古DNA 及其在生物系统与进化研究中的应用

古DNA是指从已经死亡的古代生物的遗体和遗迹中得到的DNA。本文回顾了近20年古DNA研究所经历的3个阶段, 从早期参与研究的科学家较少并主要利用克隆技术, 到后来由于PCR技术的出现以及提取化石DNA技术的成熟从而出现大量有关古DNA的报道; 近几年由于发现不少问题, 并引起激烈的争论, 科学家们因此而开始考虑古DNA的真实性问题, 并且提出了开展古DNA研究的严格标准。本文还讨论了古DNA在人类起源、系统发育重建、动植物驯化及考古研究中的重要意义以及现状, 表明古DNA的研究给某些原先的观点如人类的非洲起源说提供了重要证据, 也对某些观点提出了挑战; 古DNA研究还提供了某些已经灭绝生物的形态学和分子资料, 为从序列上确定古代材料的系统位置并有效地补充仅用现代DNA建立起来的谱系提供了来自古生物的依据。在动植物驯化及考古方面, 古DNA证据也为科学家提供了许多有价值的信息。最后, 本文还对古DNA研究的应用前景进行了展望。     

应用古DNA技术探究发酵微生物的适应、演化和驯化历史

发酵生产是人类最原始的对微生物的应用和实践,在人类历史上具有重要意义。然而,由于分子证据的匮乏,人类发酵生产的演变历程及相关发酵微生物的演化和驯化历史尚不清楚。本文以目前最常见的两类发酵食品——酒及发酵乳品为例介绍了发酵食品考古和相关发酵微生物的演化和驯化研究,以及古微生物学和发酵古微生物的研究现状,并讨论了将微生物古DNA技术应用于古代发酵微生物研究的可行性和难点,展示了古DNA捕获技术在本领域的应用潜力,为发酵微生物演化研究提供了新的思路和方法。     

古DNA单链测序文库的建立与检测北大核心CSCD

2005年问世的第二代测序技术在古DNA领域的应用,突破了第一代测序技术在绝灭或死亡生物全基因组获取手段上的局限。借助古基因组信息,研究者能够从更为系统的实时分子证据角度,解读诸如人类起源、大型绝灭哺乳动物迁移演化、动植物家养驯化以及早期人类社会生活模式等古生物学、遗传学与演化生物学问题。引入第二代测序技术之后,传统的古DNA研究方法及流程得以改变,剔除了原有的实验流程中耗时的分子克隆步骤,引入了与第二代测序技术紧密相关的古DNA单链测序文库构建环节。古DNA单链测序文库的构建,是将古DNA双链模板变性成单链后,通过向单链古DNA两端添加人工DNA片段,将古DNA分子转变成能被测序仪识别的文库分子。针对古DNA分子微量、高度片段化以及普遍存在碱基损伤的特点,古DNA单链测序文库,能够高效获取古DNA材料中的遗传信息。本文系统介绍古DNA单链测序文库建立流程,以及对文库质量进行检测的方法,为研究者运用第二代测序技术测定绝灭或死亡生物全基因组提供方法借鉴。     

古DNA单链测序文库的建立与检测

2005年问世的第二代测序技术在古DNA领域的应用,突破了第一代测序技术在绝灭或死亡生物全基因组获取手段上的局限。借助古基因组信息,研究者能够从更为系统的实时分子证据角度,解读诸如人类起源、大型绝灭哺乳动物迁移演化、动植物家养驯化以及早期人类社会生活模式等古生物学、遗传学与演化生物学问题。引入第二代测序技术之后,传统的古DNA研究方法及流程得以改变,剔除了原有的实验流程中耗时的分子克隆步骤,引入了与第二代测序技术紧密相关的古DNA单链测序文库构建环节。古DNA单链测序文库的构建,是将古DNA双链模板变性成单链后,通过向单链古DNA两端添加人工DNA片段,将古DNA分子转变成能被测序仪识别的文库分子。针对古DNA分子微量、高度片段化以及普遍存在碱基损伤的特点,古DNA单链测序文库,能够高效获取古DNA材料中的遗传信息。本文系统介绍古DNA单链测序文库建立流程,以及对文库质量进行检测的方法,为研究者运用第二代测序技术测定绝灭或死亡生物全基因组提供方法借鉴。     

广西地区野猪化石古DNA及猪的驯化初探

猪的起源驯化一直是人们关注的问题,古DNA技术可为家猪起源驯化研究提供历史分子证据。为探讨广西地区更新世晚期野猪与当地家猪的关系,我们收集了广西壮族自治区晚更新世三个不同地点的30个猪科动物化石样品,测定了这些样品的线粒体DNA细胞色素b基因的核苷酸序列。结合现代不同品种家猪和野猪DNA同源序列信息,构建Network网络结构图,进一步证实了家猪多地起源学说。研究显示广西古代野猪与现存欧洲猪的基因型最为接近,属于欧洲单倍型H1,从而推测其对该地区现代家猪的遗传贡献可能较低,对考古形态学研究中关于广西地区家猪独立起源的推测提出了质疑。     

