木本植物全基因组测序研究进展

近年来, 植物全基因组测序的结果正如雨后春笋般涌现, 木本植物全基因组测序也在紧锣密鼓地展开。但由于木本植物通常基因组较大, 基因组结构较为复杂, 在测序、测序后的组装、注释、功能分析等均存在较大的困难。在基因组测序分析的经费预算方面也存在着较大的压力。因此, 有必要对这方面的研究进展及其存在问题进行分析比较, 以提高林木全基因组研究方面的效率。文章在比较分析已经发展起来的3代基因测序技术(Sanger测序法、合成测序法和单分子测序法)的基础上, 选择4种已经公布的木本植物(杨树、葡萄、番木瓜、苹果), 从全基因组测序的研究背景、测序结果及应用的研究进展和存在问题等方面进行了述评, 对未来要开展的木本植物全基因组测序前的准备工作(材料选择、遗传图谱和连锁图谱的构建、测序技术的选择), 全基因组测序结果的生物信息学分析和应用进行了讨论。     

乳酸菌基因组学研究进展

伴随着高通量测序技术的快速发展和测序成本的降低,越来越多的微生物基因组全序列测定得以实现,从基因组学的层面了解乳酸菌的遗传结构和组成,进而分析和掌握其生物学功能已经逐渐成为可能.迄今为止已经有22株乳酸菌的基因组完成测序并公开发表,还有至少12株测序工作仍在进行中.本文在分析相关文献和生物信息数据基础上,从乳酸菌基因组特点、代谢多样性、进化及共线性四个方面对乳酸菌基因组学研究进展进行了总结,旨在为乳酸菌研究和应用提供参考.     

SNP分子标记及其在木本植物遗传育种的应用

木本植物因其生命周期长、基因组杂合度高、基因组较大、遗传背景不清晰等特性,制约了其研究进程。随着现代生物技术的发展,DNA分子标记技术在木本植物研究领域的应用越来越多,其中单核苷酸多态性(SNP)作为第三代分子标记技术以其高效、快速、稳定、可靠等诸多优点得到广泛应用。本文简述SNP标记的特点、开发方法、检测方法及其在木本植物遗传多样性和亲缘关系分析、品种鉴定、连锁图谱构建和辅助育种等方面的研究进展,为更好地应用SNP技术开展木本植物研究提供参考。     

应用生物信息学技术对植物表达序列标签(EST)的大规模分析

目前 ,一些基因组较小的植物 (如拟南芥 ,水稻等 )的全基因组已经基本完成测序 ,较大基因组的测序工作则主要集中在基因组中表达基因的测序上 ,表达序列标签 (EST)计划由此产生。研究表明 ,对EST进行大规模研究已成为功能基因组学研究的最佳途经之一。本文着重介绍和讨论应用生物信息学技术对植物EST数据的大规模分析。     

全基因组测序及其在遗传性疾病研究及诊断中的应用

最近,随着测序成本的不断降低,数据分析策略的不断提升,全基因组测序（whole-genome sequencing,WGS）已经在癌症、孟德尔遗传病、复杂疾病的致病基因检测中得到了一定运用,并逐步走向了临床诊断。全基因组测序不但可以检测编码区和非编码区的点突变（SNVs）和插入缺失（InDels）,还可以在全基因组范围内检测拷贝数变异（copy number variation,CNV）以及结构变异（structure variation,SV）。本文详细地介绍了全基因组测序的标准生物信息分析流程与方法,及其在疾病研究、临床诊断中的应用,并对全基因组测序在医学遗传学中的应用与研究进展,以及数据分析方面面临的挑战进行了概述。     

山羊基因组与遗传变异图谱研究进展

高通量测序技术和生物信息学的发展极大的促进了山羊分子生物学研究。山羊参考基因组的不断完善以及基因组重测序技术的应用,在全基因组水平上发现了大量的遗传变异信息(SNP、Indel和CNV),丰富了山羊分子群体遗传学研究利用的分子标记。综述了山羊参考基因组组装和全基因组变异图谱的构建及其在山羊上的研究进展,以期为进一步利用分子遗传标记进行山羊的各种性状的遗传基础研究和遗传资源保护利用提供科学依据和参考。     

