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The goal of this study was isolate genes that are regulated by Al treatments of tomato roots growing in vitro. For Al treatment, germinating tomato seeds were plated on a MS agar medium supplemented with 0, 350, and 1200 μM AlCl3 for 30 days. Total RNA was extracted from root tissues and separated on denature formamide gel to check their quantity and quality. Equal amount of total RNA from treatment and control was treated with DNAse I (Genhunter, TN) to remove genomic DNA contamination. cDNA was obtained by reverse transcription using all the regents in RNA Image Kit (Genhunter, TN). The cDNA was amplified using the fluorescently labeled anchor primers (Oligo dT-A, C, G) and 16 random primers. Amplification products were separated by electrophoresis in 6% nondenaturing polyacrylamide gels and DNA bands were observed by scanning the gel on a FMBIOIII scanner. After comparing the band profile on the gel image, fragments of gene that showed changes in intensity compared to control (0 μM AlCl3) were isolated from the gel manually. These bands were re-amplified with the same pair of primers as the original amplification and cloned onto PCR-Trap cloning vector (Genhunter, TN). After DNA sequence analysis and homology comparison with NCBI database, we have identified clone # C01HBa0256E08 on  L. esculentum  chromosome 01, clone # C10HBa0111D09 on chromosome 10 and clone # LE_HBa-31H5 on chromosome 4.

(286) Identification of Genes in Tomato Roots Induced by Aluminum Stress using Fluorescent cDNA Differential Display