Different acyl-CoA:diacylglycerol acyltransferases vary widely in function, and a targeted amino acid substitution enhances oil accumulation

Tomoko Hatanaka, Yoshiki Tomita, Daisuke Matsuoka, Daisuke Sasayama, Hiroshi Fukayama, Tetsushi Azuma, Mohammad Fazel Soltani Gishini, David Hildebrand

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Triacylglycerols (TAGs) are the major component of plant storage lipids such as oils. Acyl-CoA:diacylglycerol acyltransferase (DGAT) catalyzes the final step of the Kennedy pathway, and is mainly responsible for plant oil accumulation. We previously found that the activity of Vernonia DGAT1 was distinctively higher than that of Arabidopsis and soybean DGAT1 in a yeast microsome assay. In this study, the DGAT1 cDNAs of Arabidopsis, Vernonia, soybean, and castor bean were introduced into Arabidopsis. All Vernonia DGAT1-expressing lines showed a significantly higher oil content (49% mean increase compared with the wild-type) followed by soybean and castor bean. Most Arabidopsis DGAT1-overexpressing lines did not show a significant increase. In addition to these four DGAT1 genes, sunflower, Jatropha, and sesame DGAT1 genes were introduced into a TAG biosynthesis-defective yeast mutant. In the yeast expression culture, DGAT1s from Arabidopsis, castor bean, and soybean only slightly increased the TAG content; however, DGAT1s from Vernonia, sunflower, Jatropha, and sesame increased TAG content >10-fold more than the former three DGAT1s. Three amino acid residues were characteristically common in the latter four DGAT1s. Using soybean DGAT1, these amino acid substitutions were created by site-directed mutagenesis and substantially increased the TAG content.

Original languageEnglish
Pages (from-to)3030-3043
Number of pages14
JournalJournal of Experimental Botany
Volume73
Issue number9
DOIs
StatePublished - May 13 2022

Bibliographical note

Publisher Copyright:
© 2022 The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved.

Keywords

  • Acyl-CoA:diacylglycerol acyltransferase
  • Arabidopsis thaliana
  • DGAT1
  • Vernonia galamensis
  • site-directed mutagenesis
  • triacylglycerol
  • yeast strain H1246

ASJC Scopus subject areas

  • Physiology
  • Plant Science

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