豬肉嘌呤含量表型變異及其遺傳基礎(chǔ)研究
發(fā)布時(shí)間：2018-09-05 07:55
【摘要】：過去幾十年,隨著飲食結(jié)構(gòu)的改變和人均嘌呤攝入量不斷提升,中國(guó)高尿酸血癥和痛風(fēng)的患病率大約升高了十倍,因此食品中的嘌呤含量受到消費(fèi)者和食品研究者的重視。肉品的嘌呤含量較高,且豬肉占中國(guó)總?cè)馄废M(fèi)的65%左右,但目前國(guó)內(nèi)外有關(guān)豬肉嘌呤含量的表型變異程度和遺傳基礎(chǔ)研究尚為空白。本文選取杜洛克(6頭)、杜長(zhǎng)大(18頭)、巴馬香(18頭)、二花臉(18頭)、萊蕪(18頭)和嵌合家系(377頭)六個(gè)不同種群共455頭豬的背最長(zhǎng)肌樣品,測(cè)定了其4種嘌呤堿基含量(包括鳥嘌呤、腺嘌呤、次黃嘌呤和黃嘌呤)和多種肉質(zhì)性狀;并對(duì)其中377頭嵌合家系F6代個(gè)體進(jìn)行基因組全序列測(cè)序(7×深度)和40頭極端表型個(gè)體的肌肉組織轉(zhuǎn)錄組測(cè)序;另外,隨機(jī)選取嵌合家系三頭閹公豬,測(cè)定其12種組織嘌呤含量。進(jìn)而系統(tǒng)地分析了豬肉嘌呤含量在品種間及品種內(nèi)、組織間和性別間的差異,并開展了基于全基因組重測(cè)序的關(guān)聯(lián)研究(GWAS),定位了各嘌呤和總嘌呤含量的數(shù)量性狀位點(diǎn)(QTL),得到以下主要結(jié)果:(1)宰后豬肉嘌呤含量的動(dòng)態(tài)變化:宰后豬肉中腺嘌呤不斷向次黃嘌呤轉(zhuǎn)化,至24h趨于穩(wěn)定;但總嘌呤和鳥嘌呤含量基本維持恒定。(2)品種間差異顯著:除杜洛克外,萊蕪豬平均總嘌呤含量(114.15 mg/100g)極顯著低于其他四個(gè)品種(P0.01),而二花臉豬顯著低于巴馬香(P0.05)。(3)性別間差異顯著:嵌合家系中閹公豬的肌肉平均嘌呤含量(137.71 mg/100g)顯著低于母豬(139.90 mg/100g)。(4)組織間差異顯著:肺、肝和腎臟的嘌呤含量超過200 mg/100g,為高嘌呤組織;肌肉(包括心肌、骨骼肌、平滑肌)嘌呤含量大約為130 mg/100g,為中嘌呤組織;背膘、皮膚和蹄部結(jié)締組織嘌呤含量低于30 mg/100g,為低嘌呤組織。(5)總嘌呤含量與pH、肉色、肌內(nèi)脂肪含量(IMF)等多種肉質(zhì)性狀顯著的相關(guān)(P0.05),相關(guān)系數(shù)達(dá)0.2-0.4之間。相關(guān)性結(jié)果表明低嘌呤含量的豬肉具有較高的終pH、IMF和系水力,且肉色趨向鮮紅、嫩度和口感較好。因此培育低嘌呤優(yōu)質(zhì)肉種豬,不僅不會(huì)與傳統(tǒng)關(guān)注的重要肉質(zhì)性狀的選育方向發(fā)生沖突,甚至可能在一定程度上有助于改善這些肉質(zhì)性狀。(6)GWAS分析鑒別到40個(gè)與總嘌呤含量顯著關(guān)聯(lián)的SNP位點(diǎn)(P5.00×10-8),它們可能代表了11個(gè)QTLs,其中最顯著關(guān)聯(lián)的SNP rs32337327(P=2.75×10-9)位于豬8號(hào)染色體(SSC8)的LDB2和QDPR基因之間,而SSC3、SSC11和SSC14上的最強(qiáng)關(guān)聯(lián)SNPs則分別落在TGFBRAP1、MPHOSPH8和C12orf43的內(nèi)含子區(qū)域內(nèi)。同時(shí)還檢出263個(gè)與鳥嘌呤含量顯著關(guān)聯(lián)SNPs,它們分布于26個(gè)QTLs區(qū)域,其中最顯著關(guān)聯(lián)SNP rs343436977(P=9.72×10-13)位于ABCC4基因的內(nèi)含子上,另有9個(gè)QTLs區(qū)間內(nèi)的最顯著關(guān)聯(lián)SNPs也位于不同基因的內(nèi)含子上。本研究未檢測(cè)到與腺嘌呤和次黃嘌呤含量顯著關(guān)聯(lián)信號(hào),但發(fā)現(xiàn)SSC7和SSC8上可能存在影響它們的潛在QTLs(P5.00×10-6),其最強(qiáng)關(guān)聯(lián)SNPs分別位于PAQR8和SLC10A7基因內(nèi)。腺嘌呤與次黃嘌呤之和的GWAS結(jié)果和總嘌呤GWAS結(jié)果相似。(7)通過分析肌肉嘌呤含量極端個(gè)體的轉(zhuǎn)錄組數(shù)據(jù),共鑒別到25個(gè)差異表達(dá)基因或者序列標(biāo)簽,其中包含8個(gè)已注釋基因的轉(zhuǎn)錄本,分別為RYK、ORAOV1、MTERF1、MOSPD1、SLCO1A2、B3GNTL1、ZDHHC7和FAM92A。未發(fā)現(xiàn)GWAS信號(hào)落在差異表達(dá)基因區(qū)域內(nèi),提示豬肉嘌呤含量可能受復(fù)雜遺傳因子調(diào)控。(8)通過對(duì)GWAS最顯著SNP位點(diǎn)附近已注釋的70個(gè)候選基因和8個(gè)表達(dá)差異基因進(jìn)行GO分析,發(fā)現(xiàn)了11個(gè)功能富集條目,其中鈣離子結(jié)合富集條目包含8個(gè)相關(guān)基因。KEGG分析發(fā)現(xiàn)ABCC4、ADCY2和GRIA1等基因參與cAMP信號(hào)通路。此外,這78個(gè)基因編碼的蛋白中,有10個(gè)具有ATP結(jié)合位點(diǎn),有41個(gè)屬于膜蛋白。該結(jié)果提示鈣離子結(jié)合蛋白和cAMP信號(hào)通路很可能參與調(diào)控肌細(xì)胞中嘌呤物質(zhì)代謝與轉(zhuǎn)運(yùn),從而影響豬肌肉嘌呤含量。本研究首次定位了影響豬肉嘌呤含量的QTL位點(diǎn),鑒別了多個(gè)相關(guān)候選基因,并揭示鈣離子和cAMP信號(hào)通路對(duì)肌肉嘌呤含量有重要影響。這些結(jié)果為深入揭示肌肉嘌呤含量的遺傳機(jī)制奠定了堅(jiān)實(shí)基礎(chǔ),也為今后培育低嘌呤優(yōu)質(zhì)豬種提供了理論與技術(shù)支持。
