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Genome-wide association study reveals the genetic architecture of flowering time in rapeseed (Brassica napus L.) 油菜(Brassica napus L.)开花时间遗传结构全基因组关联研究
Liping Xu, Kaining Hu, Zhenqian Zhang, C. Guan, Song Chen, Wei Hua, Jiana Li, J. Wen, B. Yi, Jinxiong Shen, Chaozhi Ma, J. Tu, T. Fu
Flowering time adaptation is a major breeding goal in the allopolyploid species Brassica napus. To investigate the genetic architecture of flowering time, a genome-wide association study (GWAS) of flowering time was conducted with a diversity panel comprising 523 B. napus cultivars and inbred lines grown in eight different environments. Genotyping was performed with a Brassica 60K Illumina Infinium SNP array. A total of 41 single-nucleotide polymorphisms (SNPs) distributed on 14 chromosomes were found to be associated with flowering time, and 12 SNPs located in the confidence intervals of quantitative trait loci (QTL) identified in previous researches based on linkage analyses. Twenty-five candidate genes were orthologous to Arabidopsis thaliana flowering genes. To further our understanding of the genetic factors influencing flowering time in different environments, GWAS was performed on two derived traits, environment sensitivity and temperature sensitivity. The most significant SNPs were found near Bn-scaff_16362_1-p380982, just 13 kb away from BnaC09g41990D, which is orthologous to A. thaliana CONSTANS (CO), an important gene in the photoperiod flowering pathway. These results provide new insights into the genetic control of flowering time in B. napus and indicate that GWAS is an effective method by which to reveal natural variations of complex traits in B. napus.
开花时间适应是异源多倍体甘蓝型油菜的主要育种目标。为了研究开花时间的遗传结构,利用8种不同环境下523个甘蓝型油菜品种和自交系进行了开花时间全基因组关联研究(GWAS)。使用Brassica 60K Illumina Infinium SNP阵列进行基因分型。在14条染色体上共发现41个与开花时间相关的单核苷酸多态性(snp),其中12个snp位于前人通过连锁分析发现的数量性状位点(QTL)置信区间。25个候选基因与拟南芥开花基因同源。为了进一步了解不同环境下影响开花时间的遗传因素,对两个衍生性状——环境敏感性和温度敏感性进行了GWAS分析。在Bn-scaff_16362_1-p380982附近发现了最显著的snp,距离BnaC09g41990D仅13 kb,该基因与A. thaliana CONSTANS (CO)同源,后者是光周期开花途径的重要基因。这些结果为油菜开花时间的遗传调控提供了新的认识,表明GWAS是揭示油菜复杂性状自然变异的有效方法。
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引用次数: 127
The complete genome sequencing of Prevotella intermedia strain OMA14 and a subsequent fine-scale, intra-species genomic comparison reveal an unusual amplification of conjugative and mobile transposons and identify a novel Prevotella-lineage-specific repeat 普雷沃特菌中间菌株OMA14的全基因组测序和随后的精细尺度种内基因组比较揭示了共轭和移动转座子的异常扩增,并确定了一种新的普雷沃特菌谱系特异性重复序列
M. Naito, Yoshitoshi Ogura, T. Itoh, M. Shoji, M. Okamoto, Tetsuya Hayashi, K. Nakayama
Prevotella intermedia is a pathogenic bacterium involved in periodontal diseases. Here, we present the complete genome sequence of a clinical strain, OMA14, of this bacterium along with the results of comparative genome analysis with strain 17 of the same species whose genome has also been sequenced, but not fully analysed yet. The genomes of both strains consist of two circular chromosomes: the larger chromosomes are similar in size and exhibit a high overall linearity of gene organizations, whereas the smaller chromosomes show a significant size variation and have undergone remarkable genome rearrangements. Unique features of the Pre. intermedia genomes are the presence of a remarkable number of essential genes on the second chromosomes and the abundance of conjugative and mobilizable transposons (CTns and MTns). The CTns/MTns are particularly abundant in the second chromosomes, involved in its extensive genome rearrangement, and have introduced a number of strain-specific genes into each strain. We also found a novel 188-bp repeat sequence that has been highly amplified in Pre. intermedia and are specifically distributed among the Pre. intermedia-related species. These findings expand our understanding of the genetic features of Pre. intermedia and the roles of CTns and MTns in the evolution of bacteria.
