Identification of European species of Maja (Decapoda: Brachyura: Majidae): RFLP analyses of COI mtDNA and morphological considerations


  • Guillermo Guerao IRTA, Unitat Operativa de Cultius Aquàtics
  • Karl B. Andree IRTA, Unitat Operativa de Cultius Aquàtics
  • Carlo Froglia Istituto di Scienze Marine (CNR)
  • Carles G. Simeó IRTA, Unitat Operativa de Cultius Aquàtics
  • Guiomar Rotllant IRTA, Unitat Operativa de Cultius Aquàtics



Maja, M. squinado, M. brachydactyla, molecular identification, morfometry, COI, RFLP


Four species of crabs of the genus Maja have been described along the European coast: M. brachydactyla, M. squinado, M. goltziana and M. crispata. The commercially important species M. brachydactyla and M. squinado achieve the largest body sizes and are the most similar in morphology, and are therefore easily confused. The four species of Maja were identified using a novel morphometric index and a polymerase chain reaction followed by restriction fragment length polymorphism analysis (RFLP). The relationship between carapace length and the distance between the tips of antorbital spines was used to distinguish adults of M. brachydactyla and M. squinado. PCR-RFLP analysis of a partial sequence of the mitochondrial cytochrome oxidase type I (COI) revealed that the four species of the genus Maja can be unambiguously discriminated using the combination of restriction endonucleases enzymes HpyCH4V and Ase I. The molecular identification may be particularly useful in larvae, juvenile and young crabs, when the morphological differences found in adults are not applicable.


Download data is not yet available.


Balss, H. – 1922. Crustacea VII: Decapoda Brachyura (Oxyrhyncha und Brachyrhyncha) und geographische Übersicht über Crustacea Decapoda. In: W. Michaelsen (ed.), Beiträge zur Kenntnis der Meeresfauna Westafrikas, pp. 69-110. Friederichsen and Co., Hamburg.

Bossier, P., W. Xiaomei, F. Catania, S. Dooms, G. Van Stappen, E. Naessens and P. Sorgeloos. – 2004. An RFLP database for authentication of commercial cyst samples of the brine shrimp Artemia spp. (International Study on Artemia LXX). Aquaculture, 231: 93-112. doi:10.1016/j.aquaculture.2003.11.001

Bucklin, A., P.H. Wiebe, S.B. Smolenack, N.J. Copley, J.G. Beaudet, K.G. Bonner, J. Färber-Lorda and J.J Pierson. – 2007. DNA barcodes for species identification of euphausiids (Euphausiacea, Crustacea). J. Plank. Res., 29(6): 483-493. doi:10.1093/plankt/fbm031

Bucklin, A., R.R Hopcroft, K.N. Kosobokova, L.M. Nigro, B.D. Ortman, R.M. Jennings and C.J. Sweetman. – 2009. DNA barcoding of Artic Ocean holozooplankton for species identification and recognition. Deep-Sea Res. II, 57: 40-48. doi:10.1016/j.dsr2.2009.08.005

Clark, P.F. – 1986. The larval stages of Maja squinado (Herbst, 1788) (Crustacea: Brachyura: Majidae) reared in the laboratory. J. Nat. Hist., 20: 825-836. doi:10.1080/00222938600770591

Costa, F.O., J.R. deWaard, J. Boutillier, S. Ratnasingham, R.T. Dooh, M. Hajibabaei and P.D.N. Hebert. – 2007. Biological identifications through DNA barcodes: the case of the Crustacea. Can. J. Fish. Aquat. Sci., 64: 272-295. doi:10.1139/F07-008

Chow, S., N. Suzuki, H. Imai and T. Yoshimura. – 2006. Molecular species identification of spiny lobster phyllosoma larvae of the genus Panulirus from the northwestern Pacific. Mar. Biotechnol., 8: 260-267. doi:10.1007/s10126-005-5151-9 PMid:16555028

Cuadras, C.M. – 1991. Problemas de probabilidades y estadística. Vol. 2: Inferencia estadística. Promociones y Publicaciones Universitarias, S.A. Barcelona.

