Pure culture and reconstitution of the Anthoceros-Nostoc symbiotic association

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Abstract
The partners of the symbiotic association between Anthoceros punctatus L. and Nostoc spp. have been cultured separately in a pure state. The symbiotic association was reconstituted following dual culture in liquid Anthoceros growth medium with a variety of axenic Nostoc isolates and mutant strains. The heterocyst frequency of competent Nostoc strains increased four- to fivefold when in symbiotic association relative to free-living N2-grown cultures. Dinitrogen fixation by symbiotic Nostoc supported the growth of Anthoceros tissue, although this growth was nitrogen-limited relative to that supported by exogenous ammonium. When the association was reconstituted in the presence of two or three wild-type and mutant Nostoc strains some of these strains were found to compete in infection of Anthoceros tissue and a fraction of the symbiotic Nostoc colonies contained more than one strain. Exogenous ammonium did not affect infection, but repressed development of the symbiotic Nostoc colonies in Anthoceros tissue, and symbiotic Nostoc in N2-grown Anthoceros tissue appeared to regress from the symbiotic state in the presence of exogenous ammonium. The results show that the Anthoceros-Nostoc symbiotic association is amenable to specific experimental manipulations; their implications are discussed with respect to infection of Anthoceros tissue and control of the development of symbiotic Nostoc.