本文選題：APS　切入點：傳代轉(zhuǎn)錄　出處：《西北農(nóng)林科技大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：肉雞的遺傳選育和集約化飼養(yǎng)提高了肉雞生產(chǎn)效率,但致使肉雞抗病力降低,肉雞生產(chǎn)中需要添加抗生素類生長促進劑等免疫調(diào)節(jié)劑來改善肉雞健康狀態(tài),但易產(chǎn)生藥物殘留等問題,營養(yǎng)表觀遺傳學(xué)研究發(fā)現(xiàn)通過對父母代施加營養(yǎng)素處理可傳代調(diào)控子代的性狀。黃芪多糖(Astragalus polysaccharides,APS)是一種高效綠色免疫調(diào)節(jié)劑,外源添加可顯著改善肉仔雞的生產(chǎn)性能和免疫機能,本研究通過向父母代種公雞日糧中添加APS,研究種公雞APS調(diào)控效應(yīng)對商品代肉雞生產(chǎn)性能及傳代基因轉(zhuǎn)錄的影響。試驗一父母代種公雞日糧添加APS對父母代、商品代肉雞生長性能的影響本試驗通過向父母代種公雞日糧添加APS調(diào)控商品代肉仔雞生產(chǎn)性能,并確定商品代調(diào)控效應(yīng)產(chǎn)生所需的APS添加水平。選用160只科寶500父母代種公雞,采用單因素5水平(基礎(chǔ)日糧分別添加日糧比例0(Blank)、10^(-5)、10^(-4)、10^(-3)、10^(-2)的APS)試驗設(shè)計,每組4個重復(fù),每個重復(fù)8只種公雞。試驗結(jié)果顯示,由于種公雞飼養(yǎng)過程中進行了嚴格限飼,外源添加APS對其生長狀況無顯著影響;APS對父母代種公雞的精液品質(zhì)及種蛋孵化率均無顯著影響;10^(-2)APS可顯著提高商品代肉仔雞5、6、7、8、9日齡體重(P≤0.05),在10、11、12、13、14日齡也接近差異顯著。本研究表明父母代種公雞日糧添加10^(-2)APS可傳代影響商品代肉雞生長性能。試驗二父母代種公雞日糧添加APS的父母代及商品代肉雞傳代轉(zhuǎn)錄信息分析本試驗通過RNA-seq方法篩選父母代種公雞日糧添加不同水平APS時,其本身及其商品代肉雞后代胸腺器官的差異表達基因,結(jié)合GO和KEGG信號通路功能富集分析探索APS對胸腺器官功能狀態(tài)的影響。本試驗采用單因素5水平(基礎(chǔ)日糧分別添加日糧比例0(Blank)、10^(-5)、10^(-4)、10^(-3)、10^(-2)的APS)試驗設(shè)計,每個處理隨機選取3個重復(fù)的種公雞及其對應(yīng)的3只商品代肉雞,采集胸腺組織樣品進行RNA-seq檢測。試驗結(jié)果顯示,30個胸腺樣品的Clean Reads數(shù)量均在4600萬條以上,且Clean Reads質(zhì)量評估和序列比對結(jié)果良好,可用于后續(xù)差異表達基因篩選等生物信息分析;父母代種公雞4個APS處理組(依次為10^(-5)、10^(-4)、10^(-3)、10^(-2))與對照組相比分別篩選到了313(249個上調(diào),64個下調(diào))、177(68個上調(diào),109個下調(diào))、129(66個上調(diào),63個下調(diào))和164(84個上調(diào),80個下調(diào))個差異表達基因,商品代各APS添加組(依次為10^(-5)、10^(-4)、10^(-3)、10^(-2))與對照組相比篩選到的差異表達基因數(shù)量顯著少于父母代種公雞(P≤0.05),分別為52(30個上調(diào),22個下調(diào))、90(55個上調(diào),35個下調(diào))、82(52個上調(diào),63個下調(diào))和66(44個上調(diào),22個下調(diào))個基因,根據(jù)功能特征選取4個差異表達基因進行實時定量PCR表達驗證,發(fā)現(xiàn)這4個基因轉(zhuǎn)錄水平的表達狀況與RNA-seq獲得的RPKM值基本一致,說明RNA-seq數(shù)據(jù)和差異基因篩選的準確性較好;GO和KEGG信號通路功能富集分析發(fā)現(xiàn),篩選到的差異表達基因主要富集在抗原識別、加工及呈遞相關(guān)的免疫功能條目及細胞骨架相關(guān)功能條目,富集性功能條目相關(guān)的差異表達基因為MHC-I和細胞骨架相關(guān)基因,且這些基因在父母代種公雞和商品代肉仔雞間存在對應(yīng)性。本研究表明父母代種公雞日糧添加APS可通過調(diào)節(jié)MHC-I和細胞骨架相關(guān)基因的表達影響其本身及商品代肉雞免疫功能。
[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鍏浮4涓狝PS澶勭悊緇，

本文編號：1625843