Darling, J.A. and C.K. Tepolt. – 2008. Highly sensitive detection of invasive shore crab (Carcinus maenas and Carcinus aestuarii) larvae in mixted plankton samples using polymerase chain reaction and restriction fragment length polymorphisms (PCR-RFLP). Aquat. Invasions, 3(2): 141-152. doi:10.3391/ai.2008.3.2.4

Dharani, G., G.A. Maitrayee, S. Karthikayalu, T.S. Kumar, M. Anbarasu and M. Vijayakumaran. – 2009. Identification of Panulirus homarus puerulus larvae by restriction fragment length polymorphism of mitochondrial cytochrome oxidase I gene. Pak. J. Biol. Sci., 12(3): 281-285. doi:10.3923/pjbs.2009.281.285 PMid:19579959

Felder, D.L., J.W. Martin and J.W. Goy. – 1985. Patterns in early postlarval develpment of decapods. In: A.M. Wenner (ed.), Larval Growth. Crustacean Issues. F. Schram (series ed.) Vol. 2. pp. 163-225. Rotterdam: Balkema Press.

Folmer, O., M. Black, W. Hoeh, R. Lutz and R. Vrijenhoek. – 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol. Mar. Biol. Biotechnol., 3: 294-299. PMid:7881515

García Muñoz, J.E., M.E. Manjón-Cabeza and J.E. García Raso. – 2008. Decapod crustacean assemblages from littoral bottoms of the Alborán Sea (Spain, west Mediterranean Sea): spatial and temporal variability. Sci. Mar., 72(3): 437-449.

García Raso, J. E. – 1984. Brachyura of the coast of Southern Spain. Spixiana, 7(2): 105-113.

Guerao, G., E. Pastor, J. Martin, M. Andrés, A. Estévez, A. Grau, J. Duran and G. Rotllant. – 2008. The larval development of Maja squinado and M. brachydactyla (Decapoda, Brachyura, Majidae) described from plankton collected and laboratory-reared material. J. Nat. Hist., 42(33-34): 2257-2276. doi:10.1080/00222930802254755

Guerao, G. and G. Rotllant. – 2009. Post-larval development and sexual dimorphism of the spider crab Maja brachydactyla (Brachyura, Majidae). Sci. Mar., 73(4): 797-808. doi:10.3989/scimar.2009.73n4797

Guerao, G. and G. Rotllant. – 2010. Development and growth of the early juveniles of the spider crab Maja squinado (Brachyura, Majidae) in an individual culture system. Aquaculture, 307: 105-110. doi:10.1016/j.aquaculture.2010.06.006

Hall, T.A. – 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic. Acids. Symp. Ser., 41: 95-98.

Hisar, S.A., E. Aksakal, O. Hisar, T. Yanik and S. Mol. – 2008. Discrimination of penaeid shrimps with PCR-RFLP analysis. J. Shellf. Res., 27(4): 917-920. doi:10.2983/0730-8000(2008)27[917:DOPSWP]2.0.CO;2

Ingle, R.W. and A.L. Rice. – 1984. The juvenile stages of eight swimming crab species (Crustacea: Brachyura: Portunidae); a comparative study. Bull. Br. Mus. Nat. Hist. (Zool.), 46: 345-354.

Khamnamtong, B., S. Klinbunga and P. Menasveta. – 2005. Species identification of five penaeid shrimps using PCR-RFLP and SSCP analyses of 16S ribosomal DNA. J. Biochem. Mol. Biol., 38(4): 491-499. PMid:16053718

Lasram, F.B.R., J.A. Tomasini, M.S. Romdhane, T.D. Chi and D. Mouillot. – 2008. Historical colonization of the Mediterranean Sea by Atlantic fishes: do biological traits matter? Hydrobiologia, 607: 51-62. doi:10.1007/s10750-008-9366-4

Le Duff, A. – 1990. L’ araignée. France Eco Pêche, 350:41-45.

Lefébure, T., C.J. Douady, M. Gouy and J. Gibert. - 2006. Relationship between morphological taxonomy and molecular divergence within Crustacea: Proposal of a molecular threshold to help species delimitation. Mol. Phylogenet. Evol., 40: 435-447. doi:10.1016/j.ympev.2006.03.014 PMid:16647275

Martin, J.W., D.L. Felder and F.M. Truesdale. – 1984. A comparative study of morphology and ontogeny in juvenile stages of four western Atlantic xanthoid crabs (Crustacea: decapoda: Brachyura). Phil. Trans. R. Soc. Lond. B, 303: 537-604. doi:10.1098/rstb.1984.0001

