Polymorphisms of clip domain serine proteinase and serine proteinase homolog in the swimming crab <i>Portunus trituberculatus</i> and their association with <i>Vibrio alginolyticus</i> -- Journal of Oceanology and Limnology,2017,35(2):235-243

	Cite this paper:
	LIU Meng, LIU Yuan, HUI Min, SONG Chengwen, CUI Zhaoxia. Polymorphisms of clip domain serine proteinase and serine proteinase homolog in the swimming crab Portunus trituberculatus and their association with Vibrio alginolyticus[J]. Journal of Oceanology and Limnology, 2017, 35(2): 235-243

Polymorphisms of clip domain serine proteinase and serine proteinase homolog in the swimming crab Portunus trituberculatus and their association with Vibrio alginolyticus

LIU Meng^1,2, LIU Yuan², HUI Min², SONG Chengwen², CUI Zhaoxia^2,3,4

1 College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China;
2 Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;
3 Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China;
4 National & Local Joint Engineering Laboratory of Ecological Mariculture, Qingdao 266071, China

Abstract:

Clip domain serine proteases (cSPs) and their homologs (SPHs) play an important role in various biological processes that are essential components of extracellular signaling cascades, especially in the innate immune responses of invertebrates. Here, polymorphisms of PtcSP and PtSPH from the swimming crab Portunus trituberculatus were investigated to explore their association with resistance/susceptibility to Vibrio alginolyticus. Polymorphic loci were identified using Clustal X, and characterized with SPSS 16.0 software, and then the significance of genotype and allele frequencies between resistant and susceptible stocks was determined by a χ² test. A total of 109 and 77 single nucleotide polymorphisms (SNPs) were identified in the genomic fragments of PtcSP and PtSPH, respectively. Notably, nearly half of PtSPH polymorphisms were found in the non-coding exon 1. Fourteen SNPs investigated were significantly associated with susceptibility/resistance to V. alginolyticus (P<0.05). Among them, eight SNPs were observed in introns, and one synonymous, four non-synonymous SNPs and one ins-del were found in coding exons. In addition, five simple sequence repeats (SSRs) were detected in intron 3 of PtcSP. Although there was no statistically significant difference of allele frequencies, the SSRs showed different polymorphic alleles on the basis of the repeat number between resistant and susceptible stocks. After further validation, polymorphisms investigated here might be applied to select potential molecular markers of P. trituberculatus with resistance to V. alginolyticus.

Key words: Portunus trituberculatus|clip domain serine proteinase|serine proteinase homolog|polymorphism|susceptibility/resistance

Received: 2015-11-19 Revised: 2016-02-01

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