brief communications 8. Frei, C. et al. J. Geophys. Res. (in the press). Academic, Dordrecht, The Netherlands, 2001). useful in the laboratory for physiological, 9. Christensen, O. B., Christensen, J. H. & Botzet, M. in 10.Frei, C., Schär, C., Lüthi, D. & Davies, H. C. Geophys. Res. Lett. ecological, evolutionary, developmental Implications of Climatic Change for the Hydrological Cycle and 25, 1431–1434 (1998). for Water Management (ed. Beniston, M.) 101–112 (Kluwer Competing financial interests: declared none. and genetic studies, having the advantages of fitness, high fertility, fast growth, uni- sexuality and isogenic progeny8. The large oocytes are easily accessible for genetic Ecology crustacean group. By virtue of its partheno- manipulation, making this species a candi- genetic reproduction, the marbled crayfish date model for transgenesis9 in decapod Parthenogenesis in an emerges not only as an interesting laboratory crustaceans. The rapid reproduction of this model but also as a potential ecological crayfish might also be of interest for outsider crayfish threat in that it could outcompete native commercial farming purposes. t has been rumoured1 that an unidenti- forms should even a single specimen be Last but not least, this crayfish, which is fied decapod crustacean, a crayfish of released into European lakes and rivers. now widespread in Europe’s aquaria, could Imarbled appearance and of uncertain To determine the origin and phylogenetic become a menace to European freshwater geographical origin that was introduced position of the marbled crayfish (Fig. 1a), ecosystems, as the release of even one speci- into the German aquarium trade in we compared the sequences of sections men into the wild would be enough to the mid-1990s, is capable of unisexual of two mitochondrial genes with those of found a population that might outcompete reproduction (parthenogenesis). Here we other related species, with particular refer- native crayfish. As an American species, it confirm that this marbled crayfish (‘Mar- ence to a similar-looking cambarid species, is a potential transmitter of the infectious morkrebs’) is parthenogenetic under labora- Procambarus fallax, from Florida. The crayfish plague that almost caused the tory conditions and use morphological and sequences in both genes of the marbled extinction of the native European crayfish molecular analysis to show that it belongs crayfish differ at only a few positions from and which still threatens wild and farmed to the American Cambaridae family. those of other cambarids, which supports populations10. Although parthenogenesis is widespread their morphological similarity (the pres- Gerhard Scholtz*, Anke Braband*, among the Crustacea2, and shrimp, lobsters, ence of a spermatheca)6. Our phylogenetic Laura Tolley†, André Reimann*, crayfish and crabs are otherwise versatile in analysis indicates a particularly close Beate Mittmann*, Chris Lukhaup‡, their modes of reproduction3–5, it has not affinity with P. fallax, although the marbled Frank Steuerwald†, Günter Vogt† been reported before in decapods, the crayfish’s species identity remains to be *Humboldt-Universität zu Berlin, Institut für largest and economically most important verified (Fig. 1b). Biologie/Vergleichende Zoologie, We tested whether Marmorkrebs could 10115 Berlin, Germany a be parthenogenetic by studying a mature e-mail: [email protected] female from a laboratory population in †Universität Heidelberg, Institut für Zoologie, Berlin whose spermatheca contained no evi- INF 230, 69120 Heidelberg, Germany dence of spermatophores from copulation, ‡Gotenstrasse 16, 1336 Bittenfeld, Germany but which repeatedly laid eggs. Between two 1. Lukhaup, C. Aquaristik Aktuell 7–8, 48–51 (2001). broods, this crayfish moulted, a procedure 2. Gruner, H.-E. in Lehrbuch der Speziellen Zoologie, Band I, 4. that clears any remnants of sperm from the Teil: Arthropoda (ed. Gruner, H.-E.) 448–1030 (Fischer, spermatheca. We also sexed laboratory Jena, Germany, 1993). 