Mitochondrial DNA analysis of ancient Sampula population in Xinjiang

The archaeological site of Sampula cemetery was located about 14 km to the southwest of the Luo County in Xinjiang Khotan, China, belonging to the ancient Yutian kingdom. ¹⁴C analysis showed that this cemetery was used from 217 B.C. to 283 A.D. Ancient DNA was analyzed by 364 bp of the mitochondrial DNA hypervariable region I (mtDNA HVR-I), and by six restriction fragment length polymorphism (RFLP) sites of mtDNA coding region. We successfully extracted and sequenced intact stretches of maternally inherited mtDNA from 13 out of 16 ancient Sampula samples. The analysis of mtDNA haplogroup distribution showed that the ancient Sampula was a complex population with both European and Asian characteristics. Median joining network of U3 sub-haplogroup and multi-dimensional scaling analysis all showed that the ancient Sampula had maternal relationship with Ossetian and Iranian.     

内蒙古砧子山墓地古人的线粒体DNA多态性分析

为了研究内蒙古元上都遗址砧子山墓地古代人群的遗传结构及其可能来源,对该墓地古人的DNA进行了抽提、扩增和测序,获得了10个个体的线粒体DNA高可变一区序列.结合现代东亚、北亚、中亚和欧洲人的线粒体DNA数据进行了系统发育分析和多维尺度分析.研究结果表明:埋藏在砧子山墓地的元代居民为汉族人,主要是来自中国北方地区的汉族.本研究为揭示元代的复杂社会结构和人群历史动态提供了新的方法.     

饮牛沟墓地古人骨线粒体DNA的研究

对内蒙古饮牛沟战国时期墓地的古代人群（Yng古代人群）进行分子生物学研究, 获得了线粒体高可变一区DNA序列, 初步确定了单倍型归属并搜寻其共享序列, 与现代人群对比构建系统发育树和多维尺度分析. 结果表明, 饮牛沟古代人群与现代东亚人群在母系遗传关系上较近.     

Mitochondrial DNA analysis of Bronze Age horses recovered from Chifeng region, Inner Mongolia, China

In this study, mitochondrial DNA (mtDNA) analysis was carried out on 9 Bronze Age horses recovered from Dashanqian and Jinggouzi archaeological sites in Chifeng region, Inner Mongolia. China to explore the origin of Chinese domestic horses. Both mtDNA 16S rRNA gene and control region (D-loop) fragments of ancient horses were amplified and sequenced. The analysis of the highly conservative 16S rRNA gene sequences indicated that the burial environment of Chifeng region is suitable for the preservation of ancient DNA (aDNA). Combing 465 mtDNA D-loop sequences representing different breeds from East Asia, Central Asia, Near East and Europe, we constructed a phylogenetic network to investigate the relationship between ancient and modern horses. The phylogenetic network showed that the 9 horses were distributed into different modern horse clusters which were closely related to them representing a certain geographical distribution. Our results showed that the maternal genetic line of the ancient horses in Chifeng region was highly diversified, which contributed to the gene pool of modern domestic horses and suggested a complex origin of domestic horses in China.     

Analysis of the matrilineal genetic structure of population in the early Iron Age from Tarim Basin, Xinjiang, China

Ancient mtDNA data of human remains were analyzed from four early Iron Age Tarim Basin sites （Yuansha, Zaghunluq, Sampula and Niya） in the southern Silk Road region. Haplogroup distributions show that ancient Tarim Basin population was comprised of well-differentiated Western and Eastern matrilineal lineages. Some West lineage of Tarim Basin population originated from Near East and Iran region. Of the East lineages, North and Northeast Asia originated lineages were the main components, and a few Southeast Asian lineages also existed, which indicated a more extensive origin and a more complex admixture. The genetic structure of ancient Tarim Basin population is relatively close to the modern populations of Xinjiang, which implied that the early Iron Age is an important period during the formation of the modern Xinjiang population.     

