Generation of a Nicotiana benthamiana full-length cDNA database

((in collaboration with TIGR and Brian Staskawicz’s group) Hailing Jin, UC Berkeley)
Nicotiana benthamiana is the best solanaceous plant for performing functional studies of genes via VIGS. A collaboration with Barbara Baker, Brian Staskawicz and Robin Buell at TIGR has been formed to create a Nicotiana benthamiana database, to be added to the public resources for solanaceous plants. The growing public database will ultimately enable parallel analysis of gene functions between Nicotiana benthamiana, potato, tomato and other solanaceous plants.

RNA was isolated from Nicotiana benthamiana tissues that include callus, roots from liquid culture grown plants, heat-stressed leaves (38 C, 3 hr and 6 hr), cold-stressed leaves (5 C 3 hr, 6hr), and pathogen challenged leaves (Pseudomonas syringae pv tomato 12hr; Xanthomonas campestris pv campestris 12 hr, 18hr; Pseudomonas syringae pv phaseolicola 18hr, and Xanthomonas campestris, pv vesicatoria 18hr). RNA was isolated from these tissues and pooled in approximately equal molar amounts. The pooled RNA was used for constructing the normalized library. The gateway based normalized Nicotiana benthamiana cDNA library has been created and ten thousand clones have been sequenced from both 5’ end and 3’ end. All sequence information has been submitted to Genbank.