{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T15:15:39Z","timestamp":1761059739684,"version":"3.37.3"},"reference-count":49,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2014,6,23]],"date-time":"2014-06-23T00:00:00Z","timestamp":1403481600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001732","name":"Danish National Research Foundation","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100001732","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Danish Council for Independent Research - Natural Sciences"},{"name":"VILLUM Research Center \u2018Plant Plasticity\u2019"},{"name":"Danish Council for Independent Research Technology and Production Sciences"},{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"crossref","award":["BB\/I025891\/1"],"award-info":[{"award-number":["BB\/I025891\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Plant Journal"],"published-print":{"date-parts":[[2014,7]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p><jats:italic><jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>otus japonicus<\/jats:italic>, like several other legumes, biosynthesizes the cyanogenic \u03b1\u2013hydroxynitrile glucosides lotaustralin and linamarin. Upon tissue disruption these compounds are hydrolysed by a specific \u03b2\u2013glucosidase, resulting in the release of hydrogen cyanide. <jats:italic><jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>otus japonicus<\/jats:italic> also produces the non\u2010cyanogenic \u03b3\u2010 and \u03b2\u2013hydroxynitrile glucosides rhodiocyanoside A and D using a biosynthetic pathway that branches off from lotaustralin biosynthesis. We previously established that <jats:styled-content style=\"fixed-case\">BGD<\/jats:styled-content>2 is the only \u03b2\u2013glucosidase responsible for cyanogenesis in leaves. Here we show that the paralogous <jats:styled-content style=\"fixed-case\">BGD<\/jats:styled-content>4 has the dominant physiological role in rhodiocyanoside degradation. Structural modelling, site\u2010directed mutagenesis and activity assays establish that a glycine residue (G211) in the aglycone binding site of <jats:styled-content style=\"fixed-case\">BGD<\/jats:styled-content>2 is essential for its ability to hydrolyse the endogenous cyanogenic glucosides. The corresponding valine (V211) in <jats:styled-content style=\"fixed-case\">BGD<\/jats:styled-content>4 narrows the active site pocket, resulting in the exclusion of non\u2010flat substrates such as lotaustralin and linamarin, but not of the more planar rhodiocyanosides. Rhodiocyanosides and the <jats:italic><jats:styled-content style=\"fixed-case\">BGD<\/jats:styled-content>4<\/jats:italic> gene only occur in <jats:italic><jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>.\u00a0japonicus<\/jats:italic> and a few closely related species associated with the <jats:italic><jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>otus corniculatus<\/jats:italic> clade within the <jats:italic><jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>otus<\/jats:italic> genus. This suggests the evolutionary scenario that substrate specialization for rhodiocyanosides evolved from a promiscuous activity of a progenitor cyanogenic \u03b2\u2013glucosidase, resembling <jats:styled-content style=\"fixed-case\">BGD<\/jats:styled-content>2, and required no more than a single amino acid substitution.<\/jats:p>","DOI":"10.1111\/tpj.12561","type":"journal-article","created":{"date-parts":[[2014,5,26]],"date-time":"2014-05-26T14:23:45Z","timestamp":1401114225000},"page":"299-311","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["The evolutionary appearance of non\u2010cyanogenic hydroxynitrile glucosides in the <i><scp>L<\/scp>otus<\/i> genus is accompanied by the substrate specialization of paralogous \u03b2\u2013glucosidases resulting from a crucial amino acid substitution"],"prefix":"10.1111","volume":"79","author":[{"given":"Daniela","family":"Lai","sequence":"first","affiliation":[{"name":"Plant 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