Mitochondrial DNA analysis of Bronze Age horses recovered from Chifeng region, Inner Mongolia, China

In this study, mitochondrial DNA (mtDNA) analysis was carried out on 9 Bronze Age horses recovered from Dashanqian and Jinggouzi archaeological sites in Chifeng region, Inner Mongolia. China to explore the origin of Chinese domestic horses. Both mtDNA 16S rRNA gene and control region (D-loop) fragments of ancient horses were amplified and sequenced. The analysis of the highly conservative 16S rRNA gene sequences indicated that the burial environment of Chifeng region is suitable for the preservation of ancient DNA (aDNA). Combing 465 mtDNA D-loop sequences representing different breeds from East Asia, Central Asia, Near East and Europe, we constructed a phylogenetic network to investigate the relationship between ancient and modern horses. The phylogenetic network showed that the 9 horses were distributed into different modern horse clusters which were closely related to them representing a certain geographical distribution. Our results showed that the maternal genetic line of the ancient horses in Chifeng region was highly diversified, which contributed to the gene pool of modern domestic horses and suggested a complex origin of domestic horses in China.     

Pitfalls in the analysis of ancient human mtDNA

The retrieval of DNA from ancient human specimens is not always successful owing to DNA deterioration and contamination although it is vital to provide new insights into the genetic structure of ancient people and to reconstruct the past history. Normally, only short DNA fragments can be retrieved from the ancient specimens. How to identify the authenticity of DNA obtained and to uncover the information it contained are difficult. We employed the ancient mtDNAs reported from Central Asia (including Xinjiang, China) as an example to discern potentially extraneous DNA contamination based on the updated mtDNA phylogeny derived from mtDNA control region, coding region, as well as complete sequence information. Our results demonstrated that many mtDNAs reported are more or less problematic.Startim, from a reliable mtDNA phylogeney and combining the available modern data into analysis, one can ascertain the authenticity of the ancient DNA, distinguish the potential errors in a data set, and efficiently decipher the meager information it harbored. The reappraisal of the mtDNAs with the age of more than 2000 years from Central Asia gave support to the suggestion of extensively (pre)historical gene admixture in this region.     

内蒙古和林格尔东周时期古代人群的分子遗传学分析

对10个古代个体的线粒体DNA高可变Ⅰ区进行了扩增和测序. 基于和林格尔古代人群与现今相关欧亚人群的mtDNA序列， 进行了系统发育分析和多维尺度分析. 研究结果表明, 和林格尔古代人群在母系遗传上与现在北亚人群的亲缘关系最近. 结合考古学、 人类学以及分子生物学的研究， 推断这个古代人群是从蒙古高原以及外贝加尔地区南下迁移至今天的内蒙古和林格尔地区的游牧人.     

一种简便高效的古代动物毛皮DNA提取方法

采用试剂盒QIAamp DNA Mini Kit从距今4000年前新疆小河墓地出土的黄牛毛皮中提取出古DNA,并对黄牛线粒体D-loop区部分片段进行了PCR扩增和序列测定.结果表明:所提取的古DNA真实可信;且该方法简便、高效,适用于古代动物毛皮DNA的提取.     

新疆察吾呼沟古代居民线粒体DNA序列多态性分析

从距今2 500～3 000年的新疆察吾呼沟墓地的9例古代人骨中成功地提取出古DNA分 子. 用4对套叠引物对线粒体DNA高可变Ⅰ区进行了PCR扩增和测序, 得到9个364　bp的线粒 体序列. 从GenBank搜索其共享序列并与欧洲、 亚洲序列进行系统发育分析, 结果表明, 早在青铜至早期铁器时代, 在我国新疆天山中部地区已经有蒙古人种存在. 察吾呼沟古代 居民应是一个欧洲和东亚人种混合的古代群体.     

Molecular biological analysis of remains from Jiangjungou Cemetery in Inner Mongolia

The 364-bp nucleotide sequence in hypervariable region I (HVRI) of mitochondrial DNA is successfully amplified from 9 out of 11 individuals of an ancient population buried in the Jiangjungou Cemetery in Inner Mongolia dated back to the Warring States Period in China. Nine fragments with different variations are obtained. A phylogenetic tree and a multidimensional scaling (MDS) plot are constructed using mtDNA sequences from the ancient population and several modern Asian populations. The results show that ancient population shares a closer genetic relationship with East Asian populations than with North and Central Asian populations. The genetic and historical evidence raise the possibility that the population might be the immigrants from Zhongyuan area, sent by the King Wuling of Zhao State to guard the nation against the attack from Huns.     

