發(fā)布時(shí)間：2018-05-26 12:30

  本文選題：巴馬香豬 + 長(zhǎng)白豬��； 參考：《廣西大學(xué)》2017年博士論文

【摘要】：本研究以巴馬香豬為研究對(duì)象,利用RNA-seq技術(shù)和凝膠雙向電泳蛋白質(zhì)組學(xué)研究技術(shù)篩選30、180日齡巴馬香豬背最長(zhǎng)肌中的差異表達(dá)基因,結(jié)合肌肉代謝關(guān)鍵酶活性、血液生化指標(biāo)、相關(guān)基因表達(dá)水平方面研究,初步揭示巴馬香豬與長(zhǎng)白豬肌肉發(fā)育及其肉質(zhì)品質(zhì)差異的分子機(jī)制,研究結(jié)果如下:1.巴馬香豬和長(zhǎng)白豬肉質(zhì)性狀、代謝相關(guān)酶活及血液生化指標(biāo)的測(cè)定:(1)180日齡巴馬香豬背膘厚為4.23±0.15cm,顯著高于長(zhǎng)白豬的1.47±0.21cm(p0.05);肌肉電導(dǎo)率為2.80±0.49ms,顯著低于長(zhǎng)白豬的4.24±0.51ms(p0.05);肌肉肉色Opto值為64.64±2.80高于長(zhǎng)白豬的63.44±0.88,差異不顯著;肌肉剪切力16.13±1.26N低于長(zhǎng)白豬的17.89±0.96N,差異不顯著;眼肌面積為13.63±0.41cm2極顯著低于于長(zhǎng)白豬的36.58±2.37 cm2(p0.01)。(2)巴馬香豬和長(zhǎng)白豬代謝關(guān)鍵酶活性、血液生化指標(biāo)發(fā)育模式一致;0日齡時(shí)巴馬香豬游離脂肪酸濃度顯著高于長(zhǎng)白豬(p0.05),LDH、TC、TG、LDL、HDL濃度顯著低于長(zhǎng)白豬(p0.05);30日齡時(shí)巴馬香豬游離脂肪酸濃度、LDH、SDH、MDH、TG濃度顯著高于長(zhǎng)白豬(p0.05),TC濃度顯著低于長(zhǎng)白豬(p0.05);180日齡時(shí)游離脂肪酸濃度、LPL、SDH、MDH、TG、LDL、HDL濃度顯著高于長(zhǎng)白豬(p0.05),LDH顯著低于長(zhǎng)白豬(p0.05)。(3)巴馬香豬肌肉組織LPL、LDH、MDH、SDH與肌內(nèi)脂肪含量無(wú)顯著性相關(guān)(p0.05)。2.巴馬香豬、長(zhǎng)白豬肌肉組織樣品RNA-seq測(cè)序,結(jié)果如下:(1)30日齡和180日齡的巴馬香豬、長(zhǎng)白豬肌背最長(zhǎng)肌肉組織樣分別獲得 23,051,730、21,475,968、23,000,120、18,698,878 對(duì) clean reads,巴馬香豬共拼接得到27491個(gè)轉(zhuǎn)錄本,19655(71.49%)個(gè)Unigene被注釋到數(shù)據(jù)庫(kù)中,7836(28.51%)個(gè)Unigene沒有被注釋,長(zhǎng)白豬共拼接了 69686個(gè)轉(zhuǎn)錄本,43447(62.34%)個(gè)Unigene被注釋到數(shù)據(jù)庫(kù)中,26239(37.66%)個(gè)Unigene沒有被注釋。(2)30日齡與180日齡巴馬香豬相比,有1380個(gè)差異基因,上調(diào)基因1141個(gè),下調(diào)基因239個(gè);2873,2713,323個(gè)基因具有一個(gè)或者多個(gè)GO注釋到生物過程、細(xì)胞組成、分子功能中;206個(gè)差異基因得到KEGG Pat hway,涉及178條通路,在30日齡和180日齡巴馬香豬肌肉組織中分別發(fā)現(xiàn) 43656、37646 個(gè) SNP 位點(diǎn)。(3)30日齡與180日齡長(zhǎng)白豬之間有351個(gè)差異基因,上調(diào)基因212個(gè),下調(diào)基因139個(gè);2609,1894,892個(gè)基因具有一個(gè)或者多個(gè)GO注釋到生物過程、細(xì)胞組成、分子功能中;長(zhǎng)白豬中共有258個(gè)差異基因得到了 KEGG Pathway,涉及126條通路,長(zhǎng)白豬30日齡和180日齡肌肉組織分別發(fā)現(xiàn)10811、7762個(gè)SNP位點(diǎn)。3.巴馬香豬、長(zhǎng)白豬肌肉組織樣品2-DE測(cè)定:(1)豬肌肉蛋白點(diǎn)的等電點(diǎn)(pI)值在4-7之間,分子量在25-100 kDa范圍之內(nèi),總共有276個(gè)差異蛋白,149個(gè)蛋白點(diǎn)在30日齡時(shí)高表達(dá),118個(gè)蛋白點(diǎn)在180日齡時(shí)高表達(dá)。(2)30與180日齡巴馬香豬共鑒定成功了 67個(gè)差異倍數(shù)在3以上的差異蛋白,30日齡高表達(dá)的32個(gè),180日齡高表達(dá)的35個(gè)。(3)180日齡巴馬香豬與長(zhǎng)白豬間共鑒定成了 16個(gè)差異倍數(shù)在3以上的差異蛋白,巴馬香豬高表達(dá)的13個(gè),長(zhǎng)白豬高表達(dá)的3個(gè)。4.不同發(fā)育階段豬背最長(zhǎng)肌關(guān)鍵后選基因的表達(dá)差異分析:(1)巴馬香豬和長(zhǎng)白豬背最長(zhǎng)肌NDUFS3、NDUFAB1、IDH3A、NU6 M、ATP5B、ATP5H、HUAMLC2B、MYL1、ADPN基因 mRNA 豐度基本一致,具有顯著的年齡依賴變化,隨著年齡的增加而下降(p0.05)。(2)巴馬香豬背最長(zhǎng)肌NDUFAB1、NU6M基因mRNA表達(dá)水平與肌間脂肪含量正相關(guān),相關(guān)系數(shù)分別為:0.826(p0.05)、0.848(p0.05);A TP55H基因mRNA表達(dá)水平與肌間脂肪含量負(fù)相關(guān),相關(guān)系數(shù)為-0.894(p0.05);HUMML2CB基因mRNA表達(dá)水平與肌肉剪切力、肌肉電導(dǎo)率、肌間脂肪含量負(fù)相關(guān),相關(guān)系數(shù)分別為:-0.984(p0.01),-0.895(p0.05)、0.894(p0.05);TPI基因mRNA表達(dá)水平與肌肉剪切力、電導(dǎo)率極相關(guān)、肌間脂肪含量相關(guān),相關(guān)系數(shù)分別為:0.880(p0.01)、0.852(p0.01)、0.820(p0.05);ADPN基因mRNA表達(dá)水平與肌肉剪切力相關(guān),相關(guān)系數(shù)為:0.840(p0.05)。
