Identification of Penicillium digitatum genes putatively involved in virulence/pathogenicity towards citrus fruit
M. López-Pérez and L. González-Candelas.
Journal of Plant Pathology Volume 90 (2, Supplement) August 2008, Book of Abstract, 9th International Congress of Plant Pathology, August 24-29, 2008 Torino,Italy,. 507 pages.
2008
บทคัดย่อ
Identification of Penicillium digitatum genes putatively involved in virulence/pathogenicity towards citrus fruit
Penicillium digitatumis the major post harvest pathogen of citrusfruit, being responsible for up to 80% of the losses due to decay.This necrotrophic fungus shows a very restricted host range,infecting only citrus fruit. Despite this fact, our knoweldege of thefactors involved in pathogenicity is very scarce; therefore weaimed to study at the molecular level the mechanisms involved inpathogenicity and virulence of this fungus. We have applied theSuppression Subtractive Hybridization (SSH) technique to obtaina cDNA library enriched inP. digitatumgenes that are upregulatedduring infection using a mixture of RNAs from uninfected fruitandin vitrogrown fungus as a “driver” and RNA from infectedfruit tissue as “tester”. After two rounds of PCR amplificationsubtracted cDNA fragments were ligated into plasmid pCRII andcloned intoE. colicompetent cells. DNA inserts from 1440 clonestogether with positive and negative controls were PCR-amplifiedand spotted onto replicate nylon membranes. Using this macroarraywe have identifiedP. digitatumgenes that are upregulated duringinfection as compared toin vitrogrowth conditions and thusare good candidates to be involved in pathogenicity. The results ofthese hybridizations together with sequence analysis of a numberof clones constitute a first step to elucidate the virulence/pathogenicitydeterminants of this important citrus pathogen. We willalso present a more detailed Northern blot analysis of selectedP.digitatumgenes.