【作者】 俞亚波；

【导师】 王金玉；

【作者基本信息】 扬州大学 ， 动物遗传育种与繁殖， 2009， 博士

【摘要】 本试验用PCR-SSCP方法检测骨调素(Osteopontin, OPN)和钙调素(Calmodulin, CAM)基因的SNP。系统研究OPN和CAM基因SNP,并与京海黄鸡的生长性状、繁殖性状、300日龄蛋壳质量性状、屠宰性状和肉品质性状进行联合分析,旨在探索京海黄鸡重要经济性状早期分子标记辅助选择有效的遗传标记。研究结果如下:1.用PCR-SSCP扩增了OPN基因的全序列,在京海黄鸡的OPN基因上发现了20个SNP:A306T,A459C,G1367T,C1389G,G1399A,A1443G,T1463C,A1489G,G2219C, C2225T,G2257C,G2283C,A2288G,G3010A,G3159A,G3770A,C3830T,A3887G,A3918G和3217T的缺失导致的多态。2.在京海黄鸡群体里发现CAM的5个SNP:G56A , A57G和C96T(GenBank NO: M31605);T479C和G480A(GenBank NO: L00101)。3在OPN基因OP2座位上,h1h1型个体蛋形指数最小,显著小于h1h2型个体(P<0.05)。在公鸡群体中,h2h2基因型个体半净膛率和全净膛率显著高于h1h1基因型的个体(P<0.05)。h1h2型的个体胸肌率显著大于h2h2型的个体(P<0.05)。公鸡中h1h1型对腿肌系水力产生显著大的负向遗传贡献率,为-15.41%;h1h2型对腿肌系氺力产生显著大正向遗传贡献率，为11.82%。母鸡中h1h2型对腿肌剪切力产生显著大的正向遗传贡献率,为9.41%;h2h2型对腿肌剪切力产生显著大的负向遗传贡献率,为-22.78%。该基因座对京海黄鸡公鸡的半净膛率、全净膛率、胸肌率和腿肌系水力的MEI都大于10%;对母鸡的腿肌剪切力的MEI为12.46%。4. OPN基因OP5座位对母鸡的屠宰性状如:半净膛率、全净膛率、胸肌重和腿肌重有显著影响。对母鸡胸肌剪切力效应显著:GG和GT型对母鸡的胸肌剪切力产生显著大的遗传贡献率,分别为25.97%和-15.06%。OP5座位上,母鸡半净膛率、全净膛率、胸肌重、腿肌重和胸肌剪切力的MEI分别为13.25,15.54,13.91,18.42和16.03。5. OPN基因OP6座位对开产日龄和300日龄蛋数有显著影响:h2h2基因型对300日龄蛋数产生极显著大的负向遗传贡献率,为-9.26%。OP6座位对公母鸡的胸肌重有显著效应,MEI超过11。6. OPN基因OP9座位h2h5型对蛋壳强度有显著大的负向遗传贡献率,为-8.83%。OP9座位对母鸡的腹脂率、胸肌率、胸肌剪切力、胸肌系氺率和腿肌系水率有显著影响,相应的MEI分别为18.10,19.28,16.37,20.410和29.39。7. OPN基因OP12座位对京海黄鸡的生长性状有显著影响:h1h6是增加体重的劣势基因。OP12座位对公母鸡的活重和腿肌重有显著效应,MEI均超过11。在母鸡群体中,h3h3型个体的腹脂率最低,遗传贡献率为-19.72%。8. OPN基因OP15基因座对京海黄鸡的生长性状有显著影响:h5h5型是增加体重的劣势基因,h1h2型是增加体重的优势基因。但是h1h2型是增加产蛋数的劣势基因。h1h1型对壳钙重和壳钙百分率产生显著大的负向遗传贡献率,分别为-14.61%和-13.67%。OP15座位对母鸡的腹脂率有显著影响:h2h4型个体值显著高于h2h2型的个体,OP15基因座对母鸡腹脂率的MEI为12.44。9. CAM基因CA2座位对300日龄蛋数和66周龄蛋数有显著效应:h2h2型是增加产蛋数的劣势基因。H3h3型个体的蛋壳强度显著低于其他基因型,h3h3型对蛋壳强度产生极显著大的负向遗传贡献率,为-24.69%。CA2座位h1h3型对胸肌剪切力产生显著大的遗传贡献率,在公母鸡群体中的遗传贡献率分别为19.97和45.05;CA2座位对公母鸡胸肌剪切力的MEI分别为:21.35和33.24。10. CAM基因CA5座位不同基因型对的母鸡腹脂重有显著大的遗传贡献率,AA型对母鸡群体产生显著大的正向遗传贡献率,为32.5%; BB型对母鸡群体产生显著大的负向遗传贡献率,为-15.03%。11. OPN基因和CAM基因对京海黄鸡的0周龄体重、8周龄体重、12周龄体重、蛋壳重、蛋壳强度、蛋形指数、蛋壳百分率、成年体重、300日龄蛋数和66周龄蛋数的互作效应显著(P<0.05)或极显著(P<0.01)。更多还原