[Abstract]:In this study, the differential expression genes in the longest muscle of 30180 day old Bama pig were screened by RNA-seq and gel two-dimensional electrophoresis proteomics technology, combining the activity of key enzymes in muscle metabolism, blood biochemical indexes and the water square surface of related genes, and preliminarily revealed the Bama Xiang pig and the Changbai The molecular mechanism of pig muscle development and the difference of meat quality were as follows: 1. Bama pig and long white pork quality, metabolism related enzyme activity and blood biochemical indexes: (1) the back fat thickness of Bama pig was 4.23 + 0.15cm at 180 day age, significantly higher than 1.47 + 0.21cm (P0.05) of the long white pig, and the muscle conductivity was 2.80 + 0.49ms, significantly lower than that of the muscle. The 4.24 + 0.51ms (P0.05) of the long white pig; the muscle color Opto value was 64.64 + 2.80 higher than that of the long white pig 63.44 + 0.88, the difference was not significant; the muscle shear force 16.13 + 1.26N was lower than that of the long white pig. The difference was not significant; the area of the eye muscle was 13.63 + 0.41cm2 significantly lower than that of the long white pig, 36.58 + 2.37 cm2 (P0.01). (2) Bama Xiang pig and long white pig generation. In 0 days of age, the concentration of free fatty acids in Bama pigs was significantly higher than that of long white pigs (P0.05), LDH, TC, TG, LDL and HDL were significantly lower than that of long white pigs (P0.05), and the concentration of free fatty acids in Bama pigs at 30 days of age was significantly higher than that of long white pigs at 30 days of age, and the concentration of LDH, SDH, MDH and TG was significantly higher than that of long white pigs. White pig (P0.05); free fatty acid concentration at 180 days of age, LPL, SDH, MDH, TG, LDL, HDL concentrations were significantly higher than that of long white pigs (P0.05), LDH was significantly lower than that of long white pigs (P0.05). (3) the muscle tissue of Bama Xiang pig was LPL, LDH, and muscle tissue samples were sequenced. The results were as follows: (1) 3 0 days old and 180 days old Bama pig, the longest muscle tissue like the longest muscle tissue of the long white pig obtained 23051730,21475968,23000120,18698878 to clean reads, and Bama Xiang Pigs spliced together to get 27491 transcripts, 19655 (71.49%) Unigene were annotated to the database, 7836 (28.51%) Unigene was not annotated, and the white pigs spliced 69 together. 