本文選題：豬　切入點：脂肪沉積　出處：《中國農(nóng)業(yè)大學(xué)》2015年博士論文

【摘要】：豬的脂肪沉積的多少嚴(yán)重影響著豬肉的生產(chǎn)效率、豬肉品質(zhì)、繁殖性能以及消費者對豬肉的選擇。本研究通過B型超聲波背膘儀對松遼黑豬和長白豬群體進行活體背膘厚度的測定,篩選出具有極端背膘厚度的個體。利用高通量測序技術(shù)檢測篩選出的具有極端背膘厚度的松遼黑豬和長白豬的背部皮下脂肪(各3對)和肝臟(各2對)的轉(zhuǎn)錄組,篩選出差異表達基因,確定在脂肪組織和肝臟中調(diào)控脂肪沉積的關(guān)鍵基因。同時,針對每個品種不同極端厚度的群體(高、低)的基因組進行混池重測序,找到2組之間的基因變異。通過整合轉(zhuǎn)錄組數(shù)據(jù)、重測序數(shù)據(jù)及QTL數(shù)據(jù)庫等以及功能基因組學(xué)、生物信息學(xué)等方法,分析差異表達基因、基因變異的變化規(guī)律及相互關(guān)系,篩選、鑒定并驗證影響豬脂肪沉積的候選基因及其變異。 本研究對比了不同的品種及不同表型個體間的肉品質(zhì)。松遼黑豬與長白豬的肉色之間有著顯著的差異,同時又有著更高的肌內(nèi)脂肪含量和更低失水率。而在同一品種的不同背膘厚度的群體之間,只有肌內(nèi)脂肪含量有極顯著的差異,其他肉品質(zhì)之間并沒有顯示出差異。 通過轉(zhuǎn)錄組測序,在具有極端背膘厚度的松遼黑豬的脂肪組織和肝臟中,分別找到了188個和91個差異表達基因。在長白豬的脂肪組織和肝臟中,也分別找到了552個和72個差異表達基因。通過生物學(xué)功能分析發(fā)現(xiàn),差異表達基因參與了脂肪酸合成、脂肪合成底物的合成、脂類代謝的激活、胰島素信號通路、PPAR信號通路、MAPK信號通路、PPAR信號通路等眾多與脂肪沉積相關(guān)的生物學(xué)過程。這些與脂肪合成、代謝相關(guān)的基因如1ASN、PCK1、ME1、ELOVL6可以被認(rèn)為影響脂肪沉積多少的候選基因。同時,在脂肪組織中找到了大量的免疫相關(guān)的差異表達基因,結(jié)果表明脂肪組織作為一個免疫器官,脂肪沉積的多少直接影響著脂肪組織在免疫中的作用。 通過全基因組重測序中,在整個基因組中找到了大量基因變異,包括SNP和插入缺失,這些基因變異大部分坐落在基因間或內(nèi)含子上。同時針對含有可能影響基因表達和功能的錯義突變、移碼突變的基因進行功能富集分析,發(fā)現(xiàn)在脂肪合成、轉(zhuǎn)運、儲存、代謝等通路上含有大量的遺傳變異。大量位點的發(fā)現(xiàn)為進一步研究影響脂肪沉積提供了基礎(chǔ)。 最后整合轉(zhuǎn)錄組測序和全基因組重測序的結(jié)果,找到關(guān)鍵差異表達基因上的變異。在脂肪酸合成的關(guān)鍵基因FASN、ACACA、ME1、PCK1等上找到很多的變異,特別是存在這些基因的錯義突變、UTR突變等,都是可能影響脂肪沉積的重點突變。本研究發(fā)現(xiàn)了脂肪合成關(guān)鍵的基因如ELOVL6、LDHA、CYP2B224坐落在脂肪沉積相關(guān)的QTL上。本研究同時挑選出了部分變異位點,與背膘厚度進行關(guān)聯(lián)分析,發(fā)現(xiàn)ME1、PCK1基因UTR區(qū)上變異位點顯著影響背膘的厚度。 綜上所述,脂肪組織中的脂肪酸合成能力是影響背膘厚度的關(guān)鍵因素,同時肝臟和脂肪組織中的代謝也發(fā)揮著重要的作用。研究結(jié)果全面闡述了不同背膘厚度的松遼黑豬和長白豬脂肪組織和肝臟基因表達譜及基因組的變異,為深入研究豬脂肪沉積的分子調(diào)控機制及進一步的實驗奠定了基礎(chǔ)。
[Abstract]:How much of the fat deposition of pigs seriously affects the pork production efficiency, pork quality, reproduction performance and consumer choice of pork. In this study, determination of backfat thickness on Songliao Black Pig and Landrace populations by B type ultrasonic backfat instrument, and screened with extreme backfat thickness individuals screened by. High throughput sequencing technology has the extreme backfat thickness of Songliao Black Pig and Landrace back fat (3 pairs) and liver (2 pairs) transcriptome, we screened the differentially expressed genes, determine the key genes regulating control of fat deposition in adipose tissue and liver. At the same time, for each of the different varieties of extreme the thickness of the groups (high, low) of the genome of the mixed pool re sequencing, find the genetic variation between the 2 groups. By integrating transcriptome data, re sequencing data and QTL database and functional genomics, bioinformatics In order to identify and verify candidate genes and their variation affecting porcine fat deposition, we analyzed the change rules and relationship of differentially expressed genes and gene mutation.
This study compared the different varieties and different phenotypes between individuals of meat quality. There are significant differences between Songliao Black Pig and Landrace flesh, and has a fat content and lower water loss rate is higher in the muscle. And in the same variety of different backfat thickness between the groups, only the fat content there were significant differences in muscle, and did not show differences between other meat quality.
