De novo sequencing of hazelnut bacterial artificial chromosomes (BACs) using multiplex Illumina sequencing and targeted marker development for eastern filbert blight resistance

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• 作者：Vidyasagar R. Sathuvalli (1)
  Shawn A. Mehlenbacher (1)
  
• 关键词：Anisogramma anomala ; Filbert ; High throughput sequencing ; Linkage mapping ; Map ; based cloning ; Simple sequence repeats ; Microsatellites
• 刊名：Tree Genetics & Genomes
• 出版年：2013
• 出版时间：August 2013
• 年：2013
• 卷：9
• 期：4
• 页码：1109-1118
• 全文大小：211KB
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• 作者单位：Vidyasagar R. Sathuvalli (1)
  Shawn A. Mehlenbacher (1)
  
  1. Department of Horticulture, Oregon State University, 4017 ALS Building, Corvallis, OR, 97331, USA
  

文摘

Bacterial artificial chromosome (BAC) libraries are widely used in map-based cloning of plant genes. Eastern filbert blight (EFB), caused by the pyrenomycete Anisogramma anomala (Peck) E. Müller, is a devastating disease of European hazelnut (Corylus avellana L.) in the Pacific Northwest. A dominant allele at a single locus from the obsolete pollenizer “Gasaway-confers complete resistance. Our map-based cloning efforts use a BAC library for “Jefferson-hazelnut, which is heterozygous for resistance. Screening the library with primer pairs designed from RAPD markers closely linked to the EFB resistance locus identified 38 BACs. We sequenced 28 of these BACs using Illumina technology, by multiplexing with barcoded adapters. De novo sequence assembly using the programs Velvet and SOPRA and further alignment using CodonCode Aligner generated contigs whose length ranged from 393 to 108,194?bp. The number of contigs per BAC ranged from 1 to 19, and estimated coverage of assembled BACs ranged from 64?% to 100?%. Preliminary analysis of the sequences identified 779 simple sequence repeats (SSRs), from which we developed 23 markers. Of these, 17 were assigned to linkage group 6 adjacent to the disease resistance locus, five were placed on other linkage groups, and one could not be assigned to a linkage group. The BAC sequences and new SSR markers will be useful for our efforts at map-based cloning of the disease resistance gene.