中国荷斯坦牛FADS2基因3′端SNP突变对乳中脂肪酸组成的影响

doi:10.3864/j.issn.0578-1752.2016.11.015

摘要/Abstract

摘要： 【目的】FADS2是多不饱和脂肪酸合成中关键的限速酶之一，可催化食物中亚油酸（LNA, C18:2n6）合成γ-亚麻酸(GLA, C18:3n6)、二十碳五烯酸(EPA, C20:5n3)、二十二碳六烯酸(DHA, C22:6n3)等长链脂肪酸。试验旨在探讨中国荷斯坦牛FADS2基因3′端非编码区SNP突变对乳中脂肪酸含量的影响。【方法】随机选择20头无亲缘关系的中国荷斯坦牛样本，用直接测序列法检测FADS2基因3′端非编码区SNP突变位点。再以江苏某大型奶牛场551头中国荷斯坦牛为材料，用飞行时间质谱法对前期发现的2个SNP位点进行检测，同时利用最小二乘模型分析了SNP突变及其单倍型对乳中脂肪酸含量及其不饱和指数的影响。【结果】中国荷斯坦牛FADS2基因3′端非编码区存在3个SNP突变位点：c.1571 A＞G、c.2743 A＞G、c.2776 A＞G。c.1571 A＞G位点GG型为优势基因型，基因型频率为0.800，G为优势等位基因，基因频率为0.887。c.2776 A＞G位点AA为优势基因型，基因型频率为0.673，A为优势等位基因，基因频率为0.819。χ² 检验表明：c.2776 A＞G位点基因型分布均符合Hardy-Weinberg 平衡(P＞0.05) ，而c.1571 A＞G位点基因型分布均偏离Hardy-Weinberg 平衡(P＜0.05)。FADS2基因c.1571 A＞G位点与c.2776 A＞G位点间连锁不平衡系数r²为0.028，未达到显著水平（P＞0.05）。c.1571 A＞G位点与c.2776 A＞G位点有3种单倍型，GA、GG和AA频率分别为0.705、0.181和0.114。多因素方差分析表明：FADS2-1571对C14:1含量、C14和C18不饱和指数的影响达到极显著水平（P＜0.01），对C18:0、SFA和MUFA含量的影响达到显著水平（P＜0.05）。GG型个体乳中C14:1含量、C14和C18不饱和指数显著高于AG型（P＜0.05）。FADS2-2776位点对C16:1含量、 C16和C20不饱和指数的影响达到极显著水平（P＜0.01），对C14:1含量的影响达到显著水平（P＜0.05），GG型个体乳中C16:1含量、C16和C20不饱和指数显著高于AG型和AA型（P＜0.05）。同时，FDAS2-1571-2776单倍型对C16:1含量和C20不饱和指数的影响达到显著水平（P＜0.05），单倍型GG型个体乳中C16:1含量和C20不饱和指数显著高于GA型和AA型（P＜0.05）。【结论】FADS2基因3′端非编码区SNP突变对中国荷斯坦牛乳中脂肪酸组成有重要影响。在进一步验证其功能情况下，可作为影响中国荷斯坦奶牛乳脂肪酸组成的主效基因加以利用。