【Abstract】 PCR-SSCP method was used to study the SNP of OPN and CAM genes that might be correlated with reproduction traits and egg quality traits in Jinghai Yellow chicken, and the systematic investigation was conducted on Jinghai Yellow chicken OPN and CAM genes to evaluate the association of molecular markers with growth traits, reproduction traits, egg shell quality traits, carcass traits and meat quality traits.The main results are listed as follows:1. The full length DNA of OPN gene was obtained using the techniques of PCR-SSCP. 20 OPN SNPs were found in Jinghai Yelow chicken:A306T,A459C,G1367T,C1389G,G1399A,A1443G,T1463C,A1489G,G2219C, C2225T,G2257C,G2283C,A2288G , G3010A , G3159A , G3770A , C3830T , A3887G , A3918G, 3217T deletion/insertion。2. Five CAM SNPs were found in Jinghai Yellow chicken: G56A , A57G and C96T(GenBank NO: M31605);T479C and G480A(GenBank NO: L00101)。3. At the OP2 locus of OPN gene, birds with h1h1 genotype has the lowest value in egg shell index compared with h1h2 genotype (P<0.05). Birds with h2h2 genotype have significant higher value in semi-eviscerated weight ratio and eviscerated weight ratio than with h1h1 genotype in male birds, and birds with h1h2 has higher value in breast meat ratio than birds with h2h2 genotype (P<0.05). The contribute percentage (CP) of water holding capacity (WHC) in thigh meat with h1h1 genotype was -15.41%, and with h1h2 genotype was 11.82% in males. The contribute percentage of shear force in thigh meat with h1h2 genotype was 9.41%, and the CP with h2h2 genotype was -22.78% in females. The main effete index (MEI) of semi-eviscerated weight ratio, eviscerated weight ratio, breast meat ratio and WHC in thigh meat was all over 10% in males, and the MEI of shear force in thigh meat was 12.46.4. Significant differences were found between genotypes in OP5 locus and semi-eviscerated weight ratio, eviscerated weight ratio, breast meat weight and thigh meat weight in females. CP of GG and GT genotype were high of shear force in breast meat, and the values were 25.97% and -15.06% in females. The (MEI) of semi-eviscerated weight ratio, eviscerated weight ratio, breast meat ratio, thigh meat weight and shear force in thigh meat were 13.25%, 15.54, 13.91, 18.42 and 16.03 in females.5. Significant differences were found between genotypes in OP6 locus and age at first egg and egg numbers at 300 days. CP of h2h2 genotype was -9.26% of egg numbers at 300 day birds. The MEI of breast meat weight was over 11 in both sexes.6. At the OP2 locus of OPN gene, CP of h2h5 genotype was -8.83% of egg shell strength. The MEI of abdominal fat ratio, breast meat ratio, shear force in breast, WHC in breast meat and thigh meat were 18.10, 9.28, 16.37, 20.41 and 29.39 in females.7. Significant differences were found between genotypes in OP12 locus and growth traits: h1h6 genotype was the latest predominant genotype that increased body weight. The MEI of live weigh, and thigh meat weight were over 11% in both sexes. For abdominal fat ratio, CP of h3h3 genotype was -19.72% in females.8. Significant differences were found between genotypes in OP15 locus and growth traits: h5h5 genotype was the one lest predominant genotype that increased body weight, h1h2 genotype was the predominant genotype that increased body weight, but h1h2 genotype was the one lest predominant genotype that increased egg numbers. For calcium weight in egg shell and calcium ratio in egg shell, CP of h2h4 genotype was -14.61% and -13.67%. Significant differences were found between genotypes in OP15 locus and abdominal fat ratio: birds with h2h2 genotype have significant high value than with h2h2 genotype, and MEI of abdominal fat ratio was 12.44 in females.9. At the CA2 locus of CAM gene, significant differences were found between genotypes in and egg numbers at 300 days and egg numbers at 66 weeks: h2h2 genotype was the latest predominant genotype that increased egg numbers. Egg strength of h3h3 genotype has the lowest value than with other genotypes, and the CP was -24.69%. The MEI of live weigh, and thigh meat weight were over in both sexes. For abdominal fat ratio, CP of h3h3 genotype was -19.72% in females. CP of shear force in breast meat of h1h3 genotype was -19.97% and 45.05% in males and females respectively, and MEI of shear force in breast meat was 21.35 and 33.24 in males and females.10. At the CA5 locus of CAM gene, CP of abdominal fat ratio of AA and BB genotype was 32.50% and -15.03% respectively in females.11. Significant or very significant interaction effect of OPN and CAM were found on birth weight, body weight at 8 week, body weight at 12 week, egg shell weight, egg shell strength, egg shell index, egg shell percentage, grown up weight, eggs at 300 day and eggs at 66 week in Jinghai Yellow chicken更多还原