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Analysis of Expressed Sequence Tags from Uromyces appendiculatus Hyphae and Haustoria and Their Comparison to Sequences from Other Rust Fungi

October 2008 , Volume 98 , Number  10
Pages  1,126 - 1,135

D. P. Puthoff, A. Neelam, M. L. Ehrenfried, B. E. Scheffler, L. Ballard, Q. Song, K. B. Campbell, B. Cooper, and M. L. Tucker

First, second, third, sixth, seventh, eighth, and ninth authors: Soybean Genomics and Improvement Lab, United States Department of Agriculture-Agriculture Research Service (USDA-ARS), Beltsville, MD 20705; and fourth and fifth authors: MSA Genomics Laboratory, USDA-ARS, Stoneville, MS 38776. First and second authors contributed equally to this manuscript.

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Accepted for publication 9 July 2008.

Hyphae, 2 to 8 days postinoculation (dpi), and haustoria, 5 dpi, were isolated from Uromyces appendiculatus infected bean leaves (Phaseolus vulgaris cv. Pinto 111) and a separate cDNA library prepared for each fungal preparation. Approximately 10,000 hyphae and 2,700 haustoria clones were sequenced from both the 5′ and 3′ ends. Assembly of all of the fungal sequences yielded 3,359 contigs and 927 singletons. The U. appendiculatus sequences were compared with sequence data for other rust fungi, Phakopsora pachyrhizi, Uromyces fabae, and Puccinia graminis. The U. appendiculatus haustoria library included a large number of genes with unknown cellular function; however, summation of sequences of known cellular function suggested that haustoria at 5 dpi had fewer transcripts linked to protein synthesis in favor of energy metabolism and nutrient uptake. In addition, open reading frames in the U. appendiculatus data set with an N-terminal signal peptide were identified and compared with other proteins putatively secreted from rust fungi. In this regard, a small family of putatively secreted RTP1-like proteins was identified in U. appendiculatus and P. graminis.

Additional keywords:expressed sequence tags, gene expression, secretome.

The American Phytopathological Society, 2008