山东山羊品种血液蛋白多态性的研究
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摘要
本研究采用垂直板聚丙烯酰胺凝胶电泳法，对山东四个地方山羊品种(莱芜黑山羊、济宁青山羊、鲁北白山羊、崂山奶山羊)、波尔山羊及波黑、波奶、波鲁杂交后代血液的血红蛋白(Hb)、转铁蛋白(Tf)、血清碱性磷酸酶(Akp)、血清淀粉酶(Amy)、血清酯酶(Es)的多态性进行检测和分析，从生化遗传多态性方面分析各品种之间的遗传变异，并通过生产性能和血液理化指标的相关分析，为性状标记和连锁关系提供依据，依据品种之间的遗传距离大小不同来组合不同的杂交组合，对杂交后代及其纯种的生产性能进行统计计算，来预测杂交优势，从而为杂交亲本的选择提供简捷、可靠的理论依据。所得结果如下：
一、先对各品种体尺体重(体高、体长、胸围、管围、尻宽、尻长、体重)及血液常规指标(WBC、RBC、HGB、HCT、MCV、MCH、MCHC、W-SCR、W-LCR、W-SCC、W-LCC)进行测定，差异性分析表明：各品种间体尺体重和血液指标差异显著(P〈0.05)或极显著(P〈0.01)；相关性分析表明，体尺体重指标间除管围外相关显著(P〈0.05)或极显著(P〈0.01)，血液常规指标间有些相关不显著(P〉0.05)。
二、蛋白多态性研究结果表明：
1．所测4个地方山羊品种中，Tf、Akp及Es Ⅰ均表现出多态性，都具有两个等位基因，两种表现型，Hb只在奶山羊中表现出多态性，有两个等位基因，两种基因型；Es Ⅱ只在莱芜黑山羊中表现出遗传变异性；Amy未表现出多态性。在波尔山羊及杂交后代中，Tf在波黑、波鲁杂交一代中检测出两种基因型，波尔山羊及波鲁杂交二代具有三种基因型，波奶杂交山羊Tf只有TfAA型存在；Es Ⅰ在各品种中均具有两个等位基因，两种表现型；Akp在波奶杂交山羊中表现为无带型，其它山羊中受两个等位基因控制，两种表现型；Hb、Amy无多态性。
2．根据各基因频率和基因型频率进行计算，奶山羊Hb的Hb~A基因占绝对优势，频率为0.9667；各品种的Tf位点中Tf~A占优势，频率(顺序为LH、JO、LB、LN、B、B×LH、B×LN、B×LB_(F1)、B×LB_(F2)以下按此顺序)分别为0.9333、0.9500、0.9453、0.9583、0.7344、0.8000、1.0000、0.8889、0.7353；Akp位点中Akp~O的基因频率最高，分别为0.6708、0.7958、0.8292、0.9661、0.9574、0.9487、1.0000、0.9354、0.9393；Es Ⅰ在不同品种中基因频率不同，山东地方品种中Es Ⅰ~-占优势，频率分别(顺序为LH、JQ、LB、LN)为0.5323、0.5774、0.5728、0.6055，引进品种及杂交后代的Es Ⅰ~+占优势，频率分别(顺序为B、B×LH、B×LB_(F1)、B×LB_(F2))为0.5918、0.6465、0.5528、0.5149，B×LN的Es Ⅰ~+和Es Ⅰ~-频率相等均为0.5000；莱芜黑山羊的Es Ⅱ~+占优势，频率为0.7418。
3．所测9个品种类群中，纯种山羊崂山奶山羊的遗传变异最小，莱芜黑山羊的遗传变异最大，平均杂合度((?))分别为0.3547，0.3012、0.2892、0.1719、0.3238、
山东农业大学颐士学位臼文（2002）
0．3039、0．1667、O．2585、0．3343；等位基因有效数（Ne）为1．5497、1．43】0、1．4069、
！．2076、1．4789、1．4366、1．2000、1．3486、1．5022；多态座位百分比（Ppoly）为 0．6、
0．6、0．6、0．8、0．6、0二、0．6、0．6、0．6；Sllallllon信息指戳分别为0．7721、02702、
0．4259、0．45 15、0．6883、0．6686、0．3333、0．5953、0．6786．
4．所测五个纯种山羊基因分化系戮为0．026】4，这表明这几个品种的邀传变异
性的97．396％是由遗传多态现象引起的，而2．614％来自品种间的差异，即这几个山
羊品种在血液蛋白水平上的逅传分化程度不高。
5．根据Tf、Abp和Esl三个多态位点计算的Net氏遗传距离对四个地方品种进
行聚类分析，JQ和 LB遗传距离最近，其次为 LB和 LN，LH与 LN的逅传k＄最
0。
6．分析了 B X LH、B X LBFI和 B X LBfl的生产性能，B X oh和 B X e产生明
显的杂交优势（P＜0．05或P＜0．01），BXL巳；有一定杂交优势，但效果不明显（P＞0．05）。
7．不同血液蛋白型的血液常规指标和产羔数不同，产羔数差异不显著，血液常
规指标中，部分指标差异显著（P＜0．05或K0．01），大部分指标差异不显著（P＞0．05）。
The purpose of this study was to analyse the genetic polymorphism of blood protein of Shandong local goats (Laiwu black goat. lining grey goat. Lubei white goat, Laoshan milk goat), Boer goat and their filial generation. Five genetic markers hemoglobin, transferrin, serum alkaline phosphatase, serum amylase and serum easterase were detected by polyacrylamide gel electrophoresis method. Genetic variantions were analysed among the different breeds from the view of biochemical genetic polymorphism and was to provide scientific basis for trait marking and linkage relationship. Statistics and calculation of production performance of purebred and filial generation were used to predict heterosis so as to provide succinct and reliable theory basis for selection of crossing parents. The results were as following:
First: The body size and body weight index (Withers height, Body length. Heart girth. Cannon girth. Rump length, Rump width. Weight) and blood index (WBC. RBC, HGB, HCT.MCV.MCH,MCHC,W-SCR, W-LCR, W-SCC.W-LCC) of different breeds were measured and analysed. The results showed that there are significant difference (P<0.05 or P<0.01) on body size index and blood index among different breeds and there are significant correlation on body size index except Rump girth, but some blood index had no correlation.
Second: The blood protein polymorphism was studied, the results were as following
1. Polymorphism is found at the Tf, Akp and Esl loci of four local goats, all of which have two alleles and two phenotypes. Hb polymorphism with two alleles and two genotypes is found only in LN. Genetic variation of Es II is found only in LH. Amy is monomorphism. There are two genotypes in B X LH and B X LBpi and three in Boer goat and BXLBF2 at Tf loci. There are two alleles and two phenotypes at Esl loci of Boer goat and filial generation. Tf and Akp loci have no polymorphism in B X LN . There is two alleles and two phenotypes at Akp loci of Boer goat and other filial generation. Hb and Amy have no polymorphism.
2. The frequency of HbA of LN is 0.9667 which is the highest. At Tf loci, the
frequencies of TfA are the highest with the frequencies of (LH,JQ,LB,LN,B, B X LH,B X LN,B X LBF,,B X LBn,the same as below) 0.9333, 0.9500,0.9453, 0.9583, 0.7344, 0.8000, 1.0000,0.8889,0.7353 respectively. At Akp loci, the frequencies of Akp?are the highest with frequencies of 0.6708, 0.7958, 0.8292, 0.9661, 0.9574, 0.9487, 1.0000, 0.9354, 0.9393 respectively, The frequency of Esl is different in different breeds. The frequencies of Esl~ are the highest with frequencies of (LH, JQ, LB, LN) 0.5323, 0.5774, 0.5728. 0.6055 in local breeds while Esf are the highest with frequencies of (B, BXLH, B X LBF1,B X LBR) 0.5918, 0.5528. 0.5000. 0.6465, 0.5149 respectively in Boer goat and their filial generation. The frequency of EsPand EsPof BXLN are equal. The frequency of Es ir of LH is 0.7418 which is the highest.
3. Genetic variation of LN is the lowest but LH is the highest of purebred. The average gene heterozygosity ( H ) are 0.3547, 0.3012, 0.2892, 0.1719, 0.3238, 0.3039,0.1667.0.2585, 0.3343 and effective number of alleles (Ne) are 1.5497, 1.4310, 1.4069, 1.2076, 1.4789, 1.4366, 1.2000, 1.3486, 1.5022 respectively. The proportion of polymorphisms (Ppoly) are 0.6, 0.6, 0.6, 0.8, 0.6, 0.2, 0.6, 0.6, 0.6 respectively. Shannon information index are 0.7721, 0.2702, 0.4259, 0.4515. 0.6883, 0.6686,0.3333,0.5953, 0.6768 respectively.