关键词: 中国荷斯坦牛, FADS2, SNPs, 脂肪酸组成

Abstract: 【Objective】D-6-fatty acid desaturase 2 (FADS2) is one of the key limiting enzymes in the conversion of dietary essential 18 carbon PUFAs (18C-PUFAs) such as linoleic acid (LNA,C18:2n-6) to γ-linolenic acid (GLA, 18:3n6), eicosapentaenoic acid (EPA, C20:5n3) and docosahexaenoic acid (DHA, C22:6n3). The objective of this study was to investigate the effects of SNPs in the 3’ untranslated regions of FADS2 gene on the compositions of fatty acids in milk of Chinese Holstein.【Method】In this study, 20 Chinese Holstein cows were selected randomly for PCR amplification and sequencing of the 3’ untranslated regions of FADS2 gene used for SNP discovery. Then the Chinese Holstein cows (n =551) were genotyped using Sequenom MassARRAY (Sequenom Inc., San Diego, CA) based on the previous SNP information in this study, and the associations between SNPs or haplotypes and compositions of fatty acids, unsaturated indexes of fatty acids in milk were analyzed by the least squares method in the GLM procedure of SPSS.【Result】Three SNPs (c.1571 A＞G, c.2743 A＞G and c.2776 A＞G) were identified in the 3’ untranslated regions of FADS2 gene. The genotype GG was the dominant genotype, which the frequencies were 0.800 for c.1571 A＞G, allele G was dominant allele with the frequency of 0.887. The genotype AA was the dominant genotype, which the frequencies were 0.673 for c. 2776 A＞G. The allele A was dominant allele with the frequency of 0.819. The coefficient of disequilibrium was 0.028 between c.1571 A＞G and c.2776 A＞G, which was not significant at P＜0.05. There were 3 haplotypes for FADS2 c.1571 A＞G and c.2776 A＞G, which the frequencies were 0.705, 0.181 and 0.114 for GA, GG and AA, respectively. χ² test indicated that the c.2776 A＞G fitted with Hardy-Weinberg equilibrium in the population (P＞0.05), and the c.1571 A＞G deviated from Hardy-Weinberg equilibrium (P＜0.05). The SNP c.1571 A＞G showed a very significant association with C14:1, unsaturated indexes of C14 and C18 (P＜0.01), and significant association with C18:0, SFA and MUFA (P＜0.05). The individuals with genotype GG had higher C14:1, unsaturated indexes of C14 and C18 than genotypes AG (P＜0.05). The SNP c.2776 A＞G showed a very significant association with C16:1, unsaturated indexes of C16 and C20 (P＜0.01), and significant association with C14:1 (P＜0.05). The individuals with genotype GG had higher C16:1, unsaturated indexes of C16 and C20 than genotypes AG and AA (P＜0.05). In the meantime, the haplotype of FADS2 1571-2776 showed a significant association with C16:1, unsaturated indexes of C20 (P＜0.05). The individuals with genotype GG had higher C16:1, unsaturated indexes of C20 than genotypes GA and AA (P＜0.05). 【Conclusion】The SNPs in the 3’ untranslated regions of FADS2 gene have significant genetic effects on composition of fatty acids in milk, but further investigation will be required to elucidate the biological and practical relevance of these SNPs.

Key words: Chinese Holstein, FADS2, SNPs, composition of fatty acids

徐晨希，王梦琦，朱小瑞，张玉锋，夏海磊，刘贤慧，王小龙，张慧敏，杨章平，毛永江. 中国荷斯坦牛FADS2基因3′端SNP突变对乳中脂肪酸组成的影响[J]. 中国农业科学, 2016, 49(11): 2194-2202.

XU Chen-xi, WANG Meng-qi, ZHU Xiao-rui, ZHANG Yu-feng, XIA Hai-lei, LIU Xian-hui, WANG Xiao-long, ZHANG Hui-min, YANG Zhang-ping, MAO Yong-jiang. Effects of SNPs in the 3’ Untranslated Regions of FADS2 on the Composition of Fatty Acids in Milk of Chinese Holstein[J]. Scientia Agricultura Sinica, 2016, 49(11): 2194-2202.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2016.11.015

https://www.chinaagrisci.com/CN/Y2016/V49/I11/2194

参考文献

[1] HARVATINE K J, BOISCLAIR Y R, BAUMAN D E. Recent advances in the regulation of milk fat synthesis. Animal, 2009(3): 40-54.

[2] WOODS V B, FEARON A M. Dietary sources of unsaturated fatty acids for animals and their transfer into meat, milk and eggs: a review. Livestock Science, 2009, 126: 1-20.

[3] LEE H, PARK W J. Unsaturated fatty acids, desaturases, and human health. Journal of Medicinal Food, 2014, 17(2): 189-197.

[4] TALPUR F N, BHANGER M I, KHOOHARO A A, MENON G Z. Seasonal variation in fatty acid composition of milk from ruminants reared under the traditional feeding system of Sindh, Pakistan. Livestock Science, 2008, 118: 166-172.

[5] TALPUR F N, BHANGER M I, KHUHAWAR M Y. Comparison of fatty acid and cholesterol content in the milk of Pakistani cow breeds. Journal of Food Composition and Analysis, 2006, 19: 698-703.

[6] HADDAD I, MOZZON M, STRABIOLI R, FREGA N C. Stereospecific analysis of triacylglycerols in camel (Camelus dromedarius) milk fat. International Dairy Journal, 2010, 20: 863-867.

[7] WIKING L, NIELSEN J H, BÅVIUS A K, EDVARDSSON A, SVENNERSTEN-SJAUNJA K. Impact of milking frequencies on the level of free fatty acids in milk, fat globule size, and fatty acid composition. Journal of Dairy Science, 2006, 89: 1004-1009.

[8] GUILLOU H, RIOUX V, CATHELINE D, THIBAULT J N, BOURIEL M, JAN S, D'ANDREA S, LEGRAND P. Conversion of hexadecanoicacid to hexadecenoic acid by rat Delta 6-desaturase. The Journal of Lipid Research, 2003, 44(3): 450-454.

