Eight SNVs in NF-κB pathway genes and their different performances between subclinical mastitis and mixed Chinese Holstein cows

- Eight SNVs on NF-κB pathway were detected in Chinese Holstein cows.
- SNVs 1, 2, 3, and 4 were significantly associated with SCS/milk production traits.
- The statistical results of subclinical mastitis cows differed from the mixed cows.
- Single nucleotide difference at eight SNVs may result in alternative splicing.
- These SNVs could be used as markers for breeding of the dairy mastitis resistance.

The nuclear factor-kappa B (NF-κB) pathway proteins are key players in controlling both innate and adaptive immunity. However, the information on NF-κB pathway genes is very limited in mastitis resistance and milk production of Chinese Holstein cows. In this study, we examine the association of the NF-κB pathway gene variants with milk quality traits and somatic cell score (SCS) in Chinese Holstein cows. Eight single nucleotide variants (SNVs) were identified within the bovine NF-κB pathway genes, using DNA pooled sequencing, PCR-RFLP, and forced PCR-RFLP methods. These SNVs include SNV1: g. 536 C>T (exon 10 of Rel), SNV2: g. 94 G>A (exon 20 of p100), SNV3: g. 43 T>C (intron 6 of p105), SNV4: g. 2397 T>G (intron 9 of p105), SNV5: g. 382 G>C (intron 1 of IκBδ), SNV6: g. 21 C>T (exon 5 of IκBζ), SNV7: g. 272 G>A (intron 6 of IκBζ), and SNV8: g. 18 C>T (intron 10 of IκBζ). The association analysis in mixed Chinese Holstein population showed that SNV1 was significantly or highly significantly associated (P < 0.01 and P < 0.05) with fat rate, protein rate and SCS. Furthermore, the SNV1-CC (wild genotype) determined serine showed the significantly lower SCS and higher milk production traits compared to TT and TC. SNV2 was significantly associated (P < 0.05) with SCS; SNV3 was significantly associated (P < 0.05) with fat rate; and SNV4 was significantly associated (P < 0.05) with fat rate and SCS. In 199 subclinical mastitis Chinese Holstein cows, the statistical results absolutely differed from the mixed Chinese Holstein individuals. Splice-site prediction by SplicePort showed that single nucleotide difference at eight SNVs results in the acceptor score and donor score changing obviously that may lead to alternative splicing. In brief, SNV1, SNV2, SNV3 and SNV4 could be useful genetic markers for mastitis resistance selection and breeding in Chinese Holstein cows. Furthermore, whether these SNVs lead to alternative splicing need further research.