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1.
DNA methylation patterns at the IGF2‐H19 locus were investigated in sperm DNA from Swiss Landrace (SL) and Swiss Large White (LW) boars. The putative IGF2 differentially methylated regions (DMR) 0, 1 and 2, a quantitative trait nucleotide (QTN) region in the intron 3 and a CpG island in the intron 4 of the IGF2 gene as well as three regions around porcine CTCF binding sites within the H19 differentially methylated domain (DMD) were selected for the DNA methylation analysis. In both breeds putative IGF2 DMR0, 1, 2 and H19 DMD were hypermethylated. Significant differences in DNA methylation content were found between the two breeds in the two DMD regions proximal to the H19 gene. The IGF2 QTN region and the CpG island in the IGF2 intron 4 were hypomethylated in sperm DNA of both breeds. The methylation analysis revealed significantly more methylated CpG sites in the intron 4 of sperm from the LW breed than in that from SL. No difference was found in global DNA methylation between the two breeds. These results indicate differences in DNA methylation patterns between breeds and it remains to be established whether variation in DNA methylation patterns impacts on phenotypic traits.  相似文献   

2.
Leptin gene has been found to be associated with various economic traits including milk production and fat quality in dairy animals. In the present study, we investigated genetic variations in intron 1 region of leptin gene in riverine buffaloes (Bubalus bubalis) using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and sequencing methods and associated them with milk traits. The study revealed three SSCP variants A, B and C among a total of 301 buffaloes from nine breeds. The frequency of variant C was found invariably high among all the breeds except in Marathwada buffalo. Variant A was found to be absent in Chilika, Nili-Ravi, Nagpuri and Pandharpuri breeds and also had the lowest frequencies in Mehsana, Jaffarabadi, Murrah and Toda breeds. Sequencing of SSCP variants revealed a total of five polymorphic sites, with three haplotypes. Statistical analysis revealed significantly high fat percentage at 150?days in SSCP variant B in Mehsana buffaloes. However, the associations of SSCP variants of leptin gene with total milk yield, 305?days milk yield and total fat yield were found to be non-significant. The present study is the first report on association analysis of leptin gene polymorphisms with milk production and milk quality traits in river buffalo.  相似文献   

3.
为了探究绵羊Cdkn1c的甲基化状态,本研究对绵羊及其它3个物种的Cdkn1c mRNA序列进行了生物信息学分析,并通过亚硫酸氢盐测序法(BSP)对新生羔羊肾脏和肺脏Cdkn1c CpG岛上的CpG位点进行了甲基化分析。结果显示,整个编码区富含CG,为CpG岛;绵羊Cdkn1c所有分析位点均非甲基化,暗示该区域并非差异甲基化区域(DMR)。  相似文献   

4.
本研究旨在分析ITGB2基因在苏博美利奴羊不同细度皮肤组织中的DNA甲基化和mRNA表达水平。以苏博美利奴羊周岁母羊为试验动物,以不同细度的皮肤组织样为试验样本,对ITGB2基因(GenBank登录号:NC_040252.1)启动子区CpG岛进行预测并设计BSP引物,并对ITGB2基因(GenBank登录号:NM_001009485.1)、GAPDH基因(GenBank登录号:NM_001190390.1)mRNA序列设计引物,采用重亚硫酸盐测序法(BSP法)进行扩增纯化后将其连接pMD19-T载体,转化JM109细胞过夜培养,形成单菌落,筛选阳性克隆菌进行测序,对所获序列进行分析,分析ITGB2基因启动子区CpG岛在周岁母羊皮肤组织的甲基化模式,并运用实时荧光定量PCR检测ITGB2基因在苏博美利奴羊不同细度皮肤组织中的mRNA表达水平。结果显示,极细组苏博美利奴羊CpG岛甲基化率(94.29%)高于极粗组苏博美利奴羊的CpG岛甲基化率(87.62%),其中,极细组苏博美利奴羊CpG2、CpG3、CpG4、CpG7甲基化率(100%、100%、100%和80.00%)均高于极粗组(86.67%、93.33%、80.00%和73.33%);ITGB2基因在苏博美利奴羊极粗皮肤组织中的表达量极显著高于极细皮肤组织的表达量(P < 0.01),且ITGB2基因的DNA甲基化水平与mRNA表达量呈明显负相关。研究表明,DNA甲基化对皮肤生长发育有一定作用,可作为一个候选的表观遗传标记用于苏博美利奴羊。  相似文献   

