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The marked uniformity of mt DNA and a reduction in microsatellite allele size expansion indicates that North American pumas derive from a recent (late Pleistocene circa 10,000 years ago) replacement and recolonization by a small number of founders who themselves originated from a centrum of puma genetic diversity in eastern South America 200,000-300,000 years ago.The recolonization of North American pumas was coincident with a massive late Pleistocene extinction event that eliminated 80% of large vertebrates in North America and may have extirpated pumas from that continent as well.The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U. Genomic DNA specimens from 315 pumas of specified geographic origin (261 contemporary and 54 museum specimens) were collected for molecular genetic and phylogenetic analyses of three mitochondrial gene sequences (16S r RNA, ATPase-8, and NADH-5) plus composite microsatellite genotypes (10 feline loci).Six phylogeographic groupings or subspecies were resolved, and the entire North American population (186 individuals from 15 previously named subspecies) was genetically homogeneous [identical] [emphasis added] in overall variation relative to central and South American populations.SSCP analyses were performed on mt DNA amplification products from contemporary samples (Orita et al. The PCR product was denatured at 95C and electrophoresed on a 4.5% nondenaturing polyacrylamide gel with 10% glycerol at 60 W for 6 h.Several individuals with identical banding patterns were sequenced to confirm that each had the same allele.The MP analysis was conducted by a general heuristic search using simple sequence addition of sequences and tree bisection-reconnection branch swapping.
PCR reactions were performed using 20 ng of genomic DNA in the presence of 10 m M Tris-HCl (p H 8.3), 50 m M KC1, 1.5 m M Mg Cl2, 200 |x M each of d ATP, d CTP, d GTP, d TTP, 0.16 mg/ml BSA, 1 |jl M of each primer, and 1 unit Taq poly-merase enzyme in a volume of 10 jjl I (2.5 units in 25 jjl I for museum specimens).Some 32 separate geographic subspecies of puma have been described based on geographic and morphometric criteria (Figure 1A) (Neff 1983; Young and Goldman 1946). Captive animals were wild born and of known geographic origin.Samples were assigned to the nearest subspecies based on geographic location (Cabrera 1963; Jackson 1955; Neff 1983; Young and Goldman 1946).We would like to thank all the collaborators that collected or aided in the collection of samples for this study. © 2000 The American Genetic Association Puma concolor, a large American cat species, occupies the most extensive range of any New World terrestrial mammal, spanning 110 degrees of latitude from the Canadian Yukon to the Straits of Magellan.We also appreciate the expert technical assistance and insightful advice of Marilyn Raymond, Melody Roelke-Parker, Victor David, Clay Stephens, Janice Martenson, Stan Cevario, Carlos Driscoll, Al Roca, and Ellen Frazier. Until the recent Holocene, pumas coexisted with a diverse array of carnivores including the American lion (Panthera atrox), the North American cheetah (Miracynonyx trumani), and the saber toothed tiger (Smilodon fatalis).