中间普雷沃氏菌是一种与牙周病有关的致病菌。在这里,我们展示了这种细菌的临床菌株OMA14的完整基因组序列,以及与同一物种的菌株17的比较基因组分析结果,菌株17的基因组也已测序,但尚未完全分析。这两种菌株的基因组都由两条圆形染色体组成:较大的染色体大小相似,并表现出较高的基因组织整体线性,而较小的染色体则表现出显著的大小差异,并经历了显著的基因组重排。Pre的独特功能。中间基因组是指在第二染色体上存在大量必需基因和丰富的共轭和可移动转座子(ctn和mtn)。ctn / mtn在第二条染色体中特别丰富,参与其广泛的基因组重排,并将许多菌株特异性基因引入每个菌株。我们还发现了一个新的188bp重复序列,在Pre中被高度扩增。中间媒介,并专门分布在Pre中。intermedia-related物种。这些发现扩大了我们对Pre基因特征的理解。中间介质以及ctn和mtn在细菌进化中的作用。
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引用次数: 21
Constructing a linkage–linkage disequilibrium map using dominant-segregating markers 利用优势分离标记构建连锁-连锁不平衡图谱
Xuli Zhu, Leiming Dong, Libo Jiang, Huan Li, Lidan Sun, Hui Zhang, Weiwu Yu, Haokai Liu, Wensheng Dai, Yanru Zeng, R. Wu
The relationship between linkage disequilibrium (LD) and recombination fraction can be used to infer the pattern of genetic variation and evolutionary process in humans and other systems. We described a computational framework to construct a linkage–LD map from commonly used biallelic, single-nucleotide polymorphism (SNP) markers for outcrossing plants by which the decline of LD is visualized with genetic distance. The framework was derived from an open-pollinated (OP) design composed of plants randomly sampled from a natural population and seeds from each sampled plant, enabling simultaneous estimation of the LD in the natural population and recombination fraction due to allelic co-segregation during meiosis. We modified the framework to infer evolutionary pasts of natural populations using those marker types that are segregating in a dominant manner, given their role in creating and maintaining population genetic diversity. A sophisticated two-level EM algorithm was implemented to estimate and retrieve the missing information of segregation characterized by dominant-segregating markers such as single methylation polymorphisms. The model was applied to study the relationship between linkage and LD for a non-model outcrossing species, a gymnosperm species, Torreya grandis, naturally distributed in mountains of the southeastern China. The linkage–LD map constructed from various types of molecular markers opens a powerful gateway for studying the history of plant evolution.
