Given the central role of haustoria in the infection process,
several attempts have been made to purify them to a
degree suitable for proteomic and genetic analyses. Enrichment
has relied mainly on concavalin A sepharose affinity
chromatography [15, 16] and density gradient centrifugation
[17, 18]. Although both methods enrich haustoria to a degree,
they suffer from the drawback that the resulting enrichments
are contaminated with both plant material and hyphae, and it
is therefore difficult to identify haustoria proteins unequivocally,
although a subtractive approach can be used [16]. Here,
we present findings on the haustoria proteome using haustoria
that were purified to near homogeneity by immunoprecipitation
with monoclonal antibodies prepared specifically to
haustoria of P. triticina. We have used a high-resolution, high
mass accuracy mass spectrometer to analyze tryptic peptides
derived from these haustoria and have tentatively identified
1192 proteins with two or more peptides matched (p < 0.05).
Using bioinformatics-based algorithms and characteristics of
known effectors, 140 effector protein candidates have been
tentatively identified.
Given the central role of haustoria in the infection process,several attempts have been made to purify them to adegree suitable for proteomic and genetic analyses. Enrichmenthas relied mainly on concavalin A sepharose affinitychromatography [15, 16] and density gradient centrifugation[17, 18]. Although both methods enrich haustoria to a degree,they suffer from the drawback that the resulting enrichmentsare contaminated with both plant material and hyphae, and itis therefore difficult to identify haustoria proteins unequivocally,although a subtractive approach can be used [16]. Here,we present findings on the haustoria proteome using haustoriathat were purified to near homogeneity by immunoprecipitationwith monoclonal antibodies prepared specifically tohaustoria of P. triticina. We have used a high-resolution, highmass accuracy mass spectrometer to analyze tryptic peptidesderived from these haustoria and have tentatively identified1192 proteins with two or more peptides matched (p < 0.05).Using bioinformatics-based algorithms and characteristics ofknown effectors, 140 effector protein candidates have beententatively identified.
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