[1]段修军,孙国波,张蕾,等.基于RNA-Seq鉴定黑羽番鸭肉质风味差异的候选基因[J].江苏农业学报,2022,38(03):739-747.[doi:doi:10.3969/j.issn.1000-4440.2022.03.020]
　DUAN Xiu-jun,SUN Guo-bo,ZHANG Lei,et al.Identification of candidate genes related to meat flavor in black Muscovy duck based on RNA-Seq[J].,2022,38(03):739-747.[doi:doi:10.3969/j.issn.1000-4440.2022.03.020]
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基于RNA-Seq鉴定黑羽番鸭肉质风味差异的候选基因()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
38
期数:
2022年03期
页码:
739-747
栏目:
畜牧兽医·水产养殖
出版日期:
2022-06-30

文章信息/Info

Title:
Identification of candidate genes related to meat flavor in black Muscovy duck based on RNA-Seq
作者:
段修军; 孙国波; 张蕾; 陆艳凤; 王丽华; 田培余; 刘嘉晟; 杨孟广
（江苏农牧科技职业学院，江苏泰州225300）
Author(s):
DUAN Xiu-jun; SUN Guo-bo; ZHANG Lei; LU Yan-feng; WANG Li-hua; TIAN Pei-yu; LIU Jia-sheng; YANG Meng-guang
(Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China)
关键词:
黑羽番鸭; 挥发性风味物质; 脂肪代谢; 转录组测序
Keywords:
black Muscovy duck; volatile flavor compounds; fat metabolism; transcriptome sequencing
分类号:
S834+.89
DOI:
doi:10.3969/j.issn.1000-4440.2022.03.020
文献标志码:
A
摘要:
为挖掘黑羽番鸭肉质风味相关候选基因，通过对黑羽番鸭胸肌、腿肌中挥发性风味物质进行测定，并通过Illumina HiSeq2500高通量测序进行转录组对比分析，结合参考基因组对所获得的序列进行序列比对、基因注释和差异表达等分析，筛选出差异表达基因并进行GO富集分析。通过荧光定量 PCR（Real-time PCR，qRT-PCR）方法检测4个差异候选基因表达水平，验证测序结果的可靠性。结果表明，黑羽番鸭胸肌中挥发性风味物质辛醛、2,3-辛二酮含量显著高于腿肌，3-羟基-2-丁酮含量显著低于腿肌（P＜0.05）。通过参考基因组比对和差异表达分析，初步获得614个差异表达基因，171个差异基因在黑羽番鸭腿肌组织上调表达，443个差异基因在黑羽番鸭腿肌组织下调表达。结合GO分析和KEGG富集分析，最终获得了20个候选功能基因，它们分别参与机体氨基酸形成、糖代谢以及脂肪代谢等生物过程，其中10个差异候选基因在黑羽番鸭胸肌中参与肌内脂肪代谢过程，这些基因可能通过形成肌内挥发性风味物质进而影响肌肉风味。qRT-PCR 验证结果表明，筛选出的差异候选基因表达趋势与转录组测序中表达的趋势相似，说明测序结果可靠。在黑羽番鸭胸肌、腿肌中挥发性风味物质对比基础上，结合RNA-Seq技术对黑羽番鸭肉质风味前体物质特异性形成机制进行综合分析，筛选出ACOT11、APOA1、FABP3等脂肪代谢相关的差异基因可能与黑羽番鸭胸、腿肌肉质风味形成相关。
Abstract:
To explore the candidate genes related to meat flavor in black Muscovy duck, volatile flavor compounds in breast and leg muscles of black Muscovy duck were determined, and transcriptome analysis was performed by Illumina HiSeq2500 high-throughput sequencing. Based on reference genome, the obtained sequences were analyzed by sequence alignment, gene annotation and differential expression analysis, and differentially expressed genes were screened out and GO enrichment analysis was performed. Moreover, to verify the sequencing results, qRT-PCR was used to detect the expression levels of four differential candidate genes. Results showed that the contents of octanal and 2,3-octanedione in breast muscle were significantly higher than those in leg muscle, and the content of 3-hydroxy-2-butanone in breast muscle was significantly lower than that in leg muscle (P＜0.05). Through genome comparison and differential expression analysis, 614 differentially expressed genes were preliminarily obtained, 171 differentially expressed genes were up-regulated and 443 differentially expressed genes were down-regulated in leg muscles of black Muscovy duck. By GO and KEGG enrichment analysis, 20 functional candidate genes were finally obtained. These candidate genes were involved in biological processes, such as amino acid formation, glucose metabolism and fat metabolism. Among them, ten differentially expressed candidate genes were involved in fat metabolism in breast muscle of black Muscovy duck, and these genes may affect muscle flavor by forming intramuscular volatile flavor compounds. Results of qRT-PCR verification showed that the expression trends of screened differentially expressed candidate genes were similar to those in transcriptome sequencing, indicating that the sequencing results were reliable. Based on the comparation of volatile flavor compounds in breast and leg muscles of black Muscovy duck, the specific formation mechanism of meat flavor precursors in black Muscovy duck was comprehensively analyzed, differentially expressed genes related to fat metabolism such as ACOT11, APOA1, FABP3 may be related to the formation of meat flavor in breast and leg muscles of black Muscovy duck.