家养动物驯化起源的研究方法与进展

驯化动物伴随了人类上万年的进化历程。在驯化过程中, 家养动物表型和行为特征发生巨大改变, 并与人类生产生活的进步相适应。研究动物驯化问题对于了解遗传多样性及适应性进化, 解析复杂性状的遗传机制等具有重要的意义, 成为生物学领域的重点研究内容之一。本文专注于动物驯化的初始阶段, 首先介绍了驯化起源的时间与地点、驯化途径和驱动因、驯化后的扩散和品种选育; 其次从驯化对象的角度着重介绍了考古学、分子和群体遗传学两方面的家养动物驯化起源的研究策略、优势与不足, 以及未来的发展方向; 最后, 我们综合多方面的证据, 介绍了在中国驯化的家猪(Sus domesticus)和家鸡(Gallus gallus domesticus)以及其他主要家养动物考古和分子水平上驯化起源的研究进展。本文整合多种证据为家养动物驯化起源的研究提供了相对完整的视角和新的思路。     

家马的起源历史与品种驯化特征

在家犬(Canis lupus familiaris)、牛(Bos taurus)、猪(Sus scrofa domesticus)、绵羊(Ovis aries)、山羊(Capra hircus)等家养动物之后,家马(Equus caballus)才被人类成功驯化。虽然驯化历史很短,但其对人类社会文明发展变革的影响却最大。家马出色的负重移动能力使人类社会由固定的农耕模式向移动分享模式过渡,使历史发展进入了快车道,因此其起源驯化历史一直备受关注。然而由于家马直系同源野生种早已灭绝,加之现代品种化培育引起遗传多样性骤减,使得相关研究长期争议不断。随着测序技术的不断发展和古代样品的逐步丰富,目前对家马起源驯化过程、群体遗传结构等方面的研究越来越深入。本文从核基因、mtDNA、Y染色体、古DNA等不同层面综述了家马起源与驯化历史方面的研究进展,从品种分化状况、群体演化特征等方面讨论了现代家马品种的群体遗传结构,最后总结了马匹毛色、速度、体型等重要表型性状的遗传基础,以期为今后家马的起源驯化研究、种质资源保护与开发、品种优化方向、现代马业发展等方面提供参考。     

家养动物的起源与驯化研究进展

动物的驯化是新石器时代农业革命的主要内容之一, 导致了人类生活方式从狩猎向畜牧的转变。开展家养动物起源与驯化的研究, 不仅有助于了解人类社会的发展史和人工选择下动物的进化过程, 而且有助于开展家养动物的良种选育和遗传多样性保护, 因此该研究领域受到了人们的长期关注。近年来, 随着更多考古证据的发现与分子遗传学的发展, 人们对于家养动物的起源与驯化有了更深入的认识。本文对家养动物起源与驯化研究中一些基本问题的研究方法和进展进行了综述。文章第一部分介绍了家养动物的野生祖先、起源地、起源时间、建群者大小和扩散路线等问题; 第二部分则涉及家养动物驯化的条件、过程、性状改变及其遗传机制等方面。最后指出了家养动物起源与驯化研究中依然存在的问题, 并对未来研究发展的趋势进行了讨论。     

古DNA技术在人类墓葬遗骸研究中的应用及进展

考古工作中得到的生物遗骸由于长期的风化,自然侵蚀等因素的影响,遗骸本身含有的古代生物的DNA的大部分会分解,使得对遗骸中的生物遗传信息研究变得非常困难.可将现代生物工程的PCR技术应用到考古工作中,该技术能够对残存的微量DNA进行大量的生物体外扩增,得到更多的古代生物的遗传信息,提高时遗骸种属鉴定的准确性.通过对出土的人类遗骸中微量DNA的扩增、测试和遗骸间DNA序列的对比,在计算机软件的帮助下与已知的人类DNA序列进行比较,能确定同一墓葬中不同遗骸的亲缘关系和该遗骸群体在整个人类进化体系中的位置.对这一试验过程的一些方法、技术、研究进展和目前仍然面临的一些问题作了介绍.     