[Abstract]:Over the past few decades, the prevalence of hyperuricemia and gout in China has risen tenfold as dietary changes and per capita intake of purine have increased. As a result, the purine content in foods has attracted the attention of consumers and food researchers. The study on the phenotypic variation and genetic basis of pork purine content at home and abroad is still blank. In this paper, the samples of longissimus dorsi from 6 Duo (18 head), Bama (18), Bama (18), two face (18), Laiwu (18) and inlay families (377 *) six pigs were collected. Guanine, adenine, hypoxanthine and xanthine, and a variety of meat traits; 377 chimeric F6 generations were sequenced (7 x depth) and 40 extreme phenotypic individuals were sequenced in muscle tissue transcriptomes; in addition, three castrated boars from chimeric families were randomly selected to determine the purine content of 12 tissues. Quantitative trait loci (QTLs) of purines and total purines in pork were identified based on genome-wide sequencing (GWAS). The main results were as follows: (1) Dynamic changes of purine content in postmortem pork: Adenine content in postmortem pork However, the contents of total purine and guanine were basically constant. (2) there was a significant difference among varieties. Except for Duroc, the average total purine content (* 114.15 mg/100g) of Laiwu pigs was significantly lower than that of the other four varieties (P0.01), while two * Hualian pigs were significantly lower than those of Bama (P0.05). (3) the difference between the sexes was significant: 24h. The average purine content (137.71 mg/100g) in the muscle of castrated boars was significantly lower than that of sows (139.90 mg/100g). (4) There was significant difference between tissues: the purine content in lung, liver and kidney exceeded 200 mg/100g, which was high purine tissue; the purine content in muscle (including myocardium, skeletal muscle, smooth muscle) was about 130 mg/100g, which was middle purine tissue; the purine content in backfat, skin and kidney was about 130 mg/100g. (5) Total purine content was significantly correlated with pH, meat color, intramuscular fat content (IMF) and other meat quality traits (P 0.05), the correlation coefficient was between 0.2 and 0.4. The correlation results showed that pork with low purine content had higher final pH, IMF and waterpower, and the meat color tended to bright red. Therefore, the breeding of low purine quality broiler pigs will not conflict with the breeding direction of important meat quality traits of traditional concern, and may even help to improve these meat quality traits to a certain extent. (6) GWAS analysis identified 40 SNPs (P5.00 *10-8) which were significantly associated with total purine content, and they may be progenies. 