hex remains elusive. To gain insights into this, we have performed 5’ deletion mapping of tomato β-Hex promoter and a shorter promoter fragment (pD-200, 200 bp upstream to translational start site) is identified, which was found critical for spatio-temporal transcriptional regulation of β-Hex. Further, site specific mutagenesis in RIN and ASR1 binding sites in pD-200 provides key insights into ripening specific promoter activity. Furthermore, induction of GUS activity by ethylene, yeast one hybrid assay and EMSA identify Ethylene Response Factor SlERF.E4 as a positive regulator of β-Hex. Taken together, our study suggest that SlERF.E4 together with RIN and SlASR1 transcriptionally regulates β-Hex and all these three proteins are essential for fruit ripening specific expression of β-Hex in tomato.