4. The coefficient of genetic divergence among the five purebred goat is 0.02164.This indicated that 97.386% of genetic variations of all those five goat breeds resulted from the genetic polymorphism within breeds and 2.614% is caused by the difference among those five breeds.
5. Cluster analysis based on Nei genetic distance which were calculated according to the frequencies of Tf, Akp and Esl. The results showed that in four local breeds, the genetic distance between JQ and LB is closest, the genetic distance between LH and LN is smallest.
6.Individual production performance of B X L
一、先对各品种体尺体重(体高、体长、胸围、管围、尻宽、尻长、体重)及血液常规指标(WBC、RBC、HGB、HCT、MCV、MCH、MCHC、W-SCR、W-LCR、W-SCC、W-LCC)进行测定，差异性分析表明：各品种间体尺体重和血液指标差异显著(P〈0.05)或极显著(P〈0.01)；相关性分析表明，体尺体重指标间除管围外相关显著(P〈0.05)或极显著(P〈0.01)，血液常规指标间有些相关不显著(P〉0.05)。
二、蛋白多态性研究结果表明：
1．所测4个地方山羊品种中，Tf、Akp及Es Ⅰ均表现出多态性，都具有两个等位基因，两种表现型，Hb只在奶山羊中表现出多态性，有两个等位基因，两种基因型；Es Ⅱ只在莱芜黑山羊中表现出遗传变异性；Amy未表现出多态性。在波尔山羊及杂交后代中，Tf在波黑、波鲁杂交一代中检测出两种基因型，波尔山羊及波鲁杂交二代具有三种基因型，波奶杂交山羊Tf只有TfAA型存在；Es Ⅰ在各品种中均具有两个等位基因，两种表现型；Akp在波奶杂交山羊中表现为无带型，其它山羊中受两个等位基因控制，两种表现型；Hb、Amy无多态性。
2．根据各基因频率和基因型频率进行计算，奶山羊Hb的Hb~A基因占绝对优势，频率为0.9667；各品种的Tf位点中Tf~A占优势，频率(顺序为LH、JO、LB、LN、B、B×LH、B×LN、B×LB_(F1)、B×LB_(F2)以下按此顺序)分别为0.9333、0.9500、0.9453、0.9583、0.7344、0.8000、1.0000、0.8889、0.7353；Akp位点中Akp~O的基因频率最高，分别为0.6708、0.7958、0.8292、0.9661、0.9574、0.9487、1.0000、0.9354、0.9393；Es Ⅰ在不同品种中基因频率不同，山东地方品种中Es Ⅰ~-占优势，频率分别(顺序为LH、JQ、LB、LN)为0.5323、0.5774、0.5728、0.6055，引进品种及杂交后代的Es Ⅰ~+占优势，频率分别(顺序为B、B×LH、B×LB_(F1)、B×LB_(F2))为0.5918、0.6465、0.5528、0.5149，B×LN的Es Ⅰ~+和Es Ⅰ~-频率相等均为0.5000；莱芜黑山羊的Es Ⅱ~+占优势，频率为0.7418。
3．所测9个品种类群中，纯种山羊崂山奶山羊的遗传变异最小，莱芜黑山羊的遗传变异最大，平均杂合度((?))分别为0.3547，0.3012、0.2892、0.1719、0.3238、
山东农业大学颐士学位臼文（2002）
0．3039、0．1667、O．2585、0．3343；等位基因有效数（Ne）为1．5497、1．43】0、1．4069、
！．2076、1．4789、1．4366、1．2000、1．3486、1．5022；多态座位百分比（Ppoly）为 0．6、
0．6、0．6、0．8、0．6、0二、0．6、0．6、0．6；Sllallllon信息指戳分别为0．7721、02702、
0．4259、0．45 15、0．6883、0．6686、0．3333、0．5953、0．6786．
4．所测五个纯种山羊基因分化系戮为0．026】4，这表明这几个品种的邀传变异
性的97．396％是由遗传多态现象引起的，而2．614％来自品种间的差异，即这几个山
羊品种在血液蛋白水平上的逅传分化程度不高。
5．根据Tf、Abp和Esl三个多态位点计算的Net氏遗传距离对四个地方品种进
行聚类分析，JQ和 LB遗传距离最近，其次为 LB和 LN，LH与 LN的逅传k＄最
0。
6．分析了 B X LH、B X LBFI和 B X LBfl的生产性能，B X oh和 B X e产生明
显的杂交优势（P＜0．05或P＜0．01），BXL巳；有一定杂交优势，但效果不明显（P＞0．05）。
7．不同血液蛋白型的血液常规指标和产羔数不同，产羔数差异不显著，血液常
规指标中，部分指标差异显著（P＜0．05或K0．01），大部分指标差异不显著（P＞0．05）。
The purpose of this study was to analyse the genetic polymorphism of blood protein of Shandong local goats (Laiwu black goat. lining grey goat. Lubei white goat, Laoshan milk goat), Boer goat and their filial generation. Five genetic markers hemoglobin, transferrin, serum alkaline phosphatase, serum amylase and serum easterase were detected by polyacrylamide gel electrophoresis method. Genetic variantions were analysed among the different breeds from the view of biochemical genetic polymorphism and was to provide scientific basis for trait marking and linkage relationship. Statistics and calculation of production performance of purebred and filial generation were used to predict heterosis so as to provide succinct and reliable theory basis for selection of crossing parents. The results were as following:
