Association of SPP1 and NCAPG genes with milk production traits in Chinese Holstein cows: polymorphism and functional validation analysis

Milk production traits play an important role in dairy cattle breeding, and single nucleotide polymorphisms can be used as effective molecular markers for milk production trait marker-assisted breeding in dairy cattle. Based on the results of the preliminary GWAS, candidate genes SPP1 and NCAPG asso...

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Main Authors: Chuanchuan Wang, Yafei Chen, Jinyan Zhao, Xiaofang Feng, Ruoshuang Ma, Hua Wang, Lin Xue, Jinli Tian, Lijuan Yang, Yaling Gu, Juan Zhang
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-10-01
Series:Frontiers in Veterinary Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fvets.2024.1435128/full
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author Chuanchuan Wang
Chuanchuan Wang
Yafei Chen
Jinyan Zhao
Jinyan Zhao
Xiaofang Feng
Xiaofang Feng
Ruoshuang Ma
Ruoshuang Ma
Hua Wang
Hua Wang
Lin Xue
Lin Xue
Jinli Tian
Jinli Tian
Lijuan Yang
Lijuan Yang
Yaling Gu
Yaling Gu
Juan Zhang
Juan Zhang
author_facet Chuanchuan Wang
Chuanchuan Wang
Yafei Chen
Jinyan Zhao
Jinyan Zhao
Xiaofang Feng
Xiaofang Feng
Ruoshuang Ma
Ruoshuang Ma
Hua Wang
Hua Wang
Lin Xue
Lin Xue
Jinli Tian
Jinli Tian
Lijuan Yang
Lijuan Yang
Yaling Gu
Yaling Gu
Juan Zhang
Juan Zhang
author_sort Chuanchuan Wang
collection DOAJ
description Milk production traits play an important role in dairy cattle breeding, and single nucleotide polymorphisms can be used as effective molecular markers for milk production trait marker-assisted breeding in dairy cattle. Based on the results of the preliminary GWAS, candidate genes SPP1 and NCAPG associated with milk production traits were screened. In this study, the aim was to screen and characterize the SNPs of SPP1 and NCAPG genes about milk production traits. Two SNPs and one haplotype block of the SPP1 gene and four SNPs and one haplotype block of the NCAPG gene were obtained by amplification, sequencing and association analysis, and all six SNPs were located in the exon region. Association analysis showed that all six SNPs were significantly associated with milk protein percentage. Linkage disequilibrium analysis showed that 2 SNPs of SPP1 (g. 36,700,265 C > T and g. 36,693,596 C > A) constituted a haplotype that correlated with milk protein percentage, and the dominant haplotype was H2H2, which was CCTT. 4 SNPs of NCAPG (g. 37,342,705 C > A, g. 37,343,379 G > T, g. 37,374,314 C > A and g. 37,377,857 G > A) constituted a haplotype associated with milk protein percentage, 305-days milk protein yield and 305 days milk yield. Tissue expression profiling results revealed that SPP1 and NCAPG had the highest expression in mammary tissue. Interference with SPP1 and NCAPG inhibited the proliferation of Bovine mammary epithelial cells. (BMECs), down-regulated the expression of PCNA, CDK2 and CCND1, up-regulated the expression of BAX and BAD, and promoted apoptosis. Reduced triglyceride synthesis in BMECs, down-regulated the expression of DGAT1, DGAT2, LPIN1, and AGPAT6.SPP1 and NCAPG are involved in the synthesis of milk proteins, and interfering with SPP1 and NCAPG decreased the secretion of β-casein, κ-casein, and αs1-casein, as well as up-regulated the CSN2 and CSN3 expression. The above results indicate that the SNP loci of SPP1 and NCAPG can be used as potential molecular markers to improve milk production traits in dairy cows, laying the foundation for marker-assisted selection. It also proves that SPP1 and NCAPG can be used as candidate key genes for milk production traits in dairy cows, providing new insights into the physiological mechanisms of lactation regulation in dairy cows.
