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2025年5月7日 星期三
农业生物技术学报  2019, Vol. 27 Issue (9): 1582-1595    DOI: 10.3969/j.issn.1674-7968.2019.09.007
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
中国荷斯坦牛FBP2基因对产奶性状的遗传效应分析
李茜1, 2, 李妍3, 石丽君4, 高艳霞1, 李秋凤1, 孙东晓4, *, 李建国1, *
1 河北农业大学 动物科技学院,保定 071001;
2 河北省畜牧兽医研究所,保定 071000;
3 河北农业大学 动物医学院,保定 071001;
4 中国农业大学 动物科技学院,北京 100193
Analysis of Genetic Effects of FBP2 Gene on Milk Production Traits in Chinese Holstein (Bos taurus)
LI Qian1, 2, LI Yan3, SHI Li-Jun4, GAO Yan-Xia1, LI Qiu-Feng1, SUN Dong-Xiao4, *, LI Jian-Guo1, *
1 College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, China;
2 Hebei Animal Husbandry and Veterinary Institute, Baoding 071000, China;
3 College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China;
4 College of Animal Science and Technology, China Agricultural University, Beijing 100193, China
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摘要 对产奶性状具有较大遗传效应的功能基因及突变位点的鉴定,可为奶牛(Bos taurus)标记辅助选择提供有价值的分子信息,进而提高准确性并加快遗传进展。转录组测序(RNA-seq)是当前人类(Homo sapiens)及动植物复杂性状关键基因鉴定的重要策略。作者前期研究利用高通量第二代测序技术对中国荷斯坦牛在干奶期(产前50 d)、泌乳初期(产后15 d)和泌乳高峰期(产后60 d)的肝脏组织进行了转录组测序,鉴定到40个产奶性状候选基因,其中果糖二磷酸酶2基因(fructose-bisphosphatase 2, FBP2)在不同泌乳阶段的表达量存在显著差异,该基因编码糖异生调控酶,参与碳水化合物前体到糖原的合成。本研究旨在进一步研究FBP2基因是否对中国荷斯坦牛产奶性状具有显著遗传效应。利用DNA混池PCR产物直接测序方法检测FBP2基因编码区全长及上下游各2 000 bp序列的潜在多态位点,并利用基质辅助激光吸附飞行时间质谱法(matrix-assisted laser adsorption time of flight mass spectrometry, MALDI-TOF-MS)对1 099头北京地区中国荷斯坦母牛进行个体基因型检测,进而基于混合动物模型,采用SAS 9.2软件分析多态位点与5个产奶性状的关联程度。共计发现FBP2基因有11个单核苷酸多态位点(single nucleotide polymorphisms, SNPs),其中5'调控区7个,3'调控区4个。单标记关联分析结果显示11个SNP位点均与乳蛋白量达到显著或极显著关联(P=0.0495~<0.0001);6个SNP位点同时与乳蛋白率达到显著或极显著关联(P=0.0434~0.0004);5个SNP位点同时与产奶量达到显著或极显著关联(P=0.018~0.007);1个SNP位点同时与乳脂量达到显著关联(P=0.0143);大部分SNP位点的加性效应或等位基因替代效应显著(P<0.05)或极显著(P<0.01)。SNP位点间连锁不平衡程度(linkage disequilibrium, LD)计算结果显示:11个SNPs呈高度连锁(r2=0.48~0.99)并形成2个单倍型块。关联分析结果显示:单倍型块1与产奶量、乳脂率、乳脂量、乳蛋白率和乳脂率均显著或极显著关联(P=0.0413~<0.0001),单倍型块2与产奶量、乳脂量、乳蛋白率和乳蛋白量关联达到显著或极显著关联(P=0.0413~<0.0001),单倍型块1的H2、H3和单倍型块2的H2为优势单倍型,可分别提高产奶量、乳脂和乳蛋白。本研究结果表明,FBP2基因对中国荷斯坦牛产奶量和乳成分性状具有显著遗传效应,尤其对乳蛋白性状影响较大,为中国荷斯坦牛分子育种提供遗传标记信息。
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李茜
李妍
石丽君
高艳霞
李秋凤
孙东晓
李建国
关键词 中国荷斯坦牛果糖二磷酸酶2基因(FBP2)SNP单倍型产奶性状关联分析遗传效应    
Abstract:Identification of functional genes and mutations that have large genetic effects on milk production traits can provide molecular information for marker-assisted selection in order to increase selection accuracy and accelerate genetic gain in dairy cattle (Bos Taurus). Nowadays RNA-seq is one of strategies for identification of key genes affecting complex traits in human (Homo sapiens), domestic animals and plants. In previous RNA-seq study on the transcriptomes of liver tissues from Chinese Holstein at different lactation stages (50 d before parturition , 15 d after parturition, 60 d after parturition), 40 candidate genes for milk production traits were identified. Of them, the fructose-bisphosphatase 2 gene (FBP2) was significantly differentially expressed among different periods. It encoded a gluconeogenesis regulatory enzyme which participated in glycogen synthesis from carbohydrate precursors and played an important role in gluconeogenesis. Thus, the purpose of this study was to further validate whether the FBP2 gene showed significant genetic effects on milk production traits in dairy cattle. By re-sequencing the entire exons, and 2 000 bp of 5' and 3' flanking regions of FBP2 gene using pooled DNA, 11 single nucleotide polymorphisms (SNPs) were detected, including 7 in 5' flanking region and 4 in 3' flanking region. These identified SNPs were genotyped with matrix-assisted laser adsorption time of flight mass spectrometry (MALDI-TOF-MS) technology and tested for association with 5 milk production traits in 1 099 Chinese Holstein from Beijing region. All 11 SNPs were found to be statistically significant for protein yield (P=0.0495~<0.0001). Also, 6 of them were significantly associated with protein percentage (P=0.0434~0.0004), 5 of them were significantly associated with milk yield (P=0.018~0.007) and 1 SNP was significantly associated with fat yield (P=0.0143). The additive and allele substitution effects of the most SNPs reached significance (P<0.05, P<0.01). On the other hand, with linkage disequilibrium (LD) analysis, the 11 SNPs were found to be highly linked (r2=0.48~0.99), and 2 blocks were inferred. Haplotype-based association analysis showed that the 6 haplotype combinations in block 1 were significantly associated with the 5 milk traits (P=0.0413~<0.0001), and the haplotype combinations in block 2 were significantly associated with milk yield, fat yield, protein percentage and protein yield (P=0.0413~<0.0001). H2 and H3 in block1 and H2 in block2 were advantegous haplotypes for higher milk yield and composition. This study first indicated that the FBP2 gene had significant genetic effects on milk yield and milk composition traits in dairy cattle, especially affecting milk protein, and could be useful to provide valuable gene information for molecular breeding programs of Chinese Holstein.
Key wordsChinese Holstein    Fructose-bisphosphatase 2 (FBP2)    SNP    Haplotype    Milk production traits    Association analysis    Genetic effects
收稿日期: 2019-01-07     
ZTFLH:  S813.3  
基金资助:国家现代农业(奶牛)产业技术体系建设专项资金(CARS-36)、河北省科技计划项目(No. 16226604D)、国家自然科学基金(No. 31872330)和奶牛产业技术体系北京市创新团队(No. BAIC06-2018)
通讯作者: jgli@hebau.edu.cn;sundx@cau.edu.cn   
引用本文:   
李茜, 李妍, 石丽君, 高艳霞, 李秋凤, 孙东晓, 李建国. 中国荷斯坦牛FBP2基因对产奶性状的遗传效应分析[J]. 农业生物技术学报, 2019, 27(9): 1582-1595.
LI Qian, LI Yan, SHI Li-Jun, GAO Yan-Xia, LI Qiu-Feng, SUN Dong-Xiao, LI Jian-Guo. Analysis of Genetic Effects of FBP2 Gene on Milk Production Traits in Chinese Holstein (Bos taurus). 农业生物技术学报, 2019, 27(9): 1582-1595.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.09.007     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I9/1582
 
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