APS调控种公鸡基因传代转录分析

发布时间：2018-03-17 18:00

  本文选题：APS　切入点：传代转录　出处：《西北农林科技大学》2015年硕士论文　论文类型：学位论文

【摘要】：肉鸡的遗传选育和集约化饲养提高了肉鸡生产效率,但致使肉鸡抗病力降低,肉鸡生产中需要添加抗生素类生长促进剂等免疫调节剂来改善肉鸡健康状态,但易产生药物残留等问题,营养表观遗传学研究发现通过对父母代施加营养素处理可传代调控子代的性状。黄芪多糖(Astragalus polysaccharides,APS)是一种高效绿色免疫调节剂,外源添加可显著改善肉仔鸡的生产性能和免疫机能,本研究通过向父母代种公鸡日粮中添加APS,研究种公鸡APS调控效应对商品代肉鸡生产性能及传代基因转录的影响。试验一父母代种公鸡日粮添加APS对父母代、商品代肉鸡生长性能的影响本试验通过向父母代种公鸡日粮添加APS调控商品代肉仔鸡生产性能,并确定商品代调控效应产生所需的APS添加水平。选用160只科宝500父母代种公鸡,采用单因素5水平(基础日粮分别添加日粮比例0(Blank)、10^(-5)、10^(-4)、10^(-3)、10^(-2)的APS)试验设计,每组4个重复,每个重复8只种公鸡。试验结果显示,由于种公鸡饲养过程中进行了严格限饲,外源添加APS对其生长状况无显著影响;APS对父母代种公鸡的精液品质及种蛋孵化率均无显著影响;10^(-2)APS可显著提高商品代肉仔鸡5、6、7、8、9日龄体重(P≤0.05),在10、11、12、13、14日龄也接近差异显著。本研究表明父母代种公鸡日粮添加10^(-2)APS可传代影响商品代肉鸡生长性能。试验二父母代种公鸡日粮添加APS的父母代及商品代肉鸡传代转录信息分析本试验通过RNA-seq方法筛选父母代种公鸡日粮添加不同水平APS时,其本身及其商品代肉鸡后代胸腺器官的差异表达基因,结合GO和KEGG信号通路功能富集分析探索APS对胸腺器官功能状态的影响。本试验采用单因素5水平(基础日粮分别添加日粮比例0(Blank)、10^(-5)、10^(-4)、10^(-3)、10^(-2)的APS)试验设计,每个处理随机选取3个重复的种公鸡及其对应的3只商品代肉鸡,采集胸腺组织样品进行RNA-seq检测。试验结果显示,30个胸腺样品的Clean Reads数量均在4600万条以上,且Clean Reads质量评估和序列比对结果良好,可用于后续差异表达基因筛选等生物信息分析;父母代种公鸡4个APS处理组(依次为10^(-5)、10^(-4)、10^(-3)、10^(-2))与对照组相比分别筛选到了313(249个上调,64个下调)、177(68个上调,109个下调)、129(66个上调,63个下调)和164(84个上调,80个下调)个差异表达基因,商品代各APS添加组(依次为10^(-5)、10^(-4)、10^(-3)、10^(-2))与对照组相比筛选到的差异表达基因数量显著少于父母代种公鸡(P≤0.05),分别为52(30个上调,22个下调)、90(55个上调,35个下调)、82(52个上调,63个下调)和66(44个上调,22个下调)个基因,根据功能特征选取4个差异表达基因进行实时定量PCR表达验证,发现这4个基因转录水平的表达状况与RNA-seq获得的RPKM值基本一致,说明RNA-seq数据和差异基因筛选的准确性较好;GO和KEGG信号通路功能富集分析发现,筛选到的差异表达基因主要富集在抗原识别、加工及呈递相关的免疫功能条目及细胞骨架相关功能条目,富集性功能条目相关的差异表达基因为MHC-I和细胞骨架相关基因,且这些基因在父母代种公鸡和商品代肉仔鸡间存在对应性。本研究表明父母代种公鸡日粮添加APS可通过调节MHC-I和细胞骨架相关基因的表达影响其本身及商品代肉鸡免疫功能。
[Abstract]:Broiler genetic breeding and intensive breeding to improve the production efficiency of broilers, but the broiler resistance reduces, broiler production need to add antibiotic growth promoter immunomodulators to improve broiler health status, but easy to cause problems such as drug residues, nutrition research on apparent genetics found that hormone treatment can be passaged regulation offspring traits Applied Nutrition by means of parents generation. Astragalus Polysaccharide (Astragalus polysaccharides APS) is a kind of efficient green immunomodulator, exogenous addition can significantly improve broiler production performance and immune function, this study by adding APS to adult roosters in the diet, the effect of the APS regulation of commercial production performance and passage of broiler gene transcription. Test parents roosters of dietary APS on the parent, influence the growth performance of broiler chickens in this experiment to the parent The dietary regulation of APS commercial broiler production performance, and to determine the required APS effect of goods on behalf of regulation adding level. Select 160 Cobb 500 adult roosters, a single factor 5 levels (basal diets were added to dietary ratio of 0 (Blank), 10^ (-5), 10^ (-4), 10^ (-3), 10^ (-2) APS) experimental design, with 4 replications per group, 8 chickens in each replicate. The test results show that, due to the cock in the breeding process to carry out strict restriction, exogenous APS had no significant effect on the growth status of APS; semen quality of the parent the cock and the hatching rate had no significant effect; 10^ (-2) APS can significantly improve the goods on behalf of 5,6,7,8,9 days broiler weight (P = 0.05), at 10,11,12,13,14 days are close to significant differences. This study shows that parents roosters of dietary 10^ (-2) APS generations of commodity generation the growth performance of broilers. Two test Analysis of adult roosters dietary APS parents and broiler chickens were transcribed by RNA-seq test method for screening the adult roosters fed diets with different levels of APS, differential gene expression in broiler chickens and its offspring thymus organ, the combination of GO and KEGG signal pathway enrichment analysis to explore the effect of APS on thymus organ function. By using the single factor test 5 levels (basal diet were added to dietary ratio of 0 (Blank), 10^ (-5), 10^ (-4), 10^ (-3), 10^ (-2) APS) experimental design, each randomly selected 3 repetitions of the rooster and the corresponding the 3 commercial broiler thymus tissue samples, collected by RNA-seq. The test results show that the number of Clean Reads 30 thymus samples were above 46 million, and the Clean Reads quality evaluation and sequence alignment results can be based for subsequent differential expression 鍥犵瓫閫夌瓑鐢熺墿淇℃伅鍒嗘瀽;鐖舵瘝浠ｇ�嶅叕楦�4涓狝PS澶勭悊缁，

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