Grain Legumes Research Program, International Crops Research Institute for the Semi-Arid Tropics-ICRISAT, Hyderabad, India.
PLoS One. 2011;6(11):e27275. doi: 10.1371/journal.pone.0027275. Epub 2011 Nov 15.
Chickpea (Cicer arietinum L.) is the third most important cool season food legume, cultivated in arid and semi-arid regions of the world. The goal of this study was to develop novel molecular markers such as microsatellite or simple sequence repeat (SSR) markers from bacterial artificial chromosome (BAC)-end sequences (BESs) and diversity arrays technology (DArT) markers, and to construct a high-density genetic map based on recombinant inbred line (RIL) population ICC 4958 (C. arietinum)×PI 489777 (C. reticulatum). A BAC-library comprising 55,680 clones was constructed and 46,270 BESs were generated. Mining of these BESs provided 6,845 SSRs, and primer pairs were designed for 1,344 SSRs. In parallel, DArT arrays with ca. 15,000 clones were developed, and 5,397 clones were found polymorphic among 94 genotypes tested. Screening of newly developed BES-SSR markers and DArT arrays on the parental genotypes of the RIL mapping population showed polymorphism with 253 BES-SSR markers and 675 DArT markers. Segregation data obtained for these polymorphic markers and 494 markers data compiled from published reports or collaborators were used for constructing the genetic map. As a result, a comprehensive genetic map comprising 1,291 markers on eight linkage groups (LGs) spanning a total of 845.56 cM distance was developed (http://cmap.icrisat.ac.in/cmap/sm/cp/thudi/). The number of markers per linkage group ranged from 68 (LG 8) to 218 (LG 3) with an average inter-marker distance of 0.65 cM. While the developed resource of molecular markers will be useful for genetic diversity, genetic mapping and molecular breeding applications, the comprehensive genetic map with integrated BES-SSR markers will facilitate its anchoring to the physical map (under construction) to accelerate map-based cloning of genes in chickpea and comparative genome evolution studies in legumes.
鹰嘴豆(Cicer arietinum L.)是世界上第三大重要的冷季食用豆科植物,种植在干旱和半干旱地区。本研究的目的是从细菌人工染色体(BAC)末端序列(BES)和多样性阵列技术(DArT)标记中开发新的分子标记,如微卫星或简单序列重复(SSR)标记,并基于重组自交系(RIL)群体 ICC 4958(C. arietinum)×PI 489777(C. reticulatum)构建高密度遗传图谱。构建了一个包含 55680 个克隆的 BAC 文库,生成了 46270 个 BES。对这些 BES 的挖掘提供了 6845 个 SSR,为 1344 个 SSR 设计了引物对。同时,开发了约 15000 个克隆的 DArT 阵列,在测试的 94 个基因型中发现 5397 个克隆具有多态性。在 RIL 图谱群体的亲本基因型上筛选新开发的 BES-SSR 标记和 DArT 阵列显示出与 253 个 BES-SSR 标记和 675 个 DArT 标记的多态性。对这些多态性标记和从发表的报告或合作者处汇编的 494 个标记数据进行分离数据的获得用于构建遗传图谱。结果,在 8 个连锁群(LG)上共构建了一个包含 1291 个标记的综合遗传图谱,总长度为 845.56cM(http://cmap.icrisat.ac.in/cmap/sm/cp/thudi/)。每个连锁群的标记数从 68(LG8)到 218(LG3)不等,平均标记间距离为 0.65cM。虽然开发的分子标记资源将有助于遗传多样性、遗传图谱和分子育种应用,但与 BES-SSR 标记综合的综合遗传图谱将有助于将其锚定到物理图谱(正在构建中),以加速鹰嘴豆基因的图谱克隆和豆类的比较基因组进化研究。