全基因组测序在重要家畜上的研究进展

全基因组测序研究主要包括通过不同测序技术和组装比对方法,获得某物种的全基因组序列图谱,及在此基础上构建物种全基因组遗传变异图谱进行个体或群体遗传多样性、选择信号或起源进化等方面的研究。利用单核苷酸多态性(SNP)、插入和缺失(Indel)和拷贝数变异(CNV)等遗传变异作为分子标记,全基因组测序研究已经在家畜起源进化、驯化、适应性机制、重要经济性状候选基因、群体历史动态等方面取得了许多重要的研究成果。本文主要对近几年全基因组测序在常见家畜(猪、马、牛、羊等及其近缘物种)的取得的重要研究成果进行了综述,并讨论了全基因组测序的优势、缺点及在生产中意义。此外,对基因组测序研究的未来发展进行了归纳及展望,以期为今后家畜重要经济性状的功能基因定位和物种起源、驯化研究提供参考。     

植物基因组测序的研究进展

近年来,随着测序技术的不断发展,基因组测序技术渐趋成熟并在动物和植物基因组上获得了越来越多的成功,大量植物的基因组的草图和精细图不断地被公布出来。比较和分析了三代测序技术各自的特点,对测序前的准备、基因组组装、注释和比较基因组学等方面的研究进展进行了详细的评述,阐明了植物基因组研究的特点和难点。通过植物的全基因组测序,研究者不仅可以获得该植物基因组和重要功能基因的序列信息,为从分子水平研究植物的分子进化、基因组成和基因调控等提供了一定的依据,而且还对即将测序的植物基因组研究具有重要的借鉴意义。     

蚯蚓基因组学的研究进展: 基于全基因组及线粒体基因组

蚯蚓被喻为土壤中的“生态系统工程师”, 具有高度的多样性且在全世界都有分布, 被用作土壤健康的指示生物。蚯蚓具有极强的环境适应能力, 在不断适应的过程中促进了自身基因组的进化。本文对近年来蚯蚓全基因组以及线粒体基因组的研究进展进行了综述。蚯蚓全基因组的测序、拼装和分析为研究蚯蚓生态学、污染物对蚯蚓致毒的分子机制、免疫防御的分子机制、蚯蚓再生的分子机制等奠定基础。而线粒体基因组多应用于蚯蚓分子系统发育方面的研究, 目前已有多种蚯蚓通过线粒体基因组测序完成了物种的鉴定。本文建议今后重点开展以下几方面的研究: (1)针对现有的4种蚯蚓全基因组测序结果, 进一步进行比较基因组学、进化基因组学和功能基因组学的研究。(2)完善不同种蚯蚓的基因文库和表达序列标签。(3)建立线粒体基因组、全基因组与蚯蚓物种多样性的关联分析。     

植物内生固氮菌系统发育进化新进展

在植物内生固氮菌系统发育进化关系研究中,常用的方法有形态学与蛋白质水平法、数值分类和自动化鉴定法、化学分类法、分子遗传学方法等。本文简要介绍了常用方法的关键技术,并归纳了它们的优缺点。生物学的研究进入基因组时代后,随着高通量DNA测序技术在微生物学领域应用的迅速发展,全基因组测序被应用到微生物系统发育进化研究中,然而目前并未发现对已测全基因组序列的植物内生固氮菌进行系统总结。本文在对已测序植物内生固氮菌进行归纳的基础上,又详细研究了基于基因组数据的几种具有代表性的新方法(ANI分析法、最大唯一匹配指数法、核心基因组分析、组分矢量法、基因流动性分析),并结合目前系统发育进化研究常用方法,对植物内生固氮菌系统发育进化研究趋势进行总结和展望,旨在使植物内生固氮菌的系统发育进化关系研究在精确度、可靠性等方面有所突破。     

