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. 2019 Nov 4:7:e7672.
doi: 10.7717/peerj.7672. eCollection 2019.

A pilot study-genetic diversity and population structure of snow leopards of Gilgit-Baltistan, Pakistan, using molecular techniques

Samreen Aruge #  1 , Hafsa Batool #  1 , Fida M Khan  2 , Fakhar-I-Abbas  2 , Safia Janjua  2
Affiliations

A pilot study-genetic diversity and population structure of snow leopards of Gilgit-Baltistan, Pakistan, using molecular techniques

Samreen Aruge et al. PeerJ. .

Abstract

Background: The Hindu Kush and Karakoram mountain ranges in Pakistan's northern areas are a natural habitat of the snow leopard (Panthera uncia syn. Uncia uncia) but the ecological studies on this animal are scarce since it is human shy by nature and lives in difficult mountainous tracts. The pilot study is conducted to exploit the genetic diversity and population structure of the snow leopard in this selected natural habitat of the member of the wildcat family in Pakistan.

Method: About 50 putative scat samples of snow leopard from five localities of Gilgit-Baltistan (Pakistan) along with a control sample of zoo maintained male snow leopard were collected for comparison. Significant quality and quantity of genomic DNA was extracted from scat samples using combined Zhang-phenol-chloroform method and successful amplification of cytochrome c oxidase I gene (190 bp) using mini-barcode primers, seven simple sequence repeats (SSR) markers and Y-linked AMELY gene (200 bp) was done.

Results: Cytochrome c oxidase I gene sequencing suggested that 33/50 (66%) scat samples were of snow leopard. AMELY primer suggested that out of 33 amplified samples, 21 (63.63%) scats were from male and 12 (36.36%) from female leopards. Through successful amplification of DNA of 25 out of 33 (75.75%) scat samples using SSR markers, a total of 68 alleles on seven SSR loci were identified, showing low heterozygosity, while high gene flow between population.

Discussion: The low gene flow rate among the population results in low genetic diversity causing decreased diversification. This affects the adaptability to climatic changes, thus ultimately resulting in decreased population size of the species.

Keywords: Genetics; Molecular markers; Pakistan; Panthera uncia; Population.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1. Map of sampling sites.
Map of Gilgit-Baltistan showing locations of sampling sites (Map created using ArcGIS Desktop 10.6.1).
Figure 2
Figure 2. Representative gel plate showing isolated DNA from scat samples.
1–9: scat samples of snow leopard collected from wild, C: control sample from zoo.
Figure 3
Figure 3. Amplified COI region (190 bp).
Amplified COI region (190 bp) by using mini barcode primers; 1: DNA ladder, 2–7: Snow leopard amplified samples.
Figure 4
Figure 4. Representative gel of amplified AMELY gene for sex identification.
L: 100 bps DNA ladder; C: Control sample, Lane 1–14: amplified/non amplified PCR products.
Figure 5
Figure 5. Comparison between mean observed and mean expected heterozygosity in different populations.
Figure 6
Figure 6. Cluster analysis of five populations of snow leopard based on genetic distances.
Figure 7
Figure 7. Molecular variance (AMOVA) within and among individuals of snow leopard.
See this image and copyright information in PMC

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