黃牛生長性狀選擇的IRX3基因標記研究與應(yīng)用
發(fā)布時間：2018-08-06 17:05
【摘要】：易洛魁族同源盒3(iroquois homeobox3,IRX3)是易洛魁族同源盒家族(IRXs)中的最具代表的一員,最新報道,公認的肥胖基因FTO(fat mass and obesity associated)基因非編碼區(qū)的變異會促使基因IRX3過度表達,進而誘發(fā)肥胖,IRX3基因可有效控制褐色脂肪的形成。并且IRX3基因和FTO基因的的這種互動關(guān)系普遍存在于人類、老鼠和斑馬魚中。進而可推測IRX3基因很可能影響肉牛脂肪代謝進而影響肉牛生產(chǎn),而目前IRX3基因在肉牛上未見報道,因此分析IRX3基因標記及其應(yīng)用對于提高肉牛產(chǎn)業(yè)的經(jīng)濟效益有重要的理論和實踐意義。本試驗通過DNA混合池測序技術(shù)和PCR-RFLP技術(shù)分析研究了724個黃牛個體(晉南牛205頭、秦川牛519頭)間的遺傳變異及其對黃牛生長性狀的影響,以期找到對肉牛產(chǎn)業(yè)發(fā)展有高價值的分子標記,并提出該技術(shù)在肉牛選育中的具體應(yīng)用方案。試驗結(jié)果如下:1、黃牛IRX3基因的遺傳變異研究以30個秦川牛和晉南牛個體的DNA等量混合的DNA池為模板設(shè)計了5對引物擴增IRX3基因的全長及其啟動子區(qū),發(fā)現(xiàn)了3個多態(tài)位點,其中g(shù).-1941GA多態(tài)位點和g.-991GC多態(tài)位點位于啟動子區(qū),g.1941TG多態(tài)位點位于第二內(nèi)含子區(qū)。這些多態(tài)位點分別采用Msp1-RFLP、KpnI-RFLP和Hinf1-RFLP的分型方法檢測其在晉南牛和秦川牛的遺傳變異。對IRX3基因3個多態(tài)位點進行群體遺傳學分析、連鎖不平衡分析、單倍型分析。群體遺傳學分析結(jié)果表明兩黃牛品種間存在較大差異,SNP1位點野生型的等位基因頻率(0.647和0.601)均顯著高于突變型的等位基因頻率(0.353和0.399),SNP3位點野生型的等位基因頻率(0.916和0.849)均顯著高于突變型的等位基因頻率(0.154和0.256),而SNP2位點的突變型的等位基因頻率(0.85和0.823)較高;而且SNP3位點在晉南牛中只檢測到兩種基因型GG和GT,未發(fā)現(xiàn)純合型的突變個體。遺傳多態(tài)性指標分析表明3個位點多態(tài)信息比較豐富,育種潛力較大,除了晉南牛中SNP3位點處于低度多態(tài)(PIC0.25)以外,其他位點在群體中均處于中度多態(tài)(0.25PIC0.5)。此外卡方檢驗表明除了秦川牛在SNP2位點處于哈代-溫伯格不平衡狀態(tài)(P0.05),其它位點在兩黃牛群體中均處于哈代-溫伯格平衡狀態(tài)(P0.05)。單倍型分析結(jié)果表明相同品種間單倍型分布頻率有差異,而相同單倍型在不同品種也有差異,IRX3基因三個多態(tài)位點存在的8種單倍型,AGG單倍型的分布頻率最高,在秦川牛中可以檢測到全部的單倍型,而在晉南牛中只檢測到6種單倍型。連鎖分析結(jié)果表明不同群體間的相同兩位點間的連鎖程度仍不相同,晉南牛中的SNP1和SNP3位點的連鎖稍強,而在秦川牛中處于弱連鎖狀態(tài)。2、黃牛IRX3基因的遺傳變異與生長性狀的關(guān)聯(lián)分析IRX3基因3個多態(tài)位點與秦川牛和晉南牛的生長性狀關(guān)聯(lián)分析結(jié)果表明,SNP1位點對晉南牛的胸深有顯著影響(P0.05),純合子AA基因型的胸深顯著高于AG基因型個體;SNP2位點對晉南牛的體高、胸深、體高和體長有顯著影響(P0.05),CC基因型的個體在體高、胸深、體高和體長方面均顯著高于CG基因型個體和GG基因型個體,對秦川牛的生長性狀影響不顯著(P0.05);SNP3位點與秦川牛的胸圍直接存在顯著相關(guān)(P0.05),GG基因型個體胸圍顯著高于GT基因型,對晉南牛的生長性狀影響不顯著(P0.05)。綜合以上分析說明IRX3基因的3個多態(tài)位點可作為黃牛生長性狀選育的重要分子遺傳標記,IRX3基因可作為黃牛育種的重要候選基因。3、黃牛IRX3基因標記技術(shù)在肉牛選育中的應(yīng)用方案利用IRX3基因標記技術(shù)與多階段選擇相結(jié)合的方法通過性能測定對黃牛進行有效選育。1)理論依據(jù):找出與黃牛生長性狀顯著相關(guān)的SNP位點,確定顯著提高黃牛生長性狀的基因型及其優(yōu)勢性狀。SNP1位點對晉南牛的胸深有顯著影響(P0.05),純合子AA基因型的胸深顯著高于AG基因型個體;SNP2位點對晉南牛的體高、胸深和體長有顯著影響(P0.05),CC基因型的個體在體高、胸深和體長方面均顯著高于CG基因型個體和GG基因型個體,對秦川牛的生長性狀影響不顯著(P0.05);SNP3位點與秦川牛的胸圍直接存在顯著相關(guān)(P0.05),GG基因型個體胸圍顯著高于GT基因型,對晉南牛的生長性狀影響不顯著(P0.05)。2)斷奶前選擇:首先對犢牛編號標記,詳細記錄出生時間、父母代的信息以及體重等,以便以后的選育工作正常進行。由于斷奶時的各種性狀未表現(xiàn)出來,通過測定犢牛IRX3基因相關(guān)分子標記位點的基因型,如SNP1位點對應(yīng)的AA優(yōu)勢基因型、SNP2位點對應(yīng)的CC優(yōu)勢基因型和SNP3位點對應(yīng)的GG優(yōu)勢基因型,對犢牛進行早期選種,也可選擇能繁育具有這些優(yōu)勢基因型后代的親本進行配種;3)6月齡黃牛的主要生長性狀已經(jīng)顯現(xiàn),應(yīng)該作為主選階段。通過分子測定結(jié)合對其測定優(yōu)勢性狀如胸深、體高、體長以及胸圍,篩選優(yōu)勢性狀較為明顯的黃牛個體。
[Abstract]:The Iroquois homologous box 3 (Iroquois homeobox3, IRX3) is one of the most representative members of the Iroquois homologous box family (IRXs). The latest report shows that the variation of the noncoding region of the obese gene FTO (fat mass and obesity associated) can induce the gene IRX3 overexpression and induce obesity. The IRX3 gene can effectively control the brown fat. The interaction between the IRX3 gene and the FTO gene is common in human, mice and zebrafish. Then it can be deduced that the IRX3 gene may affect beef fat metabolism and affect beef production. At present, the IRX3 gene is not reported on beef cattle. Therefore, the IRX3 based marker and its application are analyzed to improve the beef cattle industry. The economic benefit has important theoretical and practical significance. In this experiment, the genetic variation of 724 cattle (205 cattle of Jinnan cattle, 519 heads of Qinchuan cattle) and their effects on the growth traits of yellow