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为探索钙黏蛋白13基因(cadherin 13,CDH13)甲基化在奶牛金黄色葡萄球菌乳房炎中的调控机制,本试验在已建立的金黄色葡萄球菌诱导型荷斯坦牛乳房炎模型的基础上利用重亚硫酸盐测序技术(bisulfite sequencing PCR,BSP)检测正常乳腺组织和乳房炎乳腺组织候选基因CDH13的甲基化程度,并分析其甲基化程度与基因表达水平的关系。结果显示,除金黄色葡萄球菌致病组(试验组)第12个CpG岛外,其余所有CpG岛均呈现不同程度的甲基化:-259处甲基化水平最高(50%~60%),-144、-135和-126处甲基化均较低,为5%左右。试验组和对照组总体甲基化率分别为10.13%±1.81%和14.43%±0.55%,不同位点和总体甲基化率无显著差异(P>0.05)。与正常乳腺组织相比,乳房炎感染组乳腺组织CDH13基因表达上调,CDH13基因-162和-93两个CpG岛甲基化水平与其基因表达呈显著负相关(P<0.05)。本试验结果表明,CDH13基因甲基化影响其基因表达,从而影响乳房炎的发生,为进一步探索其在金黄色葡萄球菌致病型乳房炎中的发病机制提供了参考依据。  相似文献   

7.
试验旨在获得鸡热休克蛋白90α(HSP90AA1)基因序列并分析其基因结构和相关遗传变异,检测HSP90AA1基因启动子区CpG岛的甲基化状态,初步探索HSP90AA1基因在肌肉组织生长发育中的作用。以文昌鸡和北京油鸡为试验材料,利用PCR扩增鸡HSP90AA1基因组序列;通过基因测序寻找该基因中的单核苷酸多态性(SNP)位点;使用在线软件MethPrimer预测鸡HSP90AA1基因中CpG岛的位置;应用MassArray质谱法检测鸡胸肌中HSP90AA1基因启动子区CpG岛的甲基化水平,比较分析文昌鸡和北京油鸡HSP90AA1基因的甲基化差异。结果显示,在鸡HSP90AA1基因组中共发现7个SNPs位点,分别位于启动子区(A-189G,C-109T)、第1外显子(A+6G)、第2外显子(C+343T)、第2内含子(A+634G、A+836G)和第7内含子(A+3449G);鸡HSP90AA1基因包含10个外显子和9个内含子,其启动子区存在1个CpG岛,位于-1 802~-469bp处;在HSP90AA1基因启动子区共检测了42个CpG位点的甲基化水平,文昌鸡和北京油鸡中分别有9个(CpG_16.17.18、CpG_21.22.23、CpG_32.33和CpG_57)和4个CpG位点(CpG_1、CpG_5.6和CpG_57)在胸肌生长发育过程中发生甲基化改变。结果表明,文昌鸡与北京油鸡HSP90AA1基因序列信息和启动子区CpG岛的甲基化水平不同,这可能导致两种鸡对于应激反应具有不同的耐受程度。以上试验结果将为文昌鸡和北京油鸡生长发育规律、系统选育等方面的研究提供表观遗传学依据。  相似文献   

8.
For semen suppliers, predicting the low fertility of service bull candidates before artificial insemination would help prevent economic loss; however, predicting bull fertility through in vitro assessment of semen is yet to be established. In the present study, we focused on the methylated CpG sites of sperm nuclear DNA and examined methylation levels to screen new biomarkers for predicting bull fertility. In frozen-thawed semen samples collected from Japanese Black bulls, for which the sire conception rate (SCR) was recorded, the methylation level of each CpG site was analyzed using human methylation microarray. According to regression analysis, 143 CpG sites related to SCR were significantly differentially methylated. Whole genome bisulfite sequence data were obtained from three semen samples and the differentially methylated regions (DMRs) that included the target CpG sites selected by human methylation microarray were confirmed. Using combined bisulfite restriction analysis, fertility-related methylation changes were detected in 10 DMRs. With the exception of one DMR, the methylation levels of these DMRs were significantly different between groups with high fertility (> 50%) and low fertility (< 40%). From multiple regression analysis of methylation levels and SCR, three DMRs were selected that could effectively predict bull fertility. We suggest that these fertility-related differences in spermatozoal methylation levels could be new epigenetic biomarkers for predicting bull fertility.  相似文献   

