Detection and Characterization of Amplified Fragment Length Polymorphism Markers for Clinical Mastitis in Canadian Holsteins

Abstract

Mastitis is the most frequent, complex, and costly disease in dairy cattle. Genetic improvement of milk production traits has accompanied an increased susceptibility to mastitis. To determine genome-wide quantitative trait locus-linked markers for mastitis resistance, a total of 200 cows, comprising 100 top clinical mastitis- (CM) resistant and 100 top CM-susceptible cows, were screened by selective DNA pooling and amplified fragment length polymorphism (AFLP) technique. The AFLP analysis on resistant and susceptible pools using 89 selective primer combinations revealed 27 significant AFLP markers at a false discovery rate (FDR) of <5%. The most promising AFLP marker was then selected for further characterization. Individual AFLP genotyping of the marker on all selected animals confirmed a significant difference. Sequence analysis detected a single nucleotide polymorphism (A↔G) responsible for the AFLP polymorphism, which was named CGIL4. The PCR-RFLP analysis indicated that the frequency of allele A was significantly higher in the resistant group. The logistic regression analysis demonstrated that the marker was significantly associated with somatic cell score, CM residual values, and production traits. Radiation hybrid mapping assigned the marker to Bos taurus autosome 22. The present study provides promising markers for marker-assisted selection for CM resistance. Our results also demonstrated the capability of AFLP on selective DNA pools as a method for detection of genome regions containing quantitative trait loci.