A fast sex detection method for the Whiteleg shrimp Litopenaeus vannamei by post-PCR high resolution melting (HRM)

Abstract The Pacific Whiteleg shrimp Litopenaeus vannamei is one of the most important aquaculture species globally, for which monosex female culture is desirable due to its higher weight at harvest. For this purpose, the development and validation of biotechnologies directed to manipulate sexual differentiation requires a verification protocol based on molecular sex markers in the early life stages where no secondary sexual characteristics are displayed. In the present study, the sequences of a 542 bp putatively sex-linked locus containing three single nucleotide polymorphisms (SNP) followed the homogametic (male, n = 8) and heterogametic (female, n = 4) sex-determination system. The locus is probably not located in previously reported sex-associated linkage groups. A post-PCR high resolution melt (HRM) analysis was designed to amplify and differentiate homozygotes from heterozygotes of the SNP in the locus. The expected patterns in a ZZ/ZW genetic system was confirmed in wild and cultivated shrimp samples (n = 168). These results are of interest for sex detection and for future detailed studies on the sex-determination system in L. vannamei.