CAD1 and CCR2 protein complex formation in monolignol biosynthesis in Populus trichocarpa

New Phytol. 2019 Apr;222(1):244-260. doi: 10.1111/nph.15505. Epub 2018 Oct 27.


Lignin is the major phenolic polymer in plant secondary cell walls and is polymerized from monomeric subunits, the monolignols. Eleven enzyme families are implicated in monolignol biosynthesis. Here, we studied the functions of members of the cinnamyl alcohol dehydrogenase (CAD) and cinnamoyl-CoA reductase (CCR) families in wood formation in Populus trichocarpa, including the regulatory effects of their transcripts and protein activities on monolignol biosynthesis. Enzyme activity assays from stem-differentiating xylem (SDX) proteins showed that RNAi suppression of PtrCAD1 in P. trichocarpa transgenics caused a reduction in SDX CCR activity. RNAi suppression of PtrCCR2, the only CCR member highly expressed in SDX, caused a reciprocal reduction in SDX protein CAD activities. The enzyme assays of mixed and coexpressed recombinant proteins supported physical interactions between PtrCAD1 and PtrCCR2. Biomolecular fluorescence complementation and pull-down/co-immunoprecipitation experiments supported a hypothesis of PtrCAD1/PtrCCR2 heterodimer formation. These results provide evidence for the formation of PtrCAD1/PtrCCR2 protein complexes in monolignol biosynthesis in planta.

Keywords: Populus trichocarpa; co-immunoprecipitation; enzyme activity; monolignol biosynthetic pathway; nuclear magnetic resonance (NMR); stem-differentiating xylem protein.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aldehyde Oxidoreductases / genetics
  • Aldehyde Oxidoreductases / metabolism
  • Down-Regulation / genetics
  • Gene Expression Regulation, Plant
  • Lignin / metabolism*
  • Magnetic Resonance Spectroscopy
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Plants, Genetically Modified
  • Populus / genetics
  • Populus / metabolism*
  • RNA Interference
  • Recombinant Proteins / metabolism
  • Xylem / metabolism


  • Plant Proteins
  • Recombinant Proteins
  • Lignin
  • Aldehyde Oxidoreductases
  • cinnamoyl CoA reductase