Patent applications filed by The Samuel Roberts Noble Foundation
The applications describe methods for direct plant transformation via
Agrobacterium using vacuum. In one of the disclosures,
Agrobacterium containing a vector with a gene of interest contacts the
aerial portions of a plant at flowering stage under vacuum
conditions. The vacuum applied is of sufficient strength to force the
Agrobacterium cells into intimate contact with the plant such that the
T-DNA transfer to the plant takes place.
A plant at flowering stage is defined as a plant form about the beginning of
the first flower bud formation to about the time of the last flower set. The
flowering plants are grown from vernalised seeds. These are seeds subjected to a
period of chilling before germination. The seeds are incubated at 4°C for a
period of time, preferably in the dark, and kept moist. Vernalisation is used to
speed up the formation of flowers.
In the other disclosure, the material selected for transformation is a
seedling.
As part of both disclosures, the plant material is transformed with a mix of
Agrobacterium cells containing different T-DNA to be inserted into the
plants. The transformed plant is allowed to grow into maturity and produce
seeds. Progeny from the seed is selected by the use of selectable markers and
the presence of an additional transferred gene.
Specific PCT Application Information
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PCT Application Number
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Title, Independent Claims and Summary of Claims
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Assignee
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WO
2000/037663 A2
- Earliest priority - 23 December 1998
- Filed - 23 December 1999
- OPI - 29 June 2000
- Expected expiry - N/A
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Title - Plant transformation process
Claim 1
A method for direct plant transformation using seedlings and
Agrobacterium comprising:
A) contacting at least one seedling with Agrobacterium cells, said
Agrobacterium cells harboring a vector, said vector enabling said
Agrobacterium cells to transfer T-DNA containing at least one gene or
gene fragment to said seedling;
B) applying a vacuum to said seedling in
contact with said Agrobacterium cells at a first time, said vacuum of
sufficient strength to force said Agrobacterium cells into intimate
contact with said seedling such that said Agrobacterium cells transfer
said T-DNA to cells of said seedling at a second time, wherein said first and
second time are the same or different.
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| Claim 9
A method for direct plant transformation using seedlings and
Agrobacterium comprising:
A) contacting at least one seedling with a mixture of Agrobacterium
cells, said mixture comprising cells from a Agrobacterium strain
harboring a vector with a DNA fragment and cells from said
Agrobacterium strain harboring said vector a second DNA fragment, said
vector enabling said Agrobacterium cells to transfer said T-DNA to said
seedling;
B) applying a vacuum to said seedling in contact with said
Agrobacterium cells at a first time, said vacuum of sufficient strength
to force said Agrobacterium cells into intimate contact with said
seedling such that said Agrobacterium cells transfer T-DNA to cells of
said seedling at a second time, wherein said first and second time are the same
or different.
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| Claim 18
A method for direct plant transformation using seedlings and
Agrobacterium comprising:
A) contacting at least one seedling with Agrobacterium cells, said
Agrobacterium cells harboring a vector, said vector enabling said
Agrobacterium cells to transfer T-DNA containing at least one gene or
gene fragment and a selectable marker gene to said seedling;
B) applying a
vacuum to said seedling in contact with said Agrobacterium cells at a
first time, said vacuum of sufficient strength to force said
Agrobacterium cells into intimate contact with said seedling such that
said Agrobacterium cells transfer said T-DNA to cells of said seedling
at a second time, wherein said first and second time are the same or
different;
C) allowing said transformed seedling to grow to maturity and
set seed;
D) germinating said seed to form progeny;
E) exposing said
progeny to an agent enabling detection of selectable marker gene
expression;
F) selecting for progeny expressing said selectable marker gene
and at least one gene, said expression of said selectable marker gene and at
least one gene indicating gene transfer.
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The claims as filed of the PCT application WO 2000/037663
recite:
- a method for direct transformation of seedlings with Agrobacterium
by placing the plant material with Agrobacterium having a vector with a
gene of interest or a gene fragment in its T-DNA and applying vacuum to them so
Agrobacterium enters into contact with the seedling and transfers the
T-DNA to the plant cells;
- a method as the one described above but the material is in contact with a
mixture of Agrobacterium cells harboring different DNA fragments
(aren't necessarily gene or gene fragments) in their T-DNAs; and
- a method as the one first mentioned where after the transformation plants
set seed and the progeny from these seeds is selected for the presence of a gene
of interest and a selectable marker.
Status of National phase entries listed on INPADOC:
- Australia (AU25943/00 A) - application lapsed on 28 July
2005
- Canada (CA2352488 A1) - dead application on 23 December
2004 (i.e., the application cannot be reinstated).
- Europe (EP1141356 A2) - application deemed to be withdrawn
on 2 February 2005
- Japan (JP2002533090 T) - application pending
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The Samuel Roberts Noble Foundation
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WO
2000/063400 A2
- Earliest priority - 21 April 1999
- Filed - 20 April 2000
- OPI - 26 October 2000
- Expected expiry - N/A
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Title - Plant transformation process
| Claim 1
A method for direct plant transformation using plants and
Agrobacterium comprising:
A) contacting the aerial portions of at least one plant at the time of
flowering with Agrobacterium cells, said Agrobacterium cells
harboring a vector, said vector enabling said Agrobacterium cells to
transfer T-DNA containing at least one gene or gene fragment to said plant; and
B) applying a vacuum to said plant portions in contact with said
Agrobacterium cells at a first time, said vacuum of sufficient strength
to force said Agrobacterium cells into intimate contact with said plant
such that said Agrobacterium cells transfer said T-DNA to cells of said
plant at a second time to form a transformed plant, wherein said first time and
said second time are the same or different.
