Hiroshi Ezuraa , Tsuyoshi Mizoguchia, Shin Watanabea, Sayaka Uchiia, Sun Hyon Jina, Yasutaka Kubob, Hitoshi Moric, Shunsuke Imanishid, Daisuke Shibatae aGene Research Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba-shi, Ibaraki-ken, 305-8577, - PowerPoint PPT Presentation

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Hiroshi Ezuraa , Tsuyoshi Mizoguchia, Shin Watanabea, Sayaka Uchiia, Sun Hyon Jina, Yasutaka Kubob, Hitoshi Moric, Shunsuke Imanishid, Daisuke Shibatae aGene Research Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba-shi, Ibaraki-ken, 305-8577,

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dNational Institute of Vegetable and Tea Science, NARO, 360 Kusawa, Ano, Mie-ken, ... Now M2 Plant seeds are distributed to each institutes and cultivated (Figure 2) ... – PowerPoint PPT presentation

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Title: Hiroshi Ezuraa , Tsuyoshi Mizoguchia, Shin Watanabea, Sayaka Uchiia, Sun Hyon Jina, Yasutaka Kubob, Hitoshi Moric, Shunsuke Imanishid, Daisuke Shibatae aGene Research Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba-shi, Ibaraki-ken, 305-8577,


1
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_at_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
?
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
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