9.
为验证和探索香猪卵巢转录组RNA测序检测到的视黄酸受体应答1(retinoic acid receptor responder 1,RARRES1)基因在香猪高、低产仔组之间差异表达的原因,本试验针对RARRES1基因第1外显子ATG下游富含CpG位点区段设计特异性引物,采用亚硫酸氢盐测序法(bisulfite sequencing PCR,BSP)研究卵巢RARRES1基因中的甲基化修饰水平;利用实时荧光定量PCR方法检测高、低产仔组香猪卵巢RARRES1基因的表达量,并探究其与基因甲基化水平之间的相关性。结果表明,与低产仔组相比,香猪高产仔组的甲基化水平较高;4个CpG位点均未被甲基化(CpG_7、CpG_11、CpG_12和CpG_15),另外3个CpG位点(CpG_8、CpG_17和CpG_18)基本上全部发生了甲基化。此外,与低产仔组相比,香猪高产仔组中CpG_4(P>0.05)、CpG_9(P<0.05)和CpG_16(P<0.05)位点的甲基化占比较高,而CpG_6位点在香猪低产仔组中的甲基化比例较高,两组差异极显著(P<0.01)。实时荧光定量PCR结果显示,高产仔组香猪RARRES1基因的表达水平较高(P<0.05)。Spearman相关分析结果显示,CpG_9和CpG_16两个位点的甲基化比例与RARRES1基因的表达水平高度正相关(R2=0.896,P<0.01),提示这两个位点的甲基化可能是香猪高产仔组RARRES1基因表达量较高的原因。  相似文献   

10.
DNA甲基化调控牛AQP1基因的胎盘特异性印记   总被引:1,自引:1,他引:0  
为揭示牛AQP1(aquaporin 1)基因在不同组织及胎盘中的印记状态,以及DNA甲基化修饰在印记中的调控机制,本研究采用基于SNP的PCR产物直接测序的方法,对32头健康雌性成年荷斯坦奶牛心组织及15个自然分娩后的胎盘试验样本进行检测,确定了5头杂合子个体牛和3个杂合子胎盘,对其组织(心、肝、脾、肺、肾、肌肉和脂肪)和胎盘进行AQP1等位基因表达分析及印记状态分析,利用亚硫酸氢盐测序法分析AQP1基因位于启动子和第一个外显子区的CpG岛在牛心、肝组织、2个胎盘和对应精子中的DNA甲基化状态。结果发现,在杂合子牛被检测的7个组织中,AQP1基因呈现双等位基因表达;而在胎盘中,AQP1基因为单等位基因表达。通过分析杂合子胎盘对应的亲本基因型,发现AQP1基因为母源等位基因表达,即父源印记。进一步比较分析AQP1基因启动子区CpG岛在牛组织、胎盘及对应精子中的甲基化状态,在双等位基因表达的心脏、肝脏组织中,该区域未发现差异甲基化区(differentially methylated regions,DMR);而在单等位基因表达的胎盘中,存在差异甲基化区,同时父源等位基因精子中为重甲基化状态。以上结果说明,牛AQP1基因为胎盘特异性单等位基因表达的父源印记基因,且AQP1基因位于启动子和第一个外显子区的CpG岛甲基化修饰参与调控牛胎盘的印记表达;在被检测的组织中为双等位基因表达。  相似文献   

