Neves Leandro Gomide, Davis John M, Barbazuk William B, Kirst Matias
Graduate Program in Plant Molecular and Cellular Biology, University of Florida, Gainesville, Florida 32611.
G3 (Bethesda). 2014 Jan 10;4(1):29-37. doi: 10.1534/g3.113.008714.
Loblolly pine (Pinus taeda L.) is an economically and ecologically important conifer for which a suite of genomic resources is being generated. Despite recent attempts to sequence the large genome of conifers, their assembly and the positioning of genes remains largely incomplete. The interspecific synteny in pines suggests that a gene-based map would be useful to support genome assemblies and analysis of conifers. To establish a reference gene-based genetic map, we performed exome sequencing of 14729 genes on a mapping population of 72 haploid samples, generating a resource of 7434 sequence variants segregating for 3787 genes. Most markers are single-nucleotide polymorphisms, although short insertions/deletions and multiple nucleotide polymorphisms also were used. Marker segregation in the population was used to generate a high-density, gene-based genetic map. A total of 2841 genes were mapped to pine's 12 linkage groups with an average of one marker every 0.58 cM. Capture data were used to detect gene presence/absence variations and position 65 genes on the map. We compared the marker order of genes previously mapped in loblolly pine and found high agreement. We estimated that 4123 genes had enough sequencing depth for reliable detection of markers, suggesting a high marker conversation rate of 92% (3787/4123). This is possible because a significant portion of the gene is captured and sequenced, increasing the chances of identifying a polymorphic site for characterization and mapping. This sub-centiMorgan genetic map provides a valuable resource for gene positioning on chromosomes and guide for the assembly of a reference pine genome.
火炬松(Pinus taeda L.)是一种在经济和生态方面都很重要的针叶树,目前正在为其生成一系列基因组资源。尽管最近有人尝试对针叶树的大基因组进行测序,但其组装以及基因定位仍在很大程度上不完整。松树的种间同线性表明,基于基因的图谱将有助于支持针叶树的基因组组装和分析。为了建立一个基于参考基因的遗传图谱,我们对72个单倍体样本的作图群体中的14729个基因进行了外显子组测序,生成了一个包含7434个序列变异的资源,这些变异对应3787个基因。大多数标记是单核苷酸多态性,不过也使用了短插入/缺失和多核苷酸多态性。利用群体中的标记分离来生成一个高密度的、基于基因的遗传图谱。共有2841个基因被定位到松树的12个连锁群上,平均每0.58厘摩有一个标记。捕获数据用于检测基因的存在/缺失变异,并在图谱上定位65个基因。我们比较了先前在火炬松中定位的基因的标记顺序,发现高度一致。我们估计有4123个基因有足够的测序深度来可靠地检测标记,这表明标记转化率高达92%(3787/4123)。这是可能的,因为基因的很大一部分被捕获并测序,增加了识别用于表征和定位的多态性位点的机会。这个亚厘摩遗传图谱为基因在染色体上的定位提供了宝贵的资源,并为参考松树基因组的组装提供了指导