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Comparative gene expression analysis of susceptible and resistant near-isogenic lines in common wheat infected by Puccinia triticina


Manickavelu, A; Kawaura, K; Oishi, K; Tadasu, S I; Kohara, Y; Yahiaoui, N; Keller, B; Suzuki, A; Yano, K; Ogihara, Y (2010). Comparative gene expression analysis of susceptible and resistant near-isogenic lines in common wheat infected by Puccinia triticina. DNA Research, 17(4):211-222.

Abstract

Gene expression after leaf rust infection was compared in near-isogenic wheat lines differing in the Lr10 leaf rust resistance gene. RNA from susceptible and resistant plants was used for cDNA library construction. In total, 55 008 ESTs were sequenced from the two libraries, then combined and assembled into 14 268 unigenes for further analysis. Of these ESTs, 89% encoded proteins similar to (E value of < or =10(-5)) characterized or annotated proteins from the NCBI non-redundant database representing diverse molecular functions, cellular localization and biological processes based on gene ontology classification. Further, the unigenes were classified into susceptible and resistant classes based on the EST members assembled from the respective libraries. Several genes from the resistant sample (14-3-3 protein, wali5 protein, actin-depolymerization factor and ADP-ribosylation factor) and the susceptible sample (brown plant hopper resistance protein, caffeic acid O-methyltransferase, pathogenesis-related protein and senescence-associated protein) were selected and their differential expression in the resistant and susceptible samples collected at different time points after leaf rust infection was confirmed by RT-PCR analysis. The molecular pathogenicity of leaf rust in wheat was studied and the EST data generated made a foundation for future studies.

Abstract

Gene expression after leaf rust infection was compared in near-isogenic wheat lines differing in the Lr10 leaf rust resistance gene. RNA from susceptible and resistant plants was used for cDNA library construction. In total, 55 008 ESTs were sequenced from the two libraries, then combined and assembled into 14 268 unigenes for further analysis. Of these ESTs, 89% encoded proteins similar to (E value of < or =10(-5)) characterized or annotated proteins from the NCBI non-redundant database representing diverse molecular functions, cellular localization and biological processes based on gene ontology classification. Further, the unigenes were classified into susceptible and resistant classes based on the EST members assembled from the respective libraries. Several genes from the resistant sample (14-3-3 protein, wali5 protein, actin-depolymerization factor and ADP-ribosylation factor) and the susceptible sample (brown plant hopper resistance protein, caffeic acid O-methyltransferase, pathogenesis-related protein and senescence-associated protein) were selected and their differential expression in the resistant and susceptible samples collected at different time points after leaf rust infection was confirmed by RT-PCR analysis. The molecular pathogenicity of leaf rust in wheat was studied and the EST data generated made a foundation for future studies.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Plant and Microbial Biology
Dewey Decimal Classification:580 Plants (Botany)
Uncontrolled Keywords:Basidiomycota, genetics/pathogenicity; Expressed Sequence Tags; Gene Expression Profiling, methods; Gene Expression Regulation, Fungal; Gene Expression Regulation, Plant; Genes, Plant; Molecular Sequence Data; Plant Diseases, genetics/microbiology; Plant Leaves, genetics/microbiology; Triticum, genetics/microbiology
Language:English
Date:2010
Deposited On:30 Jan 2011 11:40
Last Modified:03 Aug 2017 15:25
Publisher:Oxford University Press
ISSN:1340-2838
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1093/dnares/dsq009
PubMed ID:20360266

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