Gene expression and proteomic analysis of the formation of Phakopsora pachyrhizi appressoria

BMC Genomics

Research Article

Gary W Smythers¹  Alberto Nuñez²  Laurie L Fortis³  Christine L Stone⁴  Douglas G Luster⁴  Michael B McMahon⁴  Reid D Frederick⁴ 

[1] National Cancer Institute, Advanced Biomedical Computing Center, Building 430, 21702, Fort Detrick, MD, USA;USDA-Agricultural Research Service, Eastern Regional Research Center, 600 East Mermaid Lane, 19038, Wyndmoor, PA, USA;USDA-Agricultural Research Service, Eastern Regional Research Center, 600 East Mermaid Lane, 19038, Wyndmoor, PA, USA;USDA-National Institute of Food and Agriculture, Institute of Bioenergy, Climate, and Environment, 3245 Waterfront Centre, 800 9th Street, 20024, Southwest, Washington, District of Columbia, USA;USDA-Agricultural Research Service, Foreign Disease-Weed Science Research Unit, 1301 Ditto Avenue, 21702, Fort Detrick, MD, USA;

关键词: cDNA Library;    Spore Germination;    Germ Tube;    Suppression Subtractive Hybridization;    High Transcript Level;   

DOI  :  10.1186/1471-2164-13-269

 received in 2011-09-29, accepted in 2012-06-12,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundPhakopsora pachyrhizi is an obligate fungal pathogen causing Asian soybean rust (ASR). A dual approach was taken to examine the molecular and biochemical processes occurring during the development of appressoria, specialized infection structures by which P. pachyrhizi invades a host plant. Suppression subtractive hybridization (SSH) was utilized to generate a cDNA library enriched for transcripts expressed during appressoria formation. Two-dimensional gel electrophoresis and mass spectroscopy analysis were used to generate a partial proteome of proteins present during appressoria formation.ResultsSequence analysis of 1133 expressed sequence tags (ESTs) revealed 238 non-redundant ESTs, of which 53% had putative identities assigned. Twenty-nine of the non-redundant ESTs were found to be specific to the appressoria-enriched cDNA library, and did not occur in a previously constructed germinated urediniospore cDNA library. Analysis of proteins against a custom database of the appressoria-enriched ESTs plus Basidiomycota EST sequences available from NCBI revealed 256 proteins. Fifty-nine of these proteins were not previously identified in a partial proteome of P. pachyrhizi germinated urediniospores. Genes and proteins identified fell into functional categories of metabolism, cell cycle and DNA processing, protein fate, cellular transport, cellular communication and signal transduction, and cell rescue. However, 38% of ESTs and 24% of proteins matched only to hypothetical proteins of unknown function, or showed no similarity to sequences in the current NCBI database. Three novel Phakopsora genes were identified from the cDNA library along with six potentially rust-specific genes. Protein analysis revealed eight proteins of unknown function, which possessed classic secretion signals. Two of the extracellular proteins are reported as potential effector proteins.ConclusionsSeveral genes and proteins were identified that are expressed in P. pachyrhizi during appressoria formation. Understanding the role that these genes and proteins play in the molecular and biochemical processes in the infection process may provide insight for developing targeted control measures and novel methods of disease management.

【 授权许可】

Unknown   
© Stone et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.