Flavone synthases from Medicago truncatula are flavanone-2-hydroxylases and are important for nodulation

Plant Physiol. 2007 Jun;144(2):741-51. doi: 10.1104/pp.106.095018. Epub 2007 Apr 13.

Abstract

Flavones are important copigments found in the flowers of many higher plants and play a variety of roles in plant adaptation to stress. In Medicago species, flavones also act as signal molecules during symbiotic interaction with the diazotropic bacterium Sinorhizobium meliloti. They are the most potent nod gene inducers found in root exudates. However, flavone synthase II (FNS II), the key enzyme responsible for flavone biosynthesis, has not been characterized in Medicago species. We cloned two FNS II genes from Medicago truncatula using known FNS II sequences from other species and named them MtFNSII-1 and MtFNSII-2. Functional assays in yeast (Saccharomyces cerevisiae) suggested that the catalytic mechanisms of both cytochrome P450 monooxygenases were similar to the other known legume FNS II from licorice (Glycyrrhiza echinata). MtFNSII converted flavanones to 2-hydroxyflavanones instead of flavones whereas FNS II from the nonlegume Gerbera hybrida, converted flavanones to flavones directly. The two MtFNSII genes had distinct tissue-specific expression patterns. MtFNSII-1 was highly expressed in roots and seeds whereas MtFNSII-2 was highly expressed in flowers and siliques. In addition, MtFNSII-2 was inducible by S. meliloti and methyl jasmonate treatment, whereas MtFNSII-1 was not. Histochemical staining of transgenic hairy roots carrying the promoter-reporter constructs indicated that the MtFNSII-2 induction was tissue specific, mostly localized to vascular tissues and root hairs. RNA interference-mediated suppression of MtFNSII genes resulted in flavone depleted roots and led to significantly reduced nodulation when inoculated with S. meliloti. Our results provide genetic evidence supporting that flavones are important for nodulation in M. truncatula.

Publication types

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

MeSH terms

  • Cytochrome P-450 Enzyme System / genetics
  • Cytochrome P-450 Enzyme System / metabolism*
  • Flavanones / metabolism
  • Flavones / metabolism*
  • Gene Expression
  • Gene Silencing
  • Genes, Plant
  • Medicago truncatula / enzymology*
  • Medicago truncatula / microbiology
  • Medicago truncatula / physiology
  • Mixed Function Oxygenases / genetics
  • Mixed Function Oxygenases / metabolism*
  • Plants, Genetically Modified
  • Promoter Regions, Genetic
  • Root Nodules, Plant / enzymology*
  • Root Nodules, Plant / microbiology
  • Root Nodules, Plant / physiology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Sinorhizobium meliloti / physiology*
  • Symbiosis / physiology

Substances

  • Flavanones
  • Flavones
  • Cytochrome P-450 Enzyme System
  • Mixed Function Oxygenases
  • cytochrome P-450 CYP93B1 (Glycyrrhiza echinata)