妊娠后期饲粮蛋白质水平对藏母羊泌乳性能的影响及乳腺转录组分析  

Effects of Dietary Protein Level during Late Gestation on Lactation Performance of Tibetan Ewes and Analysis of Mammary Gland Transcriptome

作　　者：周力 姜洋洋 李蒋伟 张峰硕 杨葆春[1] 王志有[1] 桂林生 侯生珍[1] ZHOU Li;JIANG Yangyang;LI Jiangwei;ZHANG Fengshuo;YANG Baochun;WANG Zhiyou;GUI Linsheng;HOU Shengzhen(College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,China)

机构地区：[1]青海大学农牧学院,西宁810016

出　　处：《动物营养学报》2022年第7期4610-4620,共11页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：青海省农业农村厅农牧业科技创新平台(QNKP20210602);青海省林草局青海湖-祁连山保护体系(K992138);青海省乐都区基层农技推广体系改革(K151942);青海省贵南县黑藏羊高效繁育基地与产业化服务(2020GN118)。

摘　　要：本试验旨在探究妊娠后期饲粮蛋白质水平对藏母羊产奶量、乳成分及乳腺组织基因表达的影响。选取妊娠后期藏母羊60只,随机分成2组,每组30只,分别饲喂蛋白质水平为7.5%(LP组)和11.5%(HP组)的饲粮。试验期65 d(配种后第85~150天),其中预试期5 d,正试期60 d。试验结束后,通过RNA-seq技术进行乳腺转录组测序,并对差异表达基因(DEGs)进行GO功能注释、KEGG富集分析和实时荧光定量PCR(RT-qPCR)验证。结果显示:HP组羔羊的平均日增重显著高于LP组(P<0.05)。相较于LP组,HP组藏母羊的日产奶量、乳糖率、乳脂率以及乳蛋白率显著提高(P<0.05)。乳腺转录组分析显示,LP组和HP组分别产生了152766266和159469210个高质量reads,共筛选到40个DEGs,其中11个上调基因,29个下调基因;GO富集分类显示,DEGs被注释到29个条目,主要集中在细胞进程、细胞组分及结合等功能;KEGG通路分析表明,DEGs富集到30条差异信号通路,主要包括Rap1信号通路、烟酸和烟酰胺代谢、果糖和甘露糖代谢、矿物质吸收、磷脂酰肌醇-3-激酶-丝/苏氨酸蛋白激酶信号通路;通过RT-qPCR技术对2组藏母羊乳腺组织中胰岛素样生长因子1(IGF1)、细胞周期蛋白1(CCND1)、信号转导和转录激活因子3(STAT3)、蛋白络氨酸激酶(SRC)和Ⅵ型胶原蛋白A5(COL6A5)等5个调控糖、脂代谢的DEGs进行表达验证,所得结果与RNA-seq测序结果相一致。综上可知,提高妊娠后期藏母羊饲粮蛋白质水平不仅可提高羔羊的平均日增重,还能调控藏母羊乳腺糖、脂代谢,改善其泌乳性能。This study investigated the effects of dietary protein level during late gestation period on milk yield,milk composition and gene expression of mammary tissue in Tibetan ewes.Sixty late-gestation ewes were randomly divided into 2 groups with 30 ewes per group.Ewes in the 2 groups were fed diets with 7.5%(LP group)and 11.5%(HP group)protein,respectively.The experiment lasted for 65 days(days 85 to 150 after mating)with 5 days for adaptation and 60 days for test.After the experiment,RNA-seq technology was employed for mammary tissue.GO functional annotation and KEGG enrichment analysis were used to determine the functions of differentially expressed genes(DEGs),and quantitative real-time PCR(RT-qPCR)was employed for the verification of DEGs.The results showed as follows:the average daily gain of newborn lambs in HP group were significantly higher than that in LP group(P<0.05).Compared with LP group,the daily milk yield,milk lactose percentage,milk fat percentage and milk protein percentage of Tibetan ewes in HP group were significantly increased(P<0.05).Transcriptome analysis of mammary gland showed that 152766266and 159469210 clean reads were produced in LP and HP groups,respectively.A total of 40 DEGs were selected,among which 11 genes were up-regulated and 29 genes were down-regulated.The functions of these DEGs were classified into 29 GO items,and the DEGs were mainly concentrated in cellular process,cell part,binding and other functions.KEGG pathway analysis showed that 30 differentially signaling pathways were enriched(P<0.05),including Rap1 signaling pathway,nicotinate and nicotinamide metabolism,fructose and mannose metabolism,mineral absorption and phosphatidylinositol-3-kinase/serine-threonine kinase(PI3KAkt)signaling pathway.Using the RT-qPCR technology,the expression of glucose and lipid related DEGs,such as insulin-like growth factor 1(IGF1),cyclin D1(CCND1),signal transduction and transcriptional activation factor 3(STAT3),sarcoma(SRC)and typeⅣcollagen A5(COL6A5)were verified,which confirmed the rel

关 键 词：RNA-seq 高原型藏羊 妊娠期 乳腺 乳品质 粗蛋白质 生长性能 羔羊 

分 类 号：S816[农业科学—饲料科学]