De novo assembly using low-coverage short read sequence data from the rice pathogen Pseudomonas syringae pv. oryzae

Josephine A. Reinhardt, David A Baltrus, Marc T. Nishimura, William R. Jeck, Corbin D. Jones, Jeffery L. Dangl

Research output: Contribution to journalArticle

117 Citations (Scopus)

Abstract

We developed a novel approach for de novo genome assembly using only sequence data from high-throughput short read sequencing technologies. By combining data generated from 454 Life Sciences (Roche) and Illumina (formerly known as Solexa sequencing) sequencing platforms, we reliably assembled genomes into large scaffolds at a fraction of the traditional cost and without use of a reference sequence. We applied this method to two isolates of the phytopathogenic bacteria Pseudomonas syringae. Sequencing and reassembly of the well-studied tomato and Arabidopsis pathogen, PtoDC3000, facilitated development and testing of our method. Sequencing of a distantly related rice pathogen, Por1-6, demonstrated our method's efficacy for de novo assembly of novel genomes. Our assembly of Por1-6 yielded an N50 scaffold size of 531,821 bp with >75% of the predicted genome covered by scaffolds over 100,000 bp. One of the critical phenotypic differences between strains of P. syringae is the range of plant hosts they infect. This is largely determined by their complement of type III effector proteins. The genome of Por1-6 is the first sequenced for a P. syringae isolate that is a pathogen of monocots, and, as might be predicted, its complement of type III effectors differs substantially from the previously sequenced isolates of this species. The genome of Por1-6 helps to define an expansion of the P. syringae pan-genome, a corresponding contraction of the core genome, and a further diversification of the type III effector complement for this important plant pathogen species.

Original languageEnglish (US)
Pages (from-to)294-305
Number of pages12
JournalGenome Research
Volume19
Issue number2
DOIs
StatePublished - Feb 2009
Externally publishedYes

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Pseudomonas syringae
Genome
Biological Science Disciplines
Oryza
Host Specificity
Lycopersicon esculentum
Arabidopsis
Technology
Bacteria
Costs and Cost Analysis

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

De novo assembly using low-coverage short read sequence data from the rice pathogen Pseudomonas syringae pv. oryzae. / Reinhardt, Josephine A.; Baltrus, David A; Nishimura, Marc T.; Jeck, William R.; Jones, Corbin D.; Dangl, Jeffery L.

In: Genome Research, Vol. 19, No. 2, 02.2009, p. 294-305.

Research output: Contribution to journalArticle

Reinhardt, Josephine A. ; Baltrus, David A ; Nishimura, Marc T. ; Jeck, William R. ; Jones, Corbin D. ; Dangl, Jeffery L. / De novo assembly using low-coverage short read sequence data from the rice pathogen Pseudomonas syringae pv. oryzae. In: Genome Research. 2009 ; Vol. 19, No. 2. pp. 294-305.
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