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N\u2010glycans, N\u2010glycoproteins, and their processing enzymes are integral to various plant processes, including fruit ripening. Among these, \u03b1\u2010D\u2010mannosidase (\u03b1\u2010Man) and \u03b2\u2010D\u2010N\u2010acetylhexosaminidase (\u03b2\u2010Hex) are key ripening\u2010specific enzymes that modulate fruit softening. Previously, we have shown that RNAi\u2010mediated suppression of\n                    <jats:italic>\u03b1\u2010Man<\/jats:italic>\n                    or\n                    <jats:italic>\u03b2\u2010Hex<\/jats:italic>\n                    improves fruit shelf life and firmness in both climacteric and non\u2010climacteric fruits. However, the underlying molecular and biochemical basis of fruit softening regulation by\n                    <jats:italic>\u03b1\u2010Man<\/jats:italic>\n                    and\n                    <jats:italic>\u03b2\u2010Hex<\/jats:italic>\n                    was not well understood. In this study, we developed transgenic tomato (\n                    <jats:italic>Solanum lycopersicum<\/jats:italic>\n                    ) plants by silencing\n                    <jats:italic>\u03b1\u2010Man<\/jats:italic>\n                    and\n                    <jats:italic>\u03b2\u2010Hex<\/jats:italic>\n                    simultaneously using RNAi. Suppression of these enzymes reduces N\u2010glycoprotein degradation, downregulates pectin dissolution, and inhibits ripening\u2010related gene expression. RNAi fruits exhibited enhanced shelf life, greater firmness, reduced reactive oxygen species (ROS) accumulation and increased resistance against post\u2010harvest pathogens without affecting plant growth, fruit development, yield, or nutritional quality. To further explore the molecular mechanism of\n                    <jats:italic>\u03b1\u2010Man<\/jats:italic>\n                    and\n                    <jats:italic>\u03b2\u2010Hex<\/jats:italic>\n                    function, we purified and quantified N\u2010glycans in RNAi fruits and other ripening\u2010impaired mutants, identifying key N\u2010glycan species. We also carried out iTRAQ\u2010based quantitative proteome profiling to investigate the abundance of proteins in ripened fruit affected by silencing of\n                    <jats:italic>\u03b1\u2010Man<\/jats:italic>\n                    and\n                    <jats:italic>\u03b2\u2010Hex<\/jats:italic>\n                    . Molecular insights revealed that N\u2010glycan processing and degradation are key events during ripening, influencing cell wall softening, fruit redox state, and post\u2010harvest quality attributes. This study highlights the potential of co\u2010silencing\n                    <jats:italic>\u03b1\u2010Man<\/jats:italic>\n                    and\n                    <jats:italic>\u03b2\u2010Hex<\/jats:italic>\n                    as a novel approach to extending the shelf life of fruits, regardless of their climacteric behavior, without compromising quality or yield.\n                  <\/jats:p>","DOI":"10.1111\/tpj.70629","type":"journal-article","created":{"date-parts":[[2025,12,14]],"date-time":"2025-12-14T20:10:15Z","timestamp":1765743015000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["N\u2010glycan remodeling by \u03b1\u2010D\u2010mannosidase and \u03b2\u2010D\u2010N\u2010acetylhexosaminidase regulates fruit softening, redox balance, and post\u2010harvest pathogen resistance"],"prefix":"10.1111","volume":"124","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7877-5530","authenticated-orcid":false,"given":"Mohammad","family":"Irfan","sequence":"first","affiliation":[{"name":"National Institute of Plant Genome Research Aruna Asaf Ali Marg  New Delhi 110067 India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2275-9144","authenticated-orcid":false,"given":"Pankaj","family":"Kumar","sequence":"additional","affiliation":[{"name":"National Institute of Plant Genome Research Aruna Asaf Ali Marg  New Delhi 110067 India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6751-5741","authenticated-orcid":false,"given":"Vinay","family":"Kumar","sequence":"additional","affiliation":[{"name":"National Institute of Plant Genome Research Aruna Asaf Ali Marg  New Delhi 110067 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