Genetic Diversity And Phylogenetic Relationships Among Populations Of The Leopard Cat(Felis Bengalensis)

Genetic divergence and phylogenetic relationships among 6 leopard cat (Fells bengalensis) populations from 3 subspecies in China were analyzed through Random Amplified Polymorphic DNA(RAPD) and mitochondrial DNA sequencing methods. in RAPD analysis, PCR conditions including different Taq DNA polymeinse and annealing temperature were tested firstly. 36 effective primers were obtained from 40 random primers using Taq DNA polymerase produced by the BioStar Company. 900/c primers were tested valid, and on average, each primer produces 8.67 DNA bands. By contrast, only 11 effective primers were obtained from 40 random primers using Taq DNA polymerase produced by the MBI Company. 27.50% primers were tested valid, and on average, each primer produces 4.64 DNA bands. The number of DNA bands for 9 primers except P61 significantly decrease when the annealing temperature raises up 1 C. Total number of DNA bands decreased to 65 from 89, but the percent of polymorphic bands raises up to 93.85% from 87.64%. 36 primers amplified 6 mixed samples representing 6 populations respectively and 289 DNA bands were obtained, of which 231 bands, 79.93% are polymorphic. The number of DNA bands produced by single primer range from 0 to 16. 9 primers in 36 primers amplified 21 individuals from 6 populations and 65 DNA bands were obtained, of which 61 bands, 93.85% are polymorphic. 402 bp of the cytochmme b gene of the mtDNA of 16 leopard cats from 6 populations of 3 subspecies were sequenced in the study. There are 23 nucleotide substitutions (5.72%), including 15 transitions and 8 transversions. The sequence difference of the cytochrome b segments ranges from 0.000/c to 2.49% among individuals, and 0.90% to 1.34% among subspecies. The sequence difference within populations is significant. The difference within Guangxi population hits 1.99%, while within Fujiari population is down to 0.50%, which is the lowest among 6 populations. 440 bp of the control region of the mtDNA of 9 leopard cats from 5 populations of 2 subspecies and 375 bp of the control region of 1 individual were sequenced. There are 47 nucleotide substitutions (10.68%), including 39 transitions and 8 transversions. The sequence difference of the control region among the individuals ranges from 0.68% to 6.59%, and on averages 3.14%. The difference within 2 populations of F b. euptilura is 1.71%, while within 3 populations of F b. bengalensis is higher, on average 2.27%. The intersubspecies ranges from 2.73% to 6.590/c, and averages 4.0 1%. The analyses of nuclear DNA and mitochondrial DNA data are almost consistent, and V indicate considerablly abundant genetic diversity in leopard cat. The genetic differentiation is even higher within F b. bengalensis. The only lowest differentiation is found in Fujian population of F b. chiiiemLc. The control region data of the mtDNA also indicate that the genetic differentiation between F b. bengclencLr arid F b. ez鐃ziura is vely distinct It suggests that F b. bengolenris and F b. euptilura should be treated as two Evolutionarily Significant Units (ESUs), and 6 different geographic populations of the leopard cat should be treated as different Management Units (MUs). There is the genetic differentiation within F b. bengolensis. Based on genetic distances, molecular phylogenetic trees of leopard cat have been constructed by Neighbor Joining method and Unweighted Pair Group Method using Arithmetic Mean. Combined with morphological characteristics and geographical distribution, 6 popul...