Download Agrobacterium-mediated transformation technology for monocot rice

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Agrobacterium-mediated transformation
technology for monocot
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Agrobacterium
Monocot plant
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Callus (dedifferentiated tissue)
Scutellum of Immature embryo
rice
calli
immature embryo
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
maize
immature embryo
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Monocot transformation technology
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Target plants
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Corn
Rice
Wheat
Barley
Switchgrass
Miscanthus
Sorghum
Big bluestem
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
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Rye
Oat
Millet
Turf
Banana
Pineapple
Sugarcane
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Agrobacterium
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Common soil bacteria
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DNA transfer capability
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Wild-type Agrobacterium makes crown gall via
its gene transfer
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
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T-DNA transfer mechanism
Signal Molecule
Induction of
Vir Gene
Expression
P
P
Activation
Vir Region
Expression
Integration
Ti Plasmid
RB
LB
Assemble
T-DNA
Generation of
ss T-strand
Maturation of
T-complex
T-complex
Agrobacterium tumefaciens
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
T-complex
Channel Complex
Targeting to
Nucleus
Plant Cell
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Super-binary system, more efficient
S Vir
T-DNA
Hybrid vector
Vir region
T-DNA
ORI
Binary System
ori
cos
S Vir
r
oriSp
Tetr cos
Homologous
Recombination
S Vir region
T-DNA
Vir region
Acceptor Vector
ori
cos
Tetr
ORI
ori
cos
Super-binary System
Spr
Intermediate Vector
T-DNA
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
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Vectors and Strains
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Transient assay using rice calli
LBA4404
pBI
EHA101 LBA4404
pBI
pSB
Tsukinohikari
Koshihikari
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
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Strengths
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Able to introduce DNA with defined ends.
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Able to introduce relatively large fragment.
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Low copy.
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Low frequency of gene silencing.
Low frequency of somaclonal variations.
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High fertility and high quality.
Industrial standard
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
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Publications
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Articles
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Hiei Y., Ohta S., Komari T. and Kumashiro T.
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Ishida Y, Saito H, Ohta S, Hiei Y, Komari T and Kumashiro
T.
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Efficient transformation of rice (Oryza sativa L.) mediated by
Agrobacterium and sequence analysis of the boundaries of
the T-DNA. Plant J. 6: 271 (1994)
High efficiency transformation of maize (Zea mays L.)
mediated by Agrobacterium tumefaciens. Nature Biotech. 14:
745 (1996)
Hiei, Y., and Komari, T.
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Improved protocols for transformation of indica rice mediated
by Agrobacterium tumefaciens. Plant Cell, Tissue and Organ
Culture 85, 271-283 (2006)
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
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Publications
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Patents
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Callus (dedifferentiated and dedifferentiating
tissues)
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Method of transforming monocotyledon
WO 94/00977
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Immature embryo
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Method of transforming monocotyledon by using
scutellum of immature embryo.
WO 95/06722
Copyrights © 2009 Japan Tobacco Inc. All rights reserved.
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