Association between the polymorphism of CfPGRP-S1 gene and disease susceptibility/resistance of zhikong scallop (Chlamys farreri) to Listonella anguillarum challenge.
Peptidoglycan recognition protein (PGRP) is a pattern recognition receptor, playing important roles in the innate immune response against invasive pathogens. The single nucleotide polymorphism (SNP) loci in scallop PGRP gene (CfPGRP) were screened from Chlamys farreri to investigate their association with disease resistance of scallop against Listonella anguillarum. Thirteen SNP sites were identified in PGRP domain of CfPGRP, and two of them at positions 4407 and 4408 which are located in the same codon resulted in a nonsynonymous substitution. The genotype frequency of CG/CG in the resistant stock was significantly lower than that in susceptible stock (0% vs 32.4%), while that of CG/TA in the resistant stock was significantly higher than that in susceptible stock (P < 0.01). The pathogen-associated molecular patterns (PAMP) binding activity of two recombinant proteins, rCfPGRP-S1 (R) with CG variant in 4407-4408 site, rCfPGRP-S1 (Y) with TA variant in 4407-4408 site, were elucidated by examining their P/N value at 405 nm with ELISA assay. The in vitro binding activities of the two rCfPGRP-S1 variants to both lipopolysaccharide (LPS) and peptidoglycan (PGN) varied (P < 0.05) in a dose-dependent manner, and rCfPRPP-S1(Y) exhibited significantly higher affinity to PGN and LPS than that of rCfPGRP-S1(R) (P < 0.05). The growth inhibition assay was conducted to find the antibacterial activities of the two variants. Both rCfPGRP-S1(R) and rCfPGRP-S1 (Y) displayed obvious activity to suppress the growth of Escherichia coli, but there was no significant difference in suppression activity of two variants (P > 0.05). The results suggested that the polymorphism at locus 4407-4408 of CfPGRP-S1 considerably affected its PAMP binding activity, and the SNP locus 4407-4408 CG/TA was associated with disease resistance of scallop against L. anguillarum infection, which could be used as a candidate marker for future selection in zhikong scallop breeding program.
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Rd., Qingdao 266071, China.
Vinu S
VS
Siva
Chuanyan
C
Yang
Jialong
J
Yang
Lingling
L
Wang
Leilei
L
Wang
Zhi
Z
Zhou
Limei
L
Qiu
Linsheng
L
Song
D000818:Animals; D017756:Aquaculture; D001483:Base Sequence; D001947:Breeding; D002352:Carrier Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D008070:Lipopolysaccharides / Q000276:immunology; D053526:Listonella / Q000276:immunology; D008969:Molecular Sequence Data; D049897:Pectinidae / Q000235:genetics* / Q000276:immunology* / Q000382:microbiology; D010457:Peptidoglycan / Q000276:immunology; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016415:Sequence Alignment / Q000662:veterinary
D002352:Carrier Proteins; D008070:Lipopolysaccharides; D010457:Peptidoglycan; D011994:Recombinant Proteins; C118897:peptidoglycan recognition protein