The
forestry industry --which relies on woody tree species-- is a major economical
factor for several countries. Tree species like Eucalyptus and
Pinus are very important in the paper and cellulose industry. Because
of their long generation times, novel breeding methodologies are sought to
introduce disease resistance genes, influence lignin production or modulate
flowering time for example. Patents covering transformation technologies could
be of commercially strategic relevance.
Summary
- The European application assigned to the Instituto Nacional de
Investigacion y Tecnologia Agraria y Alimentaria (INIA) and the
Instituto Valenciano de Investigaciones Agrarias (IVIA), both
in Spain, is related to a United States patent analyzed in the 'Citrus' section.
The present application discloses a procedure for the transformation of
any woody species using adult plant material as primary
explant. The explant is grafted on a stock to generate shoots which are then
used as target tissue for Agrobacterium-mediated transformation.
Treated shoots are then re-grafted in vitro to select for transformed shoots,
which are then regenerated into full transgenic plants. This method addresses
the problem of how to obtain improved clones from elite woody trees from which
it is usually hard to obtain totipotent, regenerable material.
- The PCT application assigned to the Companhia Suzano de Papel e
Celulose and the University of Sao Paulo in Brazil
discloses a method to transform woody tree species starting from pre-germinated
seed material. The method disclosed was developed for transformation of
Eucalyptus spp but the claims have been worded to encompass any woody
tree species, which could include conifers. The method disclosed in the
independent claims is rather detailed in terms of timing of the multiple steps.
This timing could be very different for other tree species than
Eucalyptus, which would allow ways to design around the claim in this
patent (if granted) and thus avoid infringement.
Specific Patent Information
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Patent Number
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Tite, Independent Claims and Summary of Claims
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Assignee
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WO
2002/14463 A2
- Earliest priority - 18 August 2000
- Filed - 16 August 2001
- OPI - 21 February 2002
- International search report (A3) - 6 May 2004
- Modified first page (C1) - 1 July 2004
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Title - Method for genetic transformation of woody trees
| Claim 1
METHOD FOR GENETIC TRANSFORMATION OF WOODY TREES, which comprises the
following steps:
-sterilization and washing of seeds of woody trees;
-transference of said seeds to an appropriate medium of culture;
-germination for a period between about 2 and 17 days;
-collection of
germinated material;
-inoculation with Agrobacterium, containing
one or more genes of interest and optionally one or more marker genes, under
concentration between about 10E+7 and 10E+9 cells per ml;
-await
inoculation between about 20 and 30 hours in a liquid medium;
-transference of the material to a solid medium for a period between about 38
and 50 hours in the dark, under temperature between about 25 and 31 C and
ambient humidity;
-transference of the material to start plantlet growth in
the light;
-withdrawal of a plantlet leaf obtained between about 5 and 17
days during growth in the light;
-transference of the plantlet leaf to the
MS medium containing auxins derived from urea;
-await germination of the
plant tissue for approximately 20 days; and
-identification and selection
of the transformed plantlets.
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| Claim 2
METHOD FOR GENETIC TRANSFORMATION OF WOODY TREES, which comprises the
following steps:
-sterilization and washing of seeds of woody trees;
-transference of said seeds to an appropriate medium of culture;
-germination for a period between about 2 and 17 days;
-collection of
germinated material;
-sonication of the material as collected ; inoculation
with Agrobacterium, containing one or more genes of interest and
optionally one or various marker genes, under concentration between about 107
and 109 cells per ml;
-await inoculation between about 20 and 30 hours in a
liquid medium;
-transference of the material to a solid medium for a period
between about 38 and 50 hours in the dark, under temperature between about 25
and 31 C and ambient humidity;
-transference of the material to start the
plantlet growth in the light;
-withdrawal of a plantlet leaf obtained
between about 5 and 17 days during growth in the light;
-transference of
the plantlet leaf to the MS medium containing auxins derived from urea;
-await germination of the plant tissue for approximately 20 days and
-identification and selection of transformed plantlets.
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| Claim 9
METHOD TO OBTAIN TRANSGENIC WOODY TREE PLANTS, which comprises the following
steps:
-sterilization and washing of seeds of woody trees;
-transference of said seeds to an appropriate medium of culture;
-germination for a period between about 2 and 17 days;
-collection of
germinated material;
-inoculation with Agrobacterium, containing
one or more genes of interest and optionally one or more marker genes, under
concentration between about 107 and 109 cells per ml;
-await inoculation
between about 20 and 30 hours in a liquid medium ;
-transference of the
material to a solid medium for a period between about 38 and 50 hours in the
dark, under temperature between about 25 and 31 C and ambient humidity;
-transference of the material to start the plantlet growth step in the light;
-withdrawal of a plantlet leaf obtained between about 5 and 17 days during
growth in the light;
-transference of plantlet leaf to the MS medium
containing auxins derived from urea;
-await germination of the plant tissue
for approximately 20 days;
-identification and selection of transformed
plantlets;
-maintaining transforming plantlets in a multiplication and
elongation medium for about 20 days;
-regeneration of the final transformed
plant from one of the following regions hypocotyl; cotyledon; primary leaves and
col.
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A method for transformation of woody trees using pre-germinated seed as
target tissue for Agrobacterium-mediated transformation.
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Companhia Suzano de Papel e Celulose and Universidade De Sao Paulo
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Remarks
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- National phase entry of WO 2002/14463 in Australia (AU 2001/279510) is
pending.
- National phase entry of WO 2002/14463 in China (CN 1630723) is pending.
- National phase entry of WO 2002/14463 in Europe (EP 1448777) is pending.
- National phase entry of WO 2002/14463 in the United States
(US
2004/055041) is pending.
- Other national phase entries of WO 2002/14463 include Brazil (BR 200113455),
South Africa (ZA 200301299).
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EP
0870838 A2
- Earliest priority - 5 March 1997
- Filed - 4 March 1998
- Application pending
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Title - Procedure for the genetic transformation of adult
plants of woody species
| Claim 1
A genetic transformation procedure for adult plants of woody species which
consists of:
- inoculating explants of adult tissue of woody species, from the first shoots
of the grafts of buds of adult plants of woody species on stocks, with
appropriate vectors which carry the genes which encode the characteristics of
interest, under conditions which permit the development of transgenic shoots;
and
- micrografting in vitro said transgenic shoots, their buds or apices, on
stock cultivated in vitro and,
- subsequently, grafting the resulting micrografted plants onto other stocks
which give vigor and let the taken shoots grow to generate complete adult
transgenic plants, or directly transplanting the in vitro micrografted plants
into the soil, to give complete adult transgenic plants.
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A procedure for transformation of adult woody species with appropriate
vectors, which is not limited to Agrobacterium. Adult tissue is grafted
on stocks to generate new buds or apices that are used as target tissue for
transformation followed by in vitro micrografting of transgenic shoots.
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Remarks
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- Related United States patent
US
6103955 recites an Agrobacterium-mediated transformation method of
citrus (see section on
citrus).
- Other related patent application filed in Spain (ES 2151338).
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Note: Patent information on this page was last updated on 28 March 2006.
The information contained in this page was believed to be correct at the
time it was collated. New patents and patent applications, altered
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