Identification and analysis of C-type lectins from Helicoverpa armigera in response to the entomopathogenic fungus Metarhizium rileyi infection

Abstract

C-type lectins (CTLs) function as pattern recognition receptors (PRRs) and play an important role in the innate immunity of insects. To investigate the role of CTLs in the antifungal responses, we analyzed expression profiles of 36 CTLs of Helicoverpa armigera in the tissues (hemocytes, fat bodies, and midgut) after the infection by entomopathogenic fungus Metarhizium rileyi. The expression levels of many HaCTLs were found to be up-regulated after the infection. Four recombinant HaCTLs (rHaCTL11, rHaCTL12, rHaCTL27, and rHaCTL45) were expressed and purified. Analysis of the purified rHaCTLs revealed that rHaCTLs were able to bind to conidia and hyphal bodies of M. rileyi, and the affinity of rHaCTL11 and rHaCTL27 for hyphal bodies was weaker than for conidia. All these rHaCTLs agglutinate conidia and hyphal bodies in a calcium (Ca2+) dependent manner. Sugar specificity assays showed that d-trehalose, mannan, β-1,3-glucan, d-galactose, glucose, d-raffinose, lipopolysaccharide, and d-xylose can inhibit the binding of HaCTLs to M. rileyi. Additionally, survival assays showed that pretreatment of fungal conidia with rHaCTL11 significantly reduced the rate of host death, and knockdown of HaCTL11 significantly increased H. armigera sensitivity to fungal infection. These results suggest that HaCTLs play significant role as PRRs in the defense of H. armigera against M. rileyi infection.