Neumann, V. – 1996. Comparative investigations on the systematics and taxonomy of european Maja species (Decapoda, Brachyura, Majidae). Crustaceana, 69: 821-852. doi:10.1163/156854096X00222

Neumann, V. – 1998. A review of the Maja squinado (Crustacea: Decapoda: Brachyura) species-complex with a key to the eastern Atlantic and Mediterranean species of the genus. J. Nat. Hist., 32(10-11): 1667-1684. doi:10.1080/00222939800771191

Ng, P. K. L., D. Guinot and P.J.F. Davie. – 2008. Systema Brachyurorum: Part I. An annotated checklist of extant Brachyuran crabs of the world. Raffles Bull. Zool., Suppl. 17: 1-286.

Pascoal, A., J. Barros-Velázquez, A. Cepeda, J.M. Gallardo and P. Calo-Mata. – 2008. A polymerase chain reaction-restriction fragment length polymorphism method based on the analysis of a 16S rRNA/tRNAVal mitochondrial region for species identification of commercial penaeid shrimps (Crustacea: Decapoda: Penaeoidea) of food interest. Electrophoresis, 29: 499-509. doi:10.1002/elps.200700406 PMid:18064597

Paula, J. – 1988. The larval development of Maja goltziana d’Oliveira 1888 (Crustacea, Decapoda, Majidae) reared in the laboratory. J. Nat. Hist., 22: 1697-1708. doi:10.1080/00222938800771071

Power, A. M., S. Piyapattanakorn, R.M. O’Riordan, A. Iyengar, A.A. Myers, S.J. Hawkins, J. Delany, D. McGrath and N. Maclean.– 1999. Verification of cyprid size as a tool in the identification of two European species of Chthamalus barnacles using mtDNA-RFLP analysis. Mar. Ecol. Prog. Ser., 191: 251-256. doi:10.3354/meps191251

Rodríguez, A. – 2002. Larval development of Maja crispata Risso 1827 (Crustacea, Decapoda, Majidae) reared in the laboratory. J. Nat. Hist., 36: 1589-1600. doi:10.1080/00222930110059655

Sotelo, G., P. Morán and D. Posada. – 2008. Genetic identification of the northeastern Atlantic spiny spider crab as Maja brachydactyla Balss, 1922. J. Crustac. Biol., 28(1): 76-81. doi:10.1651/07-2875R.1

Sotelo, G., P. Morán and D. Posada. – 2009. Molecular phylogeny and biogeographic history of the European Maja spider crabs (Decapoda, Majidae). Mol. Phylogenet. Evol., 53: 314-319. doi:10.1016/j.ympev.2009.05.009 PMid:19460449

Števčić, Z. – 1974 Problemi privrednog iskoristavanja Brahiurnih rakova sjevernog Jadrana. Acta Adriat., 16(22): 397-400.

Tang, R.W.K., C. Yau and W.C. Ng. – 2010. Identification of stomatopod larvae (Crustacea: Stomatopoda) from Hong Kong waters using DNA barcodes. Mol. Ecol. Resour., 10(3): 439-448 doi:10.1111/j.1755-0998.2009.02794.x

Udekem d’Acoz, C. d’– 1999. Inventaire et distribution des crustacés décapodes de l’Atlantique nord-oriental, de la Méditerranée et des eaux continentales adjacentes au nord de 25ºN. Patrimoines naturels (M.N.H.N./S.P.N.), 40.

Van Stappen, G., H. Yu., X. Wang, S. Hoffman, K. Cooreman, P. Bossier and P. Sorgeloos. – 2007. Occurrence of allochthonous Artemia species in the Bohai Bay area, PR China, as confirmed by RFLP analysis and laboratory culture tests. Fundam. Appl. Limnol., 170: 21-28. doi:10.1127/1863-9135/2007/0170-0021

Zariquiey-Álvarez, R. – 1968. Crustáceos decápodos ibéricos. Inv. Pesq., 32: 1-510.




How to Cite

Guerao G, Andree KB, Froglia C, Simeó CG, Rotllant G. Identification of European species of Maja (Decapoda: Brachyura: Majidae): RFLP analyses of COI mtDNA and morphological considerations. Sci. mar. [Internet]. 2011Mar.30 [cited 2024May26];75(1):129-34. Available from:




Most read articles by the same author(s)

1 2 > >>