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In Arabidopsis, microRNA163 (miR163) was found to be involved in seed germination, root development, and biotic resistance. However, the regulatory roles of miR163 remain unclear. In the current study, the <jats:italic>mir163<\/jats:italic> mutant was investigated to comprehensively understand and characterize its functions in Arabidopsis. RNA\u2010sequencing and Gene Ontology enrichment analyses revealed that miR163 might be involved in \u201cresponse to stimulus\u201d and \u201cmetabolic process\u201d. Interestingly, \u201cresponse to stress\u201d, including heat, cold, and oxidative stress, was enriched under the subcategory of \u201cresponse to stimulus\u201d. We observed that miR163 and <jats:italic>PXMT<\/jats:italic> were repressed and induced under heat stress, respectively. Furthermore, the study detected significant differences in seed germination rate, hypocotyl length, and survival rate, indicating a variation in the thermotolerance between WT and <jats:italic>mir163<\/jats:italic> mutant. The results revealed that the <jats:italic>mir163<\/jats:italic> mutant had a lesser degree of germination inhibition by heat treatment than WT. In addition, the <jats:italic>mir163<\/jats:italic> mutant showed a better survival rate and longer hypocotyl length under heat treatment than the WT. The metabolomes of WT and <jats:italic>mir163<\/jats:italic> mutant were further analyzed. <jats:italic>The contents of benzene derivatives and<\/jats:italic> flavonoids were affected by miR163, which could enhance plants' defense abilities. In conclusion, miR163\/targets regulated the expression of stress\u2010responsive genes and the accumulation of defense\u2010related metabolites to alter stress tolerance.<\/jats:p>","DOI":"10.1111\/ppl.14053","type":"journal-article","created":{"date-parts":[[2023,10,25]],"date-time":"2023-10-25T03:37:06Z","timestamp":1698205026000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Regulatory roles of <scp>microRNA163<\/scp> in responses to stresses in Arabidopsis"],"prefix":"10.1111","volume":"175","author":[{"given":"Zhenyuan","family":"Kok","sequence":"first","affiliation":[{"name":"Department of Life Sciences National Chung Hsing University  Taichung Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3115-9850","authenticated-orcid":false,"given":"Yun\u2010Wei","family":"Kuo","sequence":"additional","affiliation":[{"name":"Department of Agronomy National Chung 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