Department of Environment and Agriculture, Curtin University, Kent Street, Bentley, Perth, Western Australia 6102, Australia.
Genome Biol. 2010;11(11):R109. doi: 10.1186/gb-2010-11-11-r109. Epub 2010 Nov 10.
Pyrenophora teres f. teres is a necrotrophic fungal pathogen and the cause of one of barley's most important diseases, net form of net blotch. Here we report the first genome assembly for this species based solely on short Solexa sequencing reads of isolate 0-1. The assembly was validated by comparison to BAC sequences, ESTs, orthologous genes and by PCR, and complemented by cytogenetic karyotyping and the first genome-wide genetic map for P. teres f. teres.
The total assembly was 41.95 Mbp and contains 11,799 gene models of 50 amino acids or more. Comparison against two sequenced BACs showed that complex regions with a high GC content assembled effectively. Electrophoretic karyotyping showed distinct chromosomal polymorphisms between isolates 0-1 and 15A, and cytological karyotyping confirmed the presence of at least nine chromosomes. The genetic map spans 2477.7 cM and is composed of 243 markers in 25 linkage groups, and incorporates simple sequence repeat markers developed from the assembly. Among predicted genes, non-ribosomal peptide synthetases and efflux pumps in particular appear to have undergone a P. teres f. teres-specific expansion of non-orthologous gene families.
This study demonstrates that paired-end Solexa sequencing can successfully capture coding regions of a filamentous fungal genome. The assembly contains a plethora of predicted genes that have been implicated in a necrotrophic lifestyle and pathogenicity and presents a significant resource for examining the bases for P. teres f. teres pathogenicity.
层出镰刀菌(Pyrenophora teres f. teres)是一种坏死型真菌病原体,也是大麦最重要病害之一——网斑病的病原体。本文报道了首个基于 0-1 分离株的 Solexa 短测序reads 的该物种基因组序列,该序列通过与 BAC 序列、EST 序列、直系同源基因和 PCR 比较进行了验证,并通过细胞遗传学核型分析和首个层出镰刀菌全基因组遗传图谱进行了补充。
总组装大小为 41.95 Mbp,包含 11799 个 50 个氨基酸或以上的基因模型。与两个已测序 BAC 的比较表明,高 GC 含量的复杂区域能有效组装。电泳核型分析显示,0-1 和 15A 分离株之间存在明显的染色体多态性,细胞遗传学核型分析证实至少存在 9 条染色体。遗传图谱跨越 2477.7 cM,由 25 个连锁群中的 243 个标记组成,并整合了来自组装的简单序列重复标记。在预测基因中,非核糖体肽合成酶和外排泵尤其表现出层出镰刀菌特有的非直系同源基因家族的扩张。
本研究表明,Solexa 测序可以成功捕获丝状真菌基因组的编码区。该组装包含大量与坏死型生活方式和致病性相关的预测基因,为研究层出镰刀菌致病性的基础提供了重要资源。