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Sulfonamides are herbicides that, by inhibiting the folic acid biosynthetic pathway, suppress the growth of untransformed cells. Sulfonamide resistance genes that were previously developed as selectable markers for plant transformation were based on the assumption that, in plants, the folic acid biosynthetic pathway resides in the chloroplast compartment. Consequently, the Sul resistance protein, a herbicide\u2010insensitive dihydropteroate synthase, was targeted to the chloroplast. Although these vectors produce transgenic plants, the transformation efficiencies are low compared to other markers. Here, we show that this inefficiency is due to the erroneous assumption that the folic acid pathway is located in chloroplasts. When the RbcS transit peptide was replaced by a transit peptide for protein import into mitochondria, the compartment where folic acid biosynthesis takes place in yeast, much higher resistance to sulfonamide and much higher transformation efficiencies are obtained, suggesting that current <jats:italic>sul<\/jats:italic> vectors are likely to function due to low\u2010level mistargeting of the resistance protein to mitochondria. We constructed a series of optimized transformation vectors and demonstrate that they produce transgenic events at very high frequency in both the seed plant tobacco and the green alga <jats:italic>Chlamydomonas reinhardtii<\/jats:italic>. Co\u2010transformation experiments in tobacco revealed that <jats:italic>sul<\/jats:italic> is even superior to <jats:italic>npt<jats:styled-content style=\"fixed-case\">II<\/jats:styled-content><\/jats:italic>, the currently most efficient selectable marker gene, and thus provides an attractive marker for the high\u2010throughput genetic transformation of plants and algae.<\/jats:p>","DOI":"10.1111\/pbi.13004","type":"journal-article","created":{"date-parts":[[2018,8,25]],"date-time":"2018-08-25T09:34:59Z","timestamp":1535189699000},"page":"638-649","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":41,"title":["A highly efficient sulfadiazine selection system for the generation of transgenic plants and algae"],"prefix":"10.1111","volume":"17","author":[{"given":"Iman","family":"Tabatabaei","sequence":"first","affiliation":[{"name":"Max\u2010Planck\u2010Institut f\u00fcr Molekulare Pflanzenphysiologie Potsdam\u2010Golm 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