多年生植物模式物种基因组研究的历史及进展

木本植物有许多不同于一年生草本植物的生物学特性,生物学家提出将木本植物 作为研究多年生植物的模式体系。杨属Populus树种由于研究基础较好且基因组较小,目前已 被广泛地接受作为多年生植物基因组研究的模式物种。随着杨属树种全基因组序列的测定, 杨属树种在多年生植物的功能基因组研究及一些基础科学问题的研究中将发挥重要作用。本 文综述了杨属树种基因组研究的历史、进展及将来的研究热点,旨在为我国多年生植物基因 组研究提供参考和借鉴。本文主要论述了以下几个方面的内容:（1）对杨属树种开展的细胞 遗传学研究;（2）在分子水平上对杨属树种进行的基因组研究,内容包括遗传作图、基因组 测序、物理图谱构建、基因芯片及连锁不平衡分析;（3）杨属树种基因组信息在探讨一些基 础科学问题中的潜在应用。     

History and progress of the genomics studies in the model system of perennial plant species

Strong justifications have been developed for why woody plants should be viewed as mod-el systems in plant biology. The genus Populus possesses many characteristics that are conductive to functional genomic studies,and therefore leads to its emergence as a model system in extrapolating findings in perennial plant species that are different from annual herbaceous plants. With the pro-ceeding of the whole genome sequencing,poplars will be act as a wide reference for functional ge-nomics studies in perennial plant species and will also contribute towards answering some fundamen-tal scientific questions. This paper reviewed the history and progress of the poplar genome studies and the potential keen topics in the future. The contents mainly address on: （1）the somatic genet-ics studies in Populus;（2）the genomics studies carried out in Populus,including genetic map-ping,genome sequ encing,physical map construction,microarray analysis and linkage disequilibri-um analysis;（3）the potential application of the genome information of Populus for facilitating our understanding of some basic scientific questions.     

Chromosome-level genome assembly of Japanese chestnut (Castanea crenata Sieb. et Zucc.) reveals conserved chromosomal segments in woody rosids

Japanese chestnut (Castanea crenata Sieb. et Zucc.), unlike other Castanea species, is resistant to most diseases and wasps. However, genomic data of Japanese chestnut that could be used to determine its biotic stress resistance mechanisms have not been reported to date. In this study, we employed long-read sequencing and genetic mapping to generate genome sequences of Japanese chestnut at the chromosome level. Long reads (47.7 Gb; 71.6× genome coverage) were assembled into 781 contigs, with a total length of 721.2 Mb and a contig N50 length of 1.6 Mb. Genome sequences were anchored to the chestnut genetic map, comprising 14,973 single nucleotide polymorphisms (SNPs) and covering 1,807.8 cM map distance, to establish a chromosome-level genome assembly (683.8 Mb), with 69,980 potential protein-encoding genes and 425.5 Mb repetitive sequences. Furthermore, comparative genome structure analysis revealed that Japanese chestnut shares conserved chromosomal segments with woody plants, but not with herbaceous plants, of rosids. Overall, the genome sequence data of Japanese chestnut generated in this study is expected to enhance not only its genetics and genomics but also the evolutionary genomics of woody rosids.     

泛基因组研究在遗传多样性和功能基因组学中的应用

相对于单个参考基因组仅聚焦于个体遗传信息的挖掘,泛基因组研究则能够反映整个物种或类群全部的遗传信息。随着基因组测序和分析技术的不断发展,泛基因组学逐渐成为新的研究热点,并已在植物、动物和微生物多个物种中获得了广泛应用,为全面解析物种或类群水平的遗传变异和多样性、功能基因组和系统进化重建等研究提供了强有力的工具,取得了很多显著的研究成果。尽管如此,由于泛基因组学研究尚处于发展阶段,测序费用和分析成本仍然较高,难以广泛普及; 且存在分析标准不一、数据挖掘不够全面深入、理论难以应用于生产实际等尚待解决的问题,仍有较大的发展空间。该文系统总结了泛基因组在生物遗传多样性挖掘和功能基因组学中的研究进展,主要包括其在泛基因组图谱的构建、基因组变异和有利基因的发掘、功能基因的多态性、群体遗传多样性和系统进化等多个领域中的应用和研究,探讨了其在不同领域的应用潜力。同时,讨论了目前泛基因组研究中存在的局限性和可能的解决方法,并对其将来的发展前景进行了展望。     

