Novel Polymorphic Microsatellite Markers for Panulirus ornatus and their Cross-species Primer Amplification in Panulirus homarus

Abstract

Polymorphic microsatellite loci were isolated for Panulirus ornatus using 454 GS-FLX Titanium pyrosequencing. Fifteen markers containing perfect di-, tri-, tetra-, and penta-nucleotide motifs were consistently co-amplified in five multiplexes in a panel of 91 randomly selected samples. Observed number of alleles varied from 2 to 14 per locus. Observed and expected heterozygosity ranged from 0.090 to 0.79 and 0.08 to 0.87, respectively. Ten loci deviated from Hardy-Weinberg equilibrium after sequential Bonferroni correction. Genetic linkage disequilibrium analysis between all pairs of the loci showed significant departure from the null hypothesis between 11 loci. The microsatellite markers were also amplified successfully in related Panulirus homarus species with adequate level of polymorphism. The successful cross-species primer amplification of the 15 microsatellites indicates the potential of the developed markers to be transferred to other Panulirus species. The 15 novel microsatellite markers reported in this work add to the previously characterized markers by our group, exhibit adequate levels of polymorphism for wide range of future studies investigating population structure, genetic diversity, and evolutionary relationships among Panulirus species.

Parentage analysis of tropical spiny lobster (Panulirus homarus) by microsatellite markers

“…Based on their polymorphisms, allelic sizes and PCR conditions, seven microsatellite loci, Pho ‐G06, Pho ‐G25, Pho ‐G53, Pho ‐G62, Pho ‐G74, Pho ‐G89 and Pho ‐G100, containing di‐ tri‐ and tetranucleotide motives (Delghandi et al . , Delghandi, Goddard et al ., ; Delghandi, Afzal et al . ) were selected and co‐amplified in two multiplexes (MI and MII).…”

“…, Delghandi, Goddard et al ., ; Delghandi, Afzal et al . ) were selected and co‐amplified in two multiplexes (MI and MII). For fluorescent detection and multiplexing, the forward primers were dye‐labelled (NED, VIC or 6‐FAM) (Life Science Technologies) (Table ).…”

“…This approach has been successfully applied in several marine fish species for assignment of individuals to groups of families or to estimate the relationship among individuals (Herlin, Delghandi, Wesmajervi, Taggart, McAndrew & Penman ; Vandeputte & Haffray ; Zhang, Ma, Wang, Gui & Mei ). Recently, we have isolated several microsatellite markers for tropical spiny lobster (Delghandi, Goddard, Jerry, Dao, Afzal & Al‐Jardani ; Delghandi, Afzal, Al Hinai, Al‐Breiki, Jerry & Dao ; Delghandi, Goddard, Jerry, Dao, Al Hinai, Al‐Amry & Al‐Marzouqi ). Based on genetic diversity, allelic size and polymerase chain reaction (PCR) conditions, seven loci were selected and multiplexed to assess the power of the markers for parentage assignment studies.…”

Population Genetic Structure of Ornatus Spiny Lobster (Panulirus Ornatus) in Indo-West Pacific Region

“…The present study applied five microsatellite markers selected from the study of Delghandi et al [13]. For each pair of primers, the forward primer was fluorescent labeled.…”

“…Microsatellites technique or Simple Sequence Repeats (SSR) is a powerful tool, widely applied in the identification of subspecies, life cycle linkage and population genetic structure [12]- [14].To date, a number of microsatellite markers specific to spiny lobsters have been identified and can be applied across many species and geographical regions [12], [13], [15], [16] .…”