Cloning of a cDNA probably encoding oxidosqualene cyclase associated with asiaticoside biosynthesis from Centella asiatica (L.) Urban

Plant Cell Rep. 2005 Jul;24(5):304-11. doi: 10.1007/s00299-005-0927-y. Epub 2005 Apr 15.

Abstract

A homology-based PCR method was used to clone a cDNA encoding oxidosqualene cyclase from Centella asiatica, which produces a large quantity of triterpene saponins such as asiaticoside and madecassoside. Sequence analysis of one clone found sequences related to beta-amyrin synthase. An open reading frame in the full-length clone was named CabAS (Centella asiatica putative beta-amyrin synthase). On the basis of amino acid sequence, CabAS appears to be an enzyme (beta-amyrin synthase) that synthesizes beta-amyrin. Southern analysis showed that the C. asiatica genome contains one copy of the CabAS gene. Northern blot analysis demonstrated that the CabAS gene is expressed in leaves with no detectable transcript in other plant tissues, consistent with the organ-specific accumulation of the asiaticoside. Up-regulation of expression of CabAS by methyl jasmonate in leaves was also demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / pharmacology
  • Amino Acid Sequence
  • Centella / enzymology
  • Centella / genetics
  • Centella / metabolism*
  • Cloning, Molecular
  • Cyclopentanes / pharmacology
  • DNA, Complementary / chemistry
  • Intramolecular Transferases / genetics*
  • Intramolecular Transferases / metabolism
  • Molecular Sequence Data
  • Oxylipins
  • Phylogeny
  • Plant Leaves / enzymology
  • Plant Leaves / genetics
  • Plant Leaves / metabolism*
  • RNA, Messenger / metabolism
  • Sequence Homology, Amino Acid
  • Triterpenes / metabolism*
  • Up-Regulation

Substances

  • Acetates
  • Cyclopentanes
  • DNA, Complementary
  • Oxylipins
  • RNA, Messenger
  • Triterpenes
  • methyl jasmonate
  • Intramolecular Transferases
  • 2,3-oxidosqualene-beta-amyrin-cyclase
  • lanosterol synthase
  • asiaticoside