Chromosome-scale genome assembly of the transformation-amenable common wheat cultivar 'Fielder'.

IF 3.9 2区 生物学 Q1 GENETICS & HEREDITY
Kazuhiro Sato, Fumitaka Abe, Martin Mascher, Georg Haberer, Heidrun Gundlach, Manuel Spannagl, Kenta Shirasawa, Sachiko Isobe
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引用次数: 48

Abstract

We have established a high-quality, chromosome-level genome assembly for the hexaploid common wheat cultivar 'Fielder', an American, soft, white, pastry-type wheat released in 1974 and known for its amenability to Agrobacterium tumefaciens-mediated transformation and genome editing. Accurate, long-read sequences were obtained using PacBio circular consensus sequencing with the HiFi approach. Sequence reads from 16 SMRT cells assembled using the hifiasm assembler produced assemblies with N50 greater than 20 Mb. We used the Omni-C chromosome conformation capture technique to order contigs into chromosome-level assemblies, resulting in 21 pseudomolecules with a cumulative size of 14.7 and 0.3 Gb of unanchored contigs. Mapping of published short reads from a transgenic wheat plant with an edited seed-dormancy gene, TaQsd1, identified four positions of transgene insertion into wheat chromosomes. Detection of guide RNA sequences in pseudomolecules provided candidates for off-target mutation induction. These results demonstrate the efficiency of chromosome-scale assembly using PacBio HiFi reads and their application in wheat genome-editing studies.

Abstract Image

Abstract Image

Abstract Image

可转化普通小麦品种‘菲尔德’的染色体尺度基因组组装。
我们已经建立了一个高质量的染色体水平基因组组装,用于六倍体普通小麦栽培品种'Fielder',这是一种1974年发布的美国软质,白色,糕点型小麦,以其易受农杆菌介导的转化和基因组编辑而闻名。使用PacBio循环共识测序和HiFi方法获得准确的长读序列。序列读取16个SMRT细胞,使用hifiasm汇编器组装,产生N50大于20 Mb的汇编。我们使用Omni-C染色体构象捕获技术将contigs排序到染色体水平的组装中,得到21个累积大小为14.7的假分子和0.3 Gb的未锚定contigs。对已发表的含有编辑过的种子休眠基因TaQsd1的转基因小麦植株的短序列进行定位,鉴定出转基因插入小麦染色体的四个位置。伪分子中引导RNA序列的检测为脱靶突变诱导提供了候选物。这些结果证明了使用PacBio HiFi reads进行染色体尺度组装的效率及其在小麦基因组编辑研究中的应用。
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来源期刊
DNA Research
DNA Research 生物-遗传学
CiteScore
6.00
自引率
4.90%
发文量
39
审稿时长
4.5 months
期刊介绍: DNA Research is an internationally peer-reviewed journal which aims at publishing papers of highest quality in broad aspects of DNA and genome-related research. Emphasis will be made on the following subjects: 1) Sequencing and characterization of genomes/important genomic regions, 2) Comprehensive analysis of the functions of genes, gene families and genomes, 3) Techniques and equipments useful for structural and functional analysis of genes, gene families and genomes, 4) Computer algorithms and/or their applications relevant to structural and functional analysis of genes and genomes. The journal also welcomes novel findings in other scientific disciplines related to genomes.
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