title: Molecular evidence for species-level distinctions in clouded leopards.
abstract: Among the 37 living species of Felidae, the clouded leopard (Neofelis nebulosa) is generally classified as a monotypic genus basal to the Panthera lineage of great cats. This secretive, mid-sized (16-23 kg) carnivore, now severely endangered, is traditionally subdivided into four southeast Asian subspecies (Figure 1A). We used molecular genetic methods to re-evaluate subspecies partitions and to quantify patterns of population genetic variation among 109 clouded leopards of known geographic origin (Figure 1A, Tables S1 ans S2 in the Supplemental Data available online). We found strong phylogeographic monophyly and large genetic distances between N. n. nebulosa (mainland) and N. n. diardi (Borneo; n = 3 individuals) with mtDNA (771 bp), nuclear DNA (3100 bp), and 51 microsatellite loci. Thirty-six fixed mitochondrial and nuclear nucleotide differences and 20 microsatellite loci with nonoverlapping allele-size ranges distinguished N. n. nebulosa from N. n. diardi. Along with fixed subspecies-specific chromosomal differences, this degree of differentiation is equivalent to, or greater than, comparable measures among five recognized Panthera species (lion, tiger, leopard, jaguar, and snow leopard). These distinctions increase the urgency of clouded leopard conservation efforts, and if affirmed by morphological analysis and wider sampling of N. n. diardi in Borneo and Sumatra, would support reclassification of N. n. diardi as a new species (Neofelis diardi).
authors.affiliation: Laboratory of Genomic Diversity, National Cancer Institute, Frederick, Maryland 21702, USA.
authors.forename: Valerie A
authors.initials: VA
authors.lastname: Buckley-Beason
authors.forename: Warren E
authors.initials: WE
authors.lastname: Johnson
authors.forename: Willliam G
authors.initials: WG
authors.lastname: Nash
authors.forename: Roscoe
authors.initials: R
authors.lastname: Stanyon
authors.forename: Joan C
authors.initials: JC
authors.lastname: Menninger
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Driscoll
authors.forename: JoGayle
authors.initials: J
authors.lastname: Howard
authors.forename: Mitch
authors.initials: M
authors.lastname: Bush
authors.forename: John E
authors.initials: JE
authors.lastname: Page
authors.forename: Melody E
authors.initials: ME
authors.lastname: Roelke
authors.forename: Gary
authors.initials: G
authors.lastname: Stone
authors.forename: Paolo P
authors.initials: PP
authors.lastname: Martelli
authors.forename: Ci
authors.initials: C
authors.lastname: Wen
authors.forename: Lin
authors.initials: L
authors.lastname: Ling
authors.forename: Ratna K
authors.initials: RK
authors.lastname: Duraisingam
authors.forename: Phan V
authors.initials: PV
authors.lastname: Lam
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: O'Brien
mesh_terms: D000818:Animals; D002874:Chromosome Mapping; D004272:DNA, Mitochondrial / Q000235:genetics; D045989:Felidae / Q000145:classification* / Q000235:genetics*; D014644:Genetic Variation; D018895:Microsatellite Repeats; D010802:Phylogeny; D017422:Sequence Analysis, DNA
chemical_list: D004272:DNA, Mitochondrial