链接不平衡(LD)与重组分数之间的关系可以用来推断人类和其他系统的遗传变异模式和进化过程。我们描述了一个计算框架,从常用的双等位基因,单核苷酸多态性(SNP)标记构建一个连锁- LD图谱,通过该图谱,LD的下降随着遗传距离的增加而可视化。该框架来源于一个开放授粉(OP)设计,该设计由从自然群体中随机取样的植物和每个取样植物的种子组成,可以同时估计自然群体中的LD和由于减数分裂期间等位基因共分离而导致的重组分数。我们修改了这个框架,利用那些以主导方式分离的标记类型来推断自然种群的进化历史,考虑到它们在创造和维持种群遗传多样性方面的作用。采用一种复杂的两级EM算法来估计和检索以单甲基化多态性等显性分离标记为特征的分离缺失信息。应用该模型研究了中国东南部自然分布的裸子植物大香榧(Torreya grandis)非模式异交种的连锁与遗传变异的关系。利用各种类型的分子标记构建的连锁- ld图谱为研究植物进化史打开了一个强有力的门户。
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引用次数: 15
Genotyping-by-sequencing map permits identification of clubroot resistance QTLs and revision of the reference genome assembly in cabbage (Brassica oleracea L.) 基因分型测序图谱允许白菜(Brassica oleracea L.)抗性qtl的鉴定和参考基因组组装的修订。
Jonghoon Lee, N. Izzah, Beom-Soon Choi, H. Joh, Sang-Choon Lee, Sampath Perumal, Joodeok Seo, Kyounggu Ahn, Eun Ju Jo, G. Choi, I. Nou, Yeisoo Yu, Tae-Jin Yang
Clubroot is a devastating disease caused by Plasmodiophora brassicae and results in severe losses of yield and quality in Brassica crops. Many clubroot resistance genes and markers are available in Brassica rapa but less is known in Brassica oleracea. Here, we applied the genotyping-by-sequencing (GBS) technique to construct a high-resolution genetic map and identify clubroot resistance (CR) genes. A total of 43,821 SNPs were identified using GBS data for two parental lines, one resistant and one susceptible lines to clubroot, and 18,187 of them showed >5× coverage in the GBS data. Among those, 4,103 were credibly genotyped for all 78 F2 individual plants. These markers were clustered into nine linkage groups spanning 879.9 cM with an average interval of 1.15 cM. Quantitative trait loci (QTLs) survey based on three rounds of clubroot resistance tests using F2 : 3 progenies revealed two and single major QTLs for Race 2 and Race 9 of P. brassicae, respectively. The QTLs show similar locations to the previously reported CR loci for Race 4 in B. oleracea but are in different positions from any of the CR loci found in B. rapa. We utilized two reference genome sequences in this study. The high-resolution genetic map developed herein allowed us to reposition 37 and 2 misanchored scaffolds in the 02–12 and TO1000DH genome sequences, respectively. Our data also support additional positioning of two unanchored 3.3 Mb scaffolds into the 02–12 genome sequence.
棍棒病是由芸苔菌(Plasmodiophora brassicae)引起的一种毁灭性病害,严重影响芸苔作物的产量和品质。油菜中有许多抗根茎病基因和标记,而甘蓝中所知较少。本研究采用基因分型测序(GBS)技术构建了高分辨率遗传图谱,并鉴定了俱乐部根抗性(CR)基因。2个亲本(1个抗性系和1个敏感系)的GBS数据共鉴定出43821个snp,其中18187个snp在GBS数据中覆盖率>5倍。其中,4103份对全部78株F2单株进行了可靠的基因分型。这些标记聚为9个连锁群,全长879.9 cM,平均间隔1.15 cM。利用F2: 3个子代进行三轮抗棒根病试验的数量性状位点(qtl)调查发现,芸苔属2号和9号小种分别存在2个和1个主qtl。这些qtl的位置与先前报道的甘蓝第4种的CR位点相似,但与甘蓝中发现的任何CR位点的位置不同。在本研究中我们使用了两个参考基因组序列。本文开发的高分辨率遗传图谱使我们能够分别在02-12和TO1000DH基因组序列中重新定位37个和2个错误锚定的支架。我们的数据还支持在02-12基因组序列中额外定位两个未锚定的3.3 Mb支架。
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引用次数: 84
Improvement of barley genome annotations by deciphering the Haruna Nijo genome 通过破译二条春菜基因组改进大麦基因组注释
Kazuhiro Sato, Tsuyoshi Tanaka, S. Shigenobu, Yuka Motoi, Jianzhon Wu, T. Itoh
Full-length (FL) cDNA sequences provide the most reliable evidence for the presence of genes in genomes. In this report, detailed gene structures of barley, whole genome shotgun (WGS) and additional transcript data of the cultivar Haruna Nijo were quality controlled and compared with the published Morex genome information. Haruna Nijo scaffolds have longer total sequence length with much higher N50 and fewer sequences than those in Morex WGS contigs. The longer Haruna Nijo scaffolds provided efficient FLcDNA mapping, resulting in high coverage and detection of the transcription start sites. In combination with FLcDNAs and RNA-Seq data from four different tissue samples of Haruna Nijo, we identified 51,249 gene models on 30,606 loci. Overall sequence similarity between Haruna Nijo and Morex genome was 95.99%, while that of exon regions was higher (99.71%). These sequence and annotation data of Haruna Nijo are combined with Morex genome data and released from a genome browser. The genome sequence of Haruna Nijo may provide detailed gene structures in addition to the current Morex barley genome information.
