青海三个类型藏羊血液生化遗传标记研究
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摘要
采用聚丙烯酰胺凝胶不连续垂直平板电泳、聚丙烯酰胺凝胶圆盘电泳和火焰光度法对青海三个类型藏羊(n=206):欧拉型藏羊(66只)、高原型藏羊(67只)和山谷型藏羊(73只)HB、ES、AMY、TF、S-LDH、RBC-LDH、AKP和EK 8个血液生化遗传标记的多态性进行了研究,同时对其遗传变异及亲缘关系进行了分析,得出如下结论:
1.在HB、ES、AMY、TF、S-LDH、RBC-LDH、AKP和EK 8个血液生化遗传标记中,HB、ES、AMY2、TF、AKP和EK存在多态性,而S-LDH和RBC-LDH两个位点未呈多态现象,均为单态。
2. HB位点:被检欧拉型藏羊HB位点受HB~A和HB~B两个等位基因的控制,出现HBAA、HBAB和HBBB三种基因型;而被检高原型和山谷型藏羊HB位点受HB~A、HB~B和HB~C三个等位基因的控制,出现HBAA、HBAB和HBAC三种基因型。且三个类型藏羊HB位点均以HB~A为优势等位基因和HBAA为优势基因型。
3.血清ES位点:被检三个类型藏羊血清ES位点均受ES~A和ES~a两个等位基因的控制,有ESA和ESa两种表型,三个类型藏羊ES位点均以ESA为优势表型和ESa为优势等位基因。
4.血清AMY:被检三个类型藏羊血清AMY有AMY1、AMY2和AMY3三种同工酶,但只有AMY2具有多态性;AMY2位点存在AMY2A和AMY20两种表型,均以AMY20为优势表型和AMY2~0为优势等位基因。
5.血清TF位点:被检三个类型藏羊血清TF位点具有丰富的多态性。欧拉型藏羊血清TF位点受TF~A、TF~B、TF~C、TF~D和TF~M 5个复等位基因控制,出现TFAB、TFAC、TFAD、TFBB、TFBC、TFBD、TFBM、TFCC和TFDD 9种基因型;高原型藏羊血清TF位点受TF~A、TF~B、TF~L、TF~C和TF~D 5个复等位基因控制,出现TFAC、TFBB、TFBL、TFBC、TFBD、TFCC和TFDD 7种基因型。山谷型藏羊血清TF位点受TF~A、TF~B、TF~L、TF~C、TF~D、TF~M和TF~E 7个复等位基因控制,出现TFAA、TFAB、TFAC、TFAD、TFBB、TFBL、TFBC、TFBD、TFBE、TFCC、TFCM、TFDD和TFDE 13种基因型。被检三个类型藏羊血清TF位点均以TF~B、TF~C和TF~D为优势等位基因,以TFBB、TFBC、TFBD和TFCC为优势基因型。
6. S-LDH和RBC-LDH:被检三个类型藏羊S-LDH出现LDH1、LDH2和LDH3三条区带。被检三个类型藏羊S-LDH呈单态。被检三个类型藏羊RBC-LDH仅有RBC-LDHⅠ一种表型,呈单态。
7.血清AKP:被检三个类型藏羊血清AKP经电泳分出A、B、C和D四种同工酶。但只有AKPB存在多态性,AKPB位点有AKPB和AKP0两种表型,受AKPB和AKP0一对等位基因控制;且均以AKP0为优势等位基因,被检欧拉型和高原型藏羊AKPB位点均以AKPB为优势表型,而被检山谷型藏羊则以AKP0为优势表型。
8. EK:被检欧拉型和高原型藏羊EK存在HK和LK两种表型,而被检山谷型藏羊只有HK一种表型。也即在被检欧拉型和高原型藏羊EK呈多态性,而被检山谷型藏羊EK为单态。被检三个类型藏羊EK均以HK为优势表型和Kh为优势等位基因。
9.被检三个类型藏羊的遗传变异:由被检血液生化遗传位点的平均杂合度( H )估计三个类型内遗传变异:欧拉型藏羊品种内遗传变异相对较大(0.2441),高原型藏羊次之,山谷型藏羊则小(0.2084)。对三个类型间遗传分化程度的分析结果认为,其类型内的平均杂合性(H_S)为0.2243,总群体的平均杂合性(H_T)为0.2495,类型间的平均杂合性(D_(ST))为0.0252,类型间的基因分化系数(G_(ST))为0.1010。
10.三个类型藏羊的亲缘关系:通过对三个类型藏羊的遗传距离的计算,并对其聚类分析结果表明:高原型和山谷型藏羊亲缘关系最近,高原型和欧拉型藏羊亲缘关系次之,山谷型和欧拉型藏羊亲缘关系最远。
Three types Tibetan sheep in Qinghai province(n=206)---Oura Tibetan sheep(66), Plateau Tibetan sheep(67), Mountain-Valley Tibetan sheep(73) were used to investigate the polymorphism of eight biochemical genetic markers in blood—hemoglobin(HB),serum esterase(ES), serum amylase(AMY), serum transferrin(TF), serum lactate dehydrogenase(S-LDH), red blood cell lactate dehydrogenase(RBC-LDH), serum alkaline phosphatase(AKP) and erythrocyte potassium phenotype(EK) by means of vertical non-denatured polyacrylamide gel electrophoresis(PAGE), polyacrylamide gel electrophoresis—disc and flame photometer. Meanwhile, the genetic variation and relationship among the three types Tibetan sheep in Qinghai province were analyzed. The results were as follows:
