Development of single-locus DNA microsatellite markers using 5'anchored ISSR-PCR method for the mangrove horseshoe crab, Carcinoscorpius rotundicauda (Latreille, 1802) in Peninsular Malaysia

Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.
Molecular Biology Reports (Impact Factor: 2.02). 07/2011; 39(4):3815-20. DOI: 10.1007/s11033-011-1159-6
Source: PubMed


Horseshoe crabs are said to be declining worldwide. However, there is still no published report on the status of horseshoe crabs in Malaysia. Thus, we report here eight informative microsatellite markers that were developed using the 5'-anchored ISSR-PCR enrichment procedure to diagnose the population genetic structure of the mangrove horseshoe crab, Carcinoscorpius rotundicauda from Peninsular Malaysia. This set of markers was tested on 127 samples and showed polymorphism in this species. Hence they should be useful in future essential population genetic studies of these living fossils in the Southeast Asian region.

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    • "Paran & Michelmore 1993), and ISSR for the development of single-locus co-dominant microsatellite markers (e.g. Fisher et al. 1996; Lian et al. 2001; Adibah et al. 2012). "
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    ABSTRACT: Inter-simple sequence repeats (ISSRs) are regions in the genome flanked by microsatellite sequences. PCR amplification of these regions using a single primer yields multiple amplification products that can be used as a dominant multilocus marker system for the study of genetic variation in various organisms. ISSR markers are easy to use, low-cost, and methodologically less demanding compared to other dominant markers, making it an ideal genetic marker for beginners and for organisms whose genetic information is lacking. Here, we comment upon some of the intricacies often overlooked in designing an ISSR experiment, clarify some misconceptions, and provide recommendations on using ISSR markers in genetic variation studies.
    ASM Science Journal 11/2015; 9(1):30-39.
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    ABSTRACT: Horseshoe crabs are marine arthropods that are amongst the oldest living creatures that still exist today. Among the four extant species of horseshoe crabs, Carcinoscorpius rotundicauda differs from the other species by having poisonous eggs and lays its eggs in sandy-mud areas near river mouths. With the rapid development of coastal areas worldwide, C. rotundicauda habitats are decreasing. Until now, however, there has not been any study on the species' genetic variation. Simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) markers were employed to study the genetic variation in five C. rotundicauda populations from the east and west coasts of the Malay Peninsula. Both markers showed differing levels of genetic variation, but concurred on the pattern of genetic structuring among populations of the species. This includes showing that little, although significant, genetic differentiation is present among populations, suggesting a low rate of gene flow among populations. The results also suggested that C. rotundicauda may be subjected to the land barrier effect of the Malay Peninsula, whereby gene flow is limited between populations occurring on both sides of the peninsula, increasing their genetic differentiation through time.
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