Morphogen-Based Chemical Flypaper for Agaricia humilis Coral Larvae.

Larvae of the scleractinian coral Agaricia humilis settle and metamorphose in response to chemosensory recognition of a morphogen on the surfaces of Hydrolithon boergesenii and certain other crustose coralline red algae. The requirement of the larva for this inducer apparently helps to determine the spatial pattern of recruitment in the natural environment. Previous research showed that the inducer is associated with the insoluble cell wall fraction of the recruiting algae or their microbial epibionts, and that a soluble but unstable fragment of the inducing molecule can be liberated by limited hydrolysis, either with alkali or with enzymes specific for cell wall polysaccharides. We now show that the parent morphogen can be solubilized by gentle decalcification of the algal cell walls with the chelators EGTA or EDTA, suggesting that the morphogen may be a component of the calcified recruiting alga itself, rather than a product of any noncalcified microbial epibionts. The solubilized inducer is subsequently purified by hydrophobic-interaction and DEAE chromatography. The purified, amphipathic morphogen retains activity when tightly bound to beads of a hydrophobic-interaction chromatography resin, and this activity (tested with laboratory-reared larvae) is identical in the ocean and the laboratory. We have attached the purified, resin-bound inducer to surfaces coated with a silicone adhesive and thus produced a potent artificial recruiting substratum--i.e., a morphogen-based chemical "flypaper" for A. humilis larvae. This material should prove useful in resolving the role of chemosensory recognition of morphogens in the control of substratum-specific settlement, metamorphosis, and recruitment and in the maintenance of species isolation mechanisms in the natural environment.