摘要
山东省地处黄河下游和黄淮海农区,现有崂山奶山羊、济宁青山羊、鲁北白山羊和沂蒙黑山羊等地方品种,新发现的莱芜黑山羊和牙山黑绒山羊,引进的波尔山羊及其杂交后代,共同组成了其山羊遗传资源。本试验测定了7个同域山羊种群(莱芜黑山羊(LBG)、济宁青山羊(JBG)、崂山奶山羊(LMG)、牙山黑绒山羊(YBG)、鲁北白山羊(LWG)、波尔山羊(BG)和杂交羊(HG))的mtDNA D-loop高变区部分序列,确立了不同单倍型。通过计算单倍型多样度、核苷酸多样度、群体间核苷酸分歧度、群体间净遗传距离、群体平均核苷酸差异数等参数及遗传变异AMOVA分析,研究了不同群体的遗传结构和遗传多样性水平;利用各单倍型构建了系统发育树,探讨了该区域山羊群体的起源,并对得到的几个分支进行了Mismatch分析。具体结果如下:
1、在59只个体的mtDNA D-loop高变区序列中,发现崂山奶山羊品种的一条序列存在长度为5bp的片段缺失,很可能为一串联重复单位。其余58条序列均为533bp,发现了59个多态位点,其中单一多态位点22个,占总序列的4.13%,简约信息位点37个,占总序列的6.94%。所有多态位点中出现1处颠换(100位),1处转换与颠换共存(42位),其余均为转换,具有较高的转换偏倚。
2、根据59个多态位点确立了37种单倍型,共享单倍型有H16、H21、H24和H33四种,其余33种单倍型均为各群体所特有,单倍型多样度为0.9540。Mismatch分析发现差异10个碱基的单倍型最多。
3、所有群体单倍型多样度在0.7857～0.9697之间,核苷酸多样度在0.12716～0.17170之间,表明遗传多样性水平较高。AMOVA的变异组分剖分发现,群体内变异方差组分占94.87%,远大于群体间的5.13%,说明群体分化不明显。核苷酸分歧度在1.336%～1.951%,净遗传距离在0.015%～0.665%之间,通过Da构建NJ树发现杂交羊、济宁青山羊、莱芜黑山羊、鲁北白亲缘关系较近,聚为一支,而牙山黑山羊和崂山奶山羊亲缘关系较近,聚为另一支,支持将莱芜黑山羊与牙山黑山羊划分为两个不同的品种,并及时加以保护。
4、系统发育树将37种单倍型划分为两个较大的分支,角猾羊1条序列聚入A分支,而捻角山羊则自成1支。A分支为7个群体的优势类型,B分支的影响相对较小。
5、对A、B两种类型进行Mismatch分析,发现A类型经历过群体扩张,群体扩张时间较早,B类型未经历过群体扩张。
Shandong province lies in the lower region of the Yellow River and in the farmland of the Yellow River,Huai River and Hai River.Indigenous breeds such as Laoshan milk goat and Yimeng black goat,together with new-finding Laiwu black goat,Yashan black goat and imported Boer goat as well as its hybridized descendent comprised the goat resource of the area.The genetic diversity of 6 indigenous and 1 imported goat populations(Laiwu black goat(LBG),Jining blue goat(JBG),Laoshan milk goat(LMG),Yashan black goat(YBG),Lubei white goat(LWG),Beor goat(BG) and hybridized goat(HG)) was evaluated by sequencing partial sequence of mtDNA D-loop,and defined different haplotypes. The haplotype diversity,nucleotide diversity,average nucleotide substitution per site between population,net genetic distance between population,average nucleotide difference within population were computed to evaluate the genetic structure and level of genetic diversity;meanwhile,the genetic variance was analyzed through AMOVA;dendrogram was constructed to evaluate the origin of the goat.Mismatch analysis was dealed in the resulted lineages. The results were as follows:
1. Among all the 59 sequences,a 5 bp fragment deletion was detected in one sequence from LMG,while the other sequences were 533bp long. All the sequences defined 59 polymorphic sites, among which singleton polymorphic sites are 22, parsimony informative sites are 37, account for 4.13%,6.97% of the whole sequence, respectively. Their were 1 transversion(site 100) and 1 coexistent site of transition and transversion(site 42),the others were all transition, which showed high ratio of transition to transversion.
2.59 polymorphic sites defined 37 haplotypes, population shared haplotypes were H16、H21、H24 and H33, the others were private haplotype within population, whole haplotype diversity was 0.9540.Mismatch analysis showed that 10 nucleotides differences between haplotypes was the highest.
3. Haplotype diversity and nucleotide diversity of the populations were 0.7857~0.9697 and 0.12716~0.17170 respectively, which suggested high genetic diversity. AMOVA analysis shows variation percentage within population was 94.87 % ,and among population was 5.13 % ,population differentiation was not clear. Net genetic distance between population was from 0.015%~0.665%,while the nucleotide divergence was form1.336% to 1.951%.By constructing NJ tree with Da we find that HG、JBG、LBG、and LWG clustered together, they might have close relative relationship, YBG and LMG clustered together, which shows close relationship, too. Thus LBG and YBG should be divided to two different breeds, and emergent protection is needed to YBG..
4. Dendrogram divided 37 haplotypes into two major clusters(lineage A and B), capra aegagrus clustered into lineage A, while capra falconeri clustered into other lineage. Lineage A is the predominance lineage of the 7 goat populations, while lineage B has a relatively small proportion.
5. Mismatch analysis of lineage A and B showed that lineage A experienced population expansion, and the expansion time was relatively early, while lineage B didn’t experience population expansion.
引文
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