Dual recognition activity of a rhamnose-binding lectin to pathogenic bacteria and zooxanthellae in stony coral Pocillopora damicornis

- PdRBL-1 bound LPS and Lipid A, but not LTA, β-glucan, mannose and Poly (I:C).
- PdRBL-1 bound zooxanthellae in a concentration-dependent manner.
- The PdRBL-1's binding to zooxanthellae was repressed by LPS.
- The expression of RdRBL-1 mRNA was upregulated by heat stress.

Rhamnose-binding lectin (RBL) is a type of Ca2+-independent lectin with tandem repeat carbohydrate-recognition domain, and is crucial for the innate immunity in many invertebrates. In this study, the cDNA sequence encoding RBL in coral Pocillopora damicornis (PdRBL-1) was cloned. The PdRBL-1 protein shared highest amino acid sequence similarity (55%) with the polyp of Hydra vulgaris, and contained a signal peptide and two tandem carbohydrate-recognition domains in which all cysteine residues were conserved. Surface plasmon resonance method revealed that the recombinant PdRBL-1 protein bound to LPS and Lipid A, but not to LTA, β-glucan, mannose and Poly (I:C). Results also showed that it bonded with zooxanthellae using western blotting method, and that the bound protein was detectable only at concentrations higher than 102 zooxanthellae cell mL−1. When recombinant PdRBL-1 protein was preincubated with LPS, lower amounts of protein bound to zooxanthellae compared to cells not preincubated with LPS. Furthermore, PdRBL-1 mRNA expression increased significantly at 12 h, and declined to the baseline at 24 h after heat stress at 31 °C. These results collectively suggest that PdRBL-1 could recognize not only pathogenic bacteria but also symbiotic zooxanthellae, and that the recognition of zooxanthellae by PdRBL-1 could be repressed by pathogenic bacteria through competitive binding. This information allows us to gain new insights in the mechanisms influencing the establishment and maintenance of coral-zooxanthella symbiosis in coral P. damicornis.