First: The body size and body weight index (Withers height, Body length. Heart girth. Cannon girth. Rump length, Rump width. Weight) and blood index (WBC. RBC, HGB, HCT.MCV.MCH,MCHC,W-SCR, W-LCR, W-SCC.W-LCC) of different breeds were measured and analysed. The results showed that there are significant difference (P<0.05 or P<0.01) on body size index and blood index among different breeds and there are significant correlation on body size index except Rump girth, but some blood index had no correlation.
Second: The blood protein polymorphism was studied, the results were as following
1. Polymorphism is found at the Tf, Akp and Esl loci of four local goats, all of which have two alleles and two phenotypes. Hb polymorphism with two alleles and two genotypes is found only in LN. Genetic variation of Es II is found only in LH. Amy is monomorphism. There are two genotypes in B X LH and B X LBpi and three in Boer goat and BXLBF2 at Tf loci. There are two alleles and two phenotypes at Esl loci of Boer goat and filial generation. Tf and Akp loci have no polymorphism in B X LN . There is two alleles and two phenotypes at Akp loci of Boer goat and other filial generation. Hb and Amy have no polymorphism.
2. The frequency of HbA of LN is 0.9667 which is the highest. At Tf loci, the
frequencies of TfA are the highest with the frequencies of (LH,JQ,LB,LN,B, B X LH,B X LN,B X LBF,,B X LBn,the same as below) 0.9333, 0.9500,0.9453, 0.9583, 0.7344, 0.8000, 1.0000,0.8889,0.7353 respectively. At Akp loci, the frequencies of Akp?are the highest with frequencies of 0.6708, 0.7958, 0.8292, 0.9661, 0.9574, 0.9487, 1.0000, 0.9354, 0.9393 respectively, The frequency of Esl is different in different breeds. The frequencies of Esl~ are the highest with frequencies of (LH, JQ, LB, LN) 0.5323, 0.5774, 0.5728. 0.6055 in local breeds while Esf are the highest with frequencies of (B, BXLH, B X LBF1,B X LBR) 0.5918, 0.5528. 0.5000. 0.6465, 0.5149 respectively in Boer goat and their filial generation. The frequency of EsPand EsPof BXLN are equal. The frequency of Es ir of LH is 0.7418 which is the highest.
3. Genetic variation of LN is the lowest but LH is the highest of purebred. The average gene heterozygosity ( H ) are 0.3547, 0.3012, 0.2892, 0.1719, 0.3238, 0.3039,0.1667.0.2585, 0.3343 and effective number of alleles (Ne) are 1.5497, 1.4310, 1.4069, 1.2076, 1.4789, 1.4366, 1.2000, 1.3486, 1.5022 respectively. The proportion of polymorphisms (Ppoly) are 0.6, 0.6, 0.6, 0.8, 0.6, 0.2, 0.6, 0.6, 0.6 respectively. Shannon information index are 0.7721, 0.2702, 0.4259, 0.4515. 0.6883, 0.6686,0.3333,0.5953, 0.6768 respectively.
4. The coefficient of genetic divergence among the five purebred goat is 0.02164.This indicated that 97.386% of genetic variations of all those five goat breeds resulted from the genetic polymorphism within breeds and 2.614% is caused by the difference among those five breeds.
5. Cluster analysis based on Nei genetic distance which were calculated according to the frequencies of Tf, Akp and Esl. The results showed that in four local breeds, the genetic distance between JQ and LB is closest, the genetic distance between LH and LN is smallest.
6.Individual production performance of B X L
引文
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