本文關(guān)鍵詞： 豬 肉質(zhì)性狀 全基因組關(guān)聯(lián)分析 QTL 精細(xì)定位 候選基因　出處：《江西農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：豬肉品質(zhì)是豬的重要經(jīng)濟(jì)性狀,提高豬肉品質(zhì)是現(xiàn)代種豬選育工作的重要目標(biāo)之一。然而肉質(zhì)是一個(gè)綜合性狀且遺傳力都偏低,傳統(tǒng)育種難以得到好的改良效果,因此對(duì)豬肉質(zhì)性狀分子遺傳機(jī)理的研究就顯得十分必要。本實(shí)驗(yàn)屠宰并測(cè)定了共610頭杜長(zhǎng)大個(gè)體36h肌肉酸度(pH)、剩余糖原含量(RG)、滴水損失(drip loss)、紅度(Minolta a*)、黃度(Minolta b*)、亮度(Minolta L*)、肉色(color score)、大理石紋(marbling)、水分(moisture content)等肉質(zhì)性狀表型并利用豬60K芯片進(jìn)行了全基因組關(guān)聯(lián)分析。結(jié)果共檢測(cè)到17個(gè)顯著性位點(diǎn),其中3個(gè)SNP達(dá)到了基因組顯著水平。同時(shí)在15號(hào)染色體上121 Mb處和134Mb處發(fā)現(xiàn)兩個(gè)同時(shí)影響剩余糖原含量、滴水損失和肉色的多效性數(shù)量性狀位點(diǎn)(QTL),在此命名為QTL_1和QTL_2。在QTL_1區(qū)域(129~135 Mb)內(nèi),筆者發(fā)現(xiàn)一個(gè)與RG強(qiáng)關(guān)聯(lián)的SNP SS478944394(129.80 Mb,P=1.18×10~(-11))和一個(gè)與滴水損失及肉色強(qiáng)關(guān)聯(lián)的SNP SS107833468(134.40 Mb,P10~(-5)),并推測(cè)已知的肉質(zhì)性狀主效基因PRKAG3(133.80 Mb)為其候選基因。為確定這一點(diǎn),本研究在QTL_1區(qū)域內(nèi)增加15個(gè)SNPs再做關(guān)聯(lián)分析,結(jié)果在PRKAG3基因編碼區(qū)內(nèi)發(fā)現(xiàn)一個(gè)更強(qiáng)關(guān)聯(lián)位點(diǎn)—rs42(133.80 Mb),該SNP與糖原含量(P=1.08×10~(-49))、滴水損失(P=1.95×10~(-12))、pH值(P=3.19×10~(-6))、L*值(P=1.54×10-9)、a*值(P=9.36×10~(-8))、b*值(P=7.28×10~(-13))等肉質(zhì)性狀的關(guān)聯(lián)程度都達(dá)到了基因組顯著性水平,且該位點(diǎn)解釋了杜長(zhǎng)大群體40.6%的糖原表型變異方差和8.6%的滴水損失表型變異方差。在QTL_2區(qū)域(120.78~122.78 Mb)內(nèi),SNP SS478943559(121.78 Mb)與剩余糖原含量(P=4.89×10~(-10))、滴水損失(P=2.39×10~(-5))、b*值(P=2.55×10~(-4))均顯著相關(guān),其周邊有兩個(gè)與糖代謝相關(guān)的強(qiáng)候選基因FASTKD2及MDH1B。為精細(xì)定位QTL_2,本研究在其區(qū)域內(nèi)新增了45個(gè)SNP標(biāo)記用于關(guān)聯(lián)分析,結(jié)果顯示最強(qiáng)關(guān)聯(lián)位點(diǎn)仍為SS478943559,它處在一個(gè)23 kb單倍型框(r~20.8)內(nèi)。對(duì)FASTKD2及MDH1B基因進(jìn)行cDNA測(cè)序發(fā)現(xiàn)兩個(gè)錯(cuò)義突變位點(diǎn),其中FASTKD2中錯(cuò)義突變SNP g.6742GA的次等位基因頻率為0.4,它與糖原含量、滴水損失關(guān)聯(lián)分析所得P值分別為4.47×10~(-4)、5.37×10~(-4),因此可以作為種豬肉質(zhì)選育的一個(gè)分子標(biāo)記。而MDH1B中的SNP g.3597AG突變與糖原含量、滴水損失相關(guān)性不顯著(P0.05),故排除此SNP作為因果突變的可能性。本研究加深了對(duì)杜長(zhǎng)大商品豬肉質(zhì)性狀分子遺傳機(jī)理的認(rèn)識(shí),同時(shí)為開發(fā)影響豬肉品質(zhì)性狀的分子標(biāo)記并為最終鑒別影響糖原含量及滴水損失等相關(guān)肉質(zhì)性狀的因果基因或因果突變點(diǎn)奠定了一定的工作基礎(chǔ)。
[Abstract]:Pork quality is an important economic trait of pigs, and improving pork quality is one of the important objectives of modern breeding pig breeding. However, meat quality is a comprehensive trait and the heritability is low. It is difficult to obtain good improvement effect in traditional breeding. Therefore, it is necessary to study the molecular genetic mechanism of pork quality traits. In this experiment, 610 individuals were slaughtered and their muscle acidity, pH value and residual glycogen content were measured for 36 h. The loss of drip water is the loss of drip, the degree of redness is minolta, the degree of yellowness is minolta, and the brightness is minolta. Meat color scoreen, marbling. The phenotypes of meat quality traits such as water moisture content were analyzed with porcine 60K microarray. A total of 17 significant loci were detected. Three of them reached the level of genomic significance. At the same time, at 121Mb and 134Mb on chromosome 15, two were found to affect the residual glycogen content at the same time. The quantitative trait loci of drip loss and meat color were named QTL_1 and QTL _ 2. In the QTL_1 region 129m ~ 135Mb). We find a SNP SS478944394(129.80 Mb strongly associated with RG. (1. 