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The Tomato Kinome and the Tomato Kinase Library ORFeome: Novel Resources for the Study of Kinases and Signal Transduction in Tomato and Solanaceae Species

January 2014 , Volume 27 , Number  1
Pages  7 - 17

Dharmendra K. Singh,1 Mauricio Calviño,1 Elizabeth K. Brauer,1,2 Noe Fernandez-Pozo,1 Susan Strickler,1 Roopa Yalamanchili,3 Hideyuki Suzuki,4 Koh Aoki,5 Daisuke Shibata,4 Johannes W. Stratmann,6 George V. Popescu,1,7 Lukas A. Mueller,1 and Sorina C. Popescu1

1The Boyce Thompson Institute for Plant Research, Ithaca, NY 14853 U.S.A.; 2Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, NY 14853 U.S.A.; 3Department of Plant Biology, North Carolina State University, Raleigh, NC 27607, U.S.A.; 4Kazusa DNA Research Institute, Kazusa-Kamatari 2-6-7, Kisarazu 292-0818, Japan; 5Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai 599-8531, Japan; 6Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, U.S.A.; 7National Institute for Laser, Plasma & Radiation Physics, Str. Atomistilor, Nr. 409, Magurele 077125, Bucharest, Romania

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Accepted 10 September 2013.

Protein kinase–driven phosphorylation constitutes the core of cellular signaling. Kinase components of signal transduction pathways are often targeted for inactivation by pathogens. The study of kinases and immune signal transduction in the model crop tomato (Solanum lycopersicum) would benefit from the availability of community-wide resources for large scale and systems-level experimentation. Here, we defined the tomato kinome and performed a comprehensive comparative analysis of the tomato kinome and 15 other plant species. We constructed a tomato kinase library (TOKN 1.0) of over 300 full-length open reading frames (ORF) cloned into a recombination-based vector. We developed a high-throughput pipeline to isolate and transform tomato protoplasts. A subset of the TOKN 1.0 library kinases were expressed in planta, were purified, and were used to generate a functional tomato protein microarray. All resources created were utilized to test known and novel associations between tomato kinases and Pseudomonas syringae DC3000 effectors in a large-scale format. Bsk7 was identified as a component of the plant immune response and a candidate effector target. These resources will enable comprehensive investigations of signaling pathways and host-pathogen interactions in tomato and other Solanaceae spp.

© 2014 The American Phytopathological Society