Allotriploid genotypic assignment in abalone larvae by detection of microsatellite-recombinant genotypes

Abalone species are different from most mollusks utilized in aquaculture as they are known to hybridize in laboratory-induced matings. Allotriploidization of hybrid abalone has not yet been studied, and methodology useful in verifying the genotypic condition of such allotriploids do not exist. Genotypic verification of hybridization and allotriploidization in a cross of Haliotis fulgens and Haliotis rufescens was performed utilizing 6 crossamplifying microsatellite loci. Five H. rufescens spawns were used in this experiment, dividing each spawn into control and experimental hybrid groups and further into diploids and triploids. Two microsatellite loci developed for H. fulgens and H. rufescens allowed for the genotypic identification of hybrids within diploid and triploids. To further verify the percentage of allotriploids within the genotypic hybrids in the triploid hybrid groups, microsatellite loci originally developed in Haliotis corrugata and Haliotis kamtschatkana were tested for crossamplification in H. fulgens and H. rufescens. Of 21 loci, 4 were chosen for this study based on their crossamplification, heterozygosity in the females, and centromere recombination frequencies. Allotriploids in triploid-hybrid larvae were then detected by identifying larvae with recombinant genotypes at any of those loci. One family had low success verification associated with reduced recombination frequencies for all loci in that family. These results demonstrate that allotriploid verification at larval stages is feasible but depends on the number of loci available, their crossamplification in the species, and their recombination frequencies.