第三代测序技术的研究进展与临床应用

Sanger测序法，又称第一代测序技术，作为测序金标准推动了人类基因组“工作框架图”的绘制，但通量低、成本高的缺点限制了其进一步大规模应用。第二代测序技术，又称下一代测序技术，因其通量高、成本低等优点使得基因测序在基础研究与临床诊疗中得到广泛应用，但短读长一直是其不可回避的技术短板。第三代测序技术的出现，因其具有长读长优势，为基因序列上复杂重复区域解析与高质量基因组组装提供了新的技术手段。近年来，第三代测序技术进一步发展与完善，同时在肿瘤、免疫、生殖等相关领域逐步体现出临床应用价值。本文将综述第三代测序技术的研究进展与临床应用。     

Insights into the genome structure and copy-number variation of Eimeria tenella

ABSTRACT: BACKGROUND: Eimeria is a genus of parasites in the same phylum (Apicomplexa) as human parasites such as Toxoplasma, Cryptosporidium and the malaria parasite Plasmodium. As an apicomplexan whose life-cycle involves a single host, Eimeria is a convenient model for understanding this group of organisms. Although the genomes of the Apicomplexa are diverse, that of Eimeria is unique in being composed of large alternating blocks of sequence with very different characteristics - an arrangement seen in no other organism. This arrangement has impeded efforts to fully sequence the genome of Eimeria, which remains the last of the major apicomplexans to be fully analyzed. In order to increase the value of the genome sequence data and aid in the effort to gain a better understanding of the Eimeria tenella genome, we constructed a whole genome map for the parasite. RESULTS: A total of 1245 contigs representing 70.0% of the whole genome assembly sequences (Wellcome Trust Sanger Institute) were selected and subjected to marker selection. Subsequently, 2482 HAPPY markers were developed and typed. Of these, 795 were considered as usable markers, and utilized in the construction of a HAPPY map. Markers developed from chromosomally-assigned genes were then integrated into the HAPPY map and this aided the assignment of a number of linkage groups to their respective chromosomes. BAC-end sequences and contigs from whole genome sequencing were also integrated to improve and validate the HAPPY map. This resulted in an integrated HAPPY map consisting of 60 linkage groups that covers approximately half of the estimated 60 Mb genome. Further analysis suggests that the segmental organization first seen in Chromosome 1 is present throughout the genome, with repeat-poor (P) regions alternating with repeat-rich (R) regions. Evidence of copy-number variation between strains was also uncovered. CONCLUSIONS: This paper describes the application of a whole genome mapping method to improve the assembly of the genome of E. tenella from shotgun data, and to help reveal its overall structure. A preliminary assessment of copy-number variation (extra or missing copies of genomic segments) between strains of E. tenella was also carried out. The emerging picture is of a very unusual genome architecture displaying inter-strain copy-number variation. We suggest that these features may be related to the known ability of this parasite to rapidly develop drug resistance.     

Construction of a high-density genetic map for grape using next generation restriction-site associated DNA sequencing

ABSTRACT: BACKGROUND: Genetic mapping and QTL detection are powerful methodologies in plant improvement and breeding. Construction of a high-density and high-quality genetic map would be of great benefit in the production of superior grapes to meet human demand. High throughput and low cost of the recently developed next generation sequencing (NGS) technology have resulted in its wide application in genome research. Sequencing restriction-site associated DNA (RAD) might be an efficient strategy to simplify genotyping. Combining NGS with RAD has proven to be powerful for single nucleotide polymorphism (SNP) marker development. RESULTS: An F1 population of 100 individual plants was developed. In-silico digestion-site prediction was used to select an appropriate restriction enzyme for construction of a RAD sequencing library. Next generation RAD sequencing was applied to genotype the F1 population and its parents. Applying a cluster strategy for SNP modulation, a total of 1,814 high-quality SNP markers were developed: 1,121 of these were mapped to the female genetic map, 759 to the male map, and 1,646 to the integrated map. A comparison of the genetic maps to the published Vitis vinifera genome revealed both conservation and variations. CONCLUSIONS: The applicability of next generation RAD sequencing for genotyping a grape F1 population was demonstrated, leading to the successful development of a genetic map with high density and quality using our designed SNP markers. Detailed analysis revealed that this newly developed genetic map can be used for a variety of genome investigations, such as QTL detection, sequence assembly and genome comparison. KEYWORDS: Grape; Genetic map; Next generation sequencing (NGS); Restriction-site associated DNA (RAD).