title: Mycoheterotrophy evolved from mixotrophic ancestors: evidence in Cymbidium (Orchidaceae).
abstract: OBJECTIVE
  Nutritional changes associated with the evolution of achlorophyllous, mycoheterotrophic plants have not previously been inferred with robust phylogenetic hypotheses. Variations in heterotrophy in accordance with the evolution of leaflessness were examined using a chlorophyllous-achlorophyllous species pair in Cymbidium (Orchidaceae), within a well studied phylogenetic background.
  
  
  METHODS
  To estimate the level of mycoheterotrophy in chlorophyllous and achlorophyllous Cymbidium, natural (13)C and (15)N contents (a proxy for the level of heterotrophy) were measured in four Cymbidium species and co-existing autotrophic and mycoheterotrophic plants and ectomycorrhizal fungi from two Japanese sites.
  
  
  RESULTS
  δ(13)C and δ(15)N values of the achlorophyllous C. macrorhizon and C. aberrans indicated that they are full mycoheterotrophs. δ(13)C and δ(15)N values of the chlorophyllous C. lancifolium and C. goeringii were intermediate between those of reference autotrophic and mycoheterotrophic plants; thus, they probably gain 30-50 % of their carbon resources from fungi. These data suggest that some chlorophyllous Cymbidium exhibit partial mycoheterotrophy (= mixotrophy).
  
  
  CONCLUSIONS
  It is demonstrated for the first time that mycoheterotrophy evolved after the establishment of mixotrophy rather than through direct shifts from autotrophy to mycoheterotrophy. This may be one of the principal patterns in the evolution of mycoheterotrophy. The results also suggest that the establishment of symbiosis with ectomycorrhizal fungi in the lineage leading to mixotrophic Cymbidium served as pre-adaptation to the evolution of the mycoheterotrophic species. Similar processes of nutritional innovations probably occurred in several independent orchid groups, allowing niche expansion and radiation in Orchidaceae, probably the largest plant family.
authors.affiliation: Tsukuba Botanical Garden, National Museum of Nature and Science, Tsukuba, Ibaraki, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Motomura
authors.forename: Marc-André
authors.initials: MA
authors.lastname: Selosse
authors.forename: Florent
authors.initials: F
authors.lastname: Martos
authors.forename: Akira
authors.initials: A
authors.lastname: Kagawa
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Yukawa
mesh_terms: D005075:Biological Evolution; D052836:Heterotrophic Processes / Q000502:physiology; D038821:Mycorrhizae / Q000254:growth & development; D029595:Orchidaceae / Q000145:classification / Q000378:metabolism* / Q000382:microbiology*; D010802:Phylogeny; D013559:Symbiosis