Warfarin resistance test and polymorphism screening in the VKORC1 gene in Rattus flavipectus.

The buff-breasted rat ( Rattus flavipectus) is a major agricultural pest across China. Warfarin-resistant animals have been found in several major provinces in China, and are hampering effective control. Molecular mechanisms underlying resistance to anticoagulant rodenticides have been determined for other species, but genetic information regarding resistance in R. flavipectus remains unknown. The vitamin K 2,3-epoxide reductase complex subunit 1 (VKORC1) encoded by VKORC1 gene is the molecular target of coumarin anticoagulants, and amino acid substitutions in VKORC1 coding-regions have been reported as one of the supposed mechanisms of warfarin resistance. Here, lethal feeding test in R. flavipectus ( n = 36) was conducted in Zhanjiang, China. Four animals (11%) survived the test period of 25 days and were identified as warfarin resistance. Polymorphism across the whole genome DNA sequence of the VKORC1 gene was screened out and compared with resistant and non-resistant rats. A total of nine single nucleotide polymorphisms (SNPs) were identified including seven SNPs in introns and two SNPs in exons, and the SNP (2317A > G) located in exon 3 led to the amino acid substitution (Tyr139Cys) in VKORC1 protein. Based on the characteristics of Tyr139Cys mutation of VKORC1 in humans or rats and its relationship with warfarin-resistance, Tyr139Cys mutation may be one mutation responsible for anticoagulant resistance in R. flavipectus. Given the low numbers of resistant rats in our feeding test, wider surveillance, tests of resistance development in a larger wild population and further researches on the genetic mechanisms of anticoagulant resistance in R. flavipectus are necessary. [ABSTRACT FROM AUTHOR]