Licence This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. Morphological identification and molecular confirmation of the deep-sea blue and red shrimp Aristeus antennatus larvae. Abstract The early life stages of the blue and red shrimp Aristeus antennatus Decapoda: Dendrobranchiata: Penaeoidea: Aristeidae were described by Heldt in based on plankton samples, larval rearing and assumptions of species habitat. Even with adequate keys, identification of its first larval stages remained a difficult task due to the lack of specific morphological characters which would differentiate them from other Penaeoidea species. DNA from the larvae was extracted and the molecular markers Cytochrome Oxidase I and 16S rDNA were sequenced and compared to that of adults with the objective of confirming the previous morphological description. Then, we present additional information to the morphological description of Aristeus antennatus larval stages through scanning electron microscopy and molecular analysis.
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Licence This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed.
Development and characterization of novel microsatellite markers by Next Generation Sequencing for the blue and red shrimp Aristeus antennatus.
Hypervariable microsatellite markers could be a useful tool to identify genetic stocks among geographically close fishing grounds. Potential microsatellite markers 97 identified from next-generation sequencing of an individual shrimp using a GS Junior Pyrosequencer were tested on a preliminary panel of 15 individuals representing the four worldwide genetic stocks of the species from which 35 polymorphic loci were identified and used to characterize an additional 20 individuals from the Western Mediterranean Sea.
In the Western Mediterranean sample, 32 out of 35 were polymorphic loci and the number of alleles per locus ranged from 2 to 14 and expected heterozygosity ranged from 0. No linkage disequilibrium was detected, indicating the independence of the loci. These novel microsatellites provide additional tools to address questions relating to genetic diversity, parentage studies and connectivity patterns of A. Exploited since the s, the global catch has increased annually, reaching 1, tonnes in FAO , Although studies have addressed A.
Nevertheless, microsatellites have proved their utility in identifying genetic stocks in marine penaeoid shrimps around the world Benzie , ; Brooker et al. Next-generation sequencing NGS based on pyrosequencing has enabled a high-throughput approach and has demonstrated its usefulness in the development of SSRs in non-model organisms Schoebel et al. Long reads generated by sequencing are expected to contain SSRs and their flanking regions that are needed to develop methods for SSR screening among populations.
The aim of our study was the development and characterization of novel polymorphic microsatellite loci for A. The high quality of extracted DNA was checked by resolution on a 0. The reads obtained were assembled into contigs using the GS De Novo Assembler program included in the Newbler software package v.
All contigs matching with already available A. Remaining contigs were scanned with Tandem Repeats Finder v. Alignment parameters of match, mismatch and indels were set as 2, 7 and 7, respectively; minimum alignment score to report repeats was set to 30; maximum period size to 5 and matching and indel probabilities were set as 0.
In addition, the option of flanking sequence was considered to record up to nucleotides on each side of the repeat in order to later design specific PCR primers. Contigs containing tandem repeats with at least 30 bp of flanking regions were used for primer design with Primer3 v. Verification of putative microsatellites Putative microsatellites were preliminarily screened for amplification success and polymorphism by genotyping a panel of 15 A.
This reduced genotyping costs because only a common fluorescently labelled primer was required for all putative microsatellite verifications. Table 1: Characteristics of 35 new microsatellite markers developed for A.
EPPO Global Database
Abstract Aristeus antennatus is an ecologically and economically important deep-water species in the Mediterranean Sea. In this study we investigated the genetic variability of A. By comparing our new samples with available data from the Western area, we aim to identify potential genetic stocks of A. We found two main results: i the genetic diversity values consistent with previous data within the Mediterranean and the absence of barriers to gene flow within the Mediterranean Sea; ii a constant long-term effective population size in almost all demes but a strong signature of population expansion in the pooled sample about 50, years B. We propose two explanation for our results.