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| Claim 9
A method for direct transformation of a plant comprising:
A) vernalizing and germinating initial seed to form said plant contacting the
aerial portions of said plant at the time of flowering with
Agrobacterium cells, said Agrobacterium cells harboring a
vector, said vector enabling said Agrobacterium cells to transfer T-DNA
containing at least one gene or gene fragment to said plant; and
B)
applying a vacuum to said plant portions in contact with said
Agrobacterium cells at a first time, said vacuum of sufficient strength
to force said Agrobacterium cells into intimate contact with said plant
such that said Agrobacterium cells transfer said T-DNA to cells of said
plant at a second time to form a transformed plant, wherein said first time and
said second time are the same or different.
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| Claim 17
A method for direct plant transformation using plants at the time of
flowering and Agrobacterium comprising:
A) contacting aerial
portions of at least one plant at the time of flowering with a mixture of
Agrobacterium cells, said mixture comprising cells from a
Agrobacterium strain harboring a vector with a DNA fragment and cells
from said Agrobacterium strain harboring said vector with a second DNA
fragment, said vector enabling said Agrobacterium cells to transfer
said T-DNA to said plant; and
B) applying a vacuum to said plant portions
in contact with said Agrobacterium cells at a first time, said vacuum
of sufficient strength to force said Agrobacterium cells into intimate
contact with said plant such that said Agrobacterium cells transfer
T-DNA to cells of said plant at a second time to form a transformed plant,
wherein said first time and said second time are the same or different.
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| Claim 25
A method for direct transformation of a plant at the time of flowering
comprising:
A) vernalizing and germinating initial seed to form said plant contacting
aerial portions of said plant at the time of flowering with a mixture of
Agrobacterium cells, said mixture comprising cells from a
Agrobacterium strain harboring a vector with a DNA fragment and cells
from said Agrobacterium strain harboring said vector with a second DNA
fragment, said vector enabling said Agrobacterium cells to transfer
said T-DNA to said plant; and
B) applying a vacuum to said plant portions
in contact with said Agrobacterium cells at a first time, said vacuum
of sufficient strength to force said Agrobacterium cells into intimate
contact with said plant such that said Agrobacterium cells transfer
T-DNA to cells of said plant at a second time to form a transformed plant,
wherein said first time and said second time are the same or different.
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Claim 33
A method for direct plant transformation using plants at the time of
flowering and Agrobacterium
comprising:A) contacting aerial portions of at least one plant at the time of
flowering with Agrobacterium cells, said Agrobacterium cells
harboring a vector, said vector enabling said Agrobacterium cells to
transfer T-DNA containing at least one gene or gene fragment and a selectable
marker gene to said plant;
B) applying a vacuum to said plant portions in
contact with said Agrobacterium cells at a first time, said vacuum of
sufficient strength to force said Agrobacterium cells into intimate
contact with said plant such that said Agrobacterium cells transfer
said T-DNA to cells of said plant at a second time to form a transformed plant,
wherein said first time and said second time are the same or different;
C)
allowing said transformed plant to grow to maturity and set seed;
D)
germinating said seed to form progeny;
E) exposing said progeny to an agent
enabling detection of selectable marker gene expression; and
F) selecting
for progeny expressing said selectable marker gene and at least one gene, said
expression of said selectable marker gene and at least one gene indicating gene
transfer.
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| Claim 36
A method for direct transformation of a plant at the time of flowering
comprising:
A) vernalizing and germinating initial seed to form said plant;
B)
contacting aerial portions of said plant at the time of flowering with
Agrobacterium cells, said Agrobacterium cells harboring a
vector, said vector enabling said Agrobacterium cells to transfer T-DNA
containing at least one gene or gene fragment and a selectable marker gene to
said plant;
C) applying a vacuum to said plant portions in contact with
said Agrobacterium cells at a first time, said vacuum of sufficient
strength to force said Agrobacterium cells into intimate contact with
said plant such that said Agrobacterium cells transfer said T-DNA to
cells of said plant at a second time to form a transformed plant, wherein said
first time and said second time are the same or different;
D) allowing said
transformed plant to grow to maturity and set seed; germinating said seed to
form progeny;
E) exposing said progeny to an agent enabling detection of
selectable marker gene expression; and
F) selecting for progeny expressing
said selectable marker gene and at least one gene, said expression of said
selectable marker gene and at least one gene indicating gene transfer.
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Status of National phase entries listed on INPADOC:
- Australia (AU43652/00 A) - application lapsed on 8 December
2005
- Canada (CA2370638 A1) - dead application on 20 April 2005
(i.e., the application cannot be reinstated).
- Europe (EP1171618 A2) - application deemed to be withdrawn
on 11 May 2005
- New Zealand (NZ513993 A) - application void on 30 January
2004.
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Note: Patent information on this page was last updated on 8 March 2006.
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