{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T13:09:41Z","timestamp":1789564181193,"version":"build-2803163510"},"reference-count":42,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T00:00:00Z","timestamp":1789516800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Repeated freeze\u2013thaw cycles represent a critical stress factor in frozen food processing, leading to a significant decline in the survival and fermentation performance of lactic acid bacteria.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      This study investigated the tolerance mechanism of\n                      <jats:italic>Lactiplantibacillus plantarum<\/jats:italic>\n                      QZW5, a resistant strain, after 10 cycles of alternating freezing (\u221220 \u00b0C) and thawing (20 \u00b0C). An integrated approach combining physiological and biochemical assays, environmental cryo-electron microscopy, multi-omics analysis, and qPCR validation was employed.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results and Discussion<\/jats:title>\n                    <jats:p>\n                      The results showed after freeze\u2013thaw treatment, QZW5 retained an 85.20%survival rate with a 14.80% reduction in growth rate, accompanied by gradual fermentation pH decline. Its antibacterial activity decreased markedly; for example, the inhibition zone against\n                      <jats:italic>Micrococcus luteus<\/jats:italic>\n                      decreased from 20.36 \u00b1 1.77 mm to 10.64 \u00b1 0.10 mm (\u224848% reduction),and ampicillin susceptibility increased by around 18% (inhibition zone from 24.76 \u00b1 0.35 mm to 29.11 \u00b1 0.17 mm),while its biofilm-forming capacity remained stable with OD\u2085\u2087\u2080 values of 1.90 \u00b1 0.23 (CK) and 1.98 \u00b1 0.19 (FT,\n                      <jats:italic>p<\/jats:italic>\n                      \u202f&amp;gt;\u202f0.05). Morphological alterations included serrated cell edges and intracellular granule deposition. At the molecular level, QZW5 activated a coordinated stress response: the upregulation of genes related to glyoxylate metabolism, ABC transporters, purine\/pyrimidine metabolism, and phenazine synthesis enhanced nutrient uptake and protective barrier formation; meanwhile, the downregulation of genes involved in oxidative phosphorylation and peptidoglycan synthesis reduced energy expenditure and was accompanied by programmed cell deathlike responses. Quantitative PCR (qPCR) validation confirmed that core genes such as fabZ and purN were sharply downregulated (relative expression dropped from ~2070 and ~589 to below 65 and 27 respectively), verifying that QZW5 adapts to stress by \u201cinhibiting energy consumption while enhancing repair. In conclusion, QZW5 achieves freeze\u2013thaw tolerance through the synergistic interplay of morphological adaptation, metabolic reprogramming, and gene regulation. This study provides theoretical and technical support for the development and application of QZW5 as a starter culture for frozen foods.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1873698","type":"journal-article","created":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T12:47:50Z","timestamp":1789562870000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Investigating the survival mechanisms of Lactiplantibacillus plantarum QZW5 under repeated freeze\u2013thaw stress: an integrated study using biochemical assays, environmental cryo-electron microscopy, and multi-omics approaches"],"prefix":"10.3389","volume":"17","author":[{"given":"Miao","family":"Zhang","sequence":"first","affiliation":[{"name":"Zhengzhou Research Base, National Key Laboratory of Cotton Bio-breeding and Integrated Utilization, School of Agriculture and Biomanufacturing, Zhengzhou 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