1. The polymorphism of HB, ES, AMY2, TF, AKP and EK was showed.Both S-LDH and RBC-LDH were simple.
2. There were three gene types examined which were HBAA, HBAB and HBBB controlled by two alleles including HBA and HBB on the loci of HB in Oura Tibetan sheep. There were three gene types including HBAA, HBAB and HBAC controlled by three alleles including HBA, HBB and HBC on the loci of HB in Plateau Tibetan sheep and Mountain-Valley Tibetan sheep examined. HBA was a dominant allele and HBAA was a preponderant gene type in the three types Tibetan sheep in Qinghai province.
3. There were two phenotypes including ESA and ESa controlled by two alleles including ESA and ESa on the loci of ES in the three types Tibetan sheep examined. And ESA was a dominant phenotype and ESa was a preponderant allele.
4. There were three isozymes of serum amylase: AMY1, AMY2 and AMY3, but only AMY2 has the polymorphism in the three types Tibetan sheep examined. There were two phenotypes including AMY2A and AMY20. AMY20 was dominant phenotype and AMY2~0 was preponderant allele in the three types Tibetan sheep examined.
5. On the loci of serum transferrin, there was abundant polymorphism. There were nine gene types including TFAB、TFAC、TFAD、TFBB、TFBC、TFBD、TFBM、TFCC and TFDD controlled by five alleles including TF~A、TF~B、TF~C、TF~D and TF~M on the loci of TF in Oura Tibetan sheep examined. There were seven gene types including TFAC、TFBB、TFBL、TFBC、TFBD、TFCC and TFDD controlled by five alleles including TF~A、TF~B、TF~L、TF~C and TF~D on the loci of TF in Plateau Tibetan sheep examined. There were thirteen gene types including TFAA、TFAB、TFAC、TFAD、TFBB、TFBL、TFBC、TFBD、TFBE、TFCC、TFCM、TFDD and TFDE controlled by seven alleles including TF~A、TF~B、TF~L、TF~C、TF~D、TF~M and TF~E on the loci of TF in Mountain-Valley Tibetan sheep examined. TFB、TFC and TFD were dominant alleles and TFBB、TFBC、TFBD and TFCC were preponderant gene types in the three types Tibetan sheep examined.
6. There were three zones including LDH1, LDH2 and LDH3 on the loci of S-LDH, and only RBC-LDH1 phenotype was showed on the loci of RBC-LDH in the three types Tibetan sheep examined. Both S-LDH and RBC-LDH were simple.
7. There were four isozymes of serum alkaline phosphatase including AKPA, AKPB, AKPC and AKPD, but only AKPB has the polymorphism in the three types Tibetan sheep examined. There were two phenotypes including AKPB and AKP0 controlled by a pair of alleles including AKPB and AKP0, and AKP0 was preponderant allele in the three types Tibetan sheep examined, and AKPB was a dominant phenotype in Oura Tibetan sheep and Plateau Tibetan sheep examined, but AKP0 was a preponderant phenotype in Mountain-Valley Tibetan sheep examined.
8. There were two phenotypes of EK including HK and LK in Oura Tibetan sheep and Plateau Tibetan sheep examined, but only one phenotype—HK in Mountain-Valley Tibetan sheep. HK was dominant phenotype and Kh was preponderant allele in the three types Tibetan sheep examined.
9. The genetic variability of three types Tibetan sheep examined: The degree of genetic variability estimated by mean heterozygosity for examined biochemical genetic markers in blood was relatively higher in Oura Tibetan sheep (0.2441) than in Mountain-Valley Tibetan sheep (0.2084). The average genetic diversity (H_S) and that in the total population (H_T) are 0.2243 and 0.2495 respectively. The average genetic diversity among three sheep breeds (D_(ST)) is 0.0252; the gene differentiation coefficient of total sheep population (G_(ST)) is 0.1010.
