DNA barcodes effectively identify the morphologically similar Common Opossum (Didelphis marsupialis) and Virginia Opossum (Didelphis virginiana) from areas of sympatry in Mexico

Figures & data

Figure 1.  Geographical distribution of the Virginia Opossum (D. virginiana) and the Common Opossum (D. marsupialis) in Mexico (modified from Gardner 1973). Numbers correspond to specimen localities described in Table I.

Table I.  Sequence origin of the Cox1 and the voucher museum specimen of the Virginia Opossum (D. virginiana), the Common Opossum (D. marsupialis), and other marsupial species.

Taxon	Locality number (Figure 1)	Haplotype number	Haplotype locality	Specimen number	Catalog number	Sequence source
Didelphis virginiana	–	1	University of California, Berkeley, California, USA	1		BOLD GBMA 0523-06
	–	1	Curry County, Oregon, USA	1	ASNHC 6475	Present study
	1	2	Escuinapa, Sinaloa, Mexico	3	CNMA 45120, 45121, 45122	Present study
	2	2	Tlacotalpan, Veracruz, Mexico	5	CNMA 45123, 45124, 45125, 45126, 45127	Present study
	3	2	REPSA, Distrito Federal, Mexico	5	CNMA 45114, 45115, 45116	Present study
	4	2	Tlalpan, Distrito Federal, Mexico	1	CNMA (FAC1929)	Present study
	5	2	Yetla, Guerrero, Mexico	1	CNMA 45117	Present study
	6	2	Omiltemi, Guerrero, Mexico	1	CNMA (FAC2003)	Present study
	7	2	Montecillo, Oaxaca, Mexico	2	ECO-SC-M 1862, 1867	Present study
	8	2	Mixtepec, Oaxaca, Mexico	2	CNMA 44179, 44180	Present study
	9	2	Cosoltepec, Oaxaca, Mexico	1	CNMA 45141	Present study
	10	2	Izta-Popo, Mexico, Mexico	1	CNMA 45119	Present study
	11	2	Monterrey, Nuevo León, Mexico	1	CNMA (FAC3826)	Present study
	12	3	Mérida, Yucatán, Mexico	1	FMVZ-UADY teaching	Present study
	13	3	Cozumel, Quintana Roo, Mexico	1	ASNHC 1282	Present study
	14	3	Constitución, Campeche, Mexico	3	ASNHC 6421, 6422, Tissue 2589	Present study
Didelphis marsupialis	15	4	Tuxtlas, Veracruz, Mexico	3	CNMA 45109, 45110, 45111	Present study
	16	5	Catemaco, Veracruz, Mexico	1	CNMA 43475	Present study
	17	5	Peñuela, Veracruz, Mexico	1	CNMA 45112	Present study
	18	5	Huitepec, Chiapas, Mexico	1	ECO-SC-M 744	Present study
	19	6	Ruinas Acalán, Tabasco, Mexico	1	ASNHC 1280	Present study
	20	6	Escárcega, Campeche, Mexico	2	ASNHC 1281, 6416	Present study
	21	6	Candelaria, Campeche, Mexico	2	ASNHC 1276, 6410	Present study
	14	6	Constitución, Campeche, Mexico	1	ASNHC 6413	Present study
	–	7	Puntarenas, Costa Rica	1	ROM 97295	BOLD ABSMS 535-06
	–	8	Demerara-Mahaica, Guyana	1	ROM 103921	BOLD ABSMS 548-06
Didelphis albiventris	–	9	Pichincha, Ecuador	1	ROM 104565	BOLD ABSMS 559-06
Didelphis imperfecta	–	10	Demerara-Mahaica, Guyana	1	ROM 113762	BOLD ABSMS 588-06
		10	Sipalwini, Surinam	1	ROM 117232	BOLD ABSMS 070-06
Philander opossum	–	11/12	Tuxtlas, Veracruz, Mexico	3	CNMA 45128, 45129, 45130	Present study
Philander andersoni	–	13	Napo, Ecuador	1	ROM 106102	BOLD ABSMS 569-06
Metachirus nudicaudatus	–	14	Sipaliwini, Surinam	1	ROM 117525	BOLD ABSMS 363-06

Note: ROM, Royal Ontario Museum; CNMA, Colección Nacional de Mamíferos; FAC, field collection number of Fernando A. Cervantes; ECO-SC-M, Colección Mastozoológica de El Colegio de la Frontera Sur, Unidad San Cristóbal de las Casas; FMVZ-UADY, Colección Mastozoológica de la Universidad Autónoma de Yucatán; ASNHC, Angelo State University Natural History Collections; REPSA, Reserva Ecológica del Pedregal de San Ángel.

Figure 2.  NJ tree of 49 Cox1 sequences of opossum species of the genera Didelphis, Philander, and Metachirus using K2P genetic distances. Numbers above the nodes are bootstrap support values based on 1000 replicates. *Sequence downloaded from BOLD (http://www.boldsystems.org).

Table II.  Average genetic distances between opossum species of the genera Didelphis, Philander, and Metachirus estimated according to the K2P model of nucleotide substitution for barcode sequences.

			1	2	3	4	5	6	7	8	9	10	11	12	13
1		M. nudicaudatus	–
2	P. opossum	1	0.195	–
3		2	0.193	0.002	–
4		P. andersoni	0.201	0.050	0.048	–
5		1	0.203	0.123	0.121	0.121	–
6	D. virginiana	2	0.218	0.132	0.130	0.130	0.022	–
7		3	0.216	0.130	0.128	0.128	0.020	0.005	–
8		1	0.195	0.125	0.126	0.123	0.080	0.093	0.087	–
9	D. marsupialis	2	0.193	0.126	0.125	0.121	0.078	0.091	0.086	0.002	–
10		3	0.181	0.113	0.111	0.123	0.080	0.089	0.084	0.024	0.022	–
11		4	0.181	0.117	0.115	0.125	0.084	0.093	0.087	0.027	0.025	0.006	–
12		5	0.177	0.117	0.115	0.123	0.082	0.091	0.086	0.027	0.025	0.009	0.006	–
13		D. albiventris	0.193	0.115	0.113	0.117	0.087	0.100	0.095	0.045	0.043	0.053	0.056	0.057	–
14		D. imperfecta	0.193	0.122	0.120	0.126	0.093	0.106	0.100	0.046	0.045	0.055	0.055	0.053	0.035

Gardner AL. 1973. The systematics of the genus Didelphis (Marsupialia: Didelphidae) in North and Middle America. Special Pub Mus Texas Tech Univ. 4:1–81.

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