18 脳 10 ~ (-11)) and a SNP SS107833468(134.40 MbP10 ~ (-5), which is strongly associated with drip loss and meat color. The known major gene PRKAG3(133.80 Mb for meat quality traits is a candidate gene. In this study, 15 SNPs were added to the QTL_1 region to do the association analysis, and a stronger association locus -rs42 ~ (133.80 Mb) was found in the PRKAG3 gene coding region. The SNP and the content of glycogen were 1.08 脳 10 ~ (10) ~ (-49), and the loss of water was 1.95 脳 10 ~ (10) ~ (-12) ~ (?) pH value ~ (3. 19 脳 10 ~ (-6)). The L * value is 1.54 脳 10 ~ (-9) (P ~ (9.36) 脳 10 ~ (-8)). The correlation degree of meat quality traits, such as b* value (7.28 脳 10 ~ (-13)) and so on, reached the level of genomic significance. This locus explained the variance of phenotypic variation of glycogen in population 40.6% and the variation of phenotypic variation of drop water loss of 8.6% in Du-long population. Inner. SNP SS478943559(121.78 Mb) and the content of residual glycogen P 4. 89 脳 10 ~ (10) -10 ~ (10). There was a significant correlation between water drop loss and water loss (P = 2.39 脳 10 ~ (-1) ~ (-5) P ~ (2. 55) 脳 10 ~ (-4)). There are two strong candidate genes FASTKD2 and MDH1 B. which are closely related to glucose metabolism. In this study, 45 SNP markers were added to the region for association analysis. The results showed that the strongest correlation was still SS478943559. It was located in a 23kb haplotype frame rnrm20.8). FASTKD2 and MDH1B genes were sequenced by cDNA and two missense mutation sites were found. The secondary allele frequency of missense mutation SNP g.6742GA in FASTKD2 was 0.4, which was associated with glycogen content. The P values obtained from the correlation analysis of drip loss were 4.47 脳 10 ~ (-4) and 5.37 脳 10 ~ (-4) respectively. Therefore, it can be used as a molecular marker for breeding meat quality of pig, while the mutation of SNP g.3597AG in MDH1B has no significant correlation with glycogen content and drip loss (P0.05). Therefore, the possibility of this SNP as a causal mutation was excluded. This study deepened the understanding of the molecular genetic mechanism of meat quality traits in Du-long commercial pigs. At the same time, it lays a foundation for the development of molecular markers affecting pork quality traits and for the final identification of causal genes or causal mutation sites that affect glycogen content and drip loss.
【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】：2015
【分類號(hào)】：S828

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