[9] DANDREA S, GUILLOU H, JAN S, CATHELINE D, THIBAULT J N, BOURIEL M, RIOUX V, LEGRAND P. The same rat Delta6- desaturase not only acts on 18- but also on 24-carbon fatty acids in very-long-chain polyunsaturated fatty acid biosynthesis. Biochemical Journal, 2002, 361(1): 49-55

[10] MARTINELLI N, GIRELLI D, MALERBA C, GUARINI P, ILLIG T, TRABETTI E, SANDRI M, FRISO S, PIZZOLO F, SCHAEFFER L, HEINRICH J, PIGNATTI P F, CORROCHER R, OLIVIERI O. FADS genotypes and desaturase activity estimated by the ratio of arachidonic acid to linoleic acid are associated with inflammation and coronary artery disease. The American Journal of Clinical Nutrition, 2008, 88: 941-949.

[11] KOHAMA T, OLIVERA A, EDSALL L, NAGIEC M M, DICKSON R, SPIEGEL S. Molecular cloning and functional characterization of marine sphingosine kinase. The Journal of Biological Chemistry, 1998, 273(37): 23722-23728.

[12] NWANKWO J O, SPECTOR A A, DOMANN F E. A nucleotide insertion in the transcriptional regulatory region of FADS2 gives rise to human fatty acid delta-6-desaturase deficiency. The Journal of Lipid Research, 2003, 44(12): 2311-2319.

[13] NAKAYAMA K, BAYASGALAN T, TAZOE F, YANAGISAWA Y, GOTOH T, YAMANAKA K, OGAWA A, MUNKHTULGA L, CHIMEDREGZE U, KAGAWA Y, ISHIBASHI S, IWAMOTO S. A single nucleotide polymorphism in the FADS1/FADS2 gene is associated with plasma lipid profiles in two genetically similar Asian ethnic groups with distinctive differences in lifestyle. Human Genetics, 2010, 127: 685-690.

[14] ILLIG T, GIEGER C, ZHAI G, ROMISCH-MARGL W, WANG-SATTLER R, PREHN C, ALTMAIER E, KASTENMULLER G, KATO B S, MEWES H W, MEITINGER T, DE ANGELIS M H, KRONENBERG F, SORANZO N, WICHMANN H E, SPECTOR T D, ADAMSKI J, SUHRE K. A genome-wide perspective of genetic variation in human metabolism. Nature Genetics, 2010, 42(2): 137-141.

[15] KATHIRESAN S, MELANDER O, GUIDUCCI C, SURTI A, BURTT N P, RIEDER M J, COOPER G M, ROOS C, VOIGHT B F, HAVULINNA A S, WAHLSTRAND B, HEDNER T, CORELLA D, TAI E S, ORDOVAS J M, BERGLUND G, VARTIAINEN E, JOUSILAHTI P, HEDBLAD B, TASKINEN M R, CHEH C N, SALOMAA V, PELTONEN L, GROOP L, ALTSHULER D M, ORHO-MELANDER M. Six new loci associated with blood low- density lipoprotein cholesterol, high-density lipoprotein cholesterol or triglycerides in humans. Nature Genetics, 2008, 40(2): 189-197.

[16] TANAKA T, SHEN J, ABECASIS G R, KISIALIOU A, ORDOVAS J M, GURALNIK J M, SINGLETON A, BANDINELLI S, CHERUBINI A, ARNETT D, TSAI M Y, FERRUCCI L. Genome-wide association study of plasma polyunsaturated fatty acids in the InCHIANTI study. PLoS Genetics, 2009, 5(1): e1000338.

[17] HOWARD T D, MATHIAS R A, SEEDS M C, HERRINGTON D M, HIXSON J E, SHIMMIN L C, HAWKINS G A, SELLERS M, AINSWORTH H C, SERGEANT S, MILLER L R, CHILTON F H. DNA methylation in an enhancer region of the FADS cluster is associated with FADS activity in human liver. PLoS ONE, 2014, 9(5): e97510.

[18] RENAVILLE B, PRANDI A, FANB B, SEPULCRI A, ROTHSCHILD M F , PIASENTIER E. Candidate gene marker associations with fatty acid profiles in heavy pigs. Meat Science , 2013, 93: 495-500.

[19] BOSCHETTI E, BORDON A, MELUZZI A, CASTELLINI C, DAL BOSCO A, SIRRI F. Fatty acid composition of chicken breast meat is dependent on genotype-related variation of FADS1 and FADS2 gene expression and desaturating activity. Animal, 2015, 16: 1-9.