3. Bauer, R. T. & Holt, G. J. Mar. Biol. 132, 223–235 (1998). c populations in Berlin (93 specimens from 7 4. Duffy, J. E. Nature 381, 512–514 (1996). mothers of two generations) and Heidelberg 5. Reynolds, J. D. in The Biology of Freshwater Crayfish (ed. b Astacopsis franklini (39 specimens), beginning at the earliest Holdich, D. M.) 152–191 (Blackwell Science, Oxford, 2002). Pacifastacus leniusculus stage at which sex can be determined. We 6. Scholtz, G. in The Biology of Freshwater Crayfish (ed. Holdich, Orconectes limosus D. M.) 30–52 (Blackwell Science, Oxford, 2002). 98 Procambarus clarkii found that all specimens (body length, 7. Vogt, G. in The Biology of Freshwater Crayfish (ed. Holdich, D. M.) 99 97 Parthenogenetic crayfish 0.8–8.0 cm) showed female morphology, 53–151 (Blackwell Science, Oxford, 2002). 74 Procambarus fallax which excludes protandric hermaphro- 8. Vrijenhoek, R. C. Annu. Rev. Ecol. Syst. 25, 71–96 (1994). 9. Nam, Y. K., Cho, Y. S. & Kim, D. S. Transgenic Res. 9, 97 Austropotamobius torrentium ditism as the mode of reproduction. No Austropotamobius pallipes 463–469 (2000). 76 spermatophore was detected in our study. 10. Söderhäll, K. & Cerenius, L. Freshwat. Crayfish 12, 11–35 (1999). Astacus astacus Competing financial interests: declared none. 87 Astacus leptodactylus 0.1 To rule out internal autogamy, we studied the histology and ultrastructure of the Figure 1 Characteristics of the marbled crayfish. a, Lateral view reproductive system of all 39 specimens of of the crayfish, showing the eggs attached to its pleopods. Adult the Heidelberg population from juvenile specimens are 3–10 cm in length (rostrum to telson). The basic (1.9 cm) to post-brooding (6.8 cm) stages. correction colour is brown–green, with light and dark spots. b, Phylogenetic All gonads were normal ovaries with Ultrahard polycrystalline diamond from graphite relationships of the marbled crayfish based on a comparison of oviducts, and there was no evidence of T. Irifune, A. Kurio, S. Sakamoto, T. Inoue, H. Sumiya the partial sequences of two mitochondrial genes, Cox1 and the ovotestes or male gonoducts (results not Nature 421, 599–600 (2003) gene encoding 12S ribosomal RNA; GenBank accession numbers, shown), which are usually present in In the legend to Fig. 1a of this communication, the AY151515–AY151524, AY151525–AY151534. Maximum-likelihood hermaphroditic crayfish7. Azan staining of diameter of the transparent polycrystalline diamond analysis used the Hasegawa–Kishino–Yano model, assuming rate some adult gonad sections revealed an shown is 1 mm, and not 0.1 mm as published; the heterogeneity with a log likelihood of 13075.69; the reliability of abundance of large, primary vitellogenic scale divisions represent 0.1 mm. Also, the first full branches is given as a percentage of puzzling steps where the oocytes, and proliferating clusters of small paragraph in the second column on page 600 should appropriate node arises. Parsimony analysis produced similar pre-vitellogenic oocytes in the presence of read: “Recent chemical-vapour deposition techniques results. The marbled crayfish belongs to the American Cambaridae hatchlings, indicating that a new reproduc- provided pure polycrystalline diamonds, but these (red) and is closely related to Procambarus fallax (only 2.2% of tive cycle was already under way (Fig. 1c). diamonds are not sintered and have poor intergrain base positions different). Scale bar, 0.1 nucleotide substitutions. These results provide convincing evidence adhesion. Accordingly, they have been reported to c, Azan-stained histological section of a gonad of a female with for parthenogenesis in the marbled crayfish. have a hardness of ~80–100 GPa (ref. 10), which is hatchlings exclusively composed of ovarian tissue. Arrow, pre- Our findings have several practical significantly lower than the highest value (~120 GPa; vitellogenic oocyte; arrowhead, primary vitellogenic oocyte. implications. This marbled crayfish will be ref. 4) for single-crystal diamonds.” 806 © 2003 Nature Publishing Group NATURE | VOL 421 | 20 FEBRUARY 2002 | www.nature.com/nature.