In order to identify potential avirulence factors of <i>Phytophthora infestans</i>, secreted protein profiles of six strains (IPO-0, CBS431.90, IPO-655-2A, IPO-428-2, 88069 and 90128) with different avirulence phenotypes were analysed by proteomics. The proteins from culture filtrates were visualised on 2D gels with a total number between 350 and 1000. PiAVR X was found only in avirulent strains and considered therefore as a possible avirulence determinant. Its expression increased in all strains tested when grown in Modified Henniger medium. Single nucleotide polymorphisms (SNPs) were detected within a 735 nt fragment of <i>PiAvrX</i> but none was linked to avirulence characteristics. The consensus EST database facilitated identification of approximately 50% of 45 most abundant proteins excised from strain 88069. Beta-glucosidase, Pi-NIP2 (<i>Phytophthora infestans</i> necrosis inducing protein-like protein), enoyl CoA hydratase, glutathione S transferase, peptidylprolyl isomerase, acidic chitinase and Pi-PR1 (<i>Phytophthora infestans </i>homologue of plant PR-1 protein) were among the identified proteins with a recognisable signal sequence. Enolase, quinine-oxido reductase, nucleoside di-phosphate kinase, actin depolymerisation factor, thioredoxin, ubiquitin and 14-3-3 protein were also identified but without a signal peptide. The occurrence and expression of <i>Pi-nip2, Pi-chi1 </i>and <i>Pi-pr1</i> were confirmed in the strains tested. Transformants were obtained from <i>P. infestans</i> strain T30-4 via a biolistic particle delivery approach using a single plasmid vector containing <i>Pi-pr1</i>. Detailed analysis of these transformants did not demonstrate induction of homology-dependent gene silencing. It was found that transformation rates were different among the tested <i>P. infestans</i> strains.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:421333 |
| Date | January 2004 |
| Creators | Li, Shuang |
| Publisher | University of Aberdeen |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=165891 |
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