10. relationship of three types Tibetan sheep: the results of genetic distance obtained using Nei’s formula among three types Tibetan sheep and cluster analysis showed: The tree grouped Plateau Tibetan sheep and Mountain-Valley Tibetan sheep in one branch, and then grouped Oura Tibetan sheep.
1.在HB、ES、AMY、TF、S-LDH、RBC-LDH、AKP和EK 8个血液生化遗传标记中,HB、ES、AMY2、TF、AKP和EK存在多态性,而S-LDH和RBC-LDH两个位点未呈多态现象,均为单态。
2. HB位点:被检欧拉型藏羊HB位点受HB~A和HB~B两个等位基因的控制,出现HBAA、HBAB和HBBB三种基因型;而被检高原型和山谷型藏羊HB位点受HB~A、HB~B和HB~C三个等位基因的控制,出现HBAA、HBAB和HBAC三种基因型。且三个类型藏羊HB位点均以HB~A为优势等位基因和HBAA为优势基因型。
3.血清ES位点:被检三个类型藏羊血清ES位点均受ES~A和ES~a两个等位基因的控制,有ESA和ESa两种表型,三个类型藏羊ES位点均以ESA为优势表型和ESa为优势等位基因。
4.血清AMY:被检三个类型藏羊血清AMY有AMY1、AMY2和AMY3三种同工酶,但只有AMY2具有多态性;AMY2位点存在AMY2A和AMY20两种表型,均以AMY20为优势表型和AMY2~0为优势等位基因。
5.血清TF位点:被检三个类型藏羊血清TF位点具有丰富的多态性。欧拉型藏羊血清TF位点受TF~A、TF~B、TF~C、TF~D和TF~M 5个复等位基因控制,出现TFAB、TFAC、TFAD、TFBB、TFBC、TFBD、TFBM、TFCC和TFDD 9种基因型;高原型藏羊血清TF位点受TF~A、TF~B、TF~L、TF~C和TF~D 5个复等位基因控制,出现TFAC、TFBB、TFBL、TFBC、TFBD、TFCC和TFDD 7种基因型。山谷型藏羊血清TF位点受TF~A、TF~B、TF~L、TF~C、TF~D、TF~M和TF~E 7个复等位基因控制,出现TFAA、TFAB、TFAC、TFAD、TFBB、TFBL、TFBC、TFBD、TFBE、TFCC、TFCM、TFDD和TFDE 13种基因型。被检三个类型藏羊血清TF位点均以TF~B、TF~C和TF~D为优势等位基因,以TFBB、TFBC、TFBD和TFCC为优势基因型。
6. S-LDH和RBC-LDH:被检三个类型藏羊S-LDH出现LDH1、LDH2和LDH3三条区带。被检三个类型藏羊S-LDH呈单态。被检三个类型藏羊RBC-LDH仅有RBC-LDHⅠ一种表型,呈单态。
7.血清AKP:被检三个类型藏羊血清AKP经电泳分出A、B、C和D四种同工酶。但只有AKPB存在多态性,AKPB位点有AKPB和AKP0两种表型,受AKPB和AKP0一对等位基因控制;且均以AKP0为优势等位基因,被检欧拉型和高原型藏羊AKPB位点均以AKPB为优势表型,而被检山谷型藏羊则以AKP0为优势表型。
8. EK:被检欧拉型和高原型藏羊EK存在HK和LK两种表型,而被检山谷型藏羊只有HK一种表型。也即在被检欧拉型和高原型藏羊EK呈多态性,而被检山谷型藏羊EK为单态。被检三个类型藏羊EK均以HK为优势表型和Kh为优势等位基因。
9.被检三个类型藏羊的遗传变异:由被检血液生化遗传位点的平均杂合度( H )估计三个类型内遗传变异:欧拉型藏羊品种内遗传变异相对较大(0.2441),高原型藏羊次之,山谷型藏羊则小(0.2084)。对三个类型间遗传分化程度的分析结果认为,其类型内的平均杂合性(H_S)为0.2243,总群体的平均杂合性(H_T)为0.2495,类型间的平均杂合性(D_(ST))为0.0252,类型间的基因分化系数(G_(ST))为0.1010。
10.三个类型藏羊的亲缘关系:通过对三个类型藏羊的遗传距离的计算,并对其聚类分析结果表明:高原型和山谷型藏羊亲缘关系最近,高原型和欧拉型藏羊亲缘关系次之,山谷型和欧拉型藏羊亲缘关系最远。
Three types Tibetan sheep in Qinghai province(n=206)---Oura Tibetan sheep(66), Plateau Tibetan sheep(67), Mountain-Valley Tibetan sheep(73) were used to investigate the polymorphism of eight biochemical genetic markers in blood—hemoglobin(HB),serum esterase(ES), serum amylase(AMY), serum transferrin(TF), serum lactate dehydrogenase(S-LDH), red blood cell lactate dehydrogenase(RBC-LDH), serum alkaline phosphatase(AKP) and erythrocyte potassium phenotype(EK) by means of vertical non-denatured polyacrylamide gel electrophoresis(PAGE), polyacrylamide gel electrophoresis—disc and flame photometer. Meanwhile, the genetic variation and relationship among the three types Tibetan sheep in Qinghai province were analyzed. The results were as follows:
