Plant & Animal Genomes XIV Conference

January 14-18, 2006
Town & Country Convention Center
San Diego, CA

Deep Transcriptome Analysis Of The Maize Leaf Tissue Using RL-SAGE

Malali Gowda¹ , Huameng Li¹ , Venu Reddyvarichannarayappa¹ , Weisong Gu² , Mark Casey ³ , Godfrey Asea³ , HyeRan Kim⁴ , Dave Kudrna ⁴ , Rich Pratt³ , Eric Stahlberg² , Rod Wing⁴ , Guo-Liang Wang¹

¹  Department of Plant Pathology, The Ohio State University, Columbus OH 43210

²  Ohio Supercomputer Center, The Ohio State University, Columbus, OH 43212

³  Department of Horticultural and Crop Sciences, OARDC, The Ohio State University, Wooster, Wooster OH 44691

⁴  Arizona Genomics Institute, Department of Plant Sciences, University of Arizona, AZ 85721

Using the Robust-LongSAGE (RL-SAGE) method (Gowda et al. 2004), a LongSAGE library of the maize inbred line B73 was constructed which is being used for whole genome sequencing. RNA was extracted from four week-old leaves from B73 plants. From 232,948 total tags isolated from 8,426 sequencing reads, 44,870 unique tags were identified. Among them, 22,061 are significant tags (2 or more copies). Matching analysis showed that only about 7,175 (35%) and 9,606 (48%) significant tags matched to the TIGR maize ESTs and maize genomic sequence, respectively. About 4,831 anti-sense tags were obtained when matching to the maize EST sequences. Surprisingly, only 1,007 RL-SAGE tags from maize matched to the rice ESTs or FL-cDNA sequences, and 1,699 tags matched to the sorghum ESTs. To identify the full transcription units of the expressed genes in maize, we are optimizing the procedures for construction of 5’ and 3’ LongSAGE libraries. Four 5’ and 3’ libraries will be made from fungal infected and control plants. The sequencing information of these libraries will be presented.
Gowda M. et al (2004) Robust-LongSAGE (RL-SAGE): A Substantially Improved LongSAGE Method for Gene Discovery and Transcriptome Analysis. Plant Physiology 134: 890-897.