New ancient DNA sequences suggest high genetic diversity for the woolly mammoth (Mammuthus primigenius)

Partial DNA sequences of cytochrome b gene (mtDNA) were successfully retrieved from Late Pleistocene fossil bone of Mammuthus primigenius collected from the Xiguitu County (Yakeshi), Inner Mongolia Autonomous Region and from Zhaodong, Harbin of Heilongjiang Province in northern China. Two ancient DNA fragments (109 bp and 124 bp) were authenticated by reproducible experiments in two different laboratories and by phylogenetic analysis with other Elephantidae taxa. Phylogenetic analysis using these sequences and published data in either separate or combined datasets indicate unstable relationship among the woolly mammoth and the two living elephants, Elephas and Loxodonta. In addition to the short sequences used to attempt the long independent evolution of Elephantidae terminal taxa, we suggest that a high intra-specific diversity existed in Mammuthus primigenius crossing both spatial and temporal ranges, resulting in a complex and divergent genetic background for DNA sequences so far recovered. The high genetic diversity in the extinct woolly mammoth can explain the apparent instability of Elephantidae taxa on the molecular phylogenetic trees and can reconcile the apparent paradox regarding the unresolved Elephantidae trichotomy.     

New ancient DNA sequences suggest high genetic diversity for the woolly mammoth (Mammuthus primigenius)

Partial DNA sequences of cytochrome b gene (mtDNA) were successfully retrieved from Late Pleistocene fossil bone of Mammuthus primigenius collected from the Xiguitu County ( Yakeshi), Inner Mongolia Autonomous Region and from Zhaodong, Harbin of Heilongjiang Province in northern China. Two ancient DNA fragments (109 bp and 124 bp) were authenticated by reproducible experiments in two different laboratories and by phylogenetic analysis with other Elephantidae taxa. Phylogenetic analysis using these sequences and published data in either separate or combined datasets indicate unstable relationship among the woolly mammoth and the two living elephants, Elephas and Loxodonta . In addition to the short sequences used to attempt the long independent evolution of Elephantidae terminal taxa, we suggest that a high intra-specific diversity existed in Mammuthus primigenius crossing both spatial and temporal ranges, resulting in a complex and divergent genetic background for DNA sequences so far recovered. The high genetic diversity in the extinct woolly mammoth can explain the apparent instability of Elephantidae taxa on the molecular phylogenetic trees and can reconcile the apparent paradox regarding the unresolved Elephantidae trichotomy.     

New ancient DNA sequences suggest high genetic diversity for the woolly mammoth (Mammuthus primigenius)

Partial DNA sequences of cytochrome b gene (mtDNA) were successfully retrieved from Late Pleistocene fossil bone of Mammuthus primigenius collected from the Xiguitu County (Yakeshi), Inner Mongolia Autonomous Region and from Zhaodong, Harbin of Heilongjiang Province in northern China. Two ancient DNA fragments (109 bp and 124 bp) were authenticated by reproducible experiments in two different laboratories and by phylogenetic analysis with other Elephantidae taxa. Phylogenetic analysis using these sequences and published data in either separate or combined datasets indicate unstable relationship among the woolly mammoth and the two living elephants, Elephas and Loxodonta. In addition to the short sequences used to attempt the long independent evolution of Elephantidae terminal taxa, we suggest that a high intra-specific diversity existed in Mammuthus primigenius crossing both spatial and temporal ranges, resulting in a complex and divergent genetic background for DNA sequences so far recovered. The high genetic diversity in the extinct woolly mammoth can explain the apparent instability of Elephantidae taxa on the molecular phylogenetic trees and can reconcile the apparent paradox regarding the unresolved Elephantidae trichotomy.     

新疆扎滚鲁克古代人群的线粒体DNA分析

从15例扎滚鲁克古代人牙齿样本中提取扩增线粒体DNA, 确定其单倍型归属, 并结合相关现代人群进行了系统发育分析和多维尺度分析. 结果表明,扎滚鲁克古代人群与现代中亚人群有较近的母系遗传关系, 是欧亚混合人群. 与以前的新疆古代人群研究相比, 该人群东部欧亚序列的成分明显增多, 这可能与人群所处的地理及历史时代相关.     