1. The polymorphism of HB, ES, AMY2, TF, AKP and EK was showed.Both S-LDH and RBC-LDH were simple.
2. There were three gene types examined which were HBAA, HBAB and HBBB controlled by two alleles including HBA and HBB on the loci of HB in Oura Tibetan sheep. There were three gene types including HBAA, HBAB and HBAC controlled by three alleles including HBA, HBB and HBC on the loci of HB in Plateau Tibetan sheep and Mountain-Valley Tibetan sheep examined. HBA was a dominant allele and HBAA was a preponderant gene type in the three types Tibetan sheep in Qinghai province.
3. There were two phenotypes including ESA and ESa controlled by two alleles including ESA and ESa on the loci of ES in the three types Tibetan sheep examined. And ESA was a dominant phenotype and ESa was a preponderant allele.
4. There were three isozymes of serum amylase: AMY1, AMY2 and AMY3, but only AMY2 has the polymorphism in the three types Tibetan sheep examined. There were two phenotypes including AMY2A and AMY20. AMY20 was dominant phenotype and AMY2~0 was preponderant allele in the three types Tibetan sheep examined.
5. On the loci of serum transferrin, there was abundant polymorphism. There were nine gene types including TFAB、TFAC、TFAD、TFBB、TFBC、TFBD、TFBM、TFCC and TFDD controlled by five alleles including TF~A、TF~B、TF~C、TF~D and TF~M on the loci of TF in Oura Tibetan sheep examined. There were seven gene types including TFAC、TFBB、TFBL、TFBC、TFBD、TFCC and TFDD controlled by five alleles including TF~A、TF~B、TF~L、TF~C and TF~D on the loci of TF in Plateau Tibetan sheep examined. There were thirteen gene types including TFAA、TFAB、TFAC、TFAD、TFBB、TFBL、TFBC、TFBD、TFBE、TFCC、TFCM、TFDD and TFDE controlled by seven alleles including TF~A、TF~B、TF~L、TF~C、TF~D、TF~M and TF~E on the loci of TF in Mountain-Valley Tibetan sheep examined. TFB、TFC and TFD were dominant alleles and TFBB、TFBC、TFBD and TFCC were preponderant gene types in the three types Tibetan sheep examined.
6. There were three zones including LDH1, LDH2 and LDH3 on the loci of S-LDH, and only RBC-LDH1 phenotype was showed on the loci of RBC-LDH in the three types Tibetan sheep examined. Both S-LDH and RBC-LDH were simple.
7. There were four isozymes of serum alkaline phosphatase including AKPA, AKPB, AKPC and AKPD, but only AKPB has the polymorphism in the three types Tibetan sheep examined. There were two phenotypes including AKPB and AKP0 controlled by a pair of alleles including AKPB and AKP0, and AKP0 was preponderant allele in the three types Tibetan sheep examined, and AKPB was a dominant phenotype in Oura Tibetan sheep and Plateau Tibetan sheep examined, but AKP0 was a preponderant phenotype in Mountain-Valley Tibetan sheep examined.
8. There were two phenotypes of EK including HK and LK in Oura Tibetan sheep and Plateau Tibetan sheep examined, but only one phenotype—HK in Mountain-Valley Tibetan sheep. HK was dominant phenotype and Kh was preponderant allele in the three types Tibetan sheep examined.
9. The genetic variability of three types Tibetan sheep examined: The degree of genetic variability estimated by mean heterozygosity for examined biochemical genetic markers in blood was relatively higher in Oura Tibetan sheep (0.2441) than in Mountain-Valley Tibetan sheep (0.2084). The average genetic diversity (H_S) and that in the total population (H_T) are 0.2243 and 0.2495 respectively. The average genetic diversity among three sheep breeds (D_(ST)) is 0.0252; the gene differentiation coefficient of total sheep population (G_(ST)) is 0.1010.
10. relationship of three types Tibetan sheep: the results of genetic distance obtained using Nei’s formula among three types Tibetan sheep and cluster analysis showed: The tree grouped Plateau Tibetan sheep and Mountain-Valley Tibetan sheep in one branch, and then grouped Oura Tibetan sheep.
引文
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