科学家完成古代野牛线粒体基因组全序列

<正>根据考古和遗传研究记载,家养牛是由现已灭绝的古代野牛(Bos primigenius)驯化而来,古代近东是普通牛(Bos Taurus)的起源中心。这些推论主要是通过对现代牛部分线粒体DNA序列和古代野牛样本的有限序列数据分析得来的。而近来DNA测序技术的发展,不仅为灭绝物种遗传物质的检测提供了新的机会,同     

Damage and repair of ancient DNA

Under certain conditions small amounts of DNA can survive for long periods of time and can be used as polymerase chain reaction (PCR) substrates for the study of phylogenetic relationships and population genetics of extinct plants and animals, including hominids. Because of extensive DNA degradation, these studies are limited to species that lived within the past 10(4)-10(5) years (Late Pleistocene), although DNA sequences from 10(6) years have been reported. Ancient DNA (aDNA) has been used to study phylogenetic relationships of protists, fungi, algae, plants, and higher eukaryotes such as extinct horses, cave bears, the marsupial wolf, the moa, and Neanderthal. In the past few years, this technology has been extended to the study of infectious disease in ancient Egyptian and South American mummies, the dietary habits of ancient animals, and agricultural practices and population dynamics of early native Americans. Hence, ancient DNA contains information pertinent to numerous fields of study including evolution, population genetics, ecology, climatology, medicine, archeology, and behavior. The major obstacles to the study of aDNA are its extremely low yield, contamination with modern DNA, and extensive degradation. In the course of this review, we will discuss the current aDNA literature describing the importance of aDNA studies as they relate to important biological questions and the difficulties associated with extracting useful information from highly degraded and damaged substrates derived from limited sources. In addition, we will present some of our own preliminary and published data on mechanisms of DNA degradation and some speculative thoughts on strategies for repair and restoration of aDNA.     

Mitochondrial DNA from prehistoric canids highlights relationships between dogs and South-East European wolves

The question of the origins of the dog has been much debated. The dog is descended from the wolf that at the end of the last glaciation (the archaeologically hypothesized period of dog domestication) was one of the most widespread among Holarctic mammals. Scenarios provided by genetic studies range from multiple dog-founding events to a single origin in East Asia. The earliest fossil dogs, dated approximately 17-12,000 radiocarbon ((14)C) years ago (YA), were found in Europe and in the Middle East. Ancient DNA (a-DNA) evidence could contribute to the identification of dog-founder wolf populations. To gain insight into the relationships between ancient European wolves and dogs we analyzed a 262-bp mitochondrial DNA control region fragment retrieved from five prehistoric Italian canids ranging in age from approximately 15,000 to approximately 3,000 (14)C YA. These canids were compared to a worldwide sample of 547 purebred dogs and 341 wolves. The ancient sequences were highly diverse and joined the three major clades of extant dog sequences. Phylogenetic investigations highlighted relationships between the ancient sequences and geographically widespread extant dog matrilines and between the ancient sequences and extant wolf matrilines of mainly East European origin. The results provide a-DNA support for the involvement of European wolves in the origins of the three major dog clades. Genetic data also suggest multiple independent domestication events. East European wolves may still reflect the genetic variation of ancient dog-founder populations.     

How ancient DNA may help in understanding the origin and spread of agriculture

The origin and spread of agriculture have been central questions in archaeology for the last 75 years and are increasingly being addressed by a multidisciplinary approach involving biologists, ecologists, geographers and anthropologists as well as archaeologists. Molecular genetics has the potential to make an important contribution, especially by enabling the number of times that a crop or animal was domesticated to be determined. Molecular genetics can also assign approximate dates to domestication events, identify the wild progenitor of a domesticate, and provide new forms of evidence relevant to agricultural spread. With wheat, molecular genetical studies of modern plants have suggested that einkorn was domesticated just once but that emmer might have been domesticated more than once. Ancient DNA studies of animal remains have benefited from progress made with equivalent analyses of human bones, and with plant material there have been clear demonstrations of DNA preservation in desiccated seeds. Charred remains have also been shown to contain ancient DNA but this finding is unexpected in view of the high temperatures to which these seeds have supposedly been exposed. Ancient DNA studies of wheat remains have been used in taxonomic identification and in assessment of the possible bread-making quality of the wheat grown at an Early Bronze Age site in Greece.     

中国东北第四纪晚期哺乳动物化石样品的古DNA损伤分析

二代测序技术的进步推动了古DNA研究的发展,古DNA研究在人类起源、动物演化等领域已经做出突出贡献。如何针对特定地点的古DNA样品特征,有效提取挖掘其中蕴含的古生物遗传信息,是发挥古代生物样品在诸多研究领域重要作用的前提。本研究将DNA损伤的两个主要指标(末端碱基替换率、平均片段长度)与样品的埋藏时间、所属地质时期、样品材料类型和建库方法相联系,分析不同因素对古DNA损伤的影响。结果表明:中国东北古脊椎动物样品中的古DNA分子的末端碱基替换率与埋藏点的含水量、样品埋藏时间呈正相关;不同地质时期的样品之间古DNA末端碱基替换率有显著差异;不同样品材料类型对于古DNA的末端碱基替换率未见明显影响;样品古DNA的平均片段长度与以上所研究的因素均无明显关系。研究结果为探明中国东北古脊椎动物样品的古DNA特征提供了分子依据,为有效选取不同地区的古脊椎动物样品及样品发掘后的合理保存提供了借鉴和参考。     