淡水鱼类线粒体DNA D-loop基因的引物设计和应用

 线粒体DNA测序已广泛应用于鉴定和区分种类以及解决系统进化关系问题。本文选取已测定的主要淡水鱼类的线粒体DNA D-loop基因序列进行同源性比较,寻找保守序列,利用简并性原则设计一对通用的简并引物。利用设计的引物对广东省珠江流域主要的淡水鱼类线粒体DNA D-loop控制区基因进行扩增,均能获得单一的目的DNA片断,特异性扩增产物大小为1 kb左右。经测序及与GenBank同源序列的比较,证实为包含线粒体控制区全序列的扩增产物。本研究所设计的引物和应用的方法可以快速地同时对多种鱼类进行大规模的遗传背景分析,鉴定某些难于鉴别的近缘物种,为我国鱼类的种类鉴定、地理种群鉴别及种质资源的评估提供重要的工具。     

恩施悬棺人骨mtDNA初步分析

本文以恩施宋代悬棺的骨骼或牙齿样品为对象,经过一系列的处理,获得并扩增了mtDNA的HVSI区的部分序列,通过几个实验初步认定是古DNA.同时通过与部分现代人mtDNA数据的比较参照,对所研究样品单倍群归属进行了初步确定,发现他们主要属于南方类型,但也可能有北方成分.     

珠鸡mtDNA的限制性图谱

应用AvaⅠ，BamHⅠ,BglⅠ,DraⅠ,EcoRⅠ,HpaⅠ,PvuⅡ,SacⅠ,SalⅠ,ScaⅠ和StuⅠ共11种限制酶对珠鸡（Acryllium vulturinum)mtDNA进行单酶切分析，结果表明，珠鸡mtDNA的分子量为16.7kb，另以其中除AvaⅠ和StuⅠ外的9种限制酶进行双酶切分析，构建了珠鸡mtDNA的限制性图谱，并与家鸡（Gallus gallus domesticus)比较，二者mtDNA序列歧异值为0.073。     

新疆锡伯族人群线粒体DNA的遗传学分析

分析48个锡伯人线粒体DNA高可变Ⅰ区的遗传多态性, 并与已发表的相关人群线粒体DNA数据进行比较. 结果表明, 在48个序列中共检测出43个突变位点, 界定了35种单倍型, 其线粒体DNA1序列多态性为0.982, 核苷酸多态性为0.018, 而其平均核苷酸差异为5.922. 人群比较分析表明, 锡伯人与古代拓跋鲜卑人有最近的亲缘关系, 同时也表现出与中国北方游牧民族有较近的亲缘关系, 推断锡伯人可能是拓跋鲜卑的后裔.     

茄子雄性不育株和可育株的胞质DNA和核DNA差异分析

以茄子(Solanum melongenaL)雄性不育株“正兴1号”(S)和可育株(F)的总DNA为模板,对60个随机引物进行了筛选,找到5个其RAPD(Random amplified polymorphic DNA,RAPD)扩增产物在茄子雄性不育系和对照材料间存在稳定差异的引物．将该5个引物同时扩增总DNA、核DNA和线粒体DNA(mitochondrial DNA,mtDNA)．以总DNA为模板时得到多态性片段9个,以核DNA为模板时得到5个,以mtDNA为模板时得到9个．以总DNA为模板时得到的9个扩增片段中,有5个在总DNA和mtDNA中同时出现,而在核中没有出现,即认为来自mtDNA,这5个片段中,有4个来自可育株,1个来自不育株,说明不育株和可育株在mtDNA上存在差异;有4个片段同时出现在以总DNA和核DNA为模板的扩增中,但在以mtDNA为模板的扩增中却没有出现,认为是来自核DNA,这4个片段中,有3个来自不育株,一个来自可育株,说明可育株和不育株在核DNA上也存在差异．结果初步表明：新发现的茄子雄性不育可能是核质相互作用所引起．