参考文献/References:

[1]唐现文,张响英,陆艳凤. MC4R在黑羽番鸭肌肉组织中的表达研究[J].中国家禽,2020,42(7):116-120.
[2]孙国波,王健,陆艳凤,等. 番鸭羽色与产肉性状的观测研究[J].江苏农业科学,2019,47(2):173-176.
[3]孙国波,王日君,董飚,等. 黑羽番鸭及继代选育后代的胸腿肌营养成分测定[J].贵州农业科学,2015,43(1):104-107.
[4]钱建中,段修军,卞友庆,等. 不同性别黑羽番鸭屠宰性能、常规肉品质分析[J].江苏农业科学,2014,42(2):164-166.
[5]董飚,段修军,孙国波,等. GH基因对黑羽番鸭体重和屠宰性能的遗传效应分析[J].浙江农业学报,2015,27(1):22-27.
[6]王锦锋,包小成. 樱桃谷鸭、白羽番鸭、黑羽番鸭和苏牧鸭肉品质的比较研究[J].畜牧与兽医,2010,42(2):40-42.
[7]吉文林,段修军,董飚,等. 黑羽番鸭屠宰性能及肉品质的研究[J].西南农业学报,2013,26(2):795-797.
[8]林谦,吴买生,蒋桂韬,等. 不同羽色和性别番鸭屠宰性能及肌肉成分比较研究[J].家畜生态学报,2014,35(1):30-34.
[9]章琳俐,李丽,朱志明,等. 基于RNA-seq鉴定连城白鸭肉质风味相关候选基因[J].农业生物技术学报,2021,29(4):711-722.
[10]王卫,刘文龙,覃瑶,等. 无抗放养模式有利于固始鸡挥发性风味物质的形成[J].湖南农业科学,2017(6):70-73.
[11]张蕾,章敬旗,章玮月,等. 泰州鹅开产前后肝脏组织转录组对比分析[J].福建农业学报,2020,35(9):987-996.
[12]ROBINSON M D, MCCARTHY D J, SMYTH G K. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data[J]. Bioinformatics, 2010,26(1): 139-140.
[13]YU G C, WANG L G, HAN Y Y， et al. clusterProfiler: an R package for comparing biological themes among gene clusters[J]. Omics: a Journal of Integrative Biology,2012,16(5):284-287.
[14]唐现文,张蕾,章敬旗,等.高邮鸭卵巢中双黄蛋性状相关lncRNA表达差异分析[J].甘肃农业大学学报,2021,56(2):40-49,60.
[15]孙旺斌,付琪,薛瑞林,等. 不同枣粉水平对陕北白绒山羊肉挥发性风味物质的影响[J].动物营养学报,2021,33(10):1-13.
[16]崔小燕,苟钟勇,蒋守群,等. 鸡肉风味的形成机制与调控研究进展[J].动物营养学报,2019,31(2):500-508.
[17]李柯呈,徐宝才,姚忠,等.干腌盐量对南京盐水鸭特征风味成分的影响[J].食品与发酵工业,2019,45(17):98-104.
[18]张丽,强俊,徐跑,等. 酰基辅酶A硫酯酶11基因(ACOT11)及其家族的研究进展[J].江苏农业科学,2021,49(2):12-18.
[19] ZHANG Y Z, LI Y X, MICHELE W N, et al. Targeted deletion of thioesterase superfamily member 1 promotes energy expenditure and protects against obesity and insulin resistance[J].Proceedings of the National Academy of Sciences of the United States of America,2012,109(14): 5417-5422.
[20] TAKAYUKI O, KANAKO I, RYOTA M, et al. Chronic high-fat feeding impairs adaptive induction of mitochondrial fatty acid combustion-associated proteins in brown adipose tissue of mice[J]. Biochemistry and Biophysics Reports,2017,10:32-38.