全长cDNA序列为基因在基因组中的存在提供了最可靠的证据。本文对大麦的详细基因结构、全基因组霰弹枪(WGS)和其他转录本数据进行了质量控制,并与已发表的Morex基因组信息进行了比较。与Morex WGS序列相比,Haruna Nijo支架的总序列长度更长,N50更高,序列数更少。较长的Haruna Nijo支架提供了高效的FLcDNA定位,导致转录起始位点的高覆盖率和检测。结合来自四种不同组织样本的flcdna和RNA-Seq数据,我们在30,606个位点上鉴定了51,249个基因模型。总体序列相似性为95.99%,其中外显子区域的相似性较高(99.71%)。这些二条春菜的序列和注释数据与Morex基因组数据相结合,并通过基因组浏览器发布。除了目前的Morex大麦基因组信息外,Haruna Nijo的基因组序列可能提供详细的基因结构。
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引用次数: 31
Identification of minimal eukaryotic introns through GeneBase, a user-friendly tool for parsing the NCBI Gene databank 通过GeneBase鉴定最小真核内含子,这是一个用户友好的工具,用于解析NCBI基因数据库
A. Piovesan, M. Caracausi, Marco Ricci, P. Strippoli, L. Vitale, M. C. Pelleri
We have developed GeneBase, a full parser of the National Center for Biotechnology Information (NCBI) Gene database, which generates a fully structured local database with an intuitive user-friendly graphic interface for personal computers. Features of all the annotated eukaryotic genes are accessible through three main software tables, including for each entry details such as the gene summary, the gene exon/intron structure and the specific Gene Ontology attributions. The structuring of the data, the creation of additional calculation fields and the integration with nucleotide sequences allow users to make many types of comparisons and calculations that are useful for data retrieval and analysis. We provide an original example analysis of the existing introns across all the available species, through which the classic biological problem of the ‘minimal intron’ may find a solution using available data. Based on all currently available data, we can define the shortest known eukaryotic GT-AG intron length, setting the physical limit at the 30 base pair intron belonging to the human MST1L gene. This ‘model intron’ will shed light on the minimal requirement elements of recognition used for conventional splicing functioning. Remarkably, this size is indeed consistent with the sum of the splicing consensus sequence lengths.
我们开发了GeneBase,这是国家生物技术信息中心(NCBI)基因数据库的完整解析器,它为个人计算机生成了一个具有直观用户友好图形界面的完全结构化的本地数据库。所有注释真核生物基因的特征都可以通过三个主要的软件表访问,包括每个条目的详细信息,如基因摘要、基因外显子/内含子结构和特定的基因本体属性。数据的结构化、附加计算字段的创建以及与核苷酸序列的集成允许用户进行多种类型的比较和计算,这些比较和计算对数据检索和分析非常有用。我们提供了所有可用物种中现有内含子的原始示例分析,通过该分析,“最小内含子”的经典生物学问题可能会使用可用数据找到解决方案。根据目前所有可用的数据,我们可以定义已知最短的真核生物GT-AG内含子长度,将物理限制在属于人类MST1L基因的30碱基对内含子。这个“模型内含子”将阐明用于传统拼接功能的识别的最低要求元素。值得注意的是,这个大小确实与剪接共识序列长度的总和是一致的。
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引用次数: 38
Functional and expression analyses of transcripts based on full-length cDNAs of Sorghum bicolor 高粱双色基因全长cdna转录本的功能与表达分析
Setsuko Shimada, Y. Makita, Tomoko Kuriyama-Kondou, M. Kawashima, Yoshiki Mochizuki, H. Hirakawa, Shusei Sato, Tetsuro Toyoda, M. Matsui
Sorghum bicolor is one of the most important crops for food and bioethanol production. Its small diploid genome and resistance to environmental stress make sorghum an attractive model for studying the functional genomics of the Saccharinae and other C4 grasses. We analyzed the domain-based functional annotation of the cDNAs using the gene ontology (GO) categories for molecular function to characterize all the genes cloned in the full-length cDNA library of sorghum. The sorghum cDNA library successfully captured a wide range of cDNA-encoded proteins with various functions. To characterize the protein function of newly identified cDNAs, a search of their deduced domains and comparative analyses in the Oryza sativa and Zea mays genomes were carried out. Furthermore, genes on the sense strand corresponding to antisense transcripts were classified based on the GO of molecular function. To add more information about these genes, we have analyzed the expression profiles using RNA-Seq of three tissues (spikelet, seed and stem) during the starch-filling phase. We performed functional analysis of tissue-specific genes and expression analysis of genes of starch biosynthesis enzymes. This functional analysis of sorghum full-length cDNAs and the transcriptome information will facilitate further analysis of the Saccharinae and grass families.
