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‘杰斐逊’榛子的细菌人工染色体文库构建及其与东方果核病菌抗性和花粉-柱头不亲和性相关克隆的鉴定。

A bacterial artificial chromosome library for 'Jefferson' hazelnut and identification of clones associated with eastern filbert blight resistance and pollen-stigma incompatibility.

机构信息

Department of Horticulture, Oregon State University, Corvallis, 97331, USA.

出版信息

Genome. 2011 Oct;54(10):862-7. doi: 10.1139/g11-048. Epub 2011 Sep 21.

DOI:10.1139/g11-048
PMID:21936690
Abstract

European hazelnut (Corylus avellana L.) is the only economically important nut crop in the family Betulaceae. Because of its small genome size (~385 Mb / 1C), relatively short life cycle, availability of a dense linkage map, and amenability to transformation by Agrobacterium, the European hazelnut could serve as a model plant for the Betulaceae. Here we report the construction of a bacterial artificial chromosome (BAC) library for 'Jefferson' hazelnut using the cloning enzyme MboI and the vector pECBAC1 (BamHI site). The library consists of 39,936 clones arrayed in 104,384-well microtitre plates with a mean insert size of 117 kb. The genomic coverage of the library is estimated to be about 12 genome equivalents. This library provides a valuable resource for the map-based cloning of two important genes, the resistance gene from 'Gasaway' that confers resistance to eastern filbert blight caused by the fungus Anisogramma anomala (Peck) E. Müller and the S locus that controls pollen-stigma incompatibility. Fine-resolution mapping near the two loci was carried out using random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers. Fine mapping at the disease resistance locus showed that markers W07-375 and X01-825 flanked the resistance locus at distances of 0.06 and 0.05 cM, respectively. The S locus is flanked by markers 204-950 and KG819-200 at distances of 0.14 and 0.24 cM, respectively. Assuming that 1 cM corresponds to a physical distance of 430 kb, it will take approximately two to three chromosome walks to assemble BAC contigs that span both loci.

摘要

欧洲榛(Corylus avellana L.)是山毛榉科中唯一具有重要经济价值的坚果树种。由于其基因组大小较小(~385Mb/1C)、生命周期相对较短、高密度连锁图谱可用以及易于通过根癌农杆菌进行转化,欧洲榛可作为山毛榉科的模式植物。在这里,我们报告了使用克隆酶 MboI 和载体 pECBAC1(BamHI 位点)构建‘杰斐逊’榛子的细菌人工染色体(BAC)文库。该文库包含 39936 个克隆,排列在 104384 孔微孔板中,平均插入片段大小为 117kb。该文库的基因组覆盖率估计约为 12 个基因组当量。该文库为两个重要基因的图谱克隆提供了有价值的资源,这两个基因是来自‘Gasaway’的抗性基因,赋予了对由真菌 Anisogramma anomala (Peck) E. Müller引起的东方榛枯萎病的抗性,以及控制花粉-柱头不亲和性的 S 座位。使用随机扩增多态性 DNA(RAPD)和简单序列重复(SSR)标记对两个位点进行了精细定位作图。在抗性基因座附近的精细作图表明,标记 W07-375 和 X01-825 分别以 0.06 和 0.05cM 的距离侧翼抗性基因座,S 座位由标记 204-950 和 KG819-200 以 0.14 和 0.24cM 的距离侧翼,分别。假设 1cM 对应于 430kb 的物理距离,大约需要两到三个染色体步长才能组装跨越两个基因座的 BAC 连续序列。

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