Figure 1. JSOL Micro-Tom Mutant Line

1. Figure 4. Micro-Tom Mutant Database Generation of Micro-Tom Based Mutant Lines for Tomato Genomics by Japanese Solanaceae Genomics Consortium (JSOL) Hiroshi Ezuraa , Tsuyoshi Mizoguchia, Shin Watanabea, Sayaka Uchiia, Sun Hyon Jina, Yasutaka Kubob, Hitoshi Moric, Shunsuke Imanishid, Daisuke ShibataeaGene Research Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba-shi, Ibaraki-ken, 305-8577, Japan,bGuraduate School of National Science, Okayama University, 1-1-1 Tsushima-naka, Okayama-shi, Okayama-ken, 700-8530cGuraduate School of Bioagricultural Sciences, Nagoya University, Furo-cho ChiFull Address,JapandNational Institute of Vegetable and Tea Science, NARO, 360 Kusawa, Ano, Mie-ken, 514-2392, JapaneKazusa DNA Research Institute, Kazusa-Kamatari 2-6-7, Kisarazu-shi, Chiba 292-0818, JapanEmail: ezura@gene.tsukuba.ac.jp 2. Micro-Tom Mutant Database Introduction The spotlight for most plant scientists must be post-genomic studies such as transcriptome, proteome, and metabolome. In addition, advance in bioinfomatics pioneer new category of studies like system biology. Tomato is a model plant that gives information on fruit storage, product translocation and novel metabolites. It is highly expected that other solanaceae plant will have constructive feedback from the genetic and molecular study of tomato. Moreover, the development of L. esculentum var. Micro-Tom gave us a great advantage to examine tomato plantlet in narrow room space. Our main objective is to construct an infrastructure for Micro-Tom study. Mutanegenesis has been highly effective strategy for studying the genetic bases of traits. One of the mutational approach is a chemical mutagenesis by ehylmetane sulphate (EMS) treatment which gives rise to a high mutation frequency without apparent preferences for specific genomic regions. It can also generate many alleles that enable one to get null phenotypes. At present, we are establishing Micro-Tom mutant lines by EMS treatment and a construction of the database in silico with aim to supply the experimental resource and the information for worldwide use. Focusing on both "Floral and circadian rhythm in neuter plant" and " plant hormone and fruit development", we have already set up a system for screening mutants. In addition, we have established an efficient transformation protocol for developing T-DNA tag lines in Micro-Tom. Here we report the latest progress in our work conducted by Japanese Solanaceae consortium (JSOL). To integrate various and enormous mutant information from each institutes, Micro-Tom Mutant Database are under construction in silico. This is a collaboration work with National Institute of Genetics in Japan. final object of the database is offcorse to supply the experimental resource and the information for worldwide use. Start of test operation is scheduled for during next spring. 3. Establish Micro-Tom Transformation System for T-DNA Tag Line After establishing mutant lin, it is necessary to develop T-DNA tag lines for mutant analysis. However, largest trouble delaying the study mihgt be absence of highl efficent transformation procedure for tomato species. Therefore, We have established highly efficient transformation protocol for developing T-DNA tag lines in Micro-Tom (Figure 5). We succeeded to achieve highly efficient transformation ratio (approximately 40%) by addition of acetoshringone two times during cultivation period and operation of careful rooting selection to pick up transformed cells thoroughly (Figure 7). Especially, it is effective for highly efficient transformation to repeat separate operation at rooting selection step because organs undergoing redifferentiation to form multiple shoots included both transformed and untransformed cells (Figure 6). Micro-Tom Mutant Line After organizing Japanese Solanaceae consortium (JSOL) in 2004, we immediately decided to establish Micro-Tom mutant line and to set up a system for screening mutants. We started to cultivate M1 plants at the end of 2004 (Figure 1) and we had harvested 4000 M1 Plants by this spring and obtained 2500 M2 families (Table 1). These M2 Plants have already cultivated and been ready for screening. Figure 1. JSOL Micro-Tom Mutant Line か Figure 6. Chimera both transformed and untransformed cells Figure 5. Micro-Tom Transformation System JSOL Micro-Tom Mutant Line project is a collaborate work with 5 Japanese institutes. Now M2 Plant seeds are distributed to each institutes and cultivated (Figure 2). At the same time, we are establishing TILLING SYSTEM for screening mutants. Some interesting phenotypes in M2 Plants are discribed in Figure 3. Figure 2. JSOL Micro-Tom Mutant Line Team Figure 7. Key Point for high transformation ratio 3. Conclusion JSOL Micro-Tom Mutant Line project is a collaborate work with 5 Japanese institutes. Now M2 Plant seeds are distributed to each institutes and cultivated (Figure 2). Figure 3. M2 Plants