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This study first classified the cold tolerance of sugarcane cultivars using metabolites in juice as predictor variables. The best-fit model (XGBoost discriminant analysis) estimated the higher cold tolerance of the final on-station clone progeny to the stress tolerance-inducing wild germplasm line. Stalks of the tolerant sugarcane genotype contained higher fiber for mechanical support against cellular injury, compared to susceptible varieties. Fluorescence microscopy visualized phospholipids responsible for maintaining membrane fluidity during frost in lignin surrounding the vascular bundle. Thermal imaging is proposed for real-time monitoring of spatiotemporal temperature changes, as stalk injury is initiated by ice formation at sub-freeze temperatures during winter freeze. As additional datasets for independent prediction become available, developed methods could be used to explore the biomarkers for stress resistance in simpler multivariate discriminant analysis and the distribution of specific biomarkers in cellular components by microscopic imaging, and to trace stalk injury hot spots as a function of time and relationships with resistance markers.<\/jats:p>","DOI":"10.3389\/fpls.2026.1882707","type":"journal-article","created":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T04:21:39Z","timestamp":1786076499000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Multivariate and imaging methods to classify cold tolerance of sugarcane (Saccharum spp. hybrids) cultivars and breeding clones"],"prefix":"10.3389","volume":"17","author":[{"given":"Minori","family":"Uchimiya","sequence":"first","affiliation":[{"name":"United States Department of Agriculture, Agricultural Research Service, Southern Regional Research Center","place":["New Orleans, LA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Collins","family":"Kimbeng","sequence":"additional","affiliation":[{"name":"Sugar Research Station, Louisiana State University Agricultural Center","place":["St Gabriel, LA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zachary","family":"Taylor","sequence":"additional","affiliation":[{"name":"Sugar Research Station, Louisiana State University Agricultural Center","place":["St Gabriel, LA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kerstin","family":"H\u00f6ner zu Bentrup","sequence":"additional","affiliation":[{"name":"Department of Microbiology\/Immunology, Tulane University School of Medicine","place":["New Orleans, LA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaitlyn","family":"Taylor","sequence":"additional","affiliation":[{"name":"Oak Ridge Institute for Science and Education Research Program at USDA","place":["Oak Ridge, TN, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,7]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"641","DOI":"10.1093\/aob\/mci059","article-title":"Extension growth of Impatiens glandulifera at low irradiance: Importance of nitrate and potassium accumulation","volume":"95","author":"Andrews","year":"2005","journal-title":"Ann. 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