[21]胡晓维,靳聪飞,刘新峰,等. 牛ACOT11基因克隆及其编码蛋白生物信息学分析[J].天津农学院学报,2015,22(3):1-6.
[22]艾锦新,龙安炬,罗卫星,等. 黔北麻羊FABP3基因多态性及其与生长性状的相关性研究[J].畜牧与兽医,2021,53(1):13-18.
[23] BI Y L, YUAN X Y, ZHU P F, et al. A novel long noncoding RNA, ENSGALG00000021686, regulates the intracellular transport of fatty acids by targeting the FABP3 gene in chicken[J]. Biochemical and Biophysical Research Communications,2020,528(4):706-712.
[24]HUANG Z G, XIONG L, LIU Z S, et al. The develop-mental changes and effect on IMF content of H-FABP and PPAR-PPAR-γ mRNA expression on sheep muscle[J].Asia Cenetica Sinica,2006(6):507-514.
[25]沙尔山别克·阿不地力大,舒婷,吴旭升,等. 拜城油鸡A-FABP、H-FABP和PPAR-PPAR-γ基因mRNA表达的发育性变化及其与肌内脂肪含量的相关性研究[J].中国家禽,2019,41(14):5-11.
[26]荣光辉,徐琪,徐铁山,等. 脂肪型和瘦肉型北京鸭A-FABP基因组织表达特征[J].中国畜牧杂志,2015,51(1):5-9.
[27]YI BAO, WANG J G, WANG S, et al. Overexpression of Banna mini-pig inbred line fatty acid binding protein 3 promotes adipogenesis in 3T3-L1 preadipocytes.[J]. Cell Biology International, 2014, 38(8):918-923.
[28]葛凯. 脂肪细胞大小及脂肪代谢调控巢湖鸭肌内脂肪的分子机理[D].安徽农业大学,2019.
[29]韩龙,杜翠红.葡萄糖-6-磷酸异构酶研究进展[J].药物生物技术,2012,19(6):561-564.
[30]白小娟,张丽卿.葡萄糖-6-磷酸异构酶在类风湿关节炎中的研究进展[J].山西医药杂志,2018,47(6):668-670.
[31]黄鹏,唐黎,汪爱平,等. GPI基因突变致葡萄糖磷酸异构酶缺乏症1例报告并文献复习[J].临床儿科杂志,2020,38(10):785-788.
[32] GRIESEL B A, MATSUZAKI S, BATUSHANSKY A, et al. PFKFB3-dependent glucose metabolism regulates 3T3-L1 adipocyte development[J]. The FASEB Journal, 2021, 35(7): 1-19.
[33]李骏,张倩,江东根,等. PFKFB3基因在前列腺癌中的表达及其对前列腺癌细胞糖酵解及生长的影响[J].中华腔镜泌尿外科杂志(电子版),2020,14(6):460-465.
[34]饶利栋,邓雪强,易轩,等.PFKFB3通过调控PI3K/AKT信号通路促进骨肉瘤细胞的增殖及转移[J].肿瘤,2021,41(2):77-90.
[35]牛津,张敏,王巧娟,等.PFKFB3抑制剂减轻高磷诱导的大鼠血管平滑肌细胞钙化[J].心脏杂志,2021,33(2):117-122.
[36]袁姿. 血清TLR4和PFKFB3水平与2型糖尿病及肥胖的相关性研究[D].苏州:苏州大学,2019.

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备注/Memo

备注/Memo:
收稿日期：2021-09-02基金项目：泰州市科技支撑项目（TN201915）；江苏省农业重大新品种创制项目（PZCZ201737）作者简介：段修军（1972-），男，湖南南县人，硕士，教授，研究方向为动物遗传育种与繁殖。(E-mail)sgb1981@126.com
更新日期/Last Update: 2022-07-07