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spelling doaj-art-6726dad084d645eab036b2e756bd05c42025-08-20T01:47:24ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692024-10-011110.3389/fvets.2024.14351281435128Association of SPP1 and NCAPG genes with milk production traits in Chinese Holstein cows: polymorphism and functional validation analysisChuanchuan Wang0Chuanchuan Wang1Yafei Chen2Jinyan Zhao3Jinyan Zhao4Xiaofang Feng5Xiaofang Feng6Ruoshuang Ma7Ruoshuang Ma8Hua Wang9Hua Wang10Lin Xue11Lin Xue12Jinli Tian13Jinli Tian14Lijuan Yang15Lijuan Yang16Yaling Gu17Yaling Gu18Juan Zhang19Juan Zhang20College of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaYinchuan Animal Husbandry Technical Extension and Service Center, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaCollege of Animal Science and Technology, Ningxia University, Yinchuan, ChinaKey Laboratory of Ruminant Molecular Cell Breeding in Ningxia, Ningxia University, Yinchuan, ChinaMilk production traits play an important role in dairy cattle breeding, and single nucleotide polymorphisms can be used as effective molecular markers for milk production trait marker-assisted breeding in dairy cattle. Based on the results of the preliminary GWAS, candidate genes SPP1 and NCAPG associated with milk production traits were screened. In this study, the aim was to screen and characterize the SNPs of SPP1 and NCAPG genes about milk production traits. Two SNPs and one haplotype block of the SPP1 gene and four SNPs and one haplotype block of the NCAPG gene were obtained by amplification, sequencing and association analysis, and all six SNPs were located in the exon region. Association analysis showed that all six SNPs were significantly associated with milk protein percentage. Linkage disequilibrium analysis showed that 2 SNPs of SPP1 (g. 36,700,265 C > T and g. 36,693,596 C > A) constituted a haplotype that correlated with milk protein percentage, and the dominant haplotype was H2H2, which was CCTT. 4 SNPs of NCAPG (g. 37,342,705 C > A, g. 37,343,379 G > T, g. 37,374,314 C > A and g. 37,377,857 G > A) constituted a haplotype associated with milk protein percentage, 305-days milk protein yield and 305 days milk yield. Tissue expression profiling results revealed that SPP1 and NCAPG had the highest expression in mammary tissue. Interference with SPP1 and NCAPG inhibited the proliferation of Bovine mammary epithelial cells. (BMECs), down-regulated the expression of PCNA, CDK2 and CCND1, up-regulated the expression of BAX and BAD, and promoted apoptosis. Reduced triglyceride synthesis in BMECs, down-regulated the expression of DGAT1, DGAT2, LPIN1, and AGPAT6.SPP1 and NCAPG are involved in the synthesis of milk proteins, and interfering with SPP1 and NCAPG decreased the secretion of β-casein, κ-casein, and αs1-casein, as well as up-regulated the CSN2 and CSN3 expression. The above results indicate that the SNP loci of SPP1 and NCAPG can be used as potential molecular markers to improve milk production traits in dairy cows, laying the foundation for marker-assisted selection. It also proves that SPP1 and NCAPG can be used as candidate key genes for milk production traits in dairy cows, providing new insights into the physiological mechanisms of lactation regulation in dairy cows.https://www.frontiersin.org/articles/10.3389/fvets.2024.1435128/fullSPP1NCAPGmilk production traitsassociation analysisSNPfunctional verification
spellingShingle Chuanchuan Wang
Chuanchuan Wang
Yafei Chen
Jinyan Zhao
Jinyan Zhao
Xiaofang Feng
Xiaofang Feng
Ruoshuang Ma
Ruoshuang Ma
Hua Wang
Hua Wang
Lin Xue
Lin Xue
Jinli Tian
Jinli Tian
Lijuan Yang
Lijuan Yang
Yaling Gu
Yaling Gu
Juan Zhang
Juan Zhang
Association of SPP1 and NCAPG genes with milk production traits in Chinese Holstein cows: polymorphism and functional validation analysis
Frontiers in Veterinary Science
SPP1
NCAPG
milk production traits
association analysis
SNP
functional verification
title Association of SPP1 and NCAPG genes with milk production traits in Chinese Holstein cows: polymorphism and functional validation analysis
title_full Association of SPP1 and NCAPG genes with milk production traits in Chinese Holstein cows: polymorphism and functional validation analysis
title_fullStr Association of SPP1 and NCAPG genes with milk production traits in Chinese Holstein cows: polymorphism and functional validation analysis
title_full_unstemmed Association of SPP1 and NCAPG genes with milk production traits in Chinese Holstein cows: polymorphism and functional validation analysis
title_short Association of SPP1 and NCAPG genes with milk production traits in Chinese Holstein cows: polymorphism and functional validation analysis
title_sort association of spp1 and ncapg genes with milk production traits in chinese holstein cows polymorphism and functional validation analysis
topic SPP1
NCAPG
milk production traits
association analysis
SNP
functional verification
url https://www.frontiersin.org/articles/10.3389/fvets.2024.1435128/full
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