11.
范一萍  王彦芳  陶聪 《中国畜牧兽医》2019,46(12):3627-3634
为探究解偶联蛋白3(uncoupling protein 3,UCP3)基因在巴马猪和藏猪皮下脂肪组织中的表达和甲基化水平,试验采用实时荧光定量PCR技术检测UCP3基因在巴马猪和藏猪皮下脂肪组织中的mRNA表达水平;针对猪UCP3基因启动子区域(-3 580~+920 bp),利用在线软件MethPrimer对该区域进行CpG岛预测,并采用亚硫酸氢盐测序法(bisulfite sequencing PCR,BSP)检测其甲基化水平,探究UCP3基因甲基化水平在巴马猪和藏猪中的差异。结果显示,巴马猪皮下脂肪组织UCP3基因表达量显著高于藏猪(P<0.05);在UCP3基因启动子区预测到3个CpG甲基化岛,分别是CpG island1(-3 171~-2 928 bp)、CpG island2(-154~-2 bp)和CpG island3(+648~+806 bp),其中CpG island1和CpG island3的甲基化水平在巴马猪和藏猪中差异较小,而藏猪CpG island2的甲基化水平(42.61%)高于巴马猪(24.49%)。本研究绘制了2个猪种CpG island2甲基化水平的黑白点图,其中CpG位点为4、8、9、10、11、12、15,藏猪甲基化频率分别比巴马猪高28.26%、17.39%、26.09%、26.09%、26.09%、23.91%和34.78%。在CpG island2处预测到3个转录因子结合位点(SP2、PPARγ和EGR1)。结果表明,巴马猪和藏猪皮下脂肪组织中UCP3基因mRNA水平的表达差异可能是由于CpG island2的甲基化水平不同所导致,藏猪DNA甲基化水平在一定程度上阻碍了转录因子与启动子调控区域的结合,从而抑制了UCP3基因的表达。  相似文献   

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为揭示猪肌分化因子(myogenic differentiation 1,MyoD1)基因启动子区多态性,本试验分别以野猪×从江香猪二元杂交猪、杜×长×大外三元杂交猪及贵州宗地花猪为研究对象,采用DNA池和直接测序技术,筛选MyoD1基因5'UTR及部分第1外显子区SNP位点,利用生物信息学软件预测SNP位点对核心启动子区、CpG岛和转录因子结合位点的影响。结果表明,在MyoD1基因5'UTR及部分第1外显子区筛查到3个SNPs位点,分别为A-39G、T+150C和C+227G;生物信息学软件预测发现,A-39G位点附近出现重要转录因子结合位点消失和新位点生成;CpGIslandsearcher软件分析得到多态位点突变前后CpG岛大小及GC含量发生改变,据此推测猪MyoD1基因5'UTR区域的A-39G位点对调控启动子功能元件有重要影响。  相似文献   

14.
试验旨在检测5-羟色胺-N-乙酰基转移酶(AANAT)基因在绵羊休情季节和繁殖季节(卵泡期和黄体期)卵巢组织中的转录差异,并分析转录差异是否由DNA甲基化修饰程度改变所导致。试验采用自然环境条件和饲养管理一致,且体重差异在0.5 kg范围内的空怀母滩羊作为试验动物,采集其休情期、卵泡期和黄体期(每个时期3只)的卵巢组织,采用SYBR染料法进行实时荧光定量PCR检测AANAT基因在滩羊不同繁殖时期卵巢组织中的转录水平。随后针对转录水平有差异的两个时期(休情期和卵泡期)的样本,利用MethPrimer 2.0在线软件预测AANAT基因启动子区和第一外显子区的CpG岛;用重亚硫酸盐测序法(BSP法)检测AANAT基因启动子区及第一外显子区的甲基化程度。试验结果显示,滩羊休情期卵巢组织中AANAT基因转录水平显著低于卵泡期的AANAT基因转录水平(P<0.05),休情期与黄体期滩羊卵巢组织中AANAT基因的转录水平差异不显著(P>0.05)。滩羊卵巢组织中AANAT基因启动子区上存在着一个长度为173 bp的CpG岛,第一外显子区存在着一个长度为118 bp的CG岛。然而,两个甲基化岛区内的单个CpG位点甲基化程度在滩羊休情期和卵泡期之间均不存在显著差异,暗示AANAT基因的表达受甲基化修饰外的因素调控。本研究结果可为进一步探讨AANAT基因在季节性发情和卵泡成熟中的功能提供参考资料。  相似文献   