An efficient pipeline for ancient DNA mapping and recovery of endogenous ancient DNA from whole‐genome sequencing data

Ancient DNA research has developed rapidly over the past few decades due to improvements in PCR and next‐generation sequencing (NGS) technologies, but challenges still exist. One major challenge in relation to ancient DNA research is to recover genuine endogenous ancient DNA sequences from raw sequencing data. This is often difficult due to degradation of ancient DNA and high levels of contamination, especially homologous contamination that has extremely similar genetic background with that of the real ancient DNA. In this study, we collected whole‐genome sequencing (WGS) data from 6 ancient samples to compare different mapping algorithms. To further explore more effective methods to separate endogenous DNA from homologous contaminations, we attempted to recover reads based on ancient DNA specific characteristics of deamination, depurination, and DNA fragmentation with different parameters. We propose a quick and improved pipeline for separating endogenous ancient DNA while simultaneously decreasing homologous contaminations to very low proportions. Our goal in this research was to develop useful recommendations for ancient DNA mapping and for separation of endogenous DNA to facilitate future studies of ancient DNA.     

古DNA提取技术新进展

古DNA是揭示古代生物生长状态以及生物千百万年来进化情况的最重要的信息载体,在治疗人类遗传性的疑难病症及牲畜饲养和粮食作物种植等方面都有重大的贡献。古DNA提取技术作为获得该重要的信息载体的最重要手段,长久以来受到了世界各地的考古学家以及医学研究学者们的高度重视。随着科学的发展,古DNA提取技术已经形成多种核心方法:Chelex-100法、酚-氯仿抽提法、二氧化硅(硅粒)法、NaOH法、硅离心柱法试剂盒、磁珠法试剂盒等方法。本文将根据最新的研究成果对以上提到的几种方法进行分析比较,以期能够为将来古DNA提取技术的发展创新提供新的思路与方向。     

古DNA提取技术新进展

古DNA 是揭示古代生物生长状态以及生物千百万年来进化情况的最重要的信息载体,在治疗人类遗传性的疑难病症及牲畜饲养和粮食作物种植等方面都有重大的贡献。古DNA提取技术作为获得该重要的信息载体的最重要手段,长久以来受到了世界各地的考古学家以及医学研究学者们的高度重视。随着科学的发展,古DNA提取技术已经形成多种核心方法：Chelex-100 法、酚-氯仿抽提法、二氧化硅（硅粒）法、NaOH 法、硅离心柱法试剂盒、磁珠法试剂盒等方法。本文将根据最新的研究成果对以上提到的几种方法进行分析比较,以期能够为将来古DNA提取技术的发展创新提供新的思路与方向。     

Optimization of DNA Recovery and Amplification from Non-Carbonized Archaeobotanical Remains

Ancient DNA (aDNA) recovered from archaeobotanical remains can provide key insights into many prominent archaeological research questions, including processes of domestication, past subsistence strategies, and human interactions with the environment. However, it is often difficult to isolate aDNA from ancient plant materials, and furthermore, such DNA extracts frequently contain inhibitory substances that preclude successful PCR amplification. In the age of high-throughput sequencing, this problem is even more significant because each additional endogenous aDNA molecule improves analytical resolution. Therefore, in this paper, we compare a variety of DNA extraction techniques on primarily desiccated archaeobotanical remains and identify which method consistently yields the greatest amount of purified DNA. In addition, we test five DNA polymerases to determine how well they replicate DNA extracted from non-charred ancient plant remains. Based upon the criteria of resistance to enzymatic inhibition, behavior in quantitative real-time PCR, replication fidelity, and compatibility with aDNA damage, we conclude these polymerases have nuanced properties, requiring researchers to make educated decisions as to which one to use for a given task. The experimental findings should prove useful to the aDNA and archaeological communities by guiding future research methodologies and ensuring precious archaeobotanical remains are studied in optimal ways, and may thereby yield important new perspectives on the interactions between humans and past plant communities.     

Ancient DNA: Using molecular biology to explore the past

Ancient DNA has been discovered in many types of preserved biological material, including bones, mummies, museum skins, insects in amber and plant fossils, and has become an important research tool in disciplines as diverse as archaeology, conservation biology and forensic science. In archaeology, ancient DNA can contribute both to the interpretation of individual sites and to the development of hypotheses about past populations. Site interpretation is aided by DNA-based sex typing of fragmentary human bones, and by the use of genetic techniques to assess the degree of kinship between the remains of different individuals. On a broader scale, population migrations can be traced by studying genetic markers in ancient DNA, as in recent studies of the colonisation of the Pacific islands, while ancient DNA in preserved plant remains can provide information on the development of agriculture.