cattle were analyzed by DNA mixing pool sequencing and PCR-RFLP technology, in order to find the molecular markers of high value for the development of beef cattle production and put forward the technique. The specific application in breeding beef cattle. The results are as follows: 1, the genetic variation of IRX3 gene in cattle was studied by using the DNA isometric DNA pool of 30 Qinchuan cattle and southern Jinnan cattle to design 5 pairs of primers to amplify the full length of the IRX3 gene and its promoter region, and 3 polymorphic loci were found, of which g.-1941GA polymorphic loci and g.-991GC were found. The polymorphic loci were located in the promoter region, and the g.1941TG polymorphic loci were located in the intron second. The polymorphic loci used Msp1-RFLP, KpnI-RFLP and Hinf1-RFLP to detect their genetic variation in the southern Jinnan and Qinchuan cattle. The 3 polymorphic loci of the IRX3 gene were analyzed by population genetic analysis, linkage disequilibrium analysis and haplotype analysis. The results of population genetics analysis showed that there were significant differences among the two yellow cattle. The allele frequencies of the wild type SNP1 loci (0.647 and 0.601) were significantly higher than those of the mutant allele frequencies (0.353 and 0.399). The frequencies of the allele (0.916 and 0.849) of the wild type SNP3 loci (0.916 and 0.849) were significantly higher than those of the mutant allele frequencies (0.154 and 0.25). 6), the mutant allele frequencies of the SNP2 loci (0.85 and 0.823) were higher, and only two genotypes GG and GT were detected in the southern Jinnan cattle, and no homozygous mutant individuals were found. The genetic polymorphism index analysis showed that the 3 loci polymorphism information was rich and the breeding potential was large, except that the SNP3 loci in the southern Shanxi cattle were low. Besides the polymorphism (PIC0.25), other loci were in moderate polymorphism (0.25PIC0.5). Besides, chi square test showed that in addition to the Hardy Weinberg imbalance (P0.05) at the SNP2 site of Qinchuan cattle (P0.05), the other loci were in the Hardy Weinberg equilibrium state in the two cattle population (P0.05). The haplotype analysis showed that the same varieties were among the same varieties. The haplotype distribution frequency is different, and the same haplotype is also different in different varieties. The distribution frequency of 8 haplotypes of the three polymorphic loci of IRX3 gene is the highest. All the haplotypes can be detected in the Qinchuan cattle, and only 6 haplotypes are detected in the southern Jinnan cattle. The linkage analysis results show that the different populations are among the different populations. The linkage between the same two loci is still different. The linkage between the SNP1 and SNP3 loci in the southern Jinnan cattle is slightly stronger, while the weak linkage.2 in the Qinchuan cattle and the association of the genetic variation with the growth traits of the Yellow Cattle IRX3 gene analysis the correlation analysis between the 3 polymorphic loci of the IRX3 gene and the growth traits of the Qinchuan cattle and the southern Jinnan cattle. The results show that the SNP1 locus is the site of the relationship between the IRX3 gene and the growth traits of the southern Jinnan cattle. The chest depth of the southern Jinnan cattle was significantly affected (P0.05). The thorax depth of the homozygote AA genotype was significantly higher than that of the AG genotype. The SNP2 locus