高粱是重要的粮食作物和生物乙醇生产作物之一。高粱的小二倍体基因组和对环境胁迫的抗性使其成为研究糖精科和其他C4禾本科植物功能基因组学的一个有吸引力的模型。我们利用基因本体(GO)分子功能分类对高粱全长cDNA文库中克隆的所有基因进行了基于结构域的功能注释分析。高粱cDNA文库成功捕获了一系列具有多种功能的cDNA编码蛋白。为了表征新鉴定的cdna的蛋白质功能,对其推断结构域进行了搜索,并在水稻和玉米基因组中进行了比较分析。此外,根据分子功能GO对反义转录物对应的义链上的基因进行分类。为了进一步了解这些基因,我们利用RNA-Seq分析了淀粉灌浆期三种组织(小穗、种子和茎)的表达谱。我们进行了组织特异性基因的功能分析和淀粉生物合成酶基因的表达分析。高粱全长cdna的功能分析和转录组信息将有助于进一步分析糖精科和禾本科植物。
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引用次数: 4
New insights into the interplay between codon bias determinants in plants 植物密码子偏向决定因素之间相互作用的新见解
S. Camiolo, S. Melito, A. Porceddu
Codon bias is the non-random use of synonymous codons, a phenomenon that has been observed in species as diverse as bacteria, plants and mammals. The preferential use of particular synonymous codons may reflect neutral mechanisms (e.g. mutational bias, G|C-biased gene conversion, genetic drift) and/or selection for mRNA stability, translational efficiency and accuracy. The extent to which these different factors influence codon usage is unknown, so we dissected the contribution of mutational bias and selection towards codon bias in genes from 15 eudicots, 4 monocots and 2 mosses. We analysed the frequency of mononucleotides, dinucleotides and trinucleotides and investigated whether the compositional genomic background could account for the observed codon usage profiles. Neutral forces such as mutational pressure and G|C-biased gene conversion appeared to underlie most of the observed codon bias, although there was also evidence for the selection of optimal translational efficiency and mRNA folding. Our data confirmed the compositional differences between monocots and dicots, with the former featuring in general a lower background compositional bias but a higher overall codon bias.
