Multiplex Detection of Three Cattle (Bos taurus) Polled Variants Based on KASP Assay
YAN Ze1, XIAO Wei2, ZHANG Sheng-Li1, WANG Ya-Chun1, ZHANG Yi1,*
1 College of Animal Science and Technology, China Agricultural University, Beijing 100193, China; 2 Beijing General Station of Animal Husbandry, Beijing 100107, China
Abstract：In wild animals, horns are used to fight for survival and mates. However, in the intensive farming system, polledness is a favorable trait because helps avoid the cost and protect animal welfare. Polledness is an autosomal dominant trait with multiple variants/alleles. Three types of polled variants have been reported on chromosome 1 of taurine cattle (Bos taurus) - the Friesian variant that occurred in dairy cattle breeds, the Celtic variant that was found in beef and dual-purposed breeds, and the Mongolian variant that was identified in Mongolian cattle. In this study, based on a genotyping platform that combines KASP (kompetitive allele-specific PCR) assay and microfluidic chip, a molecular diagnostic method was developed to detect the three polled variants simultaneously. The new KASP assay was further validated by using the conventional molecular experiments including PCR amplification, gel electrophoresis and Sanger sequencing, demonstrating that the new method was accurate and reliable and could be applied in large-scale testing. Interestingly, the Mongolian polled variant was detected in a bull of Sanhe cattle, an indigenous Chinese cattle breed. In addition, a previously unpublished deletion of 2 bp was identified in the Friesian variant that could be used as a marker associated with polledness. These results provide a technical basis for molecular breeding for polled cattle.
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