Construction and characterization of a full-length cDNA library for the wheat stripe rust pathogen (Puccinia striiformis f. sp. tritici).
BACKGROUND
Puccinia striiformis is a plant pathogenic fungus causing stripe rust, one of the most important diseases on cereal crops and grasses worldwide. However, little is know about its genome and genes involved in the biology and pathogenicity of the pathogen. We initiated the functional genomic research of the fungus by constructing a full-length cDNA and determined functions of the first group of genes by sequence comparison of cDNA clones to genes reported in other fungi.


RESULTS
A full-length cDNA library, consisting of 42,240 clones with an average cDNA insert of 1.9 kb, was constructed using urediniospores of race PST-78 of P. striiformis f. sp. tritici. From 196 sequenced cDNA clones, we determined functions of 73 clones (37.2%). In addition, 36 clones (18.4%) had significant homology to hypothetical proteins, 37 clones (18.9%) had some homology to genes in other fungi, and the remaining 50 clones (25.5%) did not produce any hits. From the 73 clones with functions, we identified 51 different genes encoding protein products that are involved in amino acid metabolism, cell defense, cell cycle, cell signaling, cell structure and growth, energy cycle, lipid and nucleotide metabolism, protein modification, ribosomal protein complex, sugar metabolism, transcription factor, transport metabolism, and virulence/infection.


CONCLUSIONS
The full-length cDNA library is useful in identifying functional genes of P. striiformis.
Ling
Peng
P
US Department of Agriculture, Agricultural Research Service, Wheat Genetic, Quality, Physiology and Disease Research Unit, Pullman, WA 99164-6430, USA. pling@wsu.edu <pling@wsu.edu>
Wang
Meinan
M
Chen
Xianming
X
Campbell
Kimberly Garland
KG
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D000595:Amino Acid Sequence; D020029:Base Pairing; D001487:Basidiomycota / Q000235:genetics* / Q000472:pathogenicity*; D018387:Codon, Initiator; D018076:DNA, Complementary / Q000235:genetics*; D015723:Gene Library; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014908:Triticum / Q000382:microbiology*
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D018387:Codon, Initiator; D018076:DNA, Complementary