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Unfortunately, the starchy endosperm, which is the remaining portion of the seed after polishing, contains limited amounts of micronutrients. Here, it is shown that this is particularly the case for thiamin (vitamin B1). Therefore, a tissue\u2010specific metabolic engineering approach was conducted, aimed at enhancing the level of thiamin specifically in the endosperm. To achieve this, three major thiamin biosynthesis genes, <jats:italic>THIC<\/jats:italic>, <jats:italic>THI1<\/jats:italic> and <jats:italic>TH1<\/jats:italic>, controlled by strong endosperm\u2010specific promoters, were employed to obtain engineered rice lines. The metabolic engineering approaches included ectopic expression of <jats:italic>THIC<\/jats:italic> alone, in combination with <jats:italic>THI1<\/jats:italic> (bigenic) or combined with both <jats:italic>THI1<\/jats:italic> and <jats:italic>TH1<\/jats:italic> (trigenic). Determination of thiamin and thiamin biosynthesis intermediates reveals the impact of the engineering approaches on endosperm thiamin biosynthesis. The results show an increase of thiamin in polished rice up to threefold compared to WT, and stable upon cooking. These findings confirm the potential of metabolic engineering to enhance <jats:italic>de novo<\/jats:italic> thiamin biosynthesis in rice endosperm tissue and aid in steering future biofortification endeavours.<\/jats:p>","DOI":"10.1111\/pbi.13545","type":"journal-article","created":{"date-parts":[[2021,1,15]],"date-time":"2021-01-15T23:29:54Z","timestamp":1610753394000},"page":"1253-1267","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":49,"title":["Metabolic engineering of rice endosperm towards higher vitamin B1 accumulation"],"prefix":"10.1111","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0583-9406","authenticated-orcid":false,"given":"Simon","family":"Strobbe","sequence":"first","affiliation":[{"name":"Laboratory of Functional Plant Biology Department of Biology Ghent University Gent Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2004-0312","authenticated-orcid":false,"given":"Jana","family":"Verstraete","sequence":"additional","affiliation":[{"name":"Laboratory of Toxicology Department of Bioanalysis Ghent University Gent Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7126-348X","authenticated-orcid":false,"given":"Christophe","family":"Stove","sequence":"additional","affiliation":[{"name":"Laboratory of Toxicology Department of Bioanalysis Ghent University Gent Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7755-1420","authenticated-orcid":false,"given":"Dominique","family":"Van Der Straeten","sequence":"additional","affiliation":[{"name":"Laboratory of Functional Plant Biology Department of Biology Ghent University Gent Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,2,11]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biombioe.2018.11.032"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11103-007-9205-4"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.abb.2006.11.011"},{"key":"e_1_2_8_5_1","first-page":"67","article-title":"Brown rice\u2010beyond the color reviving a lost health food\u2010a review","volume":"2","author":"Babu P.D.","year":"2009","journal-title":"Am.\u2010Eur. 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