11 QTLs were listed, among which the most significant SNP rs32337327 (P=2.75 * 10-9) was located between LDB2 and QDPR genes of porcine chromosome 8 (SSC8), while the strongest correlation between SSC3, SSC11 and SSC14 was located in the intron region of the SSC3, SSC11 and SSC14 respectively, and 263 of them were significantly associated with guanine content. Among 26 QTLs, the most significant association SNP rs343436977 (P = 9.72 *10-13) was located on the intron of ABCC4 gene, and the most significant association SNPs in nine QTLs were also located on the intron of different genes. Their potential QTLs (P5.00 *10-6), the strongest associated SNPs were located in PAQR8 and SLC10A7, respectively. The GWAS results of the sum of adenine and hypoxanthine were similar to those of the total purine GWAS. (7) Twenty-five differentially expressed genes or sequence tags were identified by analyzing the transcriptome data of individuals with extreme muscle purine content. The transcripts of the annotated genes were RYK, ORAOV1, MTERF1, MOSPD1, SLCO1A2, B3GNTL1, ZDHHC7 and FAM92A. No GWAS signals were found in the differentially expressed gene regions, suggesting that pork purine content might be regulated by complex genetic factors. (8) 70 candidate genes and 8 differentially expressed genes were annotated near the most significant SNP site of GWAS. By GO analysis, 11 functional enrichment entries were identified, of which 8 related genes were found. KEGG analysis revealed that ABCC4, ADCY2 and GRIA1 were involved in the cAMP signaling pathway. In addition, 10 of the 78 genes encoded proteins had ATP binding sites and 41 belonged to membrane proteins. The cAMP signaling pathway is likely to participate in regulating the metabolism and transport of purine * in muscle cells, thereby affecting the purine content of porcine muscles. For the first time, this study locates the QTL locus affecting pork purine content, identifies multiple candidate genes, and reveals that calcium and cAMP signaling pathways have important effects on muscle purine content. It lays a solid foundation for revealing the genetic mechanism of purine content in muscle and provides theoretical and technical support for breeding low * purine quality pig breeds in the future.