had significant influence on body height, chest depth, body height and body length (P0.05). The CC genotype individuals were significantly higher in body height, chest depth, body height and body length than CG genotypes and GG genotypes. The growth traits of Sichuan cattle were not significantly affected (P0.05), and there was a significant correlation between the SNP3 locus and the chest circumference of Qinchuan cattle (P0.05). The GG genotype was significantly higher than that of the GT genotype, and the effect on the growth traits of the southern Jinnan cattle was not significant (P0.05). The analysis of the 3 polymorphic loci of the IRX3 base could be important for the selection of the growth traits of the cattle. Molecular genetic markers, IRX3 gene can be used as an important candidate gene.3 for cattle breeding, and the application scheme of yellow cattle IRX3 gene marker technique in breeding beef cattle is based on the combination of IRX3 gene marker technique and multi stage selection method, which can be used to select.1 effectively through performance measurement for Yellow Cattle: to find out the significant correlation with the growth traits of yellow cattle. The SNP loci confirmed that the genotype and.SNP1 locus of the dominant traits significantly affected the chest depth of the southern Shanxi cattle (P0.05). The thorax depth of the homozygote AA genotype was significantly higher than that of the AG genotype, and the SNP2 locus had a significant influence on the body height, the chest depth and the body length of the southern Shanxi cattle (P0.05), and the individuals of the CC genotypes were in body height and chest depth. And the length of body length was significantly higher than that of CG genotype and GG genotype, which had no significant influence on the growth traits of Qinchuan cattle (P0.05). The SNP3 locus had a significant correlation with the chest circumference of Qinchuan cattle (P0.05), and the individual chest circumference of GG genotype was significantly higher than that of the GT genotypes, which had no significant effect on the growth traits of the southern Shanxi cattle (P0.05) before weaning. First, the number of calves was numbered to record the birth time, the information of the parent generation and the weight, so that the future breeding work was carried out normally. The genotype of the IRX3 gene related marker loci of calves, such as the AA dominant genotype at the SNP1 site, and the CC best of the SNP2 locus, were not shown in the weaning. The GG dominant genotype of the potential genotypes and SNP3 loci can be selected early and can also be selected to breed the parent of the offspring with these dominant genotypes; 3) the main growth traits of the 6 month old yellow cattle have already appeared and should be selected as the main selection stage. Long and chest circumference were used to screen yellow cattle individuals with superior traits.
【學位授予單位】：西北農(nóng)林科技大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S823.81
本文編號：2168385