686 transcripts, 43447 (62.34%) Unigene were annotated to the database, 26239 (37.66%) Unigene was not annotated. (2) 30 days old and 180 day old Bama pig, there were 1380 differential genes, 1141 up-regulated genes and 239 down-regulation, and 28732713323 genes have one or more GO annotated to biological processes, cell composition, molecular work In energy, 206 differentially genes were obtained by KEGG Pat hway, involving 178 pathways, and 4365637646 SNP loci were found in the muscle tissues of Bama pigs at 30 days of age and 180 days old. (3) there were 351 differential genes between 30 day old and 180 day old white pigs, up 212 and 139 down-regulation, and 26091894892 genes had one or more G. O was annotated to biological process, cell composition and molecular function; 258 different genes were obtained by KEGG Pathway, involving 126 pathways, 108117762 SNP loci.3. Bama pigs, and 2-DE determination of muscle tissue samples of long white pigs: (1) isoelectric point (pI) value of pig muscle protein points. In the range of 4-7, within the range of 25-100 kDa, there were 276 differential proteins, 149 protein points high at 30 days of age and high expression of 118 protein points at 180 days of age. (2) 30 and 180 days old Bama pigs were identified by 67 differences in 3 differences and 30 days of age high expressed. (3) the 180 day old Bama pig and the long white pig were identified as 16 differential proteins above 3, 13 high expression of Bama Xiang pig, and 3.4. expression of the longest muscle of the longest muscle in 3 different developmental stages of the long white pig. (1) the longest muscle in Bama and Changbai pigs was NDUFS3, NDUFAB1, IDH3A, NU6 M, ATP5 The mRNA abundances of B, ATP5H, HUAMLC2B, MYL1 and ADPN were basically the same, with a significant age dependence, and decreased with age (P0.05). (2) the longest muscle NDUFAB1 in Bama pig's back, and the expression level of NU6M gene mRNA was positively correlated with the intermuscular fat content, and the correlation coefficients were 0.826 (P0.05) and 0.848 (P0.05), respectively. The correlation coefficient was -0.894 (P0.05), and the level of mRNA expression of HUMML2CB gene was negatively correlated with muscle shear force, muscle conductivity, and intermuscular fat content, and the correlation coefficients were -0.984 (P0.01), -0.895 (P0.05), 0.894 (P0.05); TPI gene mRNA expression level was closely related to muscle shear force, electrical conductivity, and intermuscular fat content. The correlation coefficients were 0.880 (P0.01), 0.852 (P0.01), 0.820 (P0.05), and the ADPN gene mRNA expression level was correlated with muscle shear force, and the correlation coefficient was 0.840 (P0.05).
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：S828

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