Haplotype-specific single-locus multiplex PCR assay for molecular identification of sea-bob shrimp,Xiphopenaeus kroyeri (Heller, 1862), cryptic species from the Southwest Atlantic using a DNA pooling strategy for simultaneous identification of multiple samples |
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Institution: | 1. Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, National Engineering Research Center for Floriculture, College of Landscape Architecture, Beijing Forestry University, No. 35 Qinghuadong Road, Beijing 100083, PR China;2. Miki-cho Post Office, P. O. Box 7, Ikenobe 3011-2, Kagawa-ken 761-0799, Japan;1. Departamento de Farmacología, Facultad de Medicina, C. Escolar, C. Universitaria, Coyoacán, 04510 Mexico, D. F, Mexico;2. Instituto de Química, C. Exterior, C. Universitaria, Coyoacán, 04510 Mexico, D. F, Mexico;1. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;2. Shangdong University of Traditional Chinese Medicine, Medical College, Jinan 250355, China |
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Abstract: | This work describes a single-locus multiplex PCR assay based on partial COI mitochondrial gene polymorphisms for identification of two Atlantic cryptic species of the sea-bob shrimp, Xiphopenaeus kroyeri (Heller, 1862) that were recently identified using molecular approaches. Reliable identification of cryptic species of Xiphopenaeus spp. has fundamental implications for management and conservation of the sea-bob shrimp fishery stocks. The assay was developed based on sequence polymorphisms of 130 specimens of both species, comprising samples from Venezuela to the southern coast of Brazil, validated by the amplification of 368 adult shrimp samples from nine different locations and confirmed by direct sequencing. The methodology has been optimized to enable the identification of equi-molar mixtures of DNA from up to 10 individuals by PCR reaction, allowing the fast and cheap identification of many specimens for large scale studies on fisheries biology and population genetics. The DNA pooling strategy enabled the identification of a new locality of occurrence of Xiphopenaeus sp. II in the Brazilian coast, Caravelas, indicating that the species distribution may be continuous on the coast, and not disjoint as observed so far. |
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Keywords: | Fisheries stocks Sea-bob shrimp Cryptic species COI DNA pooling Systematics |
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