Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: 5/1/2005
Publication Date: 6/1/2005
Citation: Zhong, S., Szabo, L.J., Jeong, J.S., Mitchell, T.K., Dean, R.A., Cuomo, C.A. 2005. Development of full-length cDNA libraries for EST analysis of Puccinia graminis f.sp. tritici. Phytopathology. 95:S118. Interpretive Summary:
Technical Abstract: Puccinia graminis f.sp. tritici has been one of the most devastating plant pathogens in human history, causing serious epidemics of cereal crops, especially wheat. To elucidate the mechanisms of pathogenicity and biotrophic lifestyle of this rust fungus, a whole genome sequencing project has been recently initiated. Construction of full-length cDNA libraries from different stages of the fungus and generation of expressed sequence tags (ESTs) are part of the project for gene annotation and functional characterization. Total RNA was isolated from urediniospores, germinated urediniospores, and teliospores. cDNA libraries were constructed using SMART cDNA construction methodology (Clontech) and Gateway cloning system (Invitrogen). Analysis of 196 random clones showed that the average insert size is approximately 1.5 kb, with a range from 0.3 kb to 4.2 kb. Sequence analysis of these clones indicated that 95% of the inserts were full length, based on the presence of an ATG start codon within 100 bp of the 5' end. Large-scale sequencing of these cDNA libraries is underway, and the results of EST analysis will be presented.