Design and performance of a bovine 200k SNP chip developed for endangered German Black Pied cattle (DSN)
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ABSTRACT:
INSTRUMENT(S): Affymetrix
ORGANISM(S): Bos Taurus
SUBMITTER: Humboldt Universitat zu Berlin
PROVIDER: PRJEB46861 | EVA | 2021-08-09
REPOSITORIES: EVA
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Project description:BackgroundGerman Black Pied cattle (DSN) are an endangered dual-purpose breed which was largely replaced by Holstein cattle due to their lower milk yield. DSN cattle are kept as a genetic reserve with a current herd size of around 2500 animals. The ability to track sequence variants specific to DSN could help to support the conservation of DSN's genetic diversity and to provide avenues for genetic improvement.ResultsWhole-genome sequencing data of 304 DSN cattle were used to design a customized DSN200k SNP chip harboring 182,154 variants (173,569 SNPs and 8585 indels) based on ten selection categories. We included variants of interest to DSN such as DSN unique variants and variants from previous association studies in DSN, but also variants of general interest such as variants with predicted consequences of high, moderate, or low impact on the transcripts and SNPs from the Illumina BovineSNP50 BeadChip. Further, the selection of variants based on haplotype blocks ensured that the whole-genome was uniformly covered with an average variant distance of 14.4 kb on autosomes. Using 300 DSN and 162 animals from other cattle breeds including Holstein, endangered local cattle populations, and also a Bos indicus breed, performance of the SNP chip was evaluated. Altogether, 171,978 (94.31%) of the variants were successfully called in at least one of the analyzed breeds. In DSN, the number of successfully called variants was 166,563 (91.44%) while 156,684 (86.02%) were segregating at a minor allele frequency > 1%. The concordance rate between technical replicates was 99.83 ± 0.19%.ConclusionThe DSN200k SNP chip was proved useful for DSN and other Bos taurus as well as one Bos indicus breed. It is suitable for genetic diversity management and marker-assisted selection of DSN animals. Moreover, variants that were segregating in other breeds can be used for the design of breed-specific customized SNP chips. This will be of great value in the application of conservation programs for endangered local populations in the future.
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Project description:Black cattle is a new breed of beef cattle developed by combining modern biotechnologies such as somatic cell cloning and conventional breeding methods. To provide new ideas for improving meat quality and generating new breeds of cattle, the important candidate genes affecting fat deposition in two kinds of cattle were identified. Eighteen months Black cattles and Luxi cattles were randomly assigned into two environmental. The longissimus dorsi muscle were collected on Black cattle and Luxi cattle,for analyses including fatty acid determinationrs, high-throughput sequencing metagenomics, qRT-PCR expression profile and western blot.The ratio of unsaturated fatty acids to saturated fatty acids was 1.37:1 and 1.24:1 in the muscle tissues of Black cattle and Luxi cattle, respectively. The results of RNA-Seq analysis revealed 1,415 DEGs(fold change ≥ ± 2, P<0.05) between the longissimus dorsi of Black cattle and yellow cattle. A total of 939 genes were upregulated, and the other 476 genes were downregulated. With GO enrichment analysis, it was found that the identified DEGs were significantly enriched in biological regulation, regulation of the Wnt signaling pathway, negative regulation of the Wnt signaling pathway, cAMP metabolic process, fat cell differentiation, and brown fat cell differentiation, among other functions. Regulation of lipolysis in adipocytes, AMPK signaling pathway, adipocytokine signaling pathway and PPAR signaling pathway in the KEGG pathway database were significantly enriched. PPI network analysis showed that the downregulated genes FABP4, ADIPOQ, PLIN1, PLIN2 and LIPE were closely linked to other DEGs and were the key sites of multiple metabolic pathways. Combined with qRT-PCR and protein expression profile analysis, the expression level of fat acid metabolism related genes (FABP4, ADIPOQ) in black cattle was high and the difference was significant. Changes in the expression of fatty acid metabolism-related genes in Black cattle and Luxi cattle were analyzed and important candidate marker genes (such as ADIPOQ and FABP4) that affect fat deposition were identified in order to provide a genetic basis for the efficient breeding of production performance, establish a molecular marker database for local cattle breeds and support the cultivation of new breeds.
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