[Abstract]:The Iroquois homologous box 3 (Iroquois homeobox3, IRX3) is one of the most representative members of the Iroquois homologous box family (IRXs). The latest report shows that the variation of the noncoding region of the obese gene FTO (fat mass and obesity associated) can induce the gene IRX3 overexpression and induce obesity. The IRX3 gene can effectively control the brown fat. The interaction between the IRX3 gene and the FTO gene is common in human, mice and zebrafish. Then it can be deduced that the IRX3 gene may affect beef fat metabolism and affect beef production. At present, the IRX3 gene is not reported on beef cattle. Therefore, the IRX3 based marker and its application are analyzed to improve the beef cattle industry. The economic benefit has important theoretical and practical significance. In this experiment, the genetic variation of 724 cattle (205 cattle of Jinnan cattle, 519 heads of Qinchuan cattle) and their effects on the growth traits of yellow cattle were analyzed by DNA mixing pool sequencing and PCR-RFLP technology, in order to find the molecular markers of high value for the development of beef cattle production and put forward the technique. The specific application in breeding beef cattle. The results are as follows: 1, the genetic variation of IRX3 gene in cattle was studied by using the DNA isometric DNA pool of 30 Qinchuan cattle and southern Jinnan cattle to design 5 pairs of primers to amplify the full length of the IRX3 gene and its promoter region, and 3 polymorphic loci were found, of which g.-1941GA polymorphic loci and g.-991GC were found. The polymorphic loci were located in the promoter region, and the g.1941TG polymorphic loci were located in the intron second. The polymorphic loci used Msp1-RFLP, KpnI-RFLP and Hinf1-RFLP to detect their genetic variation in the southern Jinnan and Qinchuan cattle. The 3 polymorphic loci of the IRX3 gene were analyzed by population genetic analysis, linkage disequilibrium analysis and haplotype analysis. The results of population genetics analysis showed that there were significant differences among the two yellow cattle. The allele frequencies of the wild type SNP1 loci (0.647 and 0.601) were significantly higher than those of the mutant allele frequencies (0.353 and 0.399). The frequencies of the allele (0.916 and 0.849) of the wild type SNP3 loci (0.916 and 0.849) were significantly higher than those of the mutant allele frequencies (0.154 and 0.25). 6), the mutant allele frequencies of the SNP2 loci (0.85 and 0.823) were higher, and only two genotypes GG and GT were detected in the southern Jinnan cattle, and no homozygous mutant individuals were found. The genetic polymorphism index analysis showed that the 3 loci polymorphism information was rich and the breeding potential was large, except that the SNP3 loci in the southern Shanxi cattle were low. Besides the polymorphism (PIC0.25), other loci were in moderate polymorphism (0.25PIC0.5). Besides, chi square test showed that in addition to the Hardy Weinberg imbalance (P0.05) at the SNP2 site of Qinchuan cattle (P0.05), the other loci were in the Hardy Weinberg equilibrium state in the two cattle population (P0.05). The haplotype analysis showed that the same varieties were among the same varieties. The haplotype distribution frequency is different, and the same haplotype is also different in different varieties. The distribution frequency of 8 haplotypes of the three polymorphic loci of IRX3 gene is the highest. All the haplotypes can be detected in the Qinchuan cattle, and only 6 haplotypes are detected in the southern Jinnan cattle. The linkage analysis results show that the different populations are among the different populations. The linkage between the same two loci is still different. The linkage between the SNP1 and SNP3 loci in the southern Jinnan cattle is slightly stronger, while the weak linkage.2 in the Qinchuan cattle and the association of the genetic variation with the growth traits of the Yellow Cattle IRX3 gene analysis the correlation