密码子偏差是指同义密码子的非随机使用,这种现象在细菌、植物和哺乳动物等多种物种中都有观察到。特定同义密码子的优先使用可能反映了中性机制(例如突变偏倚、G / c偏倚基因转换、遗传漂变)和/或mRNA稳定性、翻译效率和准确性的选择。这些不同的因素在多大程度上影响密码子的使用尚不清楚,因此我们分析了突变偏倚和选择对密码子偏倚的贡献,这些基因来自15个单子科植物、4个单子科植物和2个苔藓植物。我们分析了单核苷酸、二核苷酸和三核苷酸的频率,并研究了组成基因组背景是否可以解释所观察到的密码子使用概况。中性力量,如突变压力和G| c偏倚基因转换似乎是大多数观察到的密码子偏倚的基础,尽管也有证据表明选择最佳的翻译效率和mRNA折叠。我们的数据证实了单子型和双子型之间的成分差异,单子型的背景成分偏差一般较低,但总体密码子偏差较高。
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引用次数: 28
Precise Sequential DNA Ligation on A Solid Substrate: Solid-Based Rapid Sequential Ligation of Multiple DNA Molecules 在固体底物上精确的序列DNA连接:多个DNA分子的基于固体的快速序列连接
E. Takita, K. Kohda, Hajime Tomatsu, S. Hanano, Kanami Moriya, T. Hosouchi, N. Sakurai, Hideyuki Suzuki, A. Shinmyō, D. Shibata
Ligation, the joining of DNA fragments, is a fundamental procedure in molecular cloning and is indispensable to the production of genetically modified organisms that can be used for basic research, the applied bios-ciences, or both. Given that many genes cooperate in various pathways, incorporating multiple gene cassettes in tandem in a transgenic DNA construct for the purpose of genetic modification is often necessary when generating organisms that produce multiple foreign gene products. Here, we describe a novel method, designated PRESSO (precise sequential DNA ligation on a solid substrate), for the tandem ligation of multiple DNA fragments. We amplified donor DNA fragments with non-palindromic ends, and ligated the fragment to acceptor DNA fragments on solid beads. After the final donor DNA fragments, which included vector sequences, were joined to the construct that contained the array of fragments, the ligation product (the construct) was thereby released from the beads via digestion with a rare-cut meganuclease; the freed linear construct was circularized via an intra-molecular ligation. PRESSO allowed us to rapidly and efficiently join multiple genes in an optimized order and orientation. This method can overcome many technical challenges in functional genomics during the post-sequencing generation.
连接DNA片段是分子克隆的一个基本步骤,对于生产可用于基础研究、应用生物科学或两者兼而有之的转基因生物是必不可少的。考虑到许多基因在不同的途径中合作,在产生产生多种外源基因产物的生物体时,为了进行基因修饰,将多个基因盒串联在转基因DNA结构中往往是必要的。在这里,我们描述了一种新的方法,指定的PRESSO(精确顺序DNA连接在固体底物上),用于串联连接多个DNA片段。我们扩增了具有非回文末端的供体DNA片段,并将片段连接到固体珠上的受体DNA片段。在最终的供体DNA片段(包括载体序列)与包含片段阵列的构建体连接后,连接产物(构建体)因此通过稀有切割的巨核酶消化从珠子中释放出来;释放的线性结构通过分子内连接循环。PRESSO允许我们以优化的顺序和方向快速有效地连接多个基因。该方法可以克服功能基因组学在测序后生成过程中的许多技术挑战。
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引用次数: 0
Genome-Wide Microsatellite Characterization and Marker Development in the Sequenced Brassica Crop Species 芸苔属作物全基因组微卫星鉴定与标记开发
Jiaqin Shi, Shunmou Huang, Jiepeng Zhan, Jingyin Yu, Xinfa Wang, Wei Hua, Shengyi Liu, Guihua Liu, Hanzhong Wang
There was an error in the first three rows of Table 1 for the column B. oleracea Repeat number: the 3 should be 12. The corrected part of the table is given below. Table 1. Number, repeat number and total repeat length of the mono-to hexanucleotide repeats or motifs of microsatellites in the assembled genomic sequences of B. rapa, B. oleracea and B. napus Motif B. rapa B. oleracea which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
在表1的前三行b列中有一个错误。重复数:3应该是12。表格的更正部分如下表1。B. rapa, B. oleracea和B. napus Motif B. rapa B. oleracea基因组序列中单至六核苷酸重复序列或微卫星序列的重复数、重复数和总重复长度,该序列允许在任何介质中不受限制地重复使用、分发和复制,前提是原始作品被适当引用。
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引用次数: 3
期刊
DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes
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