Through transcriptome sequencing, in adipose tissue and liver with extreme backfat thickness in Songliao Black pig, were found 188 and 91 differentially expressed genes in adipose tissue and liver in Landrace pigs, were also found 552 and 72 differentially expressed genes. The analysis can be found through biological function, difference the expression of genes involved in fatty acid synthesis, the synthesis of fat synthesis substrate activation, lipid metabolism, insulin signaling pathway, PPAR signaling pathway, MAPK signaling pathway, PPAR signaling pathway and many other biological processes associated with fat deposition. These fat synthesis, metabolism related genes such as 1ASN, PCK1, ME1, ELOVL6 can be considered how many of the candidate gene affecting fat deposition in adipose tissue. At the same time, to find a large number of immune related gene expression. The results suggest that adipose tissue as an immune organ, how much fat deposition It has a direct effect on the role of adipose tissue in the immune system.
Through whole genome sequencing, to find a large number of mutations in the genome, including SNP and indel, most of these mutations located in genes or introns. At the same time for containing may affect missense gene expression and function of gene mutation, mutation of shift function enrichment analysis, found in fat synthesis, transport, storage, metabolic pathway containing a large amount of genetic variation. A large number of sites that provide a basis for further study on the effects of fat deposition.
Finally, the integration of transcriptome sequencing and whole genome sequencing results, find the key differences between the expression of the mutations in key genes in fatty acid synthesis. FASN, ACACA, ME1, PCK1 and so on to find a lot of variation, especially in the presence of missense mutations in these genes, UTR mutation, are likely to influence on fat deposition mutation. This study found a fat synthesis of key genes such as ELOVL6, LDHA, CYP2B224 is located in QTL related to fat deposition. This study also selected some mutation sites for association analysis, found that ME1 and backfat thickness, ectopic point significantly affect backfat thickness PCK1 gene UTR region.
In summary, fatty acid synthesis ability in adipose tissue is a key factor affecting the backfat thickness at the same time, liver and adipose tissue metabolism also plays an important role. The research results comprehensively expounds the variation of Songliao Black Pigs and Landrace pigs adipose tissue and liver gene expression profiles of different backfat thickness and genome, laid the foundation molecular regulation mechanism research of pig fat deposition and further experiments.

【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：S828

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本文編號：1681017