analysis between the 3 polymorphic loci of the IRX3 gene and the growth traits of the Qinchuan cattle and the southern Jinnan cattle. The results show that the SNP1 locus is the site of the relationship between the IRX3 gene and the growth traits of the southern Jinnan cattle. The chest depth of the southern Jinnan cattle was significantly affected (P0.05). The thorax depth of the homozygote AA genotype was significantly higher than that of the AG genotype. The SNP2 locus had significant influence on body height, chest depth, body height and body length (P0.05). The CC genotype individuals were significantly higher in body height, chest depth, body height and body length than CG genotypes and GG genotypes. The growth traits of Sichuan cattle were not significantly affected (P0.05), and there was a significant correlation between the SNP3 locus and the chest circumference of Qinchuan cattle (P0.05). The GG genotype was significantly higher than that of the GT genotype, and the effect on the growth traits of the southern Jinnan cattle was not significant (P0.05). The analysis of the 3 polymorphic loci of the IRX3 base could be important for the selection of the growth traits of the cattle. Molecular genetic markers, IRX3 gene can be used as an important candidate gene.3 for cattle breeding, and the application scheme of yellow cattle IRX3 gene marker technique in breeding beef cattle is based on the combination of IRX3 gene marker technique and multi stage selection method, which can be used to select.1 effectively through performance measurement for Yellow Cattle: to find out the significant correlation with the growth traits of yellow cattle. The SNP loci confirmed that the genotype and.SNP1 locus of the dominant traits significantly affected the chest depth of the southern Shanxi cattle (P0.05). The thorax depth of the homozygote AA genotype was significantly higher than that of the AG genotype, and the SNP2 locus had a significant influence on the body height, the chest depth and the body length of the southern Shanxi cattle (P0.05), and the individuals of the CC genotypes were in body height and chest depth. And the length of body length was significantly higher than that of CG genotype and GG genotype, which had no significant influence on the growth traits of Qinchuan cattle (P0.05). The SNP3 locus had a significant correlation with the chest circumference of Qinchuan cattle (P0.05), and the individual chest circumference of GG genotype was significantly higher than that of the GT genotypes, which had no significant effect on the growth traits of the southern Shanxi cattle (P0.05) before weaning. First, the number of calves was numbered to record the birth time, the information of the parent generation and the weight, so that the future breeding work was carried out normally. The genotype of the IRX3 gene related marker loci of calves, such as the AA dominant genotype at the SNP1 site, and the CC best of the SNP2 locus, were not shown in the weaning. The GG dominant genotype of the potential genotypes and SNP3 loci can be selected early and can also be selected to breed the parent of the offspring with these dominant genotypes; 3) the main growth traits of the 6 month old yellow cattle have already appeared and should be selected as the main selection stage. Long and chest circumference were used to screen yellow cattle individuals with superior traits.
【學位授予單位】：西北農(nóng)林科技大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S823.81
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