【學(xué)位授予單位】：江西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S828
本文編號(hào)：2223660
[Abstract]:Over the past few decades, the prevalence of hyperuricemia and gout in China has risen tenfold as dietary changes and per capita intake of purine have increased. As a result, the purine content in foods has attracted the attention of consumers and food researchers. The study on the phenotypic variation and genetic basis of pork purine content at home and abroad is still blank. In this paper, the samples of longissimus dorsi from 6 Duo (18 head), Bama (18), Bama (18), two face (18), Laiwu (18) and inlay families (377 *) six pigs were collected. Guanine, adenine, hypoxanthine and xanthine, and a variety of meat traits; 377 chimeric F6 generations were sequenced (7 x depth) and 40 extreme phenotypic individuals were sequenced in muscle tissue transcriptomes; in addition, three castrated boars from chimeric families were randomly selected to determine the purine content of 12 tissues. Quantitative trait loci (QTLs) of purines and total purines in pork were identified based on genome-wide sequencing (GWAS). The main results were as follows: (1) Dynamic changes of purine content in postmortem pork: Adenine content in postmortem pork However, the contents of total purine and guanine were basically constant. (2) there was a significant difference among varieties. Except for Duroc, the average total purine content (* 114.15 mg/100g) of Laiwu pigs was significantly lower than that of the other four varieties (P0.01), while two * Hualian pigs were significantly lower than those of Bama (P0.05). (3) the difference between the sexes was significant: 24h. The average purine content (137.71 mg/100g) in the muscle of castrated boars was significantly lower than that of sows (139.90 mg/100g). (4) There was significant difference between tissues: the purine content in lung, liver and kidney exceeded 200 mg/100g, which was high purine tissue; the purine content in muscle (including myocardium, skeletal muscle, smooth muscle) was about 130 mg/100g, which was middle purine tissue; the purine content in backfat, skin and kidney was about 130 mg/100g. (5) Total purine content was significantly correlated with pH, meat color, intramuscular fat content (IMF) and other meat quality traits (P 0.05), the correlation coefficient was between 0.2 and 0.4. The correlation results showed that pork with low purine content had higher final pH, IMF and waterpower, and the meat color tended to bright red. Therefore, the breeding of low purine quality broiler pigs will not conflict with the breeding direction of important meat quality traits of traditional concern, and may even help to improve these meat quality traits to a certain extent. (6) GWAS analysis identified 40 SNPs (P5.00 *10-8) which were significantly associated with total purine content, and they may be progenies. 11 QTLs were listed, among which the most significant SNP rs32337327 (P=2.75 * 10-9) was located between LDB2 and QDPR genes of porcine chromosome 8 (SSC8), while the strongest correlation between SSC3, SSC11 and SSC14 was located in the intron region of the SSC3, SSC11 and SSC14 respectively, and 263 of them were significantly associated with guanine content. Among 26 QTLs, the most significant association SNP rs343436977 (P = 9.72 *10-13) was located on the intron of ABCC4 gene, and the most significant association SNPs in nine QTLs were also located on the intron of different genes. Their potential QTLs (P5.00 *10-6), the strongest associated SNPs were located in PAQR8 and SLC10A7, respectively. The GWAS results of the sum of adenine and hypoxanthine were similar to those of the total purine GWAS. (7) Twenty-five differentially expressed genes or sequence tags were identified by analyzing the transcriptome data of individuals with extreme muscle purine content. The transcripts of the annotated genes were RYK, ORAOV1, MTERF1, MOSPD1, SLCO1A2, B3GNTL1, ZDHHC7 and FAM92A. No GWAS signals were found in the differentially expressed gene regions, suggesting that pork purine content might be regulated by complex genetic factors. (8) 70 candidate genes and 8 differentially expressed genes were annotated near the most significant SNP site of GWAS. By GO analysis, 11 functional enrichment entries were identified, of which 8 related genes were found. KEGG analysis revealed that ABCC4, ADCY2 and GRIA1 were involved in the cAMP signaling pathway. In addition, 10 of the 78 genes encoded proteins had ATP binding sites and 41 belonged to membrane proteins. The cAMP signaling pathway is likely to participate in regulating the metabolism and transport of purine * in muscle cells, thereby affecting the purine content of porcine muscles. For the first time, this study locates the QTL locus affecting pork purine content, identifies multiple candidate genes, and reveals that calcium and cAMP signaling pathways have important effects on muscle purine content. It lays a solid foundation for revealing the genetic mechanism of purine content in muscle and provides theoretical and technical support for breeding low * purine quality pig breeds in the future.
【學(xué)位授予單位】：江西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S828
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