A microsatellite panel for triploid verification in the abaloneHaliotis midae

Abstract

A method for ploidy verification of triploid and diploid Haliotis midae was developed using molecular microsatellite markers. In all, 30 microsatellite loci were tested in control populations. A final micro satellite multiplex consisting of seven markers were optimised and a complete protocol is reported. This protocol was successfully applied in identifying the ploidy levels of 42 ploidy-unknown abalone, showing the utility of microsatellite markers as tools for verifying the ploidy of individual abalone. The protocol can be applied in larger animals for which the isolation of nuclei and metaphase chromosomes from tissues is difficult.

Development of Genetic Markers for Triploid Verification of the Pacific Oyster, <italic>Crassostrea gigas</italic>

“…Among the 11 microsatellite markers tested, we selected six with primer sets of the same annealing temperature and on the basis of their expected heterozygosity values, which ranged from 0.71 to 0.86 and were similar to the values suggested by Slabbert et al (2010) . These primers were polymorphic in all of the 192 diploid individuals from two wild populations and had allele numbers ranging from 30 to 39, which was much greater than the original reports, probably due to the large sample in this study.…”

“…Thus, the number of primers in the multiplex should be chosen so that a clear result can be obtained with a minimum number of primer sets. To verify triploid abalones, Slabbert et al (2010) screened 30 microsatellite makers and developed a microsatellite multiplex PCR consisting of seven markers. In our study, two multiplex panels of three primers each resulted in 100% ploidy verification.…”

“…Another direct method of triploidy verification is the use of polymorphic DNA markers, such as microsatellites ( Slabbert et al, 2010 ). Microsatellites, or simple sequence repeats (SSRs), are highly polymorphic, multi-allelic, co-dominant, reproducible, and easy to interpret molecular markers that have been used to determine individual genetic variability and to study the genetic structure of a population ( Alam et al, 2012 ; Dadi et al, 2012a ; Dadi et al, 2012b ).…”