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为了探求核移植过程中DNA甲基化重编程是否充分,运用亚硫酸氢盐测序法分别检测新生死亡克隆猪和同期正常猪心脏、肝脏、脾脏、肺脏和肾脏组织中IGF2/H19基因印迹控制区(DMR1、DMR2、DMR3)的甲基化状态。结果发现,DMR1、DMR3在克隆猪和正常猪各组织中的甲基化水平不同,但差异不显著(P>0.05)。DMR2在克隆猪肺脏组织表现为超甲基化,极显著高于正常猪(P<0.01),且10个测序克隆中存在2处连续的全甲基化CpG位点(分别为4-9位和12-17位),而在其它组织中甲基化差异不显著(P>0.05)。说明DMR2在克隆猪肺脏组织可能存在DNA甲基化重编程紊乱,这也可能是导致该克隆猪死亡的因素之一。  相似文献   

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 控制马香槟毛色的CH(Champagne gene)基因家族包含4个候选基因(SLC36A1、SLC36A2、SLC36A3、SPARC),研究发现SLC36A1基因外显子2的突变是造成马香槟毛色的关键位点。为揭示中国马SLC36A1基因遗传多态性,本研究以玉树马和德保矮马共74个样本为研究对象,以马DNA池为模板扩增SLC36A1基因的10个外显子及部分内含子序列并进行测序分析。共发现马SLC36A1基因5个SNPs,分别位于内含子3(g.26699953 A>G, g.26699851 G>C, g.26699850 G>C),外显子4(g.26699562 G>A)及外显子6(g.26697018 C>T)。利用PCR RFLP方法对74个家马样本进行基因分型,发现外显子6的SNP有基因型CC、CT;外显子4的SNP有基因型GG、GA;内含子3的g.26699850 G>C突变有基因型GG、GC;内含子3的另外2个SNPs(g.26699953 A>G, g.26699851 G>C)通过测序,发现有AA、AG、GG与GG、GC、CC基因型。所有5个马SNPs均为野生型占主要优势。由此界定了马SLC36A1基因有9种单倍型(H1 H9),其中H5是最主要的单倍型。德保矮马遗传多样度为0.4190,比玉树马(0.2228)高,表明德保矮马香槟毛色遗传多态性比玉树马更丰富。玉树马与德保矮马的平均单倍型多样度为0.3160,表明其香槟毛色遗传多态性相对较低。  相似文献   

17.
Retained heterosis in F2 cows nursing F3 progeny was evaluated in 3-, 4-, and greater than or equal to 5-yr-old cows. Traits evaluated included milk yield at three stages of lactation and 200-d weight of progeny. Breed effects were evaluated in the nine parental breeds (Red Poll [R], Hereford [H], Angus [A], Limousin [L], Braunvieh [B], Pinzgauer [P], Gelbvieh [G], Simmental [S], and Charolais [C]) that contributed to the three composite populations (MARC I = 1/4 B, 1/4 C, 1/4 L, 1/8 H, 1/8 A; MARC II = 1/4 G, 1/4 S, 1/4 H, 1/4 A; and MARC III = 1/4 R, 1/4 P, 1/4 H, 1/4 A). Breed effects were significant for 12-h milk yield, estimated 200-d milk yield, and 200-d weight of progeny. Herefords were lowest (P less than .05) for 12-h milk yield and estimated 200-d milk yield, and Braunvieh produced significantly more milk than all breed groups except Pinzgauer and Simmental, for which the difference approached significance. The correlation among breed group means (nine parental breeds and three composites) for 12-h milk yield with 200-d weight of progeny was .91. When 200-d weight was adjusted to a common estimated 200-d milk yield, Hereford, Angus, Red Poll, and Limousin did not differ (P greater than .05); all were significantly lighter than Braunvieh, Pinzgauer, Gelbvieh, Simmental, and Charolais, which did not differ (P greater than .05) from each other.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