[20] DA COSTA A S H, BESSA R J B, PIRES V M R, ROLO E A, PINTO R M A, FONTES C M G A, PRATES J A M. Is hepatic lipid metabolism of beef cattle influenced by breed and dietary silage level?. BMC Veterinary Research, 2014, 10: 65-78.

[21] IBEAGHA-AWEMU E M, AKWANJI K A, BEAUDOIN F, ZHAO X. Associations between variants of FADS genes and omega-3 and omega-6 milk fatty acids of Canadian Holstein cows. BMC Genetics, 2014, 15: 25

[22] 梅秀丽. 优质鸡不同饲养方式下FADS1-FADS2基因的表达及其对脂肪酸组成的影响[D]. 雅安: 四川农业大学, 2012.

MEI X L. Fatty acid desaturase (FADS1) and fatty acid desaturase 2(FADS2) gene expression and the effects on fatty acids content of high-quality chicken in different housing systems[D]. Ya'an: Sichuan Agricultural University, 2012. (in Chinese)

[23] 朱世康. 鸡FADS2基因5’_端SNP鉴定及启动子功能分析[D]. 郑州: 河南农业大学, 2013.

ZHU S K. Polymorphisms in the 5’ region of chicken FADS2 gene and the promoter function analysis[D]. Zhengzhou: Henan Agricultural University, 2013. (in Chinese)

[24] 洪雪莹. 鸡FADS2基因3_UTR区多态性检测及其与经济性状关联性分析[D]. 郑州: 河南农业大学, 2012.

HONG X Y. Polymorphisns in the 3’UTR region of chicken FADS2 Gene and their association with economic traits[D]. Zhengzhou: Henan Agricultural University, 2012. (in Chinese)

[25] 卢冉. 鸡Δ6脂肪酸脱氢酶基因启动子区域多态性及基因时空表达的研究[D]. 郑州: 河南农业大学, 2011.

LU R. Study of chickenΔ6 fatty acid desaturase gene’s promotor region polymorphism and gene expression[D]. Zhengzhou: Henan Agricultural University, 2011. (in Chinese)

[26] ZHU S K, TIAN Y D, ZHANG S, CHEN Q X, WANG Q Y, HAN R L, KANG X T. Adjacent SNPs in the transcriptional regulatory region of the FADS2 gene associated with fatty acid and growth traits in chickens. Genetics Molecular Research, 2014, 13(2): 3329-3336.

[27] 中华人民共和国卫生部. 食品安全国家标准-婴幼儿食品和乳品中脂肪酸的测定: GB 5413.27—2010[S/OL]. 北京: 中国标准出版社, 2010.

National Health and Family Planning Commission of the People’s Republic of China. National Food Safety Standard-Determination of Fatty Acids in Foods for Infants and Young Children, Milk and Milk Products: GB 5413.27—2010[S/OL]. Beijing: Standards Press of China, 2010. (in Chinese)

[28] 林秋萍, 李瑾, 冯书惠. 气相色谱法快速测定牛奶中脂肪酸. 食品科学, 2005, 26(8): 346-348.

LIN L P, LI J, FENG S H. Fast quantitative determination of fatty acid in milk. Food Science. 2005, 26(8): 346-348. (in Chinese)

[29] 毛永江, 常玲玲, 杨章平, 吴海涛, 陈莹, 施雪奎, 李云龙, 梁祥焕, 尹召华. 中国荷斯坦牛乳中体细胞评分与脂肪酸含量和组成的相关分析. 中国农业科学. 2011, 44(24): 5073-5082.

MAO Y J, CHANG L L, YANG Z P, WU H T, CHEN Y, SHI Y K, LI Y L, LIANG X H, YIN Z H. Correlation between the SCS and the amount and composition of fatty acids in milk of Chinese Holstein. Scientia Agricultura Sinica, 2011, 44(24): 5073-5082. (in Chinese)

[30] SHI Y Y, HE L. SHEsis, a powerful software platform for analyses of linkage disequilibrium, haplotype construction, and genetic association at polymorphism loci. Cell Research, 2005, 15(2): 97-98.

[31] SOLAKIVI T, KUNNAS T, JAAKKOLA O, RENKO J, LEHTIMÄKI T, NIKKARI S T. Delta-6-desaturase gene polymorphism is associated with lipoprotein oxidation in vitro. Lipids in Health and Disease, 2013, 12: 80.

[32] SONG Z K, CAO H Y, QIN L, JIANG Y F. A case-control study between gene polymorphisms of polyunsaturated fatty acid metabolic rate-limiting enzymes and acute coronary syndrome in Chinese Han Population. BioMed Research International, 2013, ID: 928178. http: ldx.doi.org/10.1155/2013/928178.