18.
It aimed to identify the differential methylation region (DMR) and differential methylation gene (DMG) through the analysis of the genome-wide methylation difference of the longest muscle of the sheep's back of different breeds of sheep,and laid the foundation for the analysis of the differences in sheep skeletal muscle development.In this study,one-year-old sheep(Tan sheep and Hu sheep and Tan-Hu F2) were sanned by the whole genome bisulfite sequencing method(WGBS).The degree of methylation and differentially methylated region in the whole genome DNA of longissimus dorsi(LD) muscle were studied to explore the difference of DNA methylation in level in sheep.The results showed that the methylation (mC) rates of cytosine (C) were 3.55%,3.18% and 3.56% in Tan sheep,Hu sheep and Tan-Hu F2,respectively.A total of 97 731 DMRs and 10 784 related DMGs were detected.In CG,CHH and CHG sequences,the methylation levels of the three groups were not significantly different.419 GO terms and 20 related signal pathways were detected by GO and KEGG analysis,respectively,which were showed to be significantly enriched in cellular process,cell part,binding,long-term depression,and so on.Five candidate genes,ACTA2、ROCK1、CALD1、MYH3 and MYH10 were sreened out in muscle tissues.This study provided genome-wide methylation of pattern of three groups (Tan sheep and Hu sheep and Tan-Hu F2).The results provided reference information for the epigenetic study of Tan sheep and screened candidate genes related to muscle development and meat quality.  相似文献   

19.
试验旨在检测5-羟色胺-N-乙酰基转移酶(AANAT)基因在绵羊休情季节和繁殖季节(卵泡期和黄体期)卵巢组织中的转录差异,并分析转录差异是否由DNA甲基化修饰程度改变所导致。试验采用自然环境条件和饲养管理一致,且体重差异在0.5 kg范围内的空怀母滩羊作为试验动物,采集其休情期、卵泡期和黄体期(每个时期3只)的卵巢组织,采用SYBR染料法进行实时荧光定量PCR检测AANAT基因在滩羊不同繁殖时期卵巢组织中的转录水平。随后针对转录水平有差异的两个时期(休情期和卵泡期)的样本,利用MethPrimer 2.0在线软件预测AANAT基因启动子区和第一外显子区的CpG岛;用重亚硫酸盐测序法(BSP法)检测AANAT基因启动子区及第一外显子区的甲基化程度。试验结果显示,滩羊休情期卵巢组织中AANAT基因转录水平显著低于卵泡期的AANAT基因转录水平(P<0.05),休情期与黄体期滩羊卵巢组织中AANAT基因的转录水平差异不显著(P>0.05)。滩羊卵巢组织中AANAT基因启动子区上存在着一个长度为173 bp的CpG岛,第一外显子区存在着一个长度为118 bp的CG岛。然而,两个甲基化岛区内的单个CpG位点甲基化程度在滩羊休情期和卵泡期之间均不存在显著差异,暗示AANAT基因的表达受甲基化修饰外的因素调控。本研究结果可为进一步探讨AANAT基因在季节性发情和卵泡成熟中的功能提供参考资料。  相似文献   

20.
中国6个地方鸡品种的母系起源   总被引:2,自引:2,他引:0  
通过对线粒体DNA(mtDNA)D-loop区的测序和比对,探讨了中国6个地方鸡品种的母系起源。结果表明,文昌鸡、鲁西斗鸡、寿光鸡、济宁百日鸡和莱芜黑鸡分别有5、5、5、6和7种单倍型,琅琊鸡只有3种单倍型;6个地方鸡品种聚为3个分支:分支A是1个主要分支,共有17种单倍型,有寿光鸡(10只)、鲁西斗鸡(9只)、文昌鸡(5只)、莱芜黑鸡(6只)、济宁百日鸡(6只)和琅琊鸡(2只),分别占所分析各地方鸡品种个体数的100%、90%、84%、75%、60%和20%,与分布于老挝、云南红原鸡的3个大陆亚种(G.g.gallus、G.g.jabouile和G.g.spadi-ceus)关系较近;分支B中包含8只琅琊鸡(80%)和1只济宁百日鸡(10%);分支C中有1只鲁西斗鸡(10%)、1只文昌鸡(16.7%)、2只莱芜黑鸡(25%)和3只济宁百日鸡(30%);分支B和C分别与来自红原鸡G.g.gallus亚种H19单倍型和H32、H33单倍型聚在一起。推测这6个地方鸡品种分别来自云南、老挝和越南附近地区的红原鸡大陆亚种。基因流是上述品种群体间遗传分化的主要因素。错配分布和Fu'sFs检验表明